Embed Size (px)
Citation preview
Scientific Contribution 201837 th Congress of the European Academy of Allergyand Clinical Immunology
EAACI, 26 - 30 May 2018, Munich, Germany
„Allergy in a changing world“
EAACI 18 Abstract book_Cover.indd 1 10.04.2018 11:36:52
Scienti fi c Contributi on 2018 37th Congress of the European Academy of Allergy and Clinical Immunology
EAACI, 26-30 May 2018, Munich, Germany
HAL Abstractbook 2018 A4.indd 1 17-04-18 16:07
HAL Abstractbook 2018 A4.indd 2 13-04-18 09:57
Dirk-Jan Opstelten, PhDChief Scienti fi c Offi cer
Dear Congress delegate,On behalf of HAL Allergy, it is our pleasure to welcome you to the EAACI 2018 Congress in Munich. HAL Allergy will present the latest results of its research & development program both in the format of oral and poster presentati ons in the scienti fi c program of the conference as well as by organizing a company sponsored symposium. The HAL Allergy symposium “Allergy in a changing world” will be held on Monday May 28, 2018 from 13.45h to 15.15h.
The theme for this year’s EAACI Congress is “Innovati ve soluti ons for allergy”. Innovati on lies at the heart of progress in the fi eld of allergy and will help and propel health care professionals, researchers and industry to off er allergic pati ents a bett er quality of life. As one of Europe’s leading players in the fi eld of allergen immunotherapy HAL Allergy is pleased to present its contributi on to allergy research and clinical practi ce to the parti cipants of the Congress.
At this year’s conference we will provide an update on HAL Allergy’s clinical development program to demonstrate the clinical effi cacy and safety of our immunotherapies and support their registrati on. The phase II and phase III study results obtained in the clinical program of liquid birch pollen extract for Sublingual Immunotherapy (SLIT) support its use in pati ents with allergic rhiniti s/rhinoconjuncti viti s with or without asthma.In a phase II allergen exposure chamber (AEC) trial the effi cacy of SLIT with diff erent doses of Phleum pratense liquid pollen extract in grass pollen allergic pati ents was demonstrated.Next to this in a prospecti ve non-interventi onal study an overall good tolerability of a subcutaneous rush up-dosing scheme with modifi ed allergens in pollen allergic subjects in day-to-day practi ce was shown.
Apart from the clinical research we also present interesti ng data on an IgE based assay used to determine individual reacti ons to purifi ed and modifi ed single peanut allergens as well as an IgG based assay to be used in stability studies of formulated allergoids. Posters on the producti on of recombinant Bet v1 in a CHO system and on the use of handheld pollen counters are in line with the theme of our satellite symposium ”Allergy in a changing world”. Together these data underpin HAL Allergy’s conti nuous quest to move allergy immunotherapy and, consequently, the fi eld of allergy as a whole into the future.
In summary, our presented work provides an update on HAL Allergy’s product development program, which is designed to enhance the wellbeing of allergic pati ents, by providing them high quality products with demonstrated effi cacy and safety.
We wish you a fruitf ul congress; if you require further informati on about our products or our R&D program, please be invited to meet us at our symposium or visit our Scienti fi c Corner in the HAL Allergy booth in the exhibiti on area.
Kind regards,
Harry Flore, PhD Dirk-Jan Opstelten, PhDChief Executi ve Offi cer Chief Scienti fi c Offi cer
Harry Flore, PhDChief Executi ve Offi cer
Kind regards,
Harry Flore, PhD Dirk-Jan Opstelten, PhDChief Executi ve Offi cer Chief Scienti fi c Offi cer
HAL Abstractbook 2018 A4.indd 3 13-04-18 09:57
Scientific Contribution 2018HAL Allergy Symposium
Monday 28 May, 2018, 13:45 – 15:15 | Room 5Allergy in a changing world
Chairs:Ronald van Ree, The NetherlandsDirk-Jan Opstelten, The Netherlands
Climate Change; its effect on the allergy epidemicJeroen Buters, Germany
Biologics versus AIT; Who will win?Nikos Papadopoulos, United Kingdom
AIT: fixing current and future needsMoisés Calderón, United Kingdom
HAL Abstractbook 2018 A4.indd 4 13-04-18 09:57
Scientific Contribution 2018 Contents Abstracts and poster presentations
ClinicalClinical program of a liquid birch pollen extract for SLIT: concordancewith current regulatory requirements of EMA ................................................................................................................... 6Sublingual Immunotherapy with a liquid Phleum pratense extract is effective in grass pollen allergic patients – Results of a phase II allergen exposure chamber (AEC) trial ................................................................ 8Dose response and onset of action of sublingual Immunotherapy with a liquid Phleum pratense extract in grass pollen allergic patients ........................................................................................................................ 10Efficacy of SLIT with different doses of a liquid phleum pratense extract was shown during the grass pollen season and in an Allergen Exposure Chamber (AEC) ......................................................................... 12Tolerability of a two week Rush up-dosing with modified allergens in pollen allergic subjects in the day-to-day practice ............................................................................................................................................. 14Tolerability of a two week Rush up-dosing with modified Trees, modified Grasses or modified Grasses / Trees mixture in pollen allergic subjects in the day-to-day practice ............................................................ 16
DevelopmentiELISA as a tool to measure IgE binding towards single modified peanut allergens .................................................... 18An IgG inhibition ELISA to measure the stability of alum-adsorbed grass pollen allergoids ....................................... 20Glycosylation of rBet v 1.0101 by Chinese Hamster Ovary cells affects proteolytic stability towards endo/lysosomal degradation .......................................................................................................................... 22Towards personalized pollen exposure measurements using hand held pollen samplers ......................................... 24
Contents
HAL Abstractbook 2018 A4.indd 5 13-04-18 09:57
Clinical program of a liquid birch pollen extract for SLIT: concordance with current regulatory requirements of EMA
O. Pfaar1,2, L. Klimek², D. Boot³, P.J. de Kam³, A. Narkus⁴, D. Yu⁵, A. Larionov⁵ and D.J. Opstelten⁵1Department of Otorhinolaryngology, Head and Neck Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University,
Mannheim, Germany. ²Center for Rhinology and Allergology, Wiesbaden, Germany. ³At time of clinical trial HAL Allergy BV, Leiden, The Netherlands,
⁴MC Narkus, Bleckede, Germany, ⁵HAL Allergy BV, Leiden, The Netherlands
BackgroundAccording to the current EMA guideline (CHMP/EWP/18504/2006) a dose-response relationship for clinical efficacy with different doses in several study-arms should be established e.g. by nasal challenge tests. Subsequently, confirmatory trials should be performed using a randomized placebo-controlled design with a combined symptom and medication score as primary endpoint to show a predefined and justified clinically relevant difference in the primary endpoint between test and control population. Here we report on the clinical development program of a sublingual birch immunotherapy product.
MethodsIn a first randomized, double-blind, placebo-controlled (DBPC), multi-center, dose tolerability, dose-range-finding trial, patients with birch pollen-induced rhinitis/rhinoconjunctivitis (ARC) with or without concomitant asthma (n=269, 18-60 years of age) were treated for 20 weeks with 4 doses of the liquid birch extract (3,333 AUN/ml; 10,000 AUN/ml; 20,000 AUN/ml; 40,000 AUN/ml) or placebo (ClinicalTrials.gov: NCT01639768). In a subsequent randomized, DBPC, parallel-group study, patients with birch pollen induced ARC with or without asthma (n=406, 18-65 years of age) were treated with the liquid birch pollen extract (40,000 AUN/mL) or placebo pre- and co-seasonally (ClinicalTrials.gov NCT02231307).
ResultsIn phase II the primary efficacy results demonstrated a clear dose response curve in the titrated nasal challenge (primary endpoint), with all active doses responding better than placebo and the 40,000 AUN/ml dose proved to be the most effective dose. These results were supported by the secondary efficacy parameters (Pfaar et al. Allergy 2016;71(1):99-107). In phase III a clinically relevant (32%) and statistically significant (p<0.0001) reduction in the CSMS (primary endpoint) in patients treated with the liquid birch extract (40,000 AUN/mL) compared to placebo were observed (Pfaar et al. Allergy (2016);71(suppl.102): 45(abstract 87). The secondary efficacy parameters consistently supported this finding (Pfaar et al. Allergy 2016;71(102):45 (abstract 1009).
ConclusionThe liquid birch pollen extract has been developed according to the current regulatory requirements. The optimal dose identified in phase II was mirrored by a significant and clinically relevant improvement in CSMS in phase III. The liquid birch pollen extract is an interesting new option for SLIT in patients with ARC with or without asthma.
Session number, date and time: PDS30, Tuesday 29 May 2018; 15:30-17:00Session title: Efficacy and safety of novel immunotherapy approaches
6 EAACI, 26-30 May 2018, Munich, Germany
HAL Abstractbook 2018 A4.indd 6 13-04-18 09:57
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt..
TThhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy oorr
wweerr
ee ccoo
nnttrraa
cctteedd
ffoorr tt
hhee sstt
uuddyy
bbyy HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
RReeffee
rreenncc
eess::
(1)P
faar
et a
l. Al
lerg
y 20
16;7
1(1)
:99-
107
(2)P
faar
et a
l. Al
lerg
y 20
16;7
1(10
2):4
5(ab
stra
ct 8
7)(3
)Pfa
ar e
t al.
Alle
rgy
2016
;71(
102)
:45(
abst
ract
100
9)
Figu
re 3
: Pha
se II
I stu
dy: M
ean
CSM
S du
ring
the
birc
h po
llen
seas
on fo
r pla
cebo
and
Liq
uid-
Birc
h-Ex
trac
t 40,
000
AUN
/mL
trea
ted
patie
nts
(ITT)
Mea
n (S
E)Pl
aceb
o(N
=178
)
Mea
n (S
E)Li
quid
-Birc
h-Ex
trac
t40
,000
AU
N/m
L(N
=179
)
Liqu
id-B
irch-
Extr
act 4
0,00
0 AU
N/m
L -P
lace
boDi
ffere
nce
(95
% C
I)p-
valu
e
Prim
ary
endp
oint
Com
bine
dsy
mpt
om-a
nd
med
icati
on-s
core
(CSM
S)
durin
gpo
llen
seas
on(P
S)
1,45
(0
,08)
1,00
(0
,08)
-0,4
6 (-0
,66,
-0,2
6)<0
,000
1
Seco
ndar
yen
dpoi
nts
CSM
Sdur
ing
peak
PS1,
56
(0,1
0)0,
96
(0,1
0)-0
,60
(-0,8
7, -0
,33)
<0,0
001
Sym
ptom
-sco
re d
urin
gpe
akPS
0,90
(0
,05)
0,53
(0
,05)
-0,3
7 (-0
,51,
-0,2
3)<0
,000
1
Med
icati
on-s
core
dur
ing
peak
PS0,
66
(0,0
6)0,
43
(0,0
6)-0
,23
(-0,4
0, -0
,06)
0,00
8
% w
elld
ays
durin
gPS
52,2
6 (3
,32)
67,5
3 (3
,31)
15,2
6 (7
,64,
22,
89)
0,00
01
% s
ever
eda
ysdu
ring
PS9,
47
(1,3
2)4,
04
(1,3
2)-5
,43
(-8,8
9, -1
,97)
0,00
2
Rhin
ocon
junc
tiviti
s qua
lity
of
life
ques
tionn
aire
-sco
re
(RQ
LQ-S
) dur
ing
PS
1,10
(0
,09)
0,55
(0
,09)
-0,5
5 (-0
,77,
-0,3
3)<0
,000
1
Euro
Qua
lity
ofLi
fe V
isual
An
alog
ueSc
ale
(EQ
-VAS
) du
ring
PS
-8,7
2 (1
,27)
-3,9
5 (1
,27)
4,77
(1
,68,
7,8
6)0,
0025
T abl
e 1:
Pha
se II
I stu
dy: C
linic
al re
sults
Figu
re 1
: Pha
se II
stud
y: D
ecre
ase
in L
ebel
sco
re c
ompa
red
to p
lace
bo a
fter 5
mon
ths
of
trea
tmen
t with
the
Liqu
id-B
irch-
Extr
act (
mIT
T)
F igu
re 2
: Pha
se II
stud
y: In
crea
se in
PN
IF c
ompa
red
to p
lace
bo a
fter 5
mon
ths
of tr
eatm
ent
with
the
Liq
uid-
Birc
h-Ex
trac
t
p=0.
158
p=0.
008
p<0.
001
-2,5-2
-1,5-1
-0,50
3,333AUN/ml
10,000AUN/ml
20,000AUN/ml
40,000AUN/ml
LS mean change from baseline in Lebel score compared to placebo
Trea
tmen
t gro
up
p=0.
476
p=0.
416
p=0.
385
p<0.
001
0510152025303540
3,33
3 AU
N/m
l10
,000
AU
N/m
l20
,000
AU
N/m
l40
,000
AU
N/m
l
LS meanchange frombaseline in PNIF (L/min) comparedtoplacebo
Trea
tmen
t gro
up
BBaacckk
ggrroouu
nndd&&
AAiimm
::
Acco
rdin
gto
the
curr
entE
MA
guid
elin
e(C
HM
P/EW
P/18
504/
2006
)a
dose
-res
pons
ere
latio
nshi
pfo
rcl
inic
aleffi
cacy
with
diffe
rent
dose
sin
seve
ral
stud
y-ar
ms
shou
ldbe
esta
blish
ede.
g.by
nasa
lch
alle
nge
test
s.Su
bseq
uent
ly,co
nfirm
ator
ytr
ials
shou
ldbe
perf
orm
edus
ing
ara
ndom
ized
plac
ebo-
cont
rolle
dde
sign
with
aco
mbi
ned
sym
ptom
and
med
icati
onsc
ore
aspr
imar
yen
dpoi
nta
pred
efine
dto
sho
wju
stifie
dcl
inic
ally
rele
vant
diffe
renc
ein
the
prim
ary
endp
oint
betw
een
test
and
cont
rolp
opul
ation
.Her
ew
ere
port
onth
ecl
inic
alde
velo
pmen
tpr
ogra
mof
asu
blin
gual
birc
him
mun
othe
rapy
prod
uct.
MMeett
hhooddss
::
Ina
first
rand
omize
d,do
uble
-blin
d,pl
aceb
o-co
ntro
lled
(DBP
C),
mul
ti-ce
nter
,do
seto
lera
bilit
y,do
se-r
ange
-find
ing
tria
l,pa
tient
sw
ithbi
rch
polle
n-in
duce
dal
lerg
icrh
initi
s/rh
inoc
onju
nctiv
itis
(ARC
)w
ithor
with
out
conc
omita
ntas
thm
a(n
=269
,age
:18-
60ye
ars
ofag
e)w
ere
trea
ted
for
20w
eeks
with
4do
ses
ofth
eliq
uid
birc
hex
trac
t(3
,333
AUN
/ml;
10,0
00AU
N/m
l;20
,000
AUN
/ml;
40,0
00AU
N/m
l)or
plac
ebo
(Clin
ical
Tria
ls.go
v:N
CT01
6397
68).
Ina
subs
eque
ntra
ndom
ized
,DBP
C,pa
ralle
l-gro
upst
udy,
patie
nts
with
birc
hpo
llen
indu
ced
ARC
with
orw
ithou
tast
hma
(n=4
06,1
8-65
year
sof
age)
wer
etr
eate
dw
ith
the
liqui
dbi
rch
polle
nex
trac
t(4
0,00
0AU
N/m
L)or
plac
ebo
pre-
and
co-s
easo
nally
(Clin
ical
Tria
ls.
gov
NCT
0223
1307
).
RReessuu
llttss::
Inph
ase
IIth
epr
imar
yeffi
cacy
resu
ltsde
mon
stra
ted
acl
ear
dose
resp
onse
curv
ein
the
titra
ted
nasa
lcha
lleng
e(p
rimar
yen
dpoi
nt),
with
all
activ
edo
ses
resp
ondi
ngbe
tter
than
plac
ebo
and
the
40,0
00AU
N/m
ldos
epr
oved
tobe
the
mos
teff
ectiv
edo
se.T
hese
resu
ltsw
ere
supp
orte
dby
the
seco
ndar
yeffi
cacy
para
met
ers1 .
Inph
ase
IIIa
clin
ical
lyre
leva
nt(3
2%)
and
stati
stica
llysi
gnifi
cant
(p<0
.000
1)re
ducti
onin
the
CSM
S(p
rimar
yen
dpoi
nt)
inpa
tient
str
eate
dw
ithth
eliq
uid
birc
hex
trac
t(40
,000
AUN
/mL)
com
pare
dto
plac
ebo
wer
eob
serv
ed2 .
The
seco
ndar
yeffi
cacy
para
met
ers
cons
isten
tlysu
ppor
ted
this
findi
ng3 .
CCoonncc
lluussiioo
nn::
The
liqui
dbi
rch
polle
nex
trac
tha
sbe
ende
velo
ped
acco
rdin
gto
the
curr
ent
regu
lato
ryre
quire
men
ts.T
heop
timal
dose
iden
tified
inph
ase
IIw
asm
irror
edby
asig
nific
ant
and
clin
ical
lyre
leva
ntim
prov
emen
tin
CSM
Sin
phas
eIII
.The
liqui
dbi
rch
polle
nex
trac
tis
anin
tere
sting
new
optio
nfo
rSL
ITin
patie
nts
with
ARC
with
orw
ithou
tast
hma.Clin
ical
pro
gram
of a
liqu
id b
irch
polle
n ex
trac
t for
SLI
T:
conc
orda
nce
with
cur
rent
regu
lato
ry re
quire
men
ts o
f EM
AO
. Pfa
ar1,
2 , L.
Klim
ek2 ,
D. B
oot3 ,
P.J.
de K
am3 ,
A. N
arku
s4 , D.
Yu5 ,
A. L
ario
nov5 ,
D.J.
Ops
telte
n5
1 Dep
artm
ent o
f Oto
rhin
olar
yngo
logy
, Hea
d an
d N
eck
Surg
ery,
Uni
vers
itäts
med
izin
Man
nhei
m, M
edic
al
Facu
lty M
annh
eim
, Hei
delb
erg
Uni
vers
ity,
Man
nhei
m, G
erm
any.
2 Cen
ter f
or R
hino
logy
and
Al
lerg
olog
y, W
iesb
aden
, Ger
man
y. 3 A
t tim
e of
clin
ical
tria
l HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s,4 M
C N
arku
s, B
leck
ede,
Ger
man
y, 5 H
AL A
llerg
y BV
, Lei
den,
The
Net
herla
nds
-32%
p <
0,00
01
LS-Mean CSMS
CSM
S in
pol
len
seas
on (I
TT)
Plac
ebo
SB
1,6
1,4
1,2 1
0,8
0,6
0,4
0,2 0
HAL Abstractbook 2018 A3.indd 1 13-04-18 09:54
abstract 2
PDS30 - Efficacy and safety of novel immunotherapy approaches
Clinical program of a liquid birch pollen extract for SLIT: concordance with current regulatory requirements of EMA
Sublingual Immunotherapy with a liquid Phleum pratense extract is effective in grass pollen allergic patients – Results of a phase II allergen exposure chamber (AEC) trial
E. Mantikou¹, P. Couroux², A. Salapatek², O. Pfaar3,4, A. Narkus⁵, D. Yu¹, A. Larionov¹, D.J. Opstelten¹, P.J. de Kam¹1HAL Allergy BV, Leiden, The Netherlands; 2Inflamax Research Inc., Mississauga, ON Canada; 3Department of Otorhinolaryngology, Head and Neck
Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany; 4Center for Rhinology and
Allergology, Wiesbaden, Germany; 5MC Narkus, Bleckede, Germany
BackgroundAccording to the EMA guideline on the clinical development of products for specific immunotherapy products should be tested in phase II at different doses in several study-arms to establish a dose-response relationship for clinical efficacy before confirmatory trials can be initiated. Allergen exposure in an AEC may be used as primary endpoint.
MethodsThe study was a single-center, randomized, double blind, placebo-controlled, phase II trial, treatment duration 10 months. 168 grass pollen allergic patients (18-65 years of age) with seasonal rhinitis/rhinoconjunctivitis (ARC) with (mild, GINA I) or without concomitant asthma were randomized to three different dosages of a liquid Phleum pratense extract (10,000 AUN/mL; 40,000 AUN/mL; 80,000 AUN/mL) or placebo for sublingual administration (maximum dose 5 drops, up-dosing in 4 days). The primary efficacy endpoint was the mean Total Symptom Score (TSS: sum of individual scores for eight nasal and non-nasal symptoms, rated on a scale of 0-3) at the end of the trial. For TSS assessment a validated AEC was used with a standardized controlled grass pollen exposure at an average concentration of 3500 ± 500 ppm3 for 6 hours ensuring aeroallergen exposure under controlled and reproducible conditions.
ResultsAccording to the pre-specified Emax model a borderline significant dose-response relationship for TSS was observed (p=0.050 per protocol population, 0.057 ITT). At the end of the trial all patients (ITT) treated with the different doses of the liquid Phleum Pratense extract showed an improvement in the TSS compared to placebo (LS Means (SE): placebo 10.01 points (0.80), 10.000 AUN/mL 8.06 (0.85), 40.000 AUN/mL 8.19 (0.82) and 80,000 AUN/mL 7.69 (0.83), respectively). The TSS reduction compared to placebo ranged from 18.2% to 23.2%. The treatment emergent adverse events (TEAE) were primarily local reactions, of mild intensity with a dose-response relationship. The incidence of patients developing at least one systemic reaction (all Grade I), were similar between the active treatment groups and the placebo group.
ConclusionA consistent improvement of ARC symptoms was observed with all doses of the liquid Phleum pratense extract, including the current marketed dose of 10,000 AUN/mL, compared to placebo in this AEC model. The safety profile was comparable to other sublingual immunotherapy (SLIT) products.
Session number, date and time: TPS16, Sunday 27 May 2018; 12:00-13:30Session title: Immunotherapy in the clinic 1
8 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 7
HAL Abstractbook 2018 A3.indd 2 13-04-18 09:54
Subl
ingu
al Im
mun
othe
rapy
with
a li
quid
Phl
eum
pr
aten
se e
xtra
ct is
effe
ctive
in g
rass
pol
len
alle
rgic
pati
ents
- Re
sults
of a
pha
se II
alle
rgen
exp
osur
e ch
ambe
r (AE
C) tr
ial
E. M
antik
ou1 ,
P. C
ouro
ux2 ,
A. S
alap
atek
2 , O
. Pfa
ar3,
4 , A.
Nar
kus5 ,
D. Y
u1 , A.
Lar
iono
v1 , D.
J. O
pste
lten1 ,
P.J.
de K
am1
1 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s; 2 In
flam
ax R
esea
rch
Inc.
, Miss
issau
ga, O
N C
anad
a;
3 Dep
artm
ent o
f Oto
rhin
olar
yngo
logy
, Hea
d an
d N
eck
Surg
ery,
Uni
vers
itäts
med
izin
Man
nhei
m,
Med
ical
Fac
ulty
Man
nhei
m, H
eide
lber
g U
nive
rsity
, Man
nhei
m, G
erm
any;
4 Cen
ter f
or R
hino
logy
and
Al
lerg
olog
y, W
iesb
aden
, Ger
man
y; 5 M
C N
arku
s, B
leck
ede,
Ger
man
y
BBaacckk
ggrroouu
nndd &&
AAiimm
::
Acco
rdin
gto
the
EMA
guid
elin
eon
the
clin
ical
deve
lopm
ent
ofpr
oduc
tsfo
rsp
ecifi
cim
mun
othe
rapy
prod
ucts
shou
ldbe
test
edin
phas
eII
atdi
ffere
ntdo
ses
inse
vera
lst
udy-
arm
sto
esta
blish
ado
se-r
espo
nse
rela
tions
hip
for
clin
ical
effica
cybe
fore
confi
rmat
ory
tria
lsca
nbe
initi
ated
.Alle
rgen
expo
sure
inan
AEC
may
beus
edas
am
odel
for
envi
ronm
enta
lex
posu
refo
reffi
cacy
asse
ssm
ent
ofSL
IT.
MMeett
hhooddss
::
The
stud
yw
asa
singl
e-ce
nter
,ran
dom
ized,
doub
lebl
ind,
plac
ebo-
cont
rolle
d,ph
ase
IItr
ial,
trea
tmen
tdu
ratio
n10
mon
ths.
168
gras
spo
llen
alle
rgic
patie
nts
(18-
65ye
ars
ofag
e)w
ithse
ason
alrh
initi
s/rh
inoc
onju
nctiv
itis
(ARC
)w
ith(m
ild,
GIN
AI)
orw
ithou
tco
ncom
itant
asth
ma
wer
era
ndom
ized
toth
ree
diffe
rent
dosa
ges
ofa
liqui
dPh
leum
prat
ense
extr
act
(10,
000
AUN
/mL;
40,0
00AU
N/m
L;80
,000
AUN
/mL)
orpl
aceb
ofo
rsub
lingu
alad
min
istra
tion
(max
imum
dose
5dr
ops,
up-d
osin
gin
4da
ys).
The
prim
ary
effica
cyen
dpoi
ntw
asth
em
ean
Tota
lSy
mpt
omSc
ore
(TSS
:su
mof
indi
vidu
alsc
ores
fore
ight
nasa
land
non-
nasa
lsym
ptom
s,ra
ted
ona
scal
eof
0-3)
atth
een
dof
the
tria
l.Fo
rTS
Sas
sess
men
ta
valid
ated
for
gras
spo
llen
AEC
was
used
with
ast
anda
rdize
dco
ntro
lled
gras
spo
llen
expo
sure
atan
aver
age
conc
entr
ation
of35
00±
500
ppm
3fo
r6
hour
sen
surin
gae
roal
lerg
enex
posu
r eun
derc
ontr
olle
dan
dre
prod
ucib
leco
nditi
ons.
RReessuu
llttss::
Acco
rdin
gto
the
pre-
spec
ified
Emax
mod
ela
bord
erlin
esig
nific
ant
dose
-res
pons
ere
latio
nshi
pfo
rTS
Sw
asob
serv
ed(p
=0.0
50pe
rpr
otoc
olpo
pula
tion,
0.05
7IT
T).A
tth
een
dof
the
tria
lall
patie
nts
(ITT)
trea
ted
with
the
diffe
rent
dose
sof
the
liqui
dPh
leum
Prat
ense
extr
act
show
edan
impr
ovem
ent
inth
eTS
Sco
mpa
red
topl
aceb
o(L
SM
eans
(SE)
:pla
cebo
10.0
1po
ints
(0.8
0),1
0.00
0AU
N/m
L8.
06(0
.85)
,40.
000
AUN
/mL
8.19
(0.8
2)an
d80
,000
AUN
/mL
7.69
(0.8
3),
resp
ectiv
ely)
.The
TSS
redu
ction
com
pare
dto
plac
ebo
rang
edfr
om18
.2%
to23
.2%
.Th
etr
eatm
ent
emer
gent
adve
rse
even
ts(T
EAE)
wer
epr
imar
ilylo
cal
reac
tions
,of
mild
inte
nsity
with
ado
se-
r esp
onse
rela
tions
hip.
The
inci
denc
eof
patie
nts
deve
lopi
ngat
leas
ton
esy
stem
icre
actio
n(a
llG
rade
I),w
ere
simila
rbet
wee
nth
eac
tive
trea
tmen
tgro
ups
and
the
plac
ebo
grou
p.
CCoonncc
lluussiioo
nn::
Aco
nsist
enti
mpr
ovem
ento
fARC
sym
ptom
sw
asob
serv
edw
ithal
ldo
ses
ofth
eliq
uid
Phle
umpr
aten
seex
trac
t,in
clud
ing
the
curr
ent
mar
kete
ddo
seof
10,0
00AU
N/m
L,co
mpa
red
topl
aceb
oin
this
AEC
mod
el.T
hesa
fety
profi
lew
asco
mpa
rabl
eto
othe
rsu
blin
gual
imm
unot
hera
py(S
LIT)
prod
ucts
.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt..
TThhee
pprreess
eennttee
rr iiss aa
nn eemm
ppllooyy
eeee oo
ff HHAALL
AAllllee
rrggyy..
EEAAAACC
II CCoonn
ggrreess
ss 220011
88
Tabl
e 1:
Mea
n TS
S at
Visi
t 6 (I
TT p
opul
ation
)
F igu
re 1
: Cal
cula
tion
of th
e pr
imar
y en
dpoi
nt, T
SS
Figu
re 2
: Bor
derli
ne s
igni
fican
t TSS
dos
e-re
spon
se u
sing
the
pre-
spec
ified
Em
ax m
odel
(IT
T po
pula
tion)
Para
met
er
Stati
stics
Pl
aceb
o SP
SP
SP
(N=4
0)
10,0
00 A
UN
/mL
40,0
00 A
UN
/mL
80,0
00 A
UN
/mL
(N
=35)
(N
=38)
(N
=37 )
Base
line
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
M
ean
(SD
) 14
.15
(4.2
9)
13.1
1 (4
.60)
13
.13
(3.6
8)
13.3
3 (4
.11)
M
edia
n 13
.32
13.4
5 12
.14
13.5
5
Visi
t 6
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
M
ean
(SD
) 10
.29
(5.4
3)
7.92
(5.5
9)
8.06
(4.4
6)
7.64
(5.5
3)
M
edia
n 9.
64
7.00
8.
00
6.82
Chan
ge
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
betw
een
Mea
n (S
D)
-3.8
6 (6
.19)
-5
.18
(6.9
0)
-5.0
6 (4
.22)
-5
.69
(4.7
8)
Visi
t 6 a
nd
Med
ian
-3.6
4 -3
.91
-5.6
8 -5
.82
Base
line
Tr
t E�.
(SE)
-1.9
6 (1
.17)
-1
.83
(1.1
4)
-2.3
3 (1
.15)
90
% C
I
[-3.
89 to
-0.0
3]
[-3.
72 to
0.0
6]
[-4.
23 to
-0.4
3]
Re
l. D
i�. (
%)
19
.54
18.2
4 23
.23
*The
mod
el in
clud
ed t
he m
ean
TSS
at v
isit
6 a
s a
dep
end
ent
vari
able
, tre
atm
ent
as �
xed
fact
or
and
Bas
elin
e M
ean
TSS
as c
ovar
iate
Trt
E�.=
Tre
atm
ent
e�ec
t, R
el. D
i�.=
Rel
ativ
e d
i�er
ence
.
Mild
Moderate
Severe
01
23
Daily
Sym
ptom
Sco
re
(dSS
)
red
eyes
wat
ery
eyes
itchy
/bur
ning
eye
sitc
hy e
ar/p
alat
e
runn
y no
se
cong
estio
nitc
hy n
ose
snee
zing
Non-Nasal Nasal
Adjusted Mean TSS
DO
SE
p*=
0.05
7
10.0 9.5
9.0
8.5
8.0
0 20
40
60
80
emax
HAL Abstractbook 2018 A3.indd 3 13-04-18 09:54
abstract 3
TPS16 - Immunotherapy in the clinic 1
Sublingual Immunotherapy with a liquid Phleum pratense extract is effective in grass pollen allergic patients – Results of a phase II allergen exposure chamber (AEC) trial
Dose response and onset of action of sublingual Immuno-therapy with a liquid Phleum pratense extract in grass pollen allergic patients
E. Mantikou¹, P. Couroux², A. Salapatek², O. Pfaar3,4, P.J. de Kam¹, D. Yu¹, A. Narkus⁵, A. Larionov¹, D.J. Opstelten¹1HAL Allergy BV, Leiden, The Netherlands; 2Inflamax Research Inc., Mississauga, ON Canada; ³Department of Otorhinolaryngology, Head and Neck
Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany; ⁴Center for Rhinology and
Allergology, Wiesbaden, Germany; ⁵MC Narkus, Bleckede, Germany.
BackgroundAfter establishing a tolerated dose range, studies should be performed to establish a dose-response relationship for clinical efficacy. Such studies may comprise a short-term treatment (e.g. 2-4 month) with different doses in several study-arms. Allergen exposure in allergen exposure chambers (AEC) may be used as primary endpoint.
MethodIn a single-center, randomized, double blind, placebo-controlled, phase II trial with a treatment duration of 10 months a total of 168 grass pollen allergic patients (18-65 years of age) with seasonal rhinitis/rhinoconjunctivitis (ARC) with or without concomitant asthma, were randomized to sublingual immunotherapy (SLIT) with three different dosages of a liquid Phleum pratense extract (10,000 AUN/mL; 40,000 AUN/mL; 80,000 AUN/mL) or placebo. An efficacy endpoint was the mean total symptom score (TSS: sum of individual scores for eight nasal and non-nasal symptoms, rated on a scale of 0-3) at baseline, after 5 months and 10 months of treatment. For TSS assessment a validated AEC was used with a standardized controlled grass pollen exposure at an average concentration of 3500 ± 500 ppm3 for 6 hours ensuring aeroallergen exposure under controlled and reproducible conditions.
ResultsA statistically significant improvement of the TSS after 5 months compared to baseline was shown for all groups, including placebo. The results showed a dose-response curve reaching a plateau at 40,000 AUN/mL. After 10 months of treatment with the liquid Phleum pratense extract a further improvement in TSS was seen compared to placebo. However, clear differences between the various doses of the liquid Phleum pratense extract were no longer seen at this time point.
ConclusionThe results demonstrate that the allergen exposure chamber provides a suitable model to identify a clear dose response effect of treatment with a liquid Phleum pratense extract. The treatment duration was found to be of critical importance to identify differences between the various doses, suggesting that the dose may not only impact the effect-size but also the time of onset of effect.
10 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 9
Session number, date and time: PDS30, Tuesday 29 May 2018; 15:30-17:00Session title: Efficacy and safety of novel immunotherapy approaches
HAL Abstractbook 2018 A3.indd 4 13-04-18 09:54
Dose
resp
onse
and
ons
et o
f acti
on o
f sub
lingu
al
Imm
unot
hera
py w
ith a
liqu
id P
hleu
m p
rate
nse
extr
act i
n gr
ass
polle
n al
lerg
ic p
atien
ts
E. M
antik
ou1 ,
P. C
ouro
ux2 ,
A. S
alap
atek
2 , O
. Pfa
ar3,
4 , P.
J. de
Kam
1 , D.
Yu1 ,
A. N
arku
s5 , A.
Lar
iono
v1 , D.
J. O
pste
lten1
1 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s; 2 In
flam
ax R
esea
rch
Inc.
, Miss
issau
ga, O
N C
anad
a;
3 Dep
artm
ent o
f Oto
rhin
olar
yngo
logy
, Hea
d an
d N
eck
Surg
ery,
Uni
vers
itäts
med
izin
Man
nhei
m,
Med
ical
Fac
ulty
Man
nhei
m, H
eide
lber
g U
nive
rsity
, Man
nhei
m, G
erm
any;
4Cen
ter f
or R
hino
logy
and
Al
lerg
olog
y, W
iesb
aden
, Ger
man
y; 5 M
C N
arku
s, B
leck
ede,
Ger
man
y
BBaacckk
ggrroouu
nndd &&
AAiimm
::
After
esta
blish
ing
ato
lera
ted
dose
rang
e,st
udie
ssh
ould
bepe
rfor
med
toes
tabl
isha
dose
-res
pons
ere
latio
nshi
pfo
rcl
inic
aleffi
cacy
.Suc
hst
udie
sm
ayco
mpr
isea
shor
t-te
rmtr
eatm
ent(
e.g.
2-4
mon
th)
with
diffe
rent
dose
sin
seve
ral
stud
y-ar
ms.
Alle
rgen
expo
sure
inal
lerg
enex
posu
rech
ambe
rs(A
EC)
may
beus
edas
am
odel
fore
nviro
nmen
tale
xpos
ure
fore
ffica
cyas
sess
men
tofS
LIT.
MMeett
hhooddss
::
Ina
sing
le-c
ente
r,ra
ndom
ized,
doub
lebl
ind,
plac
ebo-
cont
rolle
d,ph
ase
IItr
ialw
itha
trea
tmen
tdur
ation
of10
mon
ths
ato
talo
f168
gras
spo
llen
alle
rgic
patie
nts
(18-
65ye
ars
ofag
e)w
ithse
ason
alrh
initi
s/rh
inoc
onju
nctiv
itis
(ARC
)w
ithor
with
out
conc
omita
ntas
thm
a,w
ere
rand
omize
dto
subl
ingu
alim
mun
othe
rapy
(SLI
T)w
ithth
ree
diffe
rent
dosa
ges
ofa
liqui
dPh
leum
prat
ense
extr
act(
10,0
00AU
N/m
L;40
,000
AUN
/mL;
80,0
00AU
N/m
L)or
plac
ebo.
Aneffi
cacy
endp
oint
was
the
mea
nto
tal
sym
ptom
scor
e(T
SS:
sum
ofin
divi
dual
scor
esfo
reig
htna
sala
ndno
n-na
sals
ympt
oms,
rate
don
asc
ale
of0-
3)at
base
line,
after
5m
onth
san
d10
mon
ths
oftr
eatm
ent.
For
TSS
asse
ssm
ent
ava
lidat
edAE
Cw
asus
edw
itha
stan
dard
ized
cont
rolle
dgr
ass
polle
nex
posu
reat
anav
erag
eco
ncen
trati
onof
3500
±50
0pp
m3
for
6ho
urs
ensu
ring
aero
alle
rgen
expo
sure
unde
rco
ntro
lled
and
repr
oduc
ible
c ond
ition
s.
RReessuu
llttss::
Ast
atisti
cally
signi
fican
tim
prov
emen
tof
the
TSS
after
5m
onth
s(v
isit
4)co
mpa
red
toba
selin
ew
assh
own
for
allg
roup
s,in
clud
ing
plac
ebo.
The
resu
ltssh
owed
ado
se-r
espo
nse
curv
ere
achi
nga
plat
eau
at40
,000
AUN
/mL.
After
10m
onth
s(v
isit
6)of
trea
tmen
tw
ithth
eliq
uid
Phle
umpr
aten
seex
trac
ta
furt
her
impr
ovem
ent
inTS
Sw
asse
enco
mpa
red
topl
aceb
o.H
owev
er,
clea
rdi
ffere
nces
betw
een
the
vario
usdo
ses
ofth
eliq
uid
Phle
umpr
aten
seex
trac
tw
ere
nolo
nger
seen
atth
istim
epo
int.
CCoonncc
lluussiioo
nn::
The
resu
ltsde
mon
stra
teth
atth
eal
lerg
enex
posu
rech
ambe
rpr
ovid
esa
suita
ble
mod
elto
iden
tify
acl
ear
dose
resp
onse
effec
tof
trea
tmen
twith
aliq
uid
Phle
umpr
aten
seex
trac
t.Th
etr
eatm
ent
dura
tion
was
foun
dto
beof
criti
cal
impo
rtan
ceto
iden
tify
diffe
renc
esbe
twee
nth
eva
rious
dose
s,su
gges
ting
that
the
dose
may
not
only
impa
ctth
eeff
ect-
size
but
also
the
time
ofon
set
ofeff
ect.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt..::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ssooffHH
AALL AA
lllleerrgg
yy oorr
wweerr
ee ccoo
nnttrraa
cctteedd
ffoorr tt
hhee sstt
uuddyy
bbyy HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
Tabl
e 1:
Cha
nge
from
Bas
elin
e in
Mea
n TS
S at
Visi
ts 4
and
6, p
er tr
eatm
ent g
roup
(IT
T Po
pula
tion)
Figu
re 1
: Gra
phic
al re
pres
enta
tion
of th
e ch
ange
in th
e m
ean
TSS
from
bas
elin
e to
visi
t 4 (a
) an
d vi
sit 6
(b) i
nclu
ding
the
stan
dard
err
or o
f the
mea
n, p
er tr
eatm
ent g
roup
(ITT
po
pula
tion)
Para
met
er
Stati
stics
Pl
aceb
o SP
SP
SP
(N=4
0)
10,0
00 A
UN
/mL
40,0
00 A
UN
/mL
80,0
00 A
UN
/mL
(N
=35)
(N
=38)
(N
=37)
Base
line
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
M
ean
(SD
) 14
.15
(4.2
9)
13.1
1 (4
.60)
13
.13
(3.6
8)
13.3
3 (4
.11)
M
edia
n 13
.32
13.4
5 12
.14
13.5
5
Visi
t 4
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 35
(2)
M
ean
(SD
) 11
.34
(4.8
9)
10.2
9 (5
.31)
9.
06 (4
.23)
9.
13 (5
.66)
M
edia
n 11
.59
9.09
9.
68
8.36
Chan
ge b
etw
een
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 35
(2)
Visi
t 4 a
nd
Mea
n (S
D)
-2.8
1 (4
.81)
-2
.82
(6.9
1)
-4.0
7 (4
.50)
-4
.07
(4.2
1)
Base
line
Med
ian
-2.2
3 -2
.18
-3.6
4 -3
.82
Visi
t 6
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
M
ean
(SD
) 10
.29
(5.4
3)
7.92
(5.5
9)
8.06
(4.4
6)
7.64
(5.5
3)
M
edia
n 9.
64
7.00
8.
00
6.82
Chan
ge b
etw
een
N (m
issi
ng)
40 (0
) 35
(0)
38 (0
) 37
(0)
Visi
t 6 a
nd
Mea
n (S
D)
-3.8
6 (6
.19)
-5
.18
(6.9
0)
-5.0
6 (4
.22)
-5
.69
(4.7
8)
Base
line
Med
ian
-3.6
4 -3
.91
-5.6
8 -5
.82
a. b.
-6-5-4-3-2-10Pl
aceb
oSP
10,
000A
UN
/mL
SP 4
0,00
0AUN
/mL
SP 8
0,00
0AUN
/mL
Chan
ge o
f Mea
n TS
S fr
om B
asel
ine
to V
isit 4
as c
ompa
red
to p
lace
bo
-8-7-6-5-4-3-2-10Pl
aceb
oSP
10,
000A
UN/m
LSP
40,
000A
UN/m
LSP
80,
000A
UN/m
L
Chan
ge o
f Mea
n TS
S fr
om B
asel
ine
to V
isit
6 as
com
pare
d to
pla
cebo
LS Mean TSS LS Mean TSSp=
0,29
1
p=0,
042
p=0,
050
p=0,
051
p=0,
059
p=0,
024
HAL Abstractbook 2018 A3.indd 5 13-04-18 09:54
abstract 4
PDS30 - Efficacy and safety of novel immunotherapy approaches
Dose response and onset of action of sublingual Immunotherapy with a liquid Phleum pratense extract in grass pollen allergic patients
Efficacy of SLIT with different doses of a liquid phleum pratense extract was shown during the grass pollen season and in an Allergen Exposure Chamber (AEC)
E. Mantikou¹, P. Couroux², A. Salapatek², O. Pfaar3,4, P.J. de Kam¹, A. Narkus⁵, D. Yu¹, A. Larionov¹, D.J. Opstelten¹1HAL Allergy BV, Leiden, The Netherlands; 2Inflamax Research Inc., Mississauga, ON Canada; ³Department of Otorhinolaryngology, Head and Neck
Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany; ⁴Center for Rhinology and
Allergology, Wiesbaden, Germany; ⁵MC Narkus, Bleckede, Germany.
BackgroundIn Phase III allergen immunotherapy trials, Combined Symptom Medication Score (CSMS) is the recommended clinical outcome measure. In phase II studies, Total Symptom Score (TSS) in an AEC is accepted as primary endpoint. This study was performed both in the AEC and the field to examine whether TSS is consistent with CSMS.
MethodsIn a single-center, randomized, double blind, placebo-controlled, dose range finding trial, 168 grass pollen allergic patients (18-65 years of age) with seasonal rhinitis/rhinoconjunctivitis with or without concomitant asthma, were randomized to three different dosages of a liquid phleum pratense extract (10,000 AUN/mL, 40,000 AUN/mL, 80,000 AUN/mL) or placebo for 10 months of sublingual immunotherapy (SLIT). The total symptom score (TSS: sum of individual scores for eight nasal and non-nasal symptoms, rated on a scale of 0-3) in a validated AEC was assessed at baseline and at the end of the trial, the combined symptom medication score (CSMS: 6 nasal and non-nasal symptoms and medication score, score can range from 0-6) during the grass pollen season at month 8-10.
ResultsWhen comparing the change from baseline in TSS for each active treatment group with placebo, a statistically significant improvement in TSS was found for the 80,000 AUN/mL (p=0.024) and a trend for a reduction for both the 10,000 AUN/mL group (p=0.051) and 40,000 AUN/mL group (p=0.059), at the end of the trial. The reduction in CSMS compared to placebo was substantial for all doses of SLIT with the liquid phleum pratense extract, ranging from 27% to 38% improvement and reached statistical significance for the 10,000 AUN/mL (p=0.028) and 40,000 AUN/mL (p=0.018) groups (80,000 AUN/mL group p=0.063).
ConclusionsEfficacy of SLIT with different doses of a liquid phleum pratense extract was shown using different clinical symptom scores and clinical settings. TSS during controlled exposure to grass pollen in an AEC is consistent with CSMS measured during grass pollen season. Importantly, the clinically relevant improvement in CSMS achieved at all investigated phleum pratense dose levels, provides a strong efficacy basis for the next phase of clinical development.
12 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 11
Session number, date and time: PDS30, Tuesday 29 May 2018; 15:30-17:00Session title: Efficacy and safety of novel immunotherapy approaches
HAL Abstractbook 2018 A3.indd 6 13-04-18 09:54
Effica
cy o
f SLI
T w
ith d
iffer
ent d
oses
of a
liqu
id p
hleu
m
prat
ense
ext
ract
was
sho
wn
durin
g th
e gr
ass
polle
n se
ason
an
d in
an
Alle
rgen
Exp
osur
e Ch
ambe
r (AE
C)E.
Man
tikou
1 , P.
Cou
roux
2 , A.
Sal
apat
ek2 ,
O. P
faar
3,4 ,
P.J.
de K
am1 ,
A. N
arku
s5 , D.
Yu1 ,
A. L
ario
nov1 ,
D.J.
Ops
telte
n1
1 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s; 2 In
flam
ax R
esea
rch
Inc.
, Miss
issau
ga, O
N C
anad
a;
3 Dep
artm
ent o
f Oto
rhin
olar
yngo
logy
, Hea
d an
d N
eck
Surg
ery,
Uni
vers
itäts
med
izin
Man
nhei
m,
Med
ical
Fac
ulty
Man
nhei
m, H
eide
lber
g U
nive
rsity
, Man
nhei
m, G
erm
any;
4 Cen
ter f
or R
hino
logy
and
Al
lerg
olog
y, W
iesb
aden
, Ger
man
y; 5 M
C N
arku
s, B
leck
ede,
Ger
man
y
BBaacckk
ggrroouu
nndd &&
AAiimm
::
InPh
ase
IIIal
lerg
enim
mun
othe
rapy
tria
ls,a
Com
bine
dSy
mpt
omM
edic
ation
Scor
e(C
SMS)
isth
ere
com
men
ded
clin
ical
outc
ome
mea
sure
.In
phas
eII
stud
ies,
Tota
lSym
ptom
Scor
e(T
SS)i
nan
AEC
isac
cept
edas
prim
ary
endp
oint
.Thi
sst
udy
was
perf
orm
edbo
thin
the
AEC
and
the
field
toex
amin
ew
heth
erTS
Sis
cons
isten
tw
ithCS
MS.
MMeett
hhooddss
::
Ina
sing
le-c
ente
r,ra
ndom
ized,
doub
lebl
ind,
plac
ebo-
cont
rolle
d,do
sera
nge
findi
ngtr
ial,
168
gras
spo
llen
alle
rgic
patie
nts
(18-
65ye
ars
ofag
e)w
ithse
ason
alrh
initi
s/rh
inoc
onju
nctiv
itis
with
orw
ithou
tco
ncom
itant
asth
ma,
wer
era
ndom
ized
toth
ree
diffe
rent
dosa
ges
ofa
liqui
dph
leum
prat
ense
extr
act
(10,
000
AUN
/mL,
40,0
00AU
N/m
L,80
,000
AUN
/mL)
orpl
aceb
ofo
r10
mon
ths
ofsu
blin
gual
imm
unot
hera
py(S
LIT)
.Th
eto
tal
sym
ptom
scor
e(T
SS:
sum
ofin
divi
dual
scor
esfo
rei
ght
nasa
land
non-
nasa
lsym
ptom
s,ra
ted
ona
scal
eof
0-3)
ina
valid
ated
AEC
was
asse
ssed
atba
selin
ean
dat
the
end
ofth
etr
ial,
the
com
bine
dsy
mpt
omm
edic
ation
scor
e(C
SMS:
6na
sal
and
non-
nasa
lsy
mpt
oms
and
med
icati
onsc
ore,
scor
eca
nra
nge
from
0-6)
durin
gth
egr
ass
polle
nse
ason
atm
onth
8-10
.
RReessuu
llttss::
Whe
nco
mpa
ring
the
chan
gefr
omba
selin
ein
TSS
for
each
activ
etr
eatm
ent
grou
pw
ithpl
aceb
o,a
stati
stica
llysig
nific
ant
impr
ove-
men
tin
TSS
was
foun
dfo
rth
e80
,000
AUN
/mL
(p=0
.024
)an
da
tren
dfo
rare
ducti
onfo
rbot
hth
e10
,000
AUN
/mL
grou
p(p
=0.0
51)
and
40,0
00AU
N/m
Lgr
oup
(p=0
.059
),at
the
end
ofth
etr
ial.
The
redu
ction
inCS
MS
com
pare
dto
plac
ebo
was
subs
tanti
alfo
ral
ldo
ses
ofSL
ITw
ithth
eliq
uid
phle
umpr
aten
seex
trac
t,ra
ngin
gfr
om27
%to
38%
impr
ovem
ent
and
reac
hed
stati
stica
lsig
nific
ance
for
the
10,0
00AU
N/m
L(p
=0.0
28)
and
40,0
00AU
N/m
L(p
=0.0
18)
grou
ps(8
0,00
0AU
N/m
Lgr
oup
p=0.
063)
.
CCoonncc
lluussiioo
nn::
Effica
cyof
SLIT
with
diffe
rent
dose
sof
aliq
uid
phle
umpr
aten
seex
trac
tw
assh
own
usin
gdi
ffere
ntcl
inic
alsy
mpt
omsc
ores
and
clin
ical
setti
ngs.
TSS
durin
gco
ntro
lled
expo
sure
togr
ass
polle
nin
anAE
Cis
cons
isten
tw
ithCS
MS
mea
sure
ddu
ring
gras
spo
llen
seas
on.
Impo
rtan
tly,
the
clin
ical
lyre
leva
ntim
prov
emen
tin
CSM
Sac
hiev
edat
alli
nves
tigat
edph
leum
prat
ense
dose
leve
ls,pr
ovid
esa
stro
ngeffi
cacy
basis
fort
hene
xtph
ase
ofcl
inic
alde
velo
pmen
t.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy oorr
wweerr
ee ccoo
nnttrraa
cctteedd
ffoorr tt
hhee sstt
uuddyy
bbyy HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
Figu
re 2
: Gra
phic
al re
pres
enta
tion
of th
e m
ean
CSM
S in
clud
ing
the
stan
dard
err
or o
f the
m
ean,
per
trea
tmen
t gro
up (I
TT p
opul
ation
)
Tabl
e 1:
Mea
n CS
MS
durin
g th
e po
llen
seas
on, p
er tr
eatm
ent g
roup
(ITT
Pop
ulati
on)
F igu
re 1
: Gra
phic
al re
pres
enta
tion
of th
e ch
ange
in th
e m
ean
TSS
from
bas
elin
e to
visi
t 6
incl
udin
g th
e st
anda
rd e
rror
of t
he m
ean,
per
trea
tmen
t gro
up (I
TT p
opul
ation
)
Para
met
er
Stati
stics
Pl
aceb
o SP
SP
SP
(N=4
0)
10,0
00 A
UN
/mL
40,0
00 A
UN
/mL
80,0
00 A
UN
/mL
(N
=35)
(N
=38)
(N
=37)
Sum
mar
y n
26
31
29
32
Stat
isti
cs
Mea
n (S
D)
1.03
(0.8
1)
0.68
(0.6
6)
0.64
(0.6
1)
0.75
(0.6
8)
M
edia
n 0.
92
0.50
0.
46
0.63
Tr
t E�.
(SE)
-0.3
54 (0
.183
) -0
.394
(0.1
86)
-0.2
80 (0
.182
)
90
% C
I
[-0.
657
to -0
.050
] [-
0.70
2 to
-0.0
86]
[-0.
581
to 0
.021
]
P-
valu
e#
0.
028
0.01
8 0.
063
Re
l. D
i�. (
%)
34
.27
38.1
8 27
.12
*The
mod
el in
clud
ed t
he m
ean
CSM
S as
a d
epen
dent
var
iab
le, t
reat
men
t as
�xe
d fa
ctor
#One
-sid
ed te
st
Subj
ects
com
plet
ed≥
=75
% o
f the
ir d
iary
dat
a du
ring
the
polle
n se
ason
are
incl
uded
for t
he a
naly
sis
Trt.
E�.=
Tre
atm
ent e
�ect
, Rel
. Di�
.= R
elat
ive
di�e
renc
e.
Chan
ge o
f Mea
n TS
S fr
om B
asel
ine
to V
isit
6 as
com
pare
d to
pla
cebo
LS Mean TSS
00,20,40,60,811,21,4
Plac
ebo
SP 1
0,00
0AUN
/mL
SP 4
0,00
0AUN
/mL
SP 8
0,00
0AUN
/mL
LS Mean CSMS
Mea
n CS
MS
durin
g th
e po
llen
seas
on
p=0,
028
p=0,
018
p=0,
063
-8-7-6-5-4-3-2-10Pl
aceb
oSP
10,
000A
UN/m
LSP
40,
000A
UN/m
LSP
80,
000A
UN/m
L
p=0,
051
p=0,
059
p=0,
024
HAL Abstractbook 2018 A3.indd 7 13-04-18 09:54
abstract 5
PDS30 - Efficacy and safety of novel immunotherapy approaches
Efficacy of SLIT with different doses of a liquid phleum pratense extract was shown during the grass pollen season and in an Allergen Exposure Chamber (AEC)
Tolerability of a two week Rush up-dosing with modified allergens in pollen allergic subjects in the day-to-day practice
A. Distler¹, H. van Schijndel², D. Yu², D. Pappelendam², M. Leineweber¹, N. van Os¹1HAL Allergie GmbH, Düsseldorf, Germany; ²HAL Allergy BV, Leiden, The Netherlands.
Background & AimIn two Phase IV studies with Grass1 and Birch2 pollen preparations the tolerability of a subcutaneous Rush up-dosing (three injections in two weeks) has been tested and proven to be save in adults. In the course of a non-interventional study (NIS) the tolerability of this treatment scheme was tested in the day-to-day practice. Data from all included patients who received either one or two therapies in parallel were analysed.
MethodsThe prospective NIS was performed in 116 doctor’s offices in Germany. The Rush up-dosing (Fig. 1) with 3 injections (0.1 – 0.3 – 0.5 ml) was documented in patients getting a subcutaneous immunotherapy with modified tree or modified grass pollen allergens or a mixture of those. After each injection of initial treatment with PURETHAL® (HAL Allergy BV, Leiden, The Netherlands) the patients recorded any side effects within 24 hours after the injection in a diary. By using an electronic case report form (eCRF) every participating doctor’s office documented four injections per patient, three injections of initial treatment and one injection of maintenance treatment.
ResultsIn total data from 1069 patients were collected. 529 of the patients were male (49.5%) and the mean age of the patients was 38 years (Tab. 1). 97.9% (n = 972) of the patients with a complete documentation in the eCRF (n = 993) could reach the highest dose of 0.5 ml.Data from 1048 patients were evaluable for early phase reactions. Early local reactions were reported in 15% (n = 157) and early systemic reactions in 1.1% (n = 12) of these patients. Late phase reaction data were available for 1035 patients. Late local reactions occurred in 31.6% (n = 327) and late systemic reactions in 7.8% (n = 81) of these patients (Fig. 2). No severe systemic reactions grade III and IV were observed.34.7% of the patients with available data on side effects were mono-sensitized and 65.3% of these patients were poly-sensitized. Only with regard to early local reactions a difference was determined between these two groups (12.4% vs 16.4%) (Fig. 3). 77.5% of the patients with available data on side effects suffered from rhino-conjunctivitis and 22.5% of these patients suffered from rhino-conjunctivitis and allergic asthma. Between these two groups a difference could be observed in early local reactions (13.2% vs 21.3%), late local reactions (30.4% vs 35.6%) and late systemic reactions (6.4% vs 12.9%) (Fig. 4).A subgroup of 29 patients (2.7%) out of the whole patient population (n = 1069) was treated with two therapies (e.g. Trees 100% and Grasses 100%) in parallel. Early local reactions were observed in 13.8% (n = 4), late local reactions in 10.3% (n = 3) and late systemic reactions in 3.4% (n = 1) of these patients (Fig. 2). No early systemic reactions occurred. The maintenance dose could be administered to all patients.
ConclusionMore than 97% of the patients reached the maintenance dose of 0.5 ml and the overall tolerability was very good. In the context of this non-interventional study the data from daily practice confirmed the data that were previously obtained by two Phase IV studies.
14 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 13
Session number, date and time: TPS16, Sunday 27 May 2018; 12:00-13:30Session title: Immunotherapy in the clinic 1
HAL Abstractbook 2018 A3.indd 8 13-04-18 09:54
EEAAAACC
II CCoonn
ggrreess
ss 220011
88
Gen
der
Mal
e49
.5%
Fem
ale
50.5
%Ag
e18
–45
yea
rs o
ld66
.4%
> 45
yea
rs o
ld33
.6%
Sens
itiza
tion
Mon
o-se
nsiti
zed
34.3
%Po
ly-s
ensiti
zed
65.7
%S y
mpt
oms
Alle
rgic
rhin
o-co
njun
ctivi
tis77
.3%
Alle
rgic
rhin
o-co
njun
ctivi
tisan
d al
lerg
ic a
sthm
a 22
.7%
Tabl
e 1:
Patie
nt d
istrib
ution
0%5%10%
15%
20%
25%
30%
35%
40%
early
loca
l rea
ction
late
loca
l rea
ction
rhin
o-co
njun
ctivi
tis
0%5%10%
15%
20%
25%
30%
35%
40%
early
loca
l rea
ction
late
loca
l rea
ction
0%5%10%
15%
20%
25%
30%
35%
40%
early
loca
l rea
ction
late
loca
l rea
ction
early
syst
emic
reac
tion
late
syst
emic
reac
tion
early
syst
emic
reac
tion
late
syst
emic
reac
tion
early
syst
emic
reac
tion
late
syst
emic
reac
tion
subg
roup
two
ther
apie
sal
l pati
ents
poly
-sen
sitize
d pa
tient
s
rhin
ocon
junc
tiviti
s +
asth
ma
mon
o-se
nsiti
zed
patie
nts
Figu
re 1
:Rus
h up
-dos
ing
sche
me
Figu
re 2
:Tol
erab
ility
of R
ush
up-d
osin
g
F igu
re 3
:Tol
erab
ility
in m
ono-
sens
itize
d ve
rsus
pol
y-se
nsiti
zed
patie
nts
Figu
re 4
: Tol
erab
ility
in p
atien
ts w
ith rh
ino-
conj
uncti
vitis
ver
sus
patie
nts
with
rhin
o-co
njun
ctivi
tis a
nd a
llerg
ic a
sthm
a
Tole
rabi
lity
of a
two
wee
k Ru
sh u
p-do
sing
with
mod
ified
al
lerg
ens
in p
olle
n al
lerg
ic s
ubje
cts i
n th
e da
y-to
-day
pra
ctice
.A.
Dist
ler1 ,
H. v
an S
chijn
del2 ,
D. Y
u2 , D.
Pap
pele
ndam
2 , M
. Lei
new
eber
1 , N
. van
Os1
1 HAL
Alle
rgie
Gm
bH, D
üsse
ldor
f, G
erm
any,
2 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s
Back
grou
nd&
Aim
:In
two
Phas
eIV
stud
ies
with
Gra
ss1
and
Birc
h2po
llen
prep
arati
ons
the
tole
rabi
lity
ofa
subc
utan
eous
Rush
up-d
osin
g(t
hree
inje
ction
sin
two
wee
ks)h
asbe
ente
sted
and
prov
ento
besa
vein
adul
ts.I
nth
eco
urse
ofa
non-
inte
rven
tiona
lst
udy
(NIS
)th
eto
lera
bilit
yof
this
trea
tmen
tsc
hem
ew
aste
sted
inth
eda
y-to
-day
prac
tice.
Data
from
alli
nclu
ded
patie
nts
who
rece
ived
eith
eron
eor
two
ther
apie
sin
para
llel
wer
ean
alys
ed.
Met
hods
:Th
epr
ospe
ctive
NIS
was
perf
orm
edin
116
doct
or’s
office
sin
Ger
man
y.Th
eRu
shup
-dos
ing
(Fig
.1)w
ith3
inje
ction
s(0
.1–
0.3
–0.
5m
l)w
asdo
cum
ente
din
patie
nts
getti
nga
subc
utan
eous
imm
unot
hera
pyw
ithm
odifi
edtre
eor
mod
ified
gras
spo
llen
alle
rgen
sor
am
ixtu
reof
thos
e.Aft
erea
chin
jecti
onof
initi
altr
eatm
ent
with
PURE
THAL
®(H
ALAl
lerg
yBV
,Le
iden
,Th
eN
ethe
rland
s)th
epa
tient
sre
cord
edan
ysid
eeff
ects
with
in24
hour
saft
erth
ein
jecti
onin
adi
ary.
Byus
ing
anel
ectr
onic
case
repo
rtfo
rm(e
CRF)
ever
ypa
rtici
patin
gdo
ctor
’soffi
cedo
cu-
men
ted
four
inje
ction
spe
rpati
ent,
thre
ein
jecti
ons
ofin
itial
trea
tmen
tan
don
ein
jecti
onof
mai
nten
ance
trea
tmen
t.
Resu
lts:
Into
tald
ata
from
1069
patie
nts
wer
eco
llect
ed.5
29of
the
patie
nts
wer
em
ale
(49.
5%)
and
the
mea
nag
eof
the
patie
nts
was
38ye
ars
(Tab
.1).
97.9
%(n
=97
2)of
the
patie
nts
with
aco
mpl
ete
docu
men
t-ta
tion
inth
eeC
RF(n
=99
3)co
uld
reac
hth
ehi
ghes
tdos
eof
0.5
ml.
Data
from
1048
patie
nts
wer
eev
alua
ble
for
early
phas
ere
actio
ns.
Early
loca
lrea
ction
sw
ere
repo
rted
in15
%(n
=15
7)an
dea
rlysy
stem
icre
actio
nsin
1.1%
(n=1
2)o
fthe
sepa
tient
s.La
te p
hase
reac
tion
data
wer
eav
aila
ble
for
1035
patie
nts.
Late
loca
lre
actio
nsoc
curr
edin
31.6
%(n
=32
7)an
dla
tesy
stem
icre
actio
nsin
7.8%
(n=
81)o
fthe
sepa
tient
s(F
ig.2
).N
ose
vere
syst
emic
reac
tions
grad
eIII
and
IVw
ere
obse
rved
.
34.7
%of
the
patie
nts
with
avai
labl
eda
taon
side
effec
tsw
ere
mon
o-se
nsiti
zed
and
65.3
%of
thes
epa
tient
sw
ere
poly
-sen
sitize
d.O
nly
with
rega
rdto
early
loca
lrea
ction
sa
diffe
renc
ew
asde
term
ined
betw
een
thes
etw
ogr
oups
(12.
4%vs
16.4
%)(
Fig.
3).7
7.5%
ofth
epa
tient
sw
ithav
aila
ble
data
onsid
eeff
ects
suffe
red
from
rhin
o-co
njun
ctivi
tisan
d22
.5%
ofth
ese
patie
nts
suffe
red
from
rhin
o-co
njun
ctivi
tisan
dal
lerg
icas
thm
a.Be
twee
nth
ese
two
grou
psa
diffe
renc
eco
uld
beob
serv
edin
early
loca
lrea
ction
s(1
3.2%
v s21
.3%
),la
telo
calr
eacti
ons
(30.
4%vs
35.6
%)a
ndla
tesy
stem
icre
actio
ns(6
.4%
vs12
.9%
)(Fi
g.4)
.
Asu
bgro
upof
29pa
tient
s(2
.7%
)out
ofth
ew
hole
patie
ntpo
pula
tion
(n=
1069
)w
astr
eate
dw
ithtw
oth
erap
ies
(e.g
.Tr
ees
100%
and
Gra
sses
100%
)in
para
llel.
Early
loca
lrea
ction
sw
ere
obse
rved
in13
.8%
(n=
4),l
ate
loca
lrea
ction
sin
10.3
%(n
=3)
and
late
syst
emic
reac
tions
in3.
4%(n
=1)
ofth
ese
patie
nts
(Fig
.2).
No
early
syst
emic
reac
tions
occu
rred
.The
mai
nten
ance
dose
coul
dbe
adm
inist
ered
toal
lpati
ents
.
Conc
lusio
n:M
ore
than
97%
ofth
epa
tient
sre
ache
dth
em
aint
enan
cedo
seof
0.5
ml
and
the
over
allt
oler
abili
tyw
asve
rygo
od.I
nth
eco
ntex
tof
this
non-
inte
rven
tiona
lst
udy
the
data
from
daily
prac
tice
confi
rmed
the
data
that
wer
epr
evio
usly
obta
ined
bytw
oPh
ase
IVst
udie
s.
Refe
renc
es:
(1)
Pfaa
r et a
l.: A
ccel
erat
ed U
p-Do
sing
of S
ubcu
tane
ous
Imm
unot
hera
py w
ith a
Regi
ster
ed A
llerg
oid
Gra
ss P
olle
n Pr
epar
ation
. Int
Arch
Alle
rgy
Imm
unol
2013
; 16
0:42
0-42
4(2
)Bu
czyl
koet
al.:
Acc
eler
ated
Up-
Dosin
g of
Sub
cuta
neou
s Im
mun
othe
rapy
with
a
Regi
ster
ed A
llerg
oid
Birc
h Po
llen
Prep
arati
on. I
ntAr
ch A
llerg
y Im
mun
ol20
17;
172:
183-
186
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ssooff HH
AALL AA
lllleerrgg
yy..
Wee
ks
0
1
2
4
6
8
12
...
Initi
al tr
eatm
ent
0.1
0.3
0.5
0.5
0.5
0.5
0.5
Mai
nten
ance
trea
tmen
tIn
ject
ion
inte
rval
s: 4
±2
wee
ks
HAL Abstractbook 2018 A3.indd 9 13-04-18 09:54
abstract 6
TPS16 - Immunotherapy in the clinic 1
Tolerability of a two week Rush up-dosing with modified allergens in pollen allergic subjects in the day-to-day practice
Tolerability of a two week Rush up-dosing with modified Trees, modified Grasses or modified Grasses / Trees mixture in pollen allergic subjects in the day-to-day practice
A. Distler1, H. van Schijndel2, D. Yu2, D. Pappelendam2, M. Leineweber1, N. van Os1
¹HAL Allergie GmbH, Düsseldorf, Germany; 2HAL Allergy BV, Leiden, The Netherlands.
Background & AimTwo Phase IV studies with Grass1 and Birch² pollen preparations, respectively, have led to the authorization of a subcutaneous Rush up-dosing scheme (3 injections in 2 weeks for adults). In the Phase IV study with modified grass pollen allergens 92.8% of the patients and in the study with modified birch pollen allergens 98.4% of the patients reached the maintenance dose of 0.5 ml. In order to test the tolerability of the Rush up-dosing scheme in the day-to-day practice this non-interventional study was performed. Only data from patients who received one subcutaneous immunotherapy were analysed.
MethodsThe prospective NIS was performed in 116 doctor’s offices in Germany. The Rush up-dosing (Fig. 1) with 3 injections (0.1 – 0.3 – 0.5 ml) in two weeks was documented in patients getting a subcutaneous immunotherapy with tree or grass pollen or a mixture of those. After each injection in the initial treatment with PURETHAL® (HAL Allergy BV, Leiden, The Netherlands) the patients documented in a diary any side effects within 24 hours. By using an electronic case report form (eCRF) every participating doctor’s office documented a total of four injections per patient, three injections of initial treatment and one injection of maintenance treatment.
ResultsData from 1040 patients undergoing one therapy were evaluable. Thereof 449 (43.2%) received Trees, 435 (41.8%) Grasses and 156 (15%) a mixture of Grasses and Trees (Fig. 2). 964 patients were completely documented in the eCRF. The maintenance dose of 0.5 mL was reached by 98.8% (n = 414) of the 419 patients treated with Trees, by 97.5 % (n = 385) of the 395 patients treated with Grasses and by 96% (n = 144) of the 150 patients treated with the mixture of Grasses and Trees (Fig. 3). The tolerability was analysed according to allergens and with regard to early and late local reactions (ELR, LLR) as well as early and late systemic reactions (ESR, LSR). Data from 1019 patients were available for early phase reactions and data from 1006 patients for late phase reactions. Based on these data the breakdown shown in Figure 4 was made and the percentage of patients with at least one side effect is depicted. After injection of Grasses/Trees mixture early local reactions occurred a little more frequently compared to single allergen preparations (20.9% vs Trees 13.7% and Grasses 14.3%). The frequency of late local reactions was similar for all allergen preparations (Trees 33.8%, Grasses 31.3%, Grasses/Trees 30.3%). The same was observed for early systemic reactions (Trees 1.6%, Grasses 1.2%, GraGrasses/Trees 0%) and late systemic reactions (Trees 9.4%, Grasses 6.3%, Grasses/Trees 8.6%).Severe systemic reactions (grade III and IV) did not occur.
ConclusionIn total 97.8% of the patients reached the maintenance dose of 0.5 ml. The data of this non-interventional study show that the tolerability of the single allergen preparations Trees and Grasses and the tolerability of the Grasses/Trees mixture were similar. Due to the overall good tolerability the Rush up-dosing has proven itself in day-to-day practice.
16 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 15
ELR LLR ESR LSRTrees 13.7% 33.8% 1.6% 9.4%Grasses 14.3% 31.3% 1.2% 6.3%Mixture Grasses / Trees 20.9% 30.3% 0% 8.6%In total 15% 32.2% 1.2% 8.0%
Session number, date and time: TPS35, Monday 28 May 2018; 12:15-13:45Session title: Immunotherapy in the clinic 3
HAL Abstractbook 2018 A3.indd 10 13-04-18 09:54
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
43,2
%
41,8
%15%
Tree
sG
rass
esG
rass
es/T
rees
98,8
%97
,5%
96%
0%20%
40%
60%
80%
100%
Tree
sG
rass
esG
rass
es/T
rees
0%5%10%
15%
20%
25%
30%
35%
40%
early
loca
l rea
ction
late
loca
l rea
ction
early
syst
emic
reac
tion
late
syst
emic
reac
tion
Tree
s
Gra
sses
Gra
sses
/Tre
es
in to
tal
Figu
re 4
:Tol
erab
ility
acc
ordi
ng to
diff
eren
t alle
rgen
pre
para
tions
Figu
re 1
:Rus
h up
-dos
ing
sche
me
Figu
re 2
:Dist
ributi
on o
f tre
atm
ent w
ith d
iffer
ent a
llerg
en p
repa
ratio
ns
F igu
re 3
:Per
cent
age
of p
atien
ts w
ho re
ache
d th
e hi
ghes
t dos
e
Tole
rabi
lity
of a
two
wee
k Ru
sh u
p-do
sing
with
m
odifi
ed T
rees
, mod
ified
Gra
sses
or m
odifi
ed G
rass
es/T
rees
m
ixtu
re in
pol
len
alle
rgic
sub
ject
s in
the
day-
to-d
ay p
racti
ce.
A. D
istle
r1 , H
. van
Sch
ijnde
l2 , D.
Yu2 ,
D. P
appe
lend
am2 ,
M. L
eine
web
er1 ,
N. v
an O
s1
1 HAL
Alle
rgie
Gm
bH, D
üsse
ldor
f, G
erm
any,
2 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s
Back
grou
nd &
Aim
:Tw
oPh
ase
IVst
udie
sw
ithG
rass
1an
dBi
rch2
polle
npr
epar
ation
s,re
spec
tivel
y,ha
vele
dto
the
auth
oriza
tion
ofa
subc
utan
eous
Rush
up-
dosi
ngsc
hem
e(3
inje
ction
sin
2w
eeks
for
adul
ts).
Inth
ePh
ase
IVst
udy
with
mod
ified
gras
spo
llen
alle
rgen
s92
.8%
ofth
epa
tient
san
din
the
stud
yw
ithm
odifi
edbi
rch
polle
nal
lerg
ens
98.4
%of
the
patie
nts
reac
hed
the
mai
nten
ance
dose
of0.
5m
l.In
orde
rto
test
the
tole
rabi
lity
ofth
eRu
shup
-dos
ing
sche
me
inth
eda
y-to
-day
prac
tice
this
non-
inte
rven
tiona
lstu
dyw
aspe
rfor
med
.Onl
yda
tafr
ompa
tient
sw
hore
ceiv
edon
esu
bcut
aneo
usim
mun
othe
rapy
wer
ean
alys
ed.
Met
hods
:Th
epr
ospe
ctive
NIS
was
perf
orm
edin
116
doct
or’s
office
sin
Ger
man
y.Th
eRu
shup
-dos
ing
(Fig
.1)w
ith3
inje
ction
s(0
.1–
0.3
–0.
5m
l)in
two
wee
ksw
asdo
cum
ente
din
patie
nts
getti
nga
subc
utan
eous
imm
uno-
ther
apy
with
tree
orgr
ass
polle
nor
am
ixtu
reof
thos
e.Aft
erea
chin
jecti
onin
the
initi
altr
eatm
ent
with
PURE
THAL
®(H
ALAl
lerg
yBV
,Le
iden
,The
Net
herla
nds)
the
patie
nts
docu
men
ted
ina
diar
yan
ysi
deeff
ects
with
in24
hour
s.By
usin
gan
elec
tron
icca
sere
port
form
(eCR
F)ev
ery
parti
cipa
ting
doct
or’s
office
docu
men
ted
ato
tal
offo
urin
jecti
ons
per
patie
nt,
thre
ein
jecti
ons
ofin
itial
trea
tmen
tan
don
ein
jecti
onof
mai
nten
ance
trea
tmen
t.
Resu
lts:
Dat
afr
om10
40pa
tient
sun
derg
oing
one
ther
apy
wer
eev
alua
ble.
Ther
eof
449
(43.
2%)
rece
ived
Tree
s,43
5(4
1.8%
)G
rass
esan
d15
6(1
5%)a
mix
ture
ofG
rass
esan
dTr
ees
(Fig
.2).
964
patie
nts
wer
eco
mpl
etel
ydo
cum
ente
din
the
eCRF
.Th
em
aint
enan
cedo
seof
0.5
ml
was
reac
hed
by98
.8%
(n=
414)
ofth
e41
9pa
tient
str
eate
dw
ithTr
ees,
by97
.5%
(n=
385)
ofth
e39
5pa
tient
str
eate
dw
ithG
rass
esan
dby
96%
(n=
144)
ofth
e15
0pa
tient
str
eate
dw
ithth
em
ixtu
reof
Gra
sses
and
Tree
s(F
ig.3
).
The
tole
rabi
lity
was
anal
ysed
acco
rdin
gto
alle
rgen
san
dw
ithre
gard
toea
rlyan
dla
telo
cal
reac
tions
(ELR
,LL
R)as
wel
las
early
and
late
syst
emic
reac
tions
(ESR
,LSR
).D
ata
from
1019
patie
nts
wer
eav
aila
ble
for
early
phas
ere
actio
nsan
dda
tafr
om10
06pa
tient
sfo
rla
teph
ase
reac
tions
.Bas
edon
thes
eda
tath
ebr
eakd
own
show
nin
Figu
re4
was
mad
ean
dth
epe
rcen
tage
ofpa
tient
sw
ithat
leas
ton
esi
deeff
ect
isde
pict
ed.A
fter
inje
ction
ofG
rass
es/T
rees
mix
ture
early
loca
lrea
ction
soc
curr
eda
little
mor
efr
eque
ntly
com
pare
dto
sing
leal
lerg
enpr
epar
ation
s(2
0.9%
vsTr
ees
13.7
%an
dG
rass
es14
.3%
).Th
efr
eque
ncy
ofla
telo
cal
reac
tions
was
sim
ilar
for
all
alle
rgen
prep
arati
ons
(Tre
es33
.8%
,Gra
sses
31.3
%,G
rass
es/T
rees
30.3
%).
The
sam
ew
asob
serv
edfo
rea
rlysy
stem
icre
actio
ns(T
rees
1.6%
,Gra
sses
1.2%
,G
rass
es/T
rees
0%)
and
late
syst
emic
reac
tions
(Tre
es9.
4%,
Gra
sses
6.3%
,Gra
sses
/Tre
es8.
6%).
Seve
resy
stem
icre
actio
ns(g
rade
IIIan
dIV
)did
noto
ccur
.
Conc
lusio
n:In
tota
l97.
8%of
the
patie
nts
reac
hed
the
mai
nten
ance
dose
of0.
5m
l.Th
eda
taof
this
non-
inte
rven
tiona
lstu
dysh
owth
atth
eto
lera
bilit
yof
the
sing
leal
lerg
enpr
epar
ation
sTr
ees
and
Gra
sses
and
the
tole
rabi
lity
ofth
eG
rass
es/T
rees
mix
ture
wer
esi
mila
r.D
ueto
the
over
allg
ood
tole
rabi
lity
the
Rush
up-d
osin
gha
spr
oven
itsel
fin
day-
to-
day
prac
tice.
Refe
renc
es:
(1) P
faar
et a
l.: A
ccel
erat
ed U
p-Do
sing
of S
ubcu
tane
ous I
mm
unot
hera
py w
ith a
Reg
ister
ed A
llerg
oid
Gra
ss P
olle
n Pr
epar
ation
. Int
Arc
h Al
lerg
y Im
mun
ol 2
013;
160
:420
-424
(2) B
uczy
lko
et a
l.: A
ccel
erat
ed U
p-Do
sing
of S
ubcu
tane
ous I
mm
unot
hera
py w
ith a
Reg
ister
ed A
llerg
oid
Birc
h Po
llen
Prep
arati
on. I
nt A
rch
Alle
rgy
Imm
unol
201
7; 1
72:1
83-1
86
Wee
ks
0
1
2
4
6
8
12
...
Initi
al tr
eatm
ent
0.1
0.3
0.5
0.5
0.5
0.5
0.5
Mai
nten
ance
trea
tmen
tIn
ject
ion
inte
rval
s: 4
±2
wee
ks
HAL Abstractbook 2018 A3.indd 11 13-04-18 09:55
abstract 7
TPS35 - Immunotherapy in the clinic 3
Tolerability of a two week Rush up-dosing with modified Trees, modified Grasses or modified Grasses / Trees mixture in pollen allergic subjects in the day-to-day practice
iELISA as a tool to measure IgE binding towards single modified peanut allergens
H. van Schijndel1, N. Pantelic1, R. van den Hout1, J. Meijlis1, H. van der Kleij1, 1HAL Allergy BV, Leiden, The Netherlands.
BackgroundImmunotherapy has shown to be a potential treatment for food allergies but needs further research to improve safety. Modification of peanut allergens to reduce their allergenicity is a promising approach to develop a safe and effective immunotherapy as shown by the successful completion of a first-in–human safety and tolerability study using HAL-MPE1 in adult patients with peanut allergy (EudraCT 2013-004238-13). In order to assess the impact of modification on individual peanut allergens and to assess its impact on IgE binding by individual patient sera, we have developed peanut allergen-specific inhibition ELISAs. With this methodology we are able to identify patients with residual IgE binding to modified peanut allergens.
MethodIgE inhibition ELISAs (iELISAs) were developed and performed to test IgE binding towards purified Ara h2 and Ara h6 and their reduced and alkylated (modified) versions, using the individual responses of single patient sera.
ResultsAra h6-specific iELISAs showed that modification of Ara h6 results in >95% reduction in IgE-binding for all individual sera tested. Ara h2-specific iELISAs showed that modification of Ara h2 also results in >95% reduced IgE-binding for most of the sera, but some sera were identified which showed residual, 10%-20% IgE binding to mAra h 2. In some of the latter sera, the presence of IgE binding to a linear hydroxyproline-containing peptide could be confirmed as a possible source for the residual IgE binding to mAra h2.
ConclusionWe have developed a methodology to assess residual IgE binding to modified peanut allergens. The sensitivity of the allergen-specific iELISAs allowed us to discriminate between patient sera in which IgE binding to mAra h2 and to mAra h6 was virtually completely absent and sera in which 10-20% residual IgE binding to Ara h2 was observed. The clinical importance of these observations is yet unknown. Future clinical studies will need to reveal whether the patient-specific IgE binding profiles to individual modified peanut allergens do correlate with the adverse events profile of immunotherapy with modified peanut extract.
18 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 17
Session number, date and time: TPS16, Sunday 27 May 2018; 12:00-13.30Session title: Immunotherapy in the clinic 1
HAL Abstractbook 2018 A3.indd 12 13-04-18 09:55
iELI
SAas
a to
ol to
mea
sure
IgE
bind
ing
tow
ards
sing
le
mod
ified
pea
nut a
llerg
ens
H. v
an S
chijn
del1
, N. P
ante
lic1 ,
R. v
an d
en H
out1 ,
J. M
eijli
s1 , H
. van
der
Kle
ij1
1 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s
BBaacckk
ggrroouu
nndd &&
AAiimm
::
Imm
unot
hera
pyha
ssh
own
tobe
apo
tenti
altr
eatm
ent
for
food
alle
rgie
sbu
tnee
dsfu
rthe
rres
earc
hto
impr
ove
safe
ty.
Mod
ifica
tion
ofpe
anut
alle
rgen
sto
redu
ceth
eir
alle
rgen
icity
isa
prom
ising
appr
oach
tode
velo
pa
safe
imm
unot
hera
pyas
show
nby
the
succ
essf
ulco
mpl
etion
ofa
first
-in–h
uman
safe
tyan
dto
lera
bilit
yst
udy
usin
gH
AL-M
PE1
inad
ult
patie
nts
with
pean
utal
lerg
y(E
udra
CT20
13-0
0423
8-13
).In
orde
rto
asse
ssth
eim
pact
ofm
odifi
catio
non
IgE
bind
ing
byin
divi
dual
patie
ntse
ra,
pean
utal
lerg
en-s
peci
ficin
hibi
tion
ELIS
Asha
vebe
ende
velo
ped.
With
this
met
hodo
logy
patie
nts
with
resid
ualI
gEbi
ndin
gto
mod
ified
pean
utal
lerg
ens
can
beid
entifi
ed.
MMeett
hhooddss
::
IgE
inhi
bitio
nEL
ISAs
(iELI
SAs)
wer
ede
velo
ped
and
perf
orm
edto
test
IgE
bind
ing
tow
ards
purifi
edAr
ah2
and
Ara
h6an
dth
eir
r edu
ced
and
alky
late
d(m
odifi
ed)
vers
ions
and
tow
ards
alin
ear
pepti
de(P
EP1)
,usin
g20
sera
from
pean
ut-a
llerg
icpa
tient
s.Pl
ates
wer
eco
ated
with
the
antig
ento
bete
sted
(Ara
h2,m
Ara
h2,A
rah6
,mAr
ah6
,PEP
1).E
ight
seria
ldilu
tions
ofal
lsam
ples
wer
em
ade.
The
seru
mpo
olw
asdi
lute
dac
cord
ing
topr
evio
usly
dete
rmin
edEm
axob
tain
edin
ach
ecke
rboa
rdtit
ratio
nfo
rea
chAr
aco
at.
Dete
ction
was
carr
ied
out
bya
seco
ndar
ym
ouse
anti
hum
anIg
E-ho
rser
adish
pero
xidi
se(H
RP).
Plat
esw
ere
colo
ured
with
TMB
and
the
colo
urre
actio
nw
asst
oppe
dby
addi
ng0.
5Msu
lphu
ricac
id.
Abso
rptio
nw
asm
easu
red
at45
0nm
.A
pean
utex
trac
tin
-hou
sere
fere
nce
(IHRP
)was
used
asa
cont
rola
ndst
anda
rd.
RReessuu
llttss::
•Ar
ah6
-spe
cific
iELI
SAs
show
edth
atm
odifi
catio
nof
Ara
h6re
sults
in>9
5%re
ducti
onin
IgE-
bind
ing
for
all
indi
vidu
alse
rate
sted
(tab
le1)
.•
Ara
h2-s
peci
ficiE
LISA
ssh
owed
that
mod
ifica
tion
ofAr
ah2
also
resu
ltsin
>95%
redu
ced
IgE-
bind
ing
for
mos
tof
the
sera
,bu
tso
me
sera
wer
eid
entifi
edw
hich
show
edre
sidua
l,10
%-2
0%Ig
Ebi
ndin
gto
mAr
ah
2(t
able
1).
•In
som
eof
the
sera
show
ing
resid
ualb
indi
ngto
mAr
ah2
,the
pres
ence
ofIg
Ebi
ndin
gto
alin
ear
hydr
oxyp
rolin
e-co
ntai
ning
pepti
de(P
EP1)
coul
dbe
confi
rmed
asa
poss
ible
sour
cefo
rth
ere
sidua
lIgE
bind
ing
tom
Ara
h2(fi
g.2)
.
CCoonncc
lluussiioo
nn::
We
have
deve
lope
da
met
hodo
logy
toas
sess
resid
ualI
gEbi
ndin
gto
mod
ified
pean
utal
lerg
ens.
The
sens
itivi
tyof
the
alle
rgen
-sp
ecifi
ciE
LISA
sal
low
edus
todi
scrim
inat
ebe
twee
npa
tient
sera
inw
hich
IgE
bind
ing
tom
Ara
h2an
dto
mAr
ah6
was
virt
ually
com
plet
ely
abse
ntan
dse
rain
whi
ch10
-20%
resid
ualI
gEbi
ndin
gto
Ara
h2w
asob
serv
ed.T
hecl
inic
alim
port
ance
ofth
ese
obse
rvati
ons
isye
tun
know
n.Fu
ture
clin
ical
stud
ies
will
need
tore
veal
whe
ther
the
patie
nt-s
peci
ficIg
Ebi
ndin
gpr
ofile
sto
indi
vidu
alm
odifi
edpe
anut
alle
rgen
sdo
corr
elat
ew
ithth
ead
vers
eev
ents
profi
leof
imm
unot
hera
pyw
ithm
odifi
edpe
anut
extr
act.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
Figu
re 2
: Rel
ative
IgE-
pote
ncy
of th
e PE
P1 a
t 50%
inhi
bitio
n (R
P50%
) for
se
rum
D19
, D60
, D64
, D69
, D74
, D77
and
D10
3.
Seru
m
sam
ple
Redu
ctio
n in
IgE
bind
ing
tow
ards
mAr
ah2
(%)
Redu
ctio
n in
IgE
bind
ing
tow
ards
mAr
ah6
(%)
D19
9610
0D4
499
100
D48
9810
0D5
098
100
D60
9510
0D6
296
100
D64
8210
0D6
510
010
0D6
897
100
D69
8310
0D7
010
010
0D7
491
100
D80
8910
0D8
199
100
D98
9810
0D1
0398
100
D105
9910
0D1
0795
100
D114
9610
0D7
710
010
0
Tabl
e 1.
Red
uctio
nin
IgE
bind
ing
tow
ards
mAr
ah2
and
mAr
ah6
fo
r20
sera
from
pean
utal
lerg
icpa
tient
s.
Figu
re 1
: Typ
ical
iELI
SAre
sult
of (m
)Ara
h2
for s
erum
D19
and
D44
. Fig
ure
repr
esen
ts th
e in
hibi
tion
curv
es c
alcu
late
d us
ing
the
loga
rithm
s of
the
conc
entr
ation
(µg/
mL)
on
the
x-ax
is an
d th
e in
hibi
tion
(%) o
n th
e y-
axis.
A
ra h
2 w
as u
sed
as a
refe
renc
e fo
r the
pot
ency
of m
Ara
h2.
0,0
10,0
20,0
30,0
40,0
50,0
60,0
70,0
80,0
90,0
100,
0
-2,0
-1,5
-1,0
-0,5
0,0
0,5
1,0
1,5
2,0
2,5
% inhibition
Log
µg/m
L
Ara
h 2
mAr
a h
2
RP50
%m
Ara
h2:
0.03
5
0,0
10,0
20,0
30,0
40,0
50,0
60,0
70,0
80,0
90,0
100,
0
-3,0
-2,5
-2,0
-1,5
-1,0
-0,5
0,0
0,5
1,0
1,5
2,0
2,5
% inhibition
Log
µg/m
L
Ara
h 2
mAr
a h
2
RP50
%m
Ara
h2: 0
.009
Seru
m D
19Se
rum
D44
0
0,050,
1
0,150,
2
0,250,
3
D19
D60
D64
D69
D74
D77
D103
RP 5
0%
HAL Abstractbook 2018 A3.indd 13 13-04-18 09:55
abstract 8
TPS16 - Immunotherapy in the clinic 1
iELISA as a tool to measure IgE binding towards single modified peanut allergens
An IgG inhibition ELISA to measure the stability of alum-adsorbed grass pollen allergoids
C. Franso1, N. Sinnige1, A. Segaar1, L. Boukich1, J. Meijlis1
1HAL Allergy BV, Leiden, The Netherlands.
BackgroundAn IgG inhibition ELISA was developed for the stability determination of alum-adsorbed grass pollen allergoids. Despite the complex nature of allergoid products, this assay allows to study the stability of the allergoid Drug Product by means of allergoid specific IgG.
MethodsIgG inhibition ELISA: Rabbit IgG antibodies specific for grass allergen allergoids are pre-incubated with different concentrations of alum-adsorbed grass pollen allergoid. The mix is added to an allergoid coated microtiter plate. Unbound IgG will bind to the allergoid coat and is subsequently incubated with anti-IgG HRP labeled conjugate and stained with TMB. Results are expressed as percentage inhibition relative to the uninhibited value. The concentration of alum-adsorbed allergoid that is required to inhibit 50% IgG is used as read-out. Circular Dichroism: Far-UV CD spectra (190 – 260 nm) were recorded on a J-815 Spectropolarimeter. A cuvette with a stirring compartment was used to keep the suspension homogeneous during measurement.
ResultsThe IgG inhibition ELISA assay is specific for grass pollen allergoids (not for other allergen allergoids), has a good inter- and intra-assay precision and is robust for assay variation. Thermally stressed alum-adsorbed grass pollen allergoids were used to show that the IgG inhibition assay can be used as a stability indicating method. Severe thermal stressing resulted in a higher 50% inhibition value, indicating a loss of IgG epitopes. Furthermore, far-UV CD analyses showed that there is a close relation between the decreasing IgG binding capacity (50% inhibition values) and the loss secondary protein structures by unfolding (CD-ratio 207/222 nm values).
ConclusionThe IgG inhibition assay was demonstrated to be a valuable method to determine the stability of alum-adsorbed grass pollen allergoid preparations. In addition, a relation was shown between the IgG binding capacity and the change in secondary protein structures.
20 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 19
Session number, date and time: TPS34, Monday 28 May 2018; 12:15-13:45Session title: Immunotherapy in the clinic 2
HAL Abstractbook 2018 A3.indd 14 13-04-18 09:55
An Ig
G in
hibi
tion
ELIS
A to
mea
sure
the
stab
ility
of
Alum
-ads
orbe
d gr
ass
polle
n al
lerg
oids
C. F
rans
o1 , N
. Sin
nige
1, A
. Seg
aar1
, L. B
ouki
ch1
, J. M
eijli
s1
1 HAL
Alle
rgy
BV, L
eide
n, T
he N
ethe
rland
s
BBaacckk
ggrroouu
nndd &&
AAiimm
::Ig
Eba
sed
pote
ncy
assa
ysfo
ral
lerg
ens
are
esse
ntial
test
sfo
rm
onito
ring
stab
ility
inex
trac
ts.H
owev
er,t
hese
IgE
pote
ncy
assa
ysca
nno
tbe
used
for
pote
ncy
dete
rmin
ation
ofal
lerg
oid-
base
dpr
oduc
ts,d
ueto
thei
rre
duce
dIg
Ere
activ
ity.T
here
fore
,ast
abili
tyas
say
has
been
deve
lope
dba
sed
onth
ein
hibi
tion
ofIg
Gby
gras
spo
llen
alle
rgoi
ds(D
rug
Subs
tanc
e)an
dal
um-a
dsor
bed
alle
rgoi
ds(D
rug
Prod
uct)
.Th
efe
asib
ility
ofth
eIg
Gin
hibi
tion
assa
yas
ast
abili
tyin
dica
ting
assa
yw
asst
udie
dvi
ath
erm
alst
ress
ing
ofal
um-
adso
rbed
alle
rgoi
dsin
com
bina
tion
with
mon
itorin
gth
est
ruct
ural
chan
ges
with
Circ
ular
Dich
roism
spec
tros
copy
.
MMeett
hhooddss
::Ig
Gin
hibi
tion
assa
y:Th
ede
velo
ped
IgG
inhi
bitio
nas
say
uses
IgG
antib
odie
sth
atw
ere
raise
d(in
rabb
its)a
gain
sta
mix
edgr
ass
polle
nal
lerg
oid.
IgG
antib
odie
ssp
ecifi
cfo
rgr
ass
alle
rgen
alle
rgoi
dsar
epr
e-in
cuba
ted
with
diffe
rent
conc
entr
ation
sof
alum
-ads
orbe
dgr
ass
polle
nal
lerg
oid.
The
mix
isad
ded
toan
alle
rgoi
dco
ated
mic
rotit
erpl
ate.
Unb
ound
IgG
will
bind
toth
eal
lerg
oid
coat
and
issu
bseq
uent
lyin
cuba
ted
with
anti-
IgG
HRP
labe
led
conj
ugat
ean
dst
aine
dw
ithTM
B.Re
sults
are
expr
esse
das
perc
enta
gein
hibi
tion
rela
tive
toth
eun
inhi
bite
dva
lue.
The
conc
entr
ation
ofal
um-
adso
rbed
alle
rgoi
dth
atis
requ
ired
toin
hibi
t50
%Ig
Gis
used
asre
ad-o
ut.
Circ
ular
Dich
roism
:Fa
r-U
VCD
spec
tra
(190
–26
0nm
)w
ere
reco
rded
ona
J-81
5Sp
ectr
opol
arim
eter
.A
cuve
tte
with
asti
rrin
gco
mpa
rtm
ent
was
used
toke
epth
esu
spen
sion
hom
ogen
eous
durin
gm
easu
rem
ent.
RReessuu
llttss::
Spec
ifici
ty:
The
IgG
seru
mra
ised
agai
nst
mix
edgr
ass
polle
nal
lerg
oid
onim
mun
oblo
tdet
ecte
dpr
otei
nsin
allt
hete
nin
divi
dual
gras
spo
llen
extr
acts
(Fig
ure
1).M
ajor
alle
rgen
gras
sgr
oup
1w
asde
tect
edin
the
IgG
profi
leof
allt
este
dgr
ass
spec
ies.
Furt
herm
ore,
the
IgG
seru
mpr
oves
tobe
spec
ific
inth
eIg
GEL
ISA
for
alle
rgoi
dsof
gras
ssp
ecie
ssh
owin
ga
typi
cal
dose
-res
pons
ecu
rve
whe
reas
CCoonncc
lluussiioo
nn::Th
eIg
Gin
hibi
tion
assa
yw
asde
mon
stra
ted
tobe
ava
luab
lem
etho
dto
dete
rmin
eth
est
abili
tyof
alum
-ads
orbe
dgr
ass
polle
nal
lerg
oid
prep
arati
ons.
Inad
ditio
n,a
rela
tion
was
esta
blish
edbe
twee
nth
eIg
Gbi
ndin
gca
paci
tyan
dth
ech
ange
inse
cond
ary
prot
ein
stru
ctur
esvi
ath
erm
alst
ress
ing.
Furt
herm
ore,
the
data
show
that
alum
-ads
orbe
dal
lerg
oids
are
resis
tant
tohi
gher
tem
pera
ture
s.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
tthhee
aauutthh
oorrss aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
Figu
re1:
Imm
unob
lot
alle
rgen
profi
les
ofex
trac
tspr
epar
edfr
omdi
ffere
ntgr
ass
polle
nsp
ecie
s(la
ne2-
11),
gras
spo
llen
mix
ture
(lane
1)an
dgr
ass
polle
nal
lerg
oid
(lane
12).
Gra
ssgr
oup
1al
lerg
enis
indi
cate
dw
ithan
arro
w.
Figu
re 3
: Ig
G in
hibi
tion
curv
es (l
eft) a
nd fa
r-U
V CD
spe
ctra
(rig
ht) o
f alu
m-a
dsor
bed
mix
ed
gras
ses
alle
rgoi
d st
ored
at 5
°C (B
lue)
and
exp
osed
to 9
0°C
for 4
hou
rs (o
rang
e) a
nd 8
hou
rs
(red
).
Lane
De
scrip
tion
1Re
f. m
ixed
gra
sses
2Ph
leum
prat
ense
3Lo
lium
pere
nne
4Po
apr
aten
sis5
Seca
lece
real
e6
Agro
stis
stol
onife
ra7
Arrh
enat
heum
elati
us8
Anth
oxan
thum
odor
atum
9Fe
stuc
aru
bra
10Da
ctyl
isgl
omer
ata
11H
olcu
sla
natu
s12
Mix
ed g
rass
es a
llerg
oid
MM
arke
r
Sam
ple
50%
IgG
inhi
bitio
n v a
lue
Mix
ed g
rass
al
lerg
oid
27 A
Ueq
/mL
Alum
-ads
orbe
dm
ixed
gra
sses
al
lerg
oid
85 A
Ueq
/mL
F igu
re 4
: Rel
ation
bet
wee
n th
e de
crea
sed
IgG
bin
ding
cap
acity
and
the
chan
ge o
f se
cond
ary
stru
ctur
es (u
nfol
ding
) of 3
bat
ches
(Circ
le, t
riang
le, d
iam
ond)
alu
m-a
dsor
bed
m
ixed
gra
sses
alle
rgoi
ds e
xpos
ed to
5°C
(blu
e), 9
0°C
for 4
hou
rs (o
rang
e) a
nd 9
0°C
for
8 ho
urs
(red
).
Figu
re2:
Dose
resp
onse
curv
eof
mix
edgr
asse
sal
lerg
oid,
alum
-ads
orbe
d m
ixed
Alu
m-a
dsor
bed
mix
ed g
rass
es a
llerg
oid
Alu
m-a
dsor
bed
mite
s al
lerg
oid
gras
ses
alle
rgoi
d an
d a
alum
-ads
orbe
d m
ite a
llerg
oid.
Stab
ility
ind
icati
ng:
Seve
re t
herm
al s
tres
sing
con
ditio
ns w
ere
appl
ied
to m
onito
r st
abili
ty o
f al
lerg
oids
. Exp
osur
e to
90°
C fo
r 4
and
8 ho
urs
resu
lted
in a
hig
her 5
0% in
hibi
tion
valu
e, in
dica
ting
a lo
ss o
f IgG
epi
tope
s (F
igur
e 3)
. Far
-UV
CD s
pect
ra s
how
ed th
at a
t th
ese
tem
pera
ture
s th
e el
liptic
ity a
roun
d 20
0-21
0 nm
inc
reas
ed
whi
ch is
indi
cativ
e fo
r the
pre
senc
e of
mor
e ra
ndom
coi
l str
uctu
res
(unf
oldi
ng).
Far-
UV
CD a
naly
ses s
how
ed th
at th
ere
is a
clos
e re
latio
n be
twee
n th
e de
crea
sing
IgG
bin
ding
cap
acity
(in
crea
sing
50%
in
hibi
tion
valu
es) a
nd th
e lo
ss s
econ
dary
pro
tein
str
uctu
res
by u
n-fo
ldin
g (in
crea
sed
CD-r
atio
207/
222
nm v
alue
s) (F
igur
e 4)
.
alle
rgoi
ds fr
om m
ite p
repa
ratio
ns d
o no
t inh
ibit
IgG
bin
ding
(Fig
ure
2).
Gra
ss a
llerg
oids
ads
orbe
d on
to a
lum
iniu
m s
how
a s
imila
r do
se r
es-
pons
e cu
rve
but w
ith a
redu
ced
affini
ty d
ue to
its a
dsor
bed
stat
e.
R² =
0,8
314
50100
150
200
250
300
350 0,
500,
700,
901,
101,
301,
50
50% IgG inhibition (AUeq/mL)
CD-ra
tio 2
07/2
22 n
m
250
150
100
75 50 37 25 20 15 10
250
150
100
75 50 37 25 20 15 10
M
12
34
56
78
910
11
12
M
HAL Abstractbook 2018 A3.indd 15 13-04-18 09:55
abstract 9
TPS34 - Immunotherapy in the clinic 2
An IgG inhibition ELISA to measure the stability of alum-adsorbed grass pollen allergoids
Glycosylation of rBet v 1.0101 by Chinese Hamster Ovary cells affects proteolytic stability towards endo/lysosomal degradation
H.J.M. Warmenhoven1,3,O.E. McKenna4, J.W. van Schijndel3, G. Gadermaier4, P. Briza4, R. van Ree1,2
¹Department of Experimental Immunology, ²Otorhinolaryngology, AMC, Amsterdam, The Netherlands, 3HAL Allergy BV, Leiden, The Netherlands,
⁴Department of Molecular Biology, University of Salzburg, Austria.
BackgroundMany pharmaceutical biologicals are produced in Chinese Hamster Ovary cells (CHO) because they mimic human-like posttranslational modifications such as glycosylation. A CHO cell line stably expressing the major birch pollen allergen Bet v 1.0101 produced glycosylated (glyBet v 1 CHO) and non-glycosylated Bet v 1.0101 (Bet v 1 CHO). In this study we investigated the effect of glycosylation of Bet v 1.0101 on IgE binding, secondary structural organization and proteolytic susceptibility.
MethodsBet v 1 CHO and glyBet v 1 CHO were purified from CHO cell supernatant by concanavalin A affinity and immunoaffinity. As controls we included purified glyBet v 1 digested with PNGase F to generate deglycosylated Bet v 1 (dgBet v 1 CHO) and Bet v 1 E. coli. IgE binding capacity of Bet v 1 CHO, glyBet v 1 CHO and dgBet v 1 CHO was tested with ImmunoCAP IgE inhibition using a pool of birch pollen allergic patients sera. The secondary structure of the CHO derived Bet v 1 proteins was analyzed with far UV circular dichroism (CD) spectrometry. Proteolytic susceptibility was assessed by digesting the proteins with endo-lysosomal extracts of JAWS II cells. SDS-PAGE and mass spectrometry (MS) were used to analyze the degradation kinetics and peptide patterns.
ResultsIgE inhibition curves and CD spectra of all Bet v 1 proteins were highly similar. In contrast, glyBet v 1 CHO was degraded rapidly compared to Bet v 1 E. coli and Bet v 1 CHO which was partially restored after deglycosylation. MS revealed the presence of an N-terminal aspartate amino acid and delayed degradation of a stretch amino acids upstream of the N-linked glycosylation site (Asn83).
ConclusionGlycosylation affected the proteolytic stability of Bet v 1 by dendritic cell derived endo/lysosomal proteases which in turn may affect the display of peptides complexed with MHCII molecules leading to differential T helper cell polarization. Thus, the choice of expression host plays an important role in shaping posttranslational modifications which may alter the stability and possibly immunogenicity of the target protein.
22 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 21
Session number, date and time: PDS21, Tuesday 29 May 2018; 10:30-12:00Session title: Innate immune cell activation in type-2 inflammation
HAL Abstractbook 2018 A3.indd 16 13-04-18 09:55
Gly
cosy
latio
n of
rBet
v 1
.010
1 by
Chi
nese
Ham
ster
O
vary
cel
ls aff
ects
pro
teol
ytic
stab
ility
tow
ards
en
do-/
lyso
som
al d
egra
datio
nH
. War
men
hove
n1,3 ,
O. M
cKen
na4 ,
H. v
an S
chijn
del3 ,
G. G
ader
mai
er4 ,
P. B
riza4 ,
R. v
an R
ee1,
2
1 Dep
artm
ent o
f Exp
erim
enta
l Im
mun
olog
y, 2 O
torh
inol
aryn
golo
gy, A
MC,
Am
ster
dam
, The
Net
herla
nds,
3 H
AL A
llerg
y BV
, Lei
den,
The
Net
herla
nds,
4 Dep
artm
ent o
f Bio
scie
nces
, Uni
vers
ity o
f Sal
zbur
g, A
ustr
ia
BBaacckk
ggrroouu
nndd &&
AAiimm
::
Man
yph
arm
aceu
tical
biol
ogic
als
are
prod
uced
inCh
ines
eH
amst
erO
vary
cells
(CH
O)b
ecau
seth
eym
imic
hum
an-li
kepo
sttra
nsla
tiona
lm
odifi
catio
nssu
chas
glyc
osyl
ation
.A
CHO
cell
line
stab
lyex
pres
sing
the
maj
orbi
rch
polle
nal
lerg
enBe
tv
1.01
01pr
oduc
edgl
ycos
ylat
ed(g
lyBe
tv
1CH
O)
and
non-
glyc
osyl
ated
Bet
v1.
0101
(Bet
v1
CHO
).In
this
stud
yw
ein
vesti
gate
dth
eeff
ect
ofgl
ycos
ylati
onof
Bet
v1.
0101
onIg
Ebi
ndin
g,se
cond
ary
stru
ctur
alor
gani
zatio
nan
dpr
oteo
lytic
susc
eptib
ility
.
MMeett
hhooddss
::
IgE
bind
ing:
IgE
bind
ing
capa
city
ofBe
tv
1CH
O,g
lyBe
tv
1CH
O,
dgBe
tv1
CHO
and
Betv
1E.
coli
was
test
edw
ithIm
mun
oCAP
IgE
inhi
bitio
nus
ing
apo
olof
birc
hpo
llen
alle
rgic
patie
nts
sera
and
rBet
v1
caps
(t21
5).
Stru
ctur
ean
dpr
otea
sedi
gesti
on:
The
seco
ndar
yst
ruct
ure
ofth
eCH
Ode
rived
Bet
v1
prot
eins
was
anal
yzed
with
far
UV
circ
ular
dich
roism
(CD)
spec
trom
etry
.Pr
oteo
lytic
susc
eptib
ility
was
asse
ssed
bydi
gesti
ngth
epr
otei
nsw
ithen
do-/
lyso
som
alex
trac
tsof
JAW
SII
cells
.SDS
-PAG
Ean
dm
ass
spec
trom
etry
(MS)
wer
eus
edto
anal
yze
the
degr
adati
onki
netic
san
dpe
ptide
patt
erns
1 .
RReessuu
llttss::
IgE
bind
ing
and
CDsp
ectr
aof
all
Bet
v1
prot
eins
wer
ehi
ghly
simila
r(F
igur
e1)
.In
cont
rast
,gly
Betv
1CH
Ow
asde
grad
edra
pidl
yco
mpa
red
toBe
tv
1E.
coli
and
Bet
v1
CHO
whi
chw
aspa
rtial
lyre
stor
edaft
erde
glyc
osyl
ation
(Fig
ure
2).M
Sre
veal
edth
epr
esen
ceof
anN
-ter
min
alas
part
ate
amin
oac
id(
)an
dde
laye
dde
grad
ation
ofa
stre
tch
amin
oac
ids
upst
ream
ofth
eN
-link
edgl
ycos
ylati
onsit
e(
,Fig
ure
3).
CCoonncc
lluussiioo
nn::
Gly
cosy
latio
nde
crea
sed
the
prot
eoly
ticst
abili
tyof
Betv
1ex
pose
dto
dend
ritic
cell
deriv
eden
do-/
lyso
som
alpr
otea
ses
whi
chin
turn
may
affec
tth
edi
spla
yof
pepti
des
com
plex
edw
ithM
HCI
Im
olec
ules
lead
ing
todi
ffere
ntial
The
lper
cell
pola
rizati
on2 .
Thus
,th
ech
oice
ofex
pres
sion
host
play
san
impo
rtan
tro
lein
shap
ing
postt
rans
latio
nal
mod
ifica
tions
whi
chm
ayal
ter
the
stab
ility
and,
c ons
eque
ntly,
imm
unog
enic
ityof
the
targ
etpr
otei
n.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
HH.. WW
aarrmm
eennhhoo
vveenn
aanndd
HH.. vv
aann SS
cchhiijjnn
ddeell aa
rree ee
mmppll
ooyyeeee
ss ooff HH
AALL AA
lllleerrgg
yy..EEAA
AACCII CC
oonnggrr
eessss 22
001188
Figu
re 3
: Pep
tide
patt
erns
of d
egra
ded
Bet v
1 C
HO
(top
), gl
yBet
v 1
CH
O (m
iddl
e) a
nd d
gBet
v
1 CH
O (b
ottom
), an
alyz
ed w
ith M
S. B
ox in
dica
tes
prot
ease
resis
tant
stre
tch
of a
min
o ac
ids.
Figu
re 2
: Deg
rada
tion
of B
et v
1 E
. col
i, Be
t v 1
CH
O, g
lyBe
t v 1
CH
O a
nd d
gBet
v 1
CH
O b
y en
do/ly
soso
mal
pro
teas
es, v
isual
ized
by S
DS P
AGE.
g lyB
et v
1 C
HO
Bet v
1 C
HO
dgBe
t v 1
CH
O
RReeffee
rreenncc
eess::
(1)W
ildne
r,El
sass
eret
al.2
017
(2)D
elam
arre
etal
.200
5
Figu
re 1
left
: Inh
ibiti
on o
f IgE
bin
ding
to rB
et v
1 c
aps (
t215
) by
Bet v
1 E
. col
i, Be
t v 1
CH
O,
glyB
et v
1 C
HO
and
dgB
et v
1 C
HO
. Fig
ure
1 rig
ht: F
ar U
V CD
spe
ctra
of
Bet v
1 E
. col
i, Be
t v 1
CH
O, g
lyBe
t v 1
CH
O a
nd d
gBet
v 1
CH
O.
-15
-10-505101520
190
200
210
220
230
240
250
260
[ yti ci t pill e r al o Mθ]MRW (10 3l o md 2 mc ged-1)
Wav
elen
gth
(nm
)
Bet v
1 C
HOgl
yBet
v 1
CHO
dgBe
t v 1
CHO
Bet v
1 E
. col
iE.
col
i
Proteolytic degradation (%)
50%
deg
. = 3
-5 h
ours
Tim
e (h
rs)
0
100 80 60 40 20 0
1
0,5
3
5
8
12
24
48
Bet
v 1
CH
O
Proteolytic degradation (%)
50%
deg
. = 1
-3 h
ours
Tim
e (h
rs)
0
100 80 60 40 20 0
1
0,5
3
5
8
12
24
48
dgB
et v
1 C
HO
Proteolytic degradation (%)
50%
deg
. = 0
-0.5
hou
rs
Tim
e (h
rs)
0
100 80 60 40 20 0
1
0,5
3
5
8
12
24
48
glyB
et v
1 C
HO
Proteolytic degradation (%)
50%
deg
. = 5
hou
rs
Tim
e (h
rs)
0
100 80 60 40 20 0
1
0,5
3
5
8
12
24
48
Bet
v 1
E. c
oli
050100 0,00
0001
0,00
010,
011
100
) %( noiti bi hnI
inhi
bito
r (µM
)
HAL Abstractbook 2018 A3.indd 17 13-04-18 09:55
abstract 10
PDS21 - Innate immune cell activation in type-2 inflammation
Glycosylation of rBet v 1.0101 by Chinese Hamster Ovary cells affects proteolytic stability towards endo/lysosomal degradation
Towards personalized pollen exposure measurements using hand held pollen samplers
L.A. de Weger1, K. de Raat1,2, J. de Haan1,2, M. Mostert2,3, W. van Leeuwen2,3, R.Schelland2,3, F. Molster4, P.S. Hiemstra1
¹Department of Pulmonology, Leiden University Medical Center, ²University of Applied Sciences, ³Generade, ⁴Leidse Instrumentmakers School,
Leiden, the Netherlands
BackgroundAllergic rhinitis caused by pollen is one of the most common allergic diseases. The presence of pollen in the air is currently centrally monitored at roof top levels, and not in the direct living environment of sensitized subjects. In the current project we aimed to develop a handheld pollen sampler, called pollensniffer, that can collect pollen in the living environment of the allergic subjects. As a first step this device was validated against the standard Burkard pollen sampler and used to monitor local pollen concentrations at street level in the city of Leiden.
MethodsRooftop level pollen were monitored routinely by a Hirst type pollen sampler (Burkard, UK). The pollensniffer (6x14 cm) consists of a conical inlet and a ventilator powered by a commercial powerbank. The pollen were collected on a cellulose strip. For the validation of the pollensniffer, the device was mounted on top of the Burkard sampler. All pollen collected on the strip from the Burkard and the pollensniffer were counted microscopically. Street level pollen were monitored once every week (April-June) at three locations in the city of Leiden, during the morning, midday and evening. Statistical analyses were performed using the software package STATA 14.0 (StataCorp,TX).
ResultsThe correlation between the different pollen types collected by the pollensniffer and the Burkard sampler was high (correlation coefficient [CC] > 0.8). During the validation experiments the pollensniffer appeared to collect on average 7 times more pollen than the Burkard sampler. Street level (pollensniffer) and roof top level pollen counts (Burkard sampler) showed a very good correlation (CC>0.85). Local street level measurements in the city of Leiden showed that plane trees in one park produced pollen a week before the plane trees in another park. Grass pollen were observed at street level 3 weeks before the pollen were observed at roof top level.
ConclusionPollen numbers collected by the pollensniffer and the Burkard correlate well, but the pollensniffer collected on average 7 times more pollen than the Burkard sampler. Street level measurements showed differences in pollen loads between locations. Furthermore, street level grass pollen were detected 3 weeks before they were observed at rooftop level. These findings suggest that the pollensniffer is well suited for the measurement of pollen (and maybe other allergens) in the living environment of sensitized subjects.
AcknowledgmentThis study was supported in part by a grant from Generade and RAAK Publiek (SIA).
24 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 23
Session number, date and time: TPS44, Tuesday 29 May 2018; 12:00-13:30Session title: Pollen and other aeroallergen exposition
HAL Abstractbook 2018 A3.indd 18 13-04-18 09:55
Tow
ards
per
sona
lized
pol
len
expo
sure
m
easu
rem
ents
usin
g ha
ndhe
ld p
olle
nsam
pler
sL.
A. d
e W
eger
1 , K.
de
Raat
1,2 ,
J. de
Haa
n1,2 ,
M. M
oste
rt2,
3 , W
. van
Lee
uwen
2,3
F. M
olst
er4 ,
P.S.
Hie
mst
ra1
1 Dep
artm
ent o
f Pul
mon
olog
y, Le
iden
Uni
vers
ity M
edic
al C
ente
r, 2 U
nive
rsity
of A
pplie
d Sc
ienc
es, 3 G
ener
ade,
4 Lei
dse
Inst
rum
entm
aker
s Sch
ool,
Leid
en, t
he N
ethe
rland
s
BBaacckk
ggrroouu
nndd &&
AAiimm
::
Alle
rgic
rhin
itis
caus
edby
polle
nis
one
ofth
em
ost
com
mon
alle
rgic
dise
ases
.Th
epr
esen
ceof
polle
nin
the
air
iscu
rren
tlyce
ntra
llym
onito
red
atro
oftop
leve
ls,an
dno
tin
the
dire
ctliv
ing
envi
ronm
ent
ofse
nsiti
zed
subj
ects
.In
the
curr
ent
proj
ect
we
aim
edto
deve
lop
aha
ndhe
ldpo
llen
sam
pler
,cal
led
polle
nsni
ffer,
that
can
colle
ctpo
llen
inth
eliv
ing
envi
ronm
ent
ofth
eal
lerg
icsu
bjec
ts.
Asa
first
step
this
devi
cew
asva
lidat
edag
ains
tth
est
anda
rdBu
rkar
dpo
llen
sam
pler
and
used
tom
onito
rlo
cal
polle
nco
ncen
trati
ons
atst
reet
leve
lin
the
city
ofLe
iden
.
MMeett
hhooddss
::
Rooft
ople
vel
polle
nw
ere
mon
itore
dro
utine
lyby
aH
irst
type
polle
nsa
mpl
er(B
urka
rd,
UK)
.Fo
rth
eva
lidati
onof
the
polle
nsni
ffer
(Fig
.1)
the
devi
cew
asm
ount
edon
top
ofth
eBu
rkar
dsa
mpl
er(F
ig.3
).Al
lpol
len
colle
cted
onth
est
ripfr
omth
eBu
rkar
dan
dth
epo
llens
niffe
rw
ere
coun
ted
mic
rosc
opic
ally.
Stre
etle
velp
olle
nw
ere
mon
itore
don
ceev
ery
wee
k(A
pril-
July
)at
thre
elo
catio
nsin
the
city
ofLe
iden
(Fig
.2)
,du
ring
the
mor
ning
,mid
day
and
even
ing.
RReessuu
llttss::
The
corr
elati
onbe
twee
nth
edi
ffere
ntpo
llen
type
sco
llect
edby
the
polle
nsni
ffer
and
the
Burk
ard
sam
pler
was
high
(Fig
.4)
.Du
ring
thes
eva
lidati
onex
perim
ents
the
polle
nsni
ffer
appe
ared
toco
llect
onav
erag
e6.
5tim
esm
ore
polle
nth
anth
eBu
rkar
dsa
mpl
er.
Stre
etle
vel
(pol
lens
niffe
r)an
dro
ofto
ple
vel
polle
nco
unts
(Bur
kard
sam
pler
)sh
owed
ave
rygo
odco
rrel
ation
(Fig
.6).
Loca
lst
reet
leve
lm
easu
rem
ents
inth
eci
tyof
Leid
ensh
owed
that
plan
etr
ees
inon
epa
rkpr
oduc
edpo
llen
aw
eek
befo
reth
epl
ane
tree
sin
anot
her
park
(Fig
.7)
.G
rass
polle
nw
ere
obse
rved
atst
reet
leve
l3w
eeks
befo
reth
epo
llen
wer
eob
serv
edat
roof
top
leve
l(Fi
g.8
and
9).
CCoonncc
lluussiioo
nn::
Polle
nnu
mbe
rsco
llect
edby
the
polle
nsni
ffer
and
the
Burk
ard
corr
elat
ew
ell,
but
the
polle
nsni
ffer
colle
cted
onav
erag
e6.
5tim
esm
ore
polle
nth
anth
eBu
rkar
dsa
mpl
er.
Stre
etle
vel
mea
sure
men
tssh
owed
diffe
renc
esin
polle
nlo
ads
betw
een
loca
tions
.Fur
ther
mor
e,st
reet
leve
lgra
sspo
llen
wer
ede
tect
ed3
wee
ksbe
fore
they
wer
eob
serv
edat
rooft
ople
vel.T
hese
findi
ngs
sugg
estt
hatt
hepo
llens
niffe
ris
wel
lsui
ted
for
the
mea
sure
men
tof
polle
n(a
ndm
aybe
othe
ral
lerg
ens)
inth
eliv
ing
envi
ronm
ent
ofse
nsiti
zed
subj
ects
.
IInn rree
llaattiioo
nn ttoo
tthhiiss
pprreess
eennttaa
ttiioonn,,
II ddee
ccllaarr
ee tthh
ee ffoo
llllooww
iinngg,,
rreeaall
oorr pp
eerrccee
iivveedd
ccoonn
fflliicctt
ooff ii
nntteerr
eesstt::
nnoonnee
ooff tt
hhee aa
uutthhoo
rrss hh
aavvee
ccoonnff
lliiccttss
ooff ii
nntteerr
eesstt..
EEAAAACC
II CCoonn
ggrreess
ss 220011
88
Polle
nC
CA
LN0.
77B
ET0.
92C
AR
0.88
CU
P0.
64FR
AX
0.80
HIP
0.91
POP
0.92
SAL
0.67
ULM
0.93
Figu
re 3
: Bur
kard
pol
len
sam
pler
on
the
roof
of t
he
LUM
C w
ith th
e po
llen-
sniff
er m
ount
ed o
n th
e ra
in c
over
.02468 02468 02468
02
46
02
46
02
46
ALN
BET
CAR
CU
PFR
AX
HIP
PO
PS
ALU
LM
) gol ( REFFI NSNELL OP st nuocnell oP
polle
ncou
nts
BU
RK
AR
D (l
og)
Gra
phs
by p
olle
ntyp
e
Figu
re 4
: Sim
ulta
neou
s pol
len
coun
ts (l
og) f
rom
the
Burk
ard
polle
nsam
pler
an
d th
e po
llens
niffe
r mou
nted
on
the
rain
cov
er. I
n th
e ta
ble
the
corr
elati
on
coeffi
cien
ts b
etw
een
thes
e co
unts
are
pre
sent
ed.
F igu
re 2
: Map
of t
heci
tyof
Lei
den
indi
catin
gth
elo
catio
nof
the
polle
nsam
pler
on
the
roof
of t
heLU
MC
and
the
thre
elo
catio
nsof
the
stre
etle
velm
easu
rem
ents
. Th
e in
sert
show
s th
elo
catio
nof
the
city
of L
eide
n in
the
Net
herla
nds
Figu
re 1
: Pol
lens
niffe
r con
nect
ed to
the
pow
erba
nk
Polle
n
CC
BET
0,95
SAL
0,70
PIN
0,96
QU
E0,
93
PLAT
0,16
POA
0,83
RU
M0,
85
PLA
NT
0,38
02468
log pollencount (streetlevel, pollensniffer)
01
23
45
Log
polle
ncou
nt (r
oofto
p le
vel,
Bur
kard
)
Pla
ntag
oR
umex
Bet
ula
Que
rcus
Poa
ceae
Sal
ixP
lata
nus
Pin
acea
e
Figu
re 6
: Pol
len
coun
ts (l
og) f
rom
rooft
op le
vel (
Burk
ard)
and
st
reet
leve
l (po
llens
niffe
r ). I
n th
e ta
ble
the
corr
elati
on c
oeffi
cien
ts
betw
een
thes
e co
unts
are
pre
sent
ed
F igu
re 5
: Str
eetle
vel
(pol
lens
niffe
r, le
ft) a
nd ro
oftop
le
vel (
Burk
ard
right
) pol
len
mon
itorin
g.
075150 075150 075150
1415
1617
1819
2021
2223
2425
2627
2829
1415
1617
1819
2021
2223
2425
2627
2829
1415
1617
1819
2021
2223
2425
2627
2829
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
City
Cen
ter
Hui
g
Kw
eek
t nuoc nell op
Gra
phs
by lo
catio
n
Poac
eae
010
020
0 010
020
0 010
020
0
1415
1617
1819
2021
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
mormideve
City
Cen
ter
Hui
g
Kw
eek
st nuoc nell oP
Gra
phs
by lo
catio
n
Plat
anus
Figu
re 7
:St
reet
leve
l Pla
tanu
s pol
lenc
ount
s on
thre
e lo
catio
ns in
Lei
den.
Pla
tanu
s pol
len
are
colle
cted
in
Hui
gpar
k on
e w
eek
befo
re th
ey w
ere
colle
cted
in
Kwee
klus
t par
k.F i
gure
8:
Stre
etle
vel P
oace
ae p
olle
ncou
nts
on th
ree
loca
tions
in L
eide
n. R
ainc
loud
s ind
icat
e da
ys w
ith
rain
. Fi
gure
9: C
ompa
rison
of t
he d
aily
gra
ss p
olle
n co
unts
at r
oofto
p le
vel (
red
line)
and
the
days
um o
f the
stre
etle
vel m
easu
rem
ents
(b
lue
bars
). G
reen
arr
ows i
ndic
ate
days
that
gr
ass p
olle
n w
ere
colle
cted
at s
tree
tleve
l w
hile
no
polle
n w
ere
colle
cted
in th
e Bu
rkar
d po
llens
ampl
er a
t roo
ftop
leve
l dur
ing
the
wee
ks to
whi
ch th
ese
days
bel
ong.
A ckn
owle
dgem
ents
:Th
is st
udy
was
supp
orte
d by
a g
rant
from
G
ener
ade
(cal
l 201
6-05
) and
SIA
(pro
ject
nu
mbe
r: RA
AK.P
UB0
3.04
5)
050100150200
0002004006008
l evel t eert s t a musyad
01ap
r201
701
may
2017
01ju
n201
701
jul2
017
01au
g201
7da
te
days
um s
treet
leve
l (po
llens
niffe
r)da
ys w
ith s
treet
leve
l mea
sure
men
tsro
ofto
plev
el (B
urka
rd)
daysumat streetlevel
dailycountsat rooftoplevel
PollencountsPOLLENSNIFFER (log)
log pollencount(streetlevel, pollensniffer)
Pollen counts
pollen count
Kw
eekl
ust
Hui
gpar
k
Chu
rch
-C
ityce
nter
Buck
ard
LUM
C
HAL Abstractbook 2018 A3.indd 19 13-04-18 09:55
TPS44 - Pollen and other aeroallergen exposition
Towards personalized pollen exposure measurements using hand held pollen samplers
26 EAACI, 26-30 May 2018, Munich, GermanyEAACI, 26-30 May 2018, Munich, Germany 25
Abbreviated leaflet texts.
Basic information for PURETHAL ® PollenComposition: Suspensions for subcutaneous injection, containing 20,000 AUM/ml allergenic substances chemically modified with glutaraldehyde from Grasses, Birch, Trees, Weeds pollen or mixtures of them and absorbed onto aluminium hydroxide. Excipients: sodium chloride, phenol, aluminium hydroxide, water for injections. Indication: treatment of immediate type allergic disorders (IgE-mediated), such as allergic rhinitis, allergic conjunctivitis and allergic bronchial asthma, which are triggered by sensitization to allergenic substances from pollen in adults, adolescents and children not below the age of 5 years. Dosage and administration: Treatment starts with a subcutaneous injection of 0.05 ml, which is subsequently increased with weekly doses (conventional or rush scheme) up to a monthly maintenance dose of 0.5 ml. Treatment should be carried out over a period of 3-5 successive years. Contraindications: Acute inflammatory diseases/feverish infection at the target organ, secondary changes of the target organ (emphysema, bronchi-ectasia and others), autoimmune disorders, immune deficiencies and use of immuno-suppressants, severe uncon-trolled asthma (particularly with a FEV1 persistently below 70% of the predicted value), cardiovascular failure with increased risk if using adrenaline, use of ß-blockers (including ß-blocker containing eye preparations), clinical active malignant tumour, initiation of therapy during pregnancy, or hypersensitivity to any of the excipients. Undesirable effects: Reactions generally arise within 30 minutes after receiving the injection. However, even several hours after the injection side effects can occur. After injection local reactions at the injection site may occur. In addition systemic reactions may occur which can vary from mild sneezing to life-threatening anaphylactic shock. The most commonly reported local reactions after injection are: Swelling, oedema, hypersensitivity, erythema, urticaria, rash, pruritus and granuloma of the injection site. The most commonly reported systemic reactions are: Sneezing, cough, (allergic) rhinitis, throat irritation, asthma, dyspnoea, oropharyngeal pain, nasal congestion, nasal discomfort, nasal oedema, rhinorrhoea, nasopharyngitis, conjunctivitis, allergic rhino conjunctivitis, eye irritation, eye pruritus, lacrimation increased, angioedema, atopic dermatitis, urticaria, pruritus, erythema, swelling, fever, asthenia, nausea, abdominal pain, diarrhoea, dizziness, headache, somnolence, impaired concentration. The complete product information is available on request. HAL Allergy BV, Postbus 1204, 2302 BE Leiden, the Netherlands. Date: February 2015
Note: PURETHAL® Pollen is available as registered product or named patient prescription. In case of a registered product the locally approved product information can slightly differ from above.
Basic information for PURETHAL® MitesComposition: Suspension for subcutaneous injection containing 20,000 AUeq/ml allergenic substances chemically modified with glutaraldehyde from Dermatophagoides pteronyssinus (50%) and Dermatophagoides farina (50%) and absorbed onto aluminium hydroxide.Excipients: sodium chloride, phenol, aluminium hydroxide, water for injections. Indication: Treatment of immediate type allergic disorders (IgE-mediated), such as allergic rhinitis, allergic conjunctivitis and allergic bronchial asthma, which are triggered by sensitization to allergenic substances from house dust mites in adults, adolescents and children not below the age of 5 years. Dosage and administration: Treatment starts with a subcutaneous injection of 0.05 ml, which is subsequently increased with weekly doses up to a monthly maintenance dose of 0.5 ml. Treatment should be carried out over a period of 3-5 successive years. Contraindications: Acute inflammatory diseases/feverish infection at the target organ, secondary changes of the target organ (emphysema, bronchi-ectasia and others), autoimmune disorders, immune deficiencies and use of immuno-suppressants, severe uncontrolled asthma (particularly with a FEV1 persistently below 70% of the predicted value), cardiovascular disorders with increased risk if using adrenaline, use of ß-blockers (including ß-blocker containing eye preparations), clinical active malignant tumour, initiation of therapy during pregnancy, or hypersensitivity to any of the excipients. Undesirable effects: Reactions generally arise within 30 minutes after receiving the injection. However, even several hours after the injection side effects can be observed. After injection of PURETHAL® Mites mixture, local reactions at the injection site may occur. In addition systemic reactions may occur. These can vary from mild sneezing to life-threatening anaphylactic shock. The most commonly reported local reactions after injection are swelling, erythema, warmth, paraesthesia, pain, induration, haemorrhage, and pruritus of the injection site. The most commonly reported systemic reactions are: Hypersensitivity, headache, malaise, fatigue, nausea, conjunctivitis, eye pruritus, lacrimation increased, dyspnoea, nasal congestion, sneezing, allergic rhinitis, rhinorrhoea, erythema, pruritus, swelling, atopic eczema, angioedema. The complete product information is available on request. HAL Allergy BV, Postbus 1204, 2302 BE Leiden, The Netherlands. Date: August 2015
Note: PURETHAL® Mites mixture is available as named patient prescription.
Basic information for SUBLIVAC® and SUBLIVAC® FIXComposition: Sublingual drops, containing per ml 10,000 AU, AUN or PUN allergen extract, prepared according to the individual doctor’s prescription. Excipients: glycerol, aminocaproic acid, disodium phosphate hydrate, sodium dihydrogen phosphate dihydrate, peppermint oil, purified water.Indication: Treatment of immediate type allergic disorders (IgE-mediated) such as allergic rhinitis, allergic conjunctivitis and allergic bronchial asthma, caused by allergens.Dosage and administration: The first day of the initial treatment starts with one drop. This dose is increased every day with one drop until the highest daily dose of five drops is reached. The treatment is continued with five drops. The drops have to be administered under the tongue for at least 1 minute (preferably 2-3 minutes) before swallowing. A spoon may be used to administer the drops. It is advised to clean the dropper after use, for instance with a wet tissue. The treatment should be continued for 3 to 5 years. Contraindications: Partly or uncontrolled bronchial asthma with a FEV1 below 70%, severe autoimmune diseases, immune deficiencies and immunosuppression, malign neoplastic diseases with current symptoms, initiation of treatment during pregnancy, severe inflammation of the oral mucosa, hypersensitivity to any of the excipients. Undesirable effects: Local reactions in the mouth and throat, swelling of the lips or tongue. Reappearance of the patient specific allergic symptoms such as mild systemic reactions (itching eyes, sneezing, coughing, atopic eczema). In rare cases, intensified systemic reactions, like shortness of breath, generalized urticaria, or Quincke’s oedema, can occur. After intake, the patient might experience diarrhoea and abdominal pain. These symptoms generally arise within 30 minutes after intake of the drops, however can occur several hours after. In individual cases, anaphylactic shock has been reported. The complete product information is available upon request at HAL Allergy BV, P.O. Box 1204, 2302 BE Leiden, The Netherlands. Date: October 2016
Note: SUBLIVAC® and SUBLIVAC® FIX are available as named patient prescriptions.
HAL Abstractbook 2018 A3.indd 20 13-04-18 09:55
EAACI, 26-30 May 2018, Munich, Germany 27
Basisinformationen PURETHAL ®
Bezeichnungen und Zusammensetzung: Suspensionen zur subkutanen Injektion, enthalten an Aluminiumhydroxid adsorbierte, mit Glutaraldehyd chemisch modifizierte allergene Substanzen aus Pollen (20.000 AUM/ml) oder Milben (20.000 AUeq/ml): PURETHAL Gräser: Pollen von 10 Gräsern (inkl. Roggen); PURETHAL Gräser + Getreide: Pollen von 9 Gräsern (45%), Roggen (5%) und Weizen (50%); PURETHAL Birke: Birkenpollen; PURETHAL Bäume: Pollen von Birke, Erle, Hasel zu gleichen Teilen; PURETHAL Gräser + Birke: Pollen von 10 Gräsern (50%) und Birke (50%); PURETHAL Gräser + Bäume: Pollen von 10 Gräsern (50%) und 3 Bäumen (Birke, Erle, Hasel mit insgesamt 50%); PURETHAL Beifuß: Beifußpollen; PURETHAL Milbenmischung: Derm. pteronyssinus (50%) und Derm. farinae (50%). Sonstige Bestandteile: NaCl, Phenol, Aluminiumhydroxid, Wasser zur Injektion. Anwendungsgebiete: Spezifische Immuntherapie allergischer Erkrankungen vom Soforttyp (IgE-vermittelt), wie Heuschnupfen (allergische Rhinitis), allergische Bindehautentzündung (Konjunktivitis) und allergisches Asthma bronchiale, ausgelöst durch eine Sensibilisierung gegenüber den enthaltenen allergenen Substanzen. Gegenanzeigen, absolute: Akute Entzündungsprozesse/Infektionskrankheiten am Reaktionsorgan; Sekundärveränderungen am Reaktionsorgan (z.B. Emphysem, Bronchiektasen); Autoimmunerkrankungen; Immundefekte; gleichzeitige Anwendung von Immunsuppressiva; schweres, unkontrollierbares Asthma bronchiale, insbesondere bei einem persistierenden FEV1 unter 70% Sollwert; Erkrankungen mit Kontraindikationen gegen die Anwendung von Adrenalin; gleichzeitige Behandlung mit ß-Blockern (auch ß-Blocker enthaltende Augentropfen); maligne Tumorerkrankungen mit aktuellem Krankheitswert; Einleitung der Behandlung nicht während der Schwangerschaft; Sensibilisierung gegenüber einem der sonstigen Bestandteile; relative: Schwangerschaft und Stillzeit; akute allergische Beschwerden; nicht für Kinder unter 5 Jahren. Zum zeitlichen Intervall zu Schutzimpfungen und für weiterführende Informationen siehe Fachinformation. Nebenwirkungen: Allergische Lokal- und/oder Allgemeinreaktionen. Überempfindlichkeit, anaphylaktischer Schock. Grippe, Otitis externa, Sinobronchitis, Sinusitis, Staphylokokkenpharyngitis. Essstörung. Kopfschmerz, Schwindelgefühl, Schläfrigkeit, Parästhesie, Geschmacksstörung, Aufmerksamkeitsstörungen. Schwellung des Auges, Konjunktivitis, Rhinokonjunktivitis, Augenreizung, Augenjucken, Tränensekretion verstärkt. Schwindel, Ohrschwellung, Ohrenjucken. Arrhythmie, Tachykardie. Kreislaufkollaps, Hitzegefühl, Hämatom. Rhinitis, Nasenverstopfung, Nasenödem, Rhinorrhoe, Niesen, Asthma, Atemnot, Husten, Bronchitis, Halstrockenheit, Rachenreizung, Rachenschmerzen, Kehlkopfirritation, Nasopharyngitis, Nasenbluten. Abdominalschmerzen, Gastritis, Übelkeit, Diarrhoe. Angioödem, Urtikaria, Erythem, Juckreiz, Ekzem, atopische Dermatitis, Ausschlag, Akne, Hautreizung. Muskuloskelettale Beschwerden. Ödem, Schwellung, Ermüdung, Schwäche, Brustschmerz, Blässe, Fieber. An der Injektionstelle: meist vorübergehende Granulome, Verhärtung, Schwellung, Urtikaria, Erythem, Überempfindlichkeit, Juckreiz, Schmerz. Patienten nach der Injektion mindestens 30 Minuten überwachen, eine Schockapotheke muss bereitgestellt sein. Nebenreaktionen können auch noch zu einem späteren Zeitpunkt auftreten. Für weiterführende Informationen und zur Behandlung von Nebenwirkungen siehe Fachinformation. In seltenen Fällen kann nach der Injektion leichte Müdigkeit auftreten, was beim Führen von Kraftfahrzeugen oder beim Bedienen von Maschinen zu berücksichtigen ist. Hinweis: Rezept- und apothekenpflichtig. Weitere Angaben zu Warnhinweisen und Vorsichtsmaßnahmen für die Anwendung, Wechselwirkungen mit anderen Arzneimitteln und sonstigen Wechselwirkungen, Schwangerschaft und Stillzeit und Nebenwirkungen sind der veröffentlichten Fachinformation zu entnehmen. (Stand: 04/2018)
Basisinformationen SUBLIVAC® FIX / SUBLIVAC® PollenpräparateBezeichnungen und Zusammensetzung: Allergenlösungen zur sublingualen Immuntherapie. 1 ml enthält 10.000 AUN oder PUN. SUBLIVAC FIX Bäume: Pollen von Birke, Erle, Hasel zu gleichen Teilen; SUBLIVAC FIX Birke: Birkenpollen; SUBLIVAC FIX Gräser: Pollen von 3 Gräsern; SUBLIVAC FIX Gräser+Roggen: Pollen von 3 Gräsern (50%) und Roggen (50%); SUBLIVAC FIX Lieschgras: Pollen aus Lieschgras. SUBLIVAC enthält Allergenextrakte nach individueller ärztlicher Rezeptur. Bezeichnung und Stärke der Allergene s. Etikett. Sonstige Bestandteile: Glycerol, Wasser, 6-Aminohexansäure (ε-Amino-Capronsäure/EACA), Dinatrium-hydrogenphosphat, Natrium dihydrogenphosphat, Pfefferminzöl. Anwendungsgebiete: Spezifische Immuntherapie allergischer Erkrankungen vom Soforttyp (IgE-vermittelt). Gegenanzeigen: akute Entzündungsprozesse/Infektionskrankheiten am Reaktionsorgan; Schwere Autoimmunerkrankungen; Immundefekte (auch durch Immunsuppressiva induziert); Krebserkrankungen mit aktuellem Krankheitswert; schweres oder unzureichend behandeltes Asthma (FEV1 < 70% vom Sollwert); Überempfindlichkeit gegenüber einem der sonstigen Bestandteile; Infektionen des Mund-/Rachenraumes; nach zahnärztlicher Behandlung (z.B. Zahnentfernung). Eine Hyposensibilisierungsbehandlung soll nicht während der Schwangerschaft begonnen werden. Zum zeitlichen Intervall zu Schutzimpfungen und für weiterführende Informationen siehe Fachinformation. Nebenwirkungen: Lokale Beschwerden, Schwellungen oder Juckreiz des Mundes, der Lippen und der Zunge, Mundtrockenheit, Geschwüre im Mund, Aphthen. Allergische Reaktionen wie Niesen, laufende oder verstopfte Nase, Nasenbeschwerden; Juckreiz oder Beschwerden des Ohrs; Juckreiz, Rötung, Tränen oder Brennen der Augen, Augenentzündung; Halsreizungen, Husten, Atemnot. Schwierigkeiten beim Schlucken, Magenbeschwerden, Erbrechen, Übelkeit, Bauchschmerzen, Appetitlosigkeit, Verdauungsstörungen. Ekzeme, Hautausschlag, Nesselsucht, schnelle oder starke Schwellung der Haut. In sehr seltenen Fällen anaphylaktische Reaktionen. Hinweis: Rezept- und apothekenpflichtig. Weitere Angaben zu Warnhinweisen und Vorsichtsmaßnahmen für die Anwendung, Wechselwirkungen mit anderen Arzneimitteln und sonstigen Wechselwirkungen, Schwangerschaft und Stillzeit und Nebenwirkungen sind der veröffentlichten Fachinformation zu entnehmen. (Stand: 04/2018)
HAL Allergie GmbH, Poststraße 5 – 6, D-40213 DüsseldorfHAL Allergy Handelsgesellschaft mbH, Johnstraße 4, A-1150 Wien
HAL Abstractbook 2018 A4.indd 7 13-04-18 09:57
Notes
HAL Abstractbook 2018 A4.indd 8 13-04-18 09:57
Notes
HAL Abstractbook 2018 A4.indd 9 13-04-18 09:57
Notes
HAL Abstractbook 2018 A4.indd 10 13-04-18 09:57
International Offices
Headquarters
The NetherlandsHAL Allergy GroupJ.H.Oortweg 15-17 2333 CH Leiden
P.O. Box 12042302 BE Leiden
Tel.: +31-(0)88-1959 000www.hal-allergy.com
Offices
GermanyHAL Allergie GmbHPoststraße 5-640213 DüsseldorfTel.: +49-(0)211-977650www.hal-allergie.de
AustriaHAL Allergy Handels-GmbHJohnstraße 41150 ViennaTel.: +43 (0)1-9859880www.hal-allergy.at
PolandHAL Allergy Sp. z o.o. Ul. Rumiana 6502-956 WarsawTel.: +48-(0)22-8581614www.hal-allergy.pl
SpainHAL Allergy S.L.UParque Empresarial Mas Blau IIAvda. Les Garrigues 4608820-El Prat de LlobregatBarcelonaTel.: +34-902-110-686www.hal-allergy.es
ItalyHAL Allergy s.r.l.Piazzale Asia, 21Scala B, Piano 4, Interno 21, 00144 RomaTel: +39-06-97243570www.hal-allergy.it
Belgium / BeneluxHAL Allergy Benelux B.V.J.H. Oortweg 15-172333 CH LeidenThe NetherlandsTel.: +31-(0)88-1959 000www.hal-allergy.nl
Distributors
EstoniaBalti Intermed OÜTartu mnt 84d10112 TallinnTel.: +372-601-4140
GreeceAlfamedica S.A.Sofokleous 1bElliniko 16777Tel.: +30-210-6728318/19
PortugalA.M.D. Passos, Lda.Rue Cidade de Coimbra no. 15A 1,Parede, 2775 – 180 LisbonTel.: +351-21-4578087
HungaryHungaropharma Zrt.1061 BudapestKirály u.12Tel.: +36-1-327-6700
SloveniaIRIS Mednarodna Trgovina D.O.O.Cesta v Gorice 81000 LjubljanaTel.: +386-1-200-6684
HAL Abstractbook 2018 A4.indd 11 17-04-18 14:29
www.hal-allergy.com © HAL Allergy 2018 | MAB48094/01
Rising to the ch a l lenge of improving a l l e r g y treatmentsRising to the c
HAL Abstractbook 2018 A4.indd 12 13-04-18 09:57
