Managua 2014 Institute of Influenza, Saint-Petersburg, (Research Institute of Influenza, Saint-Petersburg, WHO National Influenza Center of RUSSIA) Vaccination

Managua2014

(Research Institute of Influenza, Saint-Petersburg,WHO National Influenza Center of RUSSIA)

Vaccination and the development of cross-reactive

vaccines in Russia

L.M. Tsybalova

Vaccine type Vaccinetitles

Manufacturing companies

Vaccine composition

Live influenza vaccine Ultravak Microgen (Russia)

Contains 15 µg of each influenza virus strain

To be used since 3 years of age

Inactivated whole virion vaccine

GrippovacSPb NIIVS (Russia) Contains 10 µg of each influenza

virus A strains and 13 µg of influenza virus B strain

Split vaccines

Ultrix

FluarixVaxigripBegrivacFluvaxinInflexal V

SPb NIIVS (Russia)

GSK-BiomedSanofi

NovartisChina

Berna Biotech(Switzerland)

Contains 15 µg of influenza virus strains and virus wall

reactogenic lipoproteins

Subunit vaccines

Grippol

Grippol plus

Sovigripp

Microgen

Pharmaceutical Company «PETROVAX PHARMA

Microgen

Contains 5 µg of each influenza virus strain and adjuvants

polyoxidonium and sovidon

Influvac

Agrippal

Abbott Products (former Solvay)Novartis

Contains 15 µg of each influenza virus strain

Vaccines used for influenza prophylaxis in Russia

Production of influenza vaccines in Russia (thousands of doses)

Blue – live vaccinesRed – whole virion vaccinesGreen – subunit vaccine

Vaccination against Influenza in compliancewith the National Immunization Schedule

Order № 125Н as of 21.03.14 of the Ministry of Health of Russia

• Children since 6 months; schoolchildren of 1-11 classes; • Students of technical colleges and High Educational Schools (Institutes, Universities); • Adults at high professional risk of infection (personnel of medical and educational institutions, transport and community services); • Pregnant women; • Adults over 60 years; • Persons liable for military service;• Persons with chronic pathology(pulmonary and cardio-vascular diseases, metabolic disorders and obesity).

25,08

18

27,46 27,9531,9 33,6

6,13

8,7

7,00 6,25

5,24,1

0

5

10

15

20

25

30

35

40

2007 2008 2009 2010 2011 2012

Национальный проект другие источники

мл

н.

чел

.

37,737,1134,2034,4626,7031,21

IMMUNIZATION COVERAGE in the RUSSIAN FEDERATION (MLN of POPULATION)

Pre-morbid background of the persons died of Influenza during 2009-2010 epidemic season

0

2

4

6

8

10

1210,2

6,7

9,6

4,7

8,3

7,0

3,5

2,1

%

Efficacy of Grippol-plus vaccinein patients with chronic pulmonary pathology (n=43)

Bronchial asthma (BA) • Morbidity rate: 5-7% of adult population • Influenza-related exacerbations in 70% of cases•WHO recommendations: annual vaccination against influenza is mandatory for all BA patients

Chronic obstructive pulmonary disease (COPD) • Registered number of patients: 2,4 mln• Disease exacerbation caused by influenza: 30% of cases• Lethal outcomes caused by ARVI and pneumonia: 20% of cases

Investigated indexPrior to

vaccination

6 months post-

vaccination

Fold decrease

Frequency of the underlying disease exacerbations 1,47±0,12 0,70±0,11* 2,1

Average number of antibiotic treatment courses per capita 0,7±0,1 0,4±0,1* 1,8

Frequency of out-patient visits 6,22±0,49 3,13±0,48* 1,9

Mid-value for hospitalization rates 1,23±0,11 0,42±0,09* 2,9

Source: Chebykina A.V. et al. New opportunities for influenza prevention ... / Russian Medical Journal, September, 2010

4%

86%

10%

2010-2011

4%

90%

6%

vaccinatednon vaccinatedno data

2011-2012

1%

97%

3%

2012-2013

PROPORTION OF VACCINATED PERSONS AMONG SARI CASES WITH INFLUENZA ETIOLOGY,

SEASONS 2010-2013

Research Institute of Influenza

Vaccination of pregnant women • During epidemics a total of 25,5% of pregnant women are diagnosed withinfluenza and ARI* • Influenza complications among pregnant women: 804 per 10 000 influenza patients• Hospitalization rates among pregnant women infected during the III trimester of pregnancy, being consistent with hospitalization level among chronically ill persons aged 15-44 years (5,6 - 11,0 per 10 000)*• During epidemics hospitalization rates of pregnant (III trimester) female influenza patients with disorders of cardio- vascular and pulmonary systems are estimated as 21,76 per 10 000, with diabetes mellitus: 44,9 per 10 000*

A number of clinical signs Grippol plus (n=32) abs.

valuesPlacebo (n=29) abs. values

Hyperemia at the injection site 4 0

Increase fatigue 6 12

Mialgias 0 3 p<0,05

Effect on homeostasis and pregnancy course of vaccination against influenza during pregnancy correlates

with values in the group of placebo recipients **

Sources: * L. Dodds et al., 2007, W. Swing et al., 2000, K. Neuzil et al., 1998 ** M.P.Kostinov, A.P.Cherdantsev et al., 2010

Dynamics of post-vaccination protective antibody level against influenza А(H1N1)pdm09 virus in pregnant women

p > 0,05 differences between comparison groups

M.P.Kostinov, A.P.Cherdantsev et al., 2010 -2014

Baseline level

After 1 month

After 3-4 months

DELIVERY 3 months post-

delivery

6 months post-

delivery

Grippol plus

Agripal

Non-pregnant females

Influenza and ARVI morbidity (per 1000 population) in the study and control groups during 2013-2014 epidemic season

Selkova E.P. et al. G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology under Rospotrebnadzor “Studies to evaluate epidemiological effectiveness and safety of Ultrix® inactivated split influenza vaccine”, 2014

Closed communities

Medical personnel

Population of Timashevsk city

Vaccinated

Non-vaccinated

Average duration (days) of influenza/ARVI cases and % of complications in patients immunized with ULTRIX® vaccine and non-immunized persons

Vaccinated

Non-vaccinated

Duration, days

Selkova E.P. et al. G.N. Gabrichevsky Research Institute for Epidemiology and Microbiology under Rospotrebnadzor “Studies to evaluate epidemiological effectiveness and safety of Ultrix® inactivated split influenza vaccine”, 2014

Epidemiological efficacy of Ultrix ® vaccine

Volunteers (male and female) aged 18 through 60 years – 5 611

3107 persons were vaccinated by Ultrix® vaccine, 2504 volunteers were not vaccinated

• Ultrix® vaccine good tolerability and not-reactogenicity was established

• Ultrix® vaccine application safety based on clinical and laboratory results was established

• Specific antibody long-lasting activity (HI test data) in immunized persons in 6 months post-vaccination was demonstrated

• Ultrix® vaccine efficacy coefficient in relation to laboratory confirmed is estimated as 86,5 %, and efficacy index as 7,4

Developmentof cross-protective

(universal) vaccines

Schematic Diagram Summarizing the Pathway of Vaccine Development Starting from Reverse Vaccinology

(8) The approved vaccine is then commercialized and used in large scale. At this point, phase IV clinical studies confirm safety.

(7) Policy-making bodies, such as the ACIP and equivalent bodies from other nations, make the recommendation on how the vaccine should be used.

(6) Scientific, clinical, and technical information is then analyzed and approved by regulatory agencies, such as the Food and Drug Administration (FDA) or the European Medicinal Agency (EMA).

(5) Finally, selected antigens are manufactured in large scale for clinical trials, and candidate vaccines are tested for safety and protective immunity in humans using established correlates of protection or efficacy studies.

(4) Protective antigens are then tested for their presence and conservation in a collection of strains representative of the species (molecular epidemiology).

(3) The selected antigens are then used to immunize animals and test whether immunization induces a protective response.

(2) Then the identified antigens are screened for expression by the pathogen and for immunogenicity during infection.

(1) First, computer analysis of the whole genome identifies the genes coding for predicted antigens and eliminates antigens with homologies to human proteins.

Subunit vaccines

Recombinant vaccines

Virus proteins included into composition of traditional and “universal” vaccines

NP

Influenza virus surface proteins М2е and NA

SLLTEVETPI – HLA-A 2 (0201-0210) 48% *

SSDLSSLL – HLA-A 1 19.1% *

SSDLSSLLTEV – HLA-A 1 19.1% *

DSSDLSSLLTEV – HLA-A 26 7.4% *

GELSSLLTEV – HLA-B 44 н/д

A human consSLLTEVETPIRNEWGCRCNDSSDL SLLTEVETPIRNEWGCRCNDSSD

* - frequency in the population

Presentation of the M2e peptide by antigens HLA I class

RECOMBINANT VACCINES DEVELOPED at the RESEARCH INSTITUTE of INFLUENZA (based on M2e protein)

Expression systems

Bacterial(E coli) Plant (Nicotiana Benthamiana)

M2e HBcM2e+HBc

M2e flagellin M2e+flagellin

M2e HBc TMV М2е

Variations:• number of М2е copies • М2е type • insertion site

Electronic photography of virus-like particles HBc (A) and recombinant protein 4M2hHBc (В)

А

B


Longevity of M2e specific antibody(IgG) in mice

Series10

100000

200000

300000

400000

500000

600000

700000

80000042 days 162 days 192 days

G-11-1 (H5N1) G-26 A(H1N1)pdm09

titer

s of

ser

um a

nti-M

2e Ig

G

p<0.05

p<0.01


Titers of IgG antibodies against M2e in BAL upon immunization of mice with candidate vaccines (50μg recombinant protein+100 μg derinatx3). BAL

of mice were collected on 14th day after third immunization.* Differences between immunized and control groups were significant (p ˂

0.01)

G-370

200

400

600

800

1000

1200

1400

1600

1800

1200*

50

PBS

4M2e(h)HBc

G-11-1 (H5N1) G-26 (H1N1pdm)0

200

400

600

800

1000

1200

1400

1600

1800

1520*1600*

50 50

4M2e(k)HBc


Longevity of M2e specific T-cell responses of 4M2e(k)HBc immunized mice

CD4 – IL-4

CD4 – IFNɣ

CD8 – IFNɣ


Dynamics of body weight and death of mice immunized with the candidate vaccine 4M2e(h)HВc+derinat (50 μg x 3 i/m) post-challenge with lethal

dose (5LD50) of influenza virus APR/8/34 or A/Aichi/1/68

0 2 4 6 8 10 12 1450

60

70

80

90

100

110

4M2e(h)HBc control Day after challenge

Body

w

eigt

h (%

)

50

60

70

80

90

100

110

0 2 4 6 8 10 12 14

Bo

dy

w

eig

th (

%)

Day after challenge

4M2e(h)HBc control

0

20

40

60

80

100

120

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Pe

rce

nt

su

rviv

al

(%)

Day after challenge

4M2e(h)HBc control

0

20

40

60

80

100

120

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Pe

rce

nt

su

rviv

al

(%

)

Day after challenge

4M2e(h)HBc control HBc

APR/8/34

APR/8/34

A/Aichi/1/68

A/Aichi/1/68


Dynamics of body weight and death of mice immunized with the candidate vaccine 4M2e(k)HBc derinat(50 μg x 3 i/m) post-challenge with 5LD50 of

influenza virus А/Kurgan/05/05 (Н5N1) or A/California/07/09

50

60

70

80

90

100

110

0 2 4 6 8 10 12 14

Bo

dy

w

eig

th

(%)

Day after challenge

4 M2e(k)НВс control

А/Kurgan/05/05

0

20

40

60

80

100

120

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Pe

rce

nt

su

rviv

al

(%)

Day after challenge

4 M2e(k)НВс control

А/Kurgan/05/05

50

60

70

80

90

100

110

0 2 4 6 8 10 12 14

Bo

dy

w

eig

th

(%)

Day after challenge

4 M2e(k)HBc control

0

20

40

60

80

100

120

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Pe

rce

nt s

urv

iva

l (%

)

Day after challenge4 M2e(k)НВс control

A/California/07/09

A/California/07/09


Determination of lung virus titres


PLAPROVA KBBE-2008-3-1-04

WP6. Development and Expression of influenza immunogens in plants

Saint-Petersburg


WHO National Center of Influenza

Candidate vaccine – chimeric virus TMV-M2e

CP TMVМ2E

SLLTEVETPIRNEWECRCNDSSDSLLTEVETPIRNEWESRSNDSSDSLLTEVETPIRNEWEARANDSSD

TMV-М2E =TMV-M2E-ser =

3‘NTRReplicase LBRB nosAct2 30K

CPUAG

amber

TMV-M2E-ala =

Ser156

Gly155

Moscow State University

1 2 3 4 5 6 7 8 9 1011121314150

20

40

60

80

100

120

M2e(human)BTM 1 LdM2e(human)BTM 5 Ldконтроль 1 Ldконтроль 5 Ld

0 2 4 6 8 10 12 14 16102030405060708090

100

M2e (human) BTM 5LD

Контроль 5LD

Post challenge days

% o

f su

rviv

ed m

ice

B

0 1 2 3 4 5 6 7 8 9 10111213140

20

40

60

80

100

120

ТМV-М2е-ala 1LDcontrol 1LDТМV-М2е-ala 5LDcontrol 5LD

C0 1 2 3 4 5 6 7 8 9 1011121314

0

20

40

60

80

100

120

ТМV-М2е-ala 5LD

control 5LD

Post challenge days

% o

f sur

vive

d m

ice

D

Survival of BALB/c mice vaccinated with TMV-М2е-ser, after challenge with 5 LD50 and 1 LD50 A/PR/8/34 (A) and A/California/04/2009 (H1N1) 5LD/50 (B). Mice were immunized with 50 µg, i/p * 3 times

Survival of BALB/c mice vaccinated with ТМV-М2е-ala, after challenge with 5 LD50 A/PR/8/34 (C) and A/California/04/09 (D). Mice were immunized with 50 µg, i/p * 3 times

A

Differences between mice survival in trail and control groups in fig. A,C, D are significant p<0,05 Research Institute of Influenza

Development of candidate vaccines based

on the HA2

А В

Influenza A virus molecular structure (А – trimer molecule, В – monomer)

А В

E. Vareckova 2013

Main antigenic HA2 sites:

Sites аа Mab

I 1-38 CF2

II, IV

125-175125-175

11F4FC12/FE1

III 38-112 CB8

Monomer HA2 before (А) and after (В) pH-dependent chain conformation

Flag HA2 35-102

Flag 4M2eh, k, h, k HA2 76-130

H2

H3

Vaccine preparations containing НА2(Research Institute of Influenza )


IgG titers to the carrier (Flg), recombinant protein (FlgHA2(35-107) and synthetic peptides after

intranasal immunization of mice Balb/c with FlgHA


Series10

2000

4000

6000

8000

10000

12000

14000

16000

18000

20000

pH=7.2 pH=5.0

H2N2 H1N1 H5N1 H3N2

Ти

тры

Ig

G

Binding of serum НА2 antibodies with influenza viruses A/Singapore (H2N2), A/PR/8/34 (H1N1), A/Aichi (H3N2), A/Kurgan (H5N1) at рН7,2 and 5,0. Intranasal immunization of mice Balb/c with FlgHA2(35-107) protein


Flg4M_HA PBS0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

0.50

3.57

log

10

Virus isolation from lungs at day 6 after challenge 1LD/50 of influenza virus A/Aichi/2/68 H3N2


Thank you for your attention!

АнтигенФенотипическая частота %

кавказоиды негры ориентыА1 14,7 3,3 ---

А2 45,7 27,3 43,2

А3 23,0 14,2 1,1

А11 11,8 1,1 17,2

А23 4,6 20,4 1,1

А29(w19) 7,6 12,3 0,4

A30 (w19) 4,8 28,3 0,3

А32 8,3 3,0 0,1

А33 3,3 9,0 13,1

B7 17,3 17,0 11,4

B8 16,1 5,8 0,2

B14 6,8 8,0 0,2

B27 7,6 3,0 0,8

B35 17,6 12,1 14,1

Cw1 7,5 1,1 32,1

Cw2 9,7 22,5 0,7

Cw3 20,1 17,5 46,5 *по материалам 8 workshop

Частота некоторых антигенов HLA I класса у представителей разных рас*

Technology of Ultrix vaccine creation provides presentation of surface antigens in a virosome form and that of internal antigens in a form of micelles while at most preserving their antigenic activity. Benefits: • Formation of systemic B- and T-cell immune response • Formation of cross-protection against influenza virus drift variants.

Ultrix - inactivated split influenza vaccine

Application of ULTRIX® vaccine 45µg in all age groups from 6 to 60 years and older was not accompanied by increase of general IgE production, also, decrease of general IgE was observed in persons with latent sensibilization thus indicating no any allergizing effect of ULTRIX®.

Immunogenic activity of ULTRIX® vaccine 45µg

due to seroconversion level to influenza virus

A(H1N1) up to 94,0% A(H3N2) up to 86,% B up to 90,0%

due to fold rise in antibodies

A(H1N1) up to 21,9 A(H3N2) до 12,6 B up to 7,5

due to seroprotection level

A(H1N1) up to 95,0% A(H3N2) up to 90,0% B up to 78,0%

Longevity of humoral immunity (45 µg of Ultrix vaccine)

Documents

Managua 2014 Institute of Influenza, Saint-Petersburg, (Research Institute of Influenza, Saint-Petersburg, WHO National Influenza Center of RUSSIA) Vaccination