Lyme and metals – triggers of inflammation causing non-specific multi-symptoms Vera Stejskal, PhD Associate Professor University of Stockholm, Sweden ©

Lyme and metals – triggers of inflammation causing non-specific multi-symptoms

Vera Stejskal, PhD Associate Professor

University of Stockholm, Sweden

© Copyright MELISA Medica Foundation

Inflammation is present in

many chronic diseases

T cell

cytokines

Inflammation

cytokines

cytokines

cytokines

Cytokines

reactive

oxygen

intermediates

Cytokines reactive oxygen intermediates

glycoproteinsreactive oxygen

intermediate

B cell

MacrophageNeutrophil


Causes of inflammation• Viruses• Microbes• Bacteria

• Lyme• Foods• Metals


Lyme diseasenon-specific clinical symptoms

skin conditions heart palpitations dizzinessheadaches numbness neck stiffness Bell’s palsy fever impaired memorypersonality changes anxiety difficulty concentratingjoint swelling pain in joints chillssleep disturbances shooting pains pain in limbsdepression fatigue lesions


Clinical relevance of laboratory testingTest Detection Sensitivit

ySpecifity Reproducible Literature

Cell culture Bacteria low - med.

45 - 71%

high NA Liveris: J Clin Microbiol, 2011

PCR DNA < 30% high NA Brettschneider: J Clin Microbiol, 1998

LDA Borrelia antigensin urine

30% 89% NA Harris & Stephens: J Spiro Tick-Borne Dis, 1995

ELISA/IFA IgM / IgG 0 - high low poor Aguero-Rosenfeld: Clin Microbiol Rev, 2005

Western Blot IgM / IgG 0 - higher low poor Aguero-Rosenfeld: Clin Microbiol Rev, 2005

C6 LPE Antibody to C6 of VlsE 0 - high low NA Aguero-Rosenfeld: Clin Microbiol Rev, 2005

ELiSpot Cytokines NA NA NA AID; Schwarzbach: Borreliosis Center Augsburg

CXCL13 in CSF Chemokines 99% 96% NA Tjernberg: J Infect, 2011

LTT (≤ 4 antigens)

Memory T cell activity (Europe)

91% 94% NA Van Baehr: J Lab Med, 2007

LTT-MELISA®

(8 antigens)Memory T cell activity (Europe)

91% 97% 93% Valentine-Thon: Diag Microbiol Infect Dis, 2007

LTT-MELISA®(10 antigens)

Memory T cell activity (USA/Canada)

91% 97% 93% Valentine-Thon & Gordon: Clin Chem Lab Med, 2010

Courtesy of Dr Elizabeth Valentine-Thon

100%

Improved detection of Lyme disease

• The lymphocyte transformation test used – MELISA – is standardized, validated and widely published

• MELISA Lyme testing uses well defined Borrelia-specific antigens


MELISA assay• White blood cells are exposed to antigens based on

patient’s current or future exposure• Lymphocyte reactivity is measured in two ways:

1) Uptake of radioisotopes by dividing lymphocytes2) Morphology - evaluation under microscope

• Results are produced as a value on a Stimulation Index

Lymphoblast

Dividing lymphoblast Macrophage


MELISA versus standard Lymphocyte Transformation Test (LTT)

MELISA uses:• A higher number of lymphocytes per test • Metal concentrations that are non-mitogenic and

non-toxic• Partial depletion of macrophages which restores the

lymphocyte-monocyte balance so that it is similar to the blood

• Morphological examination, in addition to the lymphocyte proliferation by radiolabeled thymidine

MELISA has been validated by independent research© Copyright MELISA Medica Foundation

100%

MELISA Lyme testing in Europe• 4 recombinant Borrelia antigens tested on 244 patients with

suspected Lyme disease o Osp C (afzelii)o p41-internal fragment-1 (garinii)o p42-internal fragment-2 (afzelii)o p100 (afzelii)

• 90 patients (37%) were positive to one or more antigens• Reproducibility – 93%• Specificity – 97%

o 30 healthy laboratory workers with no history of tick bites and seronegative in ELISA and WB, (1 responded positively)

• Clinical relevance: Follow up performed in 54 patients o More than 85% reduction in reactivity following treatment and clinical

improvementValentine-Thon E, Ilsemann K, Sandkamp M. A novel lymphocyte transformation test (LTT-MELISA) for Lyme borreliosis. Diagn Microbiol Infect Dis. 2007 Jan;57(1):27-34.

100%

MELISA Lyme testing in USA• 72 patients from USA and 26 patients from Europe (but

with prior residence in the USA) o Mainly arthritic symptoms

• Tested with 10 antigens:o 4 standard recombinant antigens listed previously from B.afzelii

and gariniio 4 new recombinant antigens derived from B.b.sensu strictoo full antigen lysate from SS and peptide mix

• Results:o 60% of US patients were positive to B.b.sensu stricto strain

antigens

but none responded to B.afzelii and B.gariniio European patients responded to both groups

Valentine-Thon E and Gordon E. Improved detection of Lyme disease with extended panel of recombinant Borrelia-specific antigens. Clinical Chemistry and Laboratory Medicine, 48(5):A26, 2010.

Lyme Case 1 (M 48 yrs old)

Valentine-Thon et al, Diagn Microbiol Infect Dis, 2007

• European patient• Bitten by a tick• Developed erythema

migrans• Positive in serology and

MELISA Lyme test • Diagnosed with stage 1 • Treated with

doxycycline (DOX)• Asymptomatic• Negative follow-up

MELISA Lyme testOspC p41-1 p41-2 p100

0

2

4

6

8

10

12

Sti

mu

lati

on

in

de

x

06/2005 before DOX, symptomatic08/2005 after DOX, asymptomatic

Lyme Case 2 (F 44 yrs old)

Valentine-Thon et al, Diagn Microbiol Infect Dis, 2007

• European patient• Bitten by a tick, developed

erythema migrans• Serologically positive

• Amoxycillin (AMX) treatment but symptoms persisted

• Doctor ordered test with MELISA Lyme

• MELISA Lyme test was positive

• Doxycycline (DOX) treatment and symptoms disappeared

• Negative Follow-up MELISA Lyme test OspC p41-1 p41-2 p100

0

4

8

12

16

20

24

Sti

mu

lati

on

in

de

x

12/03 after AMX, symptomatic03/04 still symptomatic04/04 after DOX, asymptomatic

Chronic Lyme Disease or Metal Induced Inflammation?

10-20% of antibiotic-treated patients continue to have symptoms1 • Penicillin and other sulphur antibiotics will bind

metals and release them from body depots• The metabolite of penicillins in vivo is D-penicillamine,

which is a chelator of mercury and other metals such as nickel2

• Lymphocytes of metal-hypersensitive patients react to mobilized metals by cytokine release which cause non-specific multi-symptoms

References: 1) www.cdc.gov/lyme/postLDS/index.html 2) Swaran, Pachauri. Chelation in Metal Intoxication. Int J Environ Res Public Health. 2010 July; 7(7): 2745–2788.


Swedish Lyme study (2007)

34 patients with suspected Lyme underwent MELISA Lyme testing

• 12% tested positive to two or more borrelia antigens

• 18% tested positive to one antigen only

27 patients were tested for metal hypersensitivity • 74% tested positive to one or more metals • 11% tested weakly positive to one or more metals


Swedish patients with suspected Lyme disease

Lyme Metals 0

10

20

30

40

50

60

70

80

90% weakly positive

% positive

Po

siti

ve (

%)


Exposure to metals

Dental: Hg, Au, Pd, Sn, Ag, Ni Medication: Fe, TiO2

Cosmetics: TiO2, Cd, Pb, Hg Foodstuffs: Hg, TiO2, Au, Ni, Fe

Vaccines: Al, Hg Smoking: Ni, As, Cd, Pb

Body implants: Co, Cr, TiO2, Ni


Courtesy of Dr Dagmar Magnusson, DDS

Silver (E174)Titanium dioxide (E171)

Nickel

Arsenic & Molybdenum

Inorganic and Methyl-mercury

Lead, Cadmium, Beryllium

Gold (E175)

Metals in food and

drink


FDA recognises mercury allergy• June 2008: The US Food and Drug administration put

a warning on their website regarding the safety of amalgam fillings:

• ”If you are allergic to any of the metals in dental amalgam, you should not get amalgam fillings.”

• ”If you believe you have an allergy or sensitivity to mercury or any of the other metals in dental amalgam (such as silver, tin, or copper), you should discuss treatment options with your dentist.”

www.fda.gov/MedicalDevices/ProductsandMedicalProcedures/DentalProducts/DentalAmalgam/ucm171094.htm


Effects of mercury in the cardiovascular system (apart from possible allergy)

1. Oxidative stress2. Inflammation3. Thrombosis4. Vascular smooth muscle proliferation and migration5. Endothelial dysfunction6. Dyslipidemia (oxidation of high-density lipoprotein and paraxonase)7. Immune dysfunction8. Mitochondrial dysfunction

Ref: Houston, Role of mercury toxicity in hypertension, cardiovascular disease and stroke.

J Clinical Hypertension, v 13, p.621-627


Deposition of radioactive Hg in various organs of a mouse after injection

Deposited Hg

White areas: deposited mercury Courtesy of Dr Seo

Accumulation of metals in breast cancer and healthy breast tissue biopsies

Fe Ni Cr Zn Hg Cd0

50

100

150

200

250

300

350

40053174 995 816 17075

6.9

42

10937

2139

3741

2.116

53,174 10,937 995 816

17,075 3,741

(µg/kg)

Blue: Breast cancerRed: Healthy tissue

Increased levels of transition metals in breast cancer tissue.Ionescu JG et al Neuro Endocrinol Lett. 2006 Dec;27 Suppl 1:36-9.

Differentiate between

Toxic effects• higher doses• single exposure• lower specificity• lower genetic influence

Immunological effects

• lower doses• chronic exposure• higher specificity• higher genetic influence, only

certain individuals are affected


Genetics determine metal susceptibility

Healthy

Skin allergy, cardiovascular disease, thyroiditis

Autoimmune diseases; MS, RA, psoriasis


Skin allergy (dermatitis, oral lichen)

Yellow canaries

How to diagnose susceptible groups?© Copyright MELISA Medica Foundation

Diagnostic tests

• Blood mineral analysis (serum or whole blood)• Urine (pre- or post-chelation) • Hair mineral analysis screening• MELISA testing for hypersensitivity


Metal allergy (cellular hypersensitivity)

Positive response:Cell growth and division

Negative response :No cell growth or division

Positive response

Patch Test

Ni

Hg

MELISA®


SHS-Hg, S-Ag, S-Au, S-Ni, S-TiHg, Ag, Au, Ni, Ti

Own cells

Tolerated by immune system

“Foreign” cells

Attacked by immune system

→ Allergy→ Autoimmunity

Zzz… 007… Withlicense to kill!

Metals bind to SH groups – changing enzyme and protein structure

How metals cause autoimmunity


Fatigue is frequent in chronic diseases

STRESS

Hippocampus

Pituitary Hypothalamus

Adrenals

Glucocorticoids

Pituitary Hormones

ACTH

Stimulation of hypothalamus-pituitary-adrenal (HPA) axis can be caused by inflammation products, so called cytokines.


Most frequent metal allergens 3,162 patients with CFS, 116 healthy subjects

Posi

tive

test

s %

Metal-specific lymphocytes: biomarkers of sensitivity in manStejskal, V, et al. Neuroendo Lett 1999; 20:289-298 © Copyright MELISA Medica Foundation

Reactivity to metals, CFS group and control subjects

MELISA in 111 patients and 116 controls

• Inorganic mercury, phenylmercury and gold: highly significant differences (P<0.001)

• Cadmium, titanium, lead and palladium:significant differences (P<0.01)

• Methylmercury and silver significant differences (P<0.05)

2% worsened

78% improved

20% unchanged


Health after dental metal replacement in metal-allergic

patients with CFS

Metal-specific lymphocytes: biomarkers of sensitivity in manStejskal, V, et al. Neuroendo Lett 1999; 20:289-298

Cellular hypersensitivity to metals might trigger FM

Lymphocyte responses in patients with FM and in healthy controls

0%

10%

20%

30%

40%

50%

60%

70%

80%Healthy Controls SI ≥ 3Healthy Controls SI ≥ 5Patients SI ≥3Patients SI ≥ 5

Case 1: MBM 53 yr F with fibromyalgia

Gold Palladium Nickel0

2

4

6

8

10

12

14

16

1994 1997 After removal 2005 After removal

Stim

ulati

on in

dex

Case 2: IS 69 yr F with fibromyalgia

Inorganic mercury Phenyl mercury Cadmium Nickel0

2

4

6

8

10

12

1994 1997 Post removal 2005 Post removal

Stim

ulati

on in

dex

Case 3: SC 53 yr F with fibromyalgia/CFS

• Born 1956 (female)• Aged 10: first amalgam fillings were placed • Urticaria and rashes throughout teenage years –

suspected nickel allergy• Aged 35: Underwent tubal ligation with a Filshie clips

(containing nickel)• Developed gallstones and gall bladder removed

(gallstones contained Ni)• Metal crowns were placed (contained 75% nickel)• Developed intense abdominal pains and diagnosed with

Irritable Bowel Syndrome


Case 3: SC• After additional crowns and bridgework

experienced muscle pain and weakness – diagnosis of fibromyalgia

• Aged 53: MELISA testing


Case 3: MELISA test Stimulation

Index


Crowns made up of 75% nickel

Case 3: MELISA test Stimulation

Index


• 2009/2010: Tubal clips removed, all dental work replaced with composite and zirconium

• August 2010: Dramatic health improvement, no IBS symptoms, no fibromyalgia symptoms


Case 3: MELISA treatment

Patient questionnaire

www.melisa.org/patient-questionnaire.php © Copyright MELISA Medica Foundation

Case 4: PM 43 yr M with rheumatoid arthritis

• Medical doctor aged 40+ years• Aortic valve operation with standard surgical steel wires

o Acute arthritis in wrists, knees and feet – unable to dress himself

o Blinding auras and visual disturbances• Two months later: Diagnosed with Rheumatoid arthritis • Developed rash after wearing new spectacles• Arthritis flare after eating fish with high nickel content• MELISA testing


Metal MELISA SI ResultMolybdenum 36.0 Strongly positiveNickel 10.1 Strongly positiveManganese 10.1 Strongly positive

Surgical steel wires contained: Nickel 18%Molybendum 3%Manganese 2%

Case 4: MELISA test


Case 4• 1 year after operation: Chest wires removed by surgeon who

insisted there was no link with symptoms• After three months returned to gym, not fully recovered but

massive improvement • 1,5 years later stopped taking methotrexate for RA and

adopted low nickel diet• Completely symptom-free• Surgeon has added “nickel” allergy to PM’s notes


Multiple sclerosis affects 2.5 million people worldwide

Plaques (demyelination)

Role of oligodendrocytes in the demyelinationprocess

Myelin

Neuron

Oligodendrocyte


Multiple sclerosis study

• 106 patients with multiple sclerosiso 82 F and 24 M

• 144 healthy controlso 80 F and 64 M

• Results:o Highly significant difference in lymphocyte reactivity to

following metals: MeHg, Au, Pd, Pb, Ti, Ni


Case 5: DF 32 yr F with MS

• Young female • Non-smoker• Orthodontic braces• 1 cobalt chrome nickel crown• Metal taste in the mouth• Bleeding gums• 1 root-filling• Clinical nickel allergy• MS diagnosed aged 31


Case 5

• MELISA test strongly positive to nickel• Removed nickel-containing metal crown

MRI 2011 MRI 2012Lesions are gone


• Optical neuritis at the age of 27• Patch test positive to mercury but negative to nickel, despite well

known clinical metal allergy• MELISA: Mercury +, Nickel +• Treatment: removal of amalgams under strict protection,

replaced with non-metallic ceramics. • Heavy anti-oxidant therapy, still ongoing• Outcome: Symptom-free for last 15 yrs, MRI normalized,

neurologist considers the patient as healthy

Stejskal et al. Neuroendo. Lett, 2006; 27:7-11(Suppl.1)

Case 6: BB 47 yr F with MS


Increased reactivity to metals has been found in the following diseases:

• Multiple Sclerosis (Prochazkova 2003, 2006), (Stejskal 2006)

• Chronic Fatigue Syndrome (Stejskal 1994, 1999)

• Rheumatoid Arthritis (Prochazkova 2003, Stejskal 2006)

• Crohn's Disease • Fibromyalgia (Öckert 2006)

• Amyotropic Lateral Sclerosis (Pleva 2000)

• Cardiovascular disease• Breast cancer (Ionescu 2006, • Stejskal unpublished)

• Lupus Erythematosis (Prochazkova 2003)

• Oral Lichen Planus (Stejskal 1996)

• Oral burning and itching (Stejskal 2006)

• Skin diseases such as eczema or psoriasis, (Prochazkova 2003; Venclikova 2003) (Kohdera, Ionescu)

• Sjögren's syndrome (Prochazkova 2003)

• Autoimmune thyroiditis (Sterzl 1999, Prochazkova 2003, 2006, Hybenova 2010)


Thank you for your attention!

[email protected] Skype: verastejskal

www.melisa.org

Documents

Lyme and metals – triggers of inflammation causing non-specific multi-symptoms Vera Stejskal, PhD Associate Professor University of Stockholm, Sweden ©