Download Lyme and metals – triggers of inflammation causing

Lyme and metals – triggers of
inflammation causing non-specific
multi-symptoms
Vera Stejskal, PhD
Associate Professor
University of Stockholm, Sweden
© Copyright MELISA Medica Foundation
Neutrophil
Macrophage
Cytokines
reactive oxygen intermediates
glycoproteins
cytokines
Inflammation
B cell
T cell
cytokines
Inflammation is present in
many chronic diseases
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Causes of inflammation
• Viruses
• Microbes
• Bacteria
• Lyme
• Foods
• Metals
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Lyme disease
non-specific clinical symptoms
skin conditions
heart palpitations
dizziness
headaches
numbness
neck stiffness
Bell’s palsy
fever
impaired memory
personality changes
anxiety
difficulty concentrating
joint swelling
pain in joints
chills
sleep disturbances
shooting pains
pain in limbs
depression
fatigue
lesions
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Clinical relevance of laboratory testing
Test
Detection
Sensitivity
Specifity
Reproducible
Literature
low med.
45 - 71%
< 30%
high
NA
Liveris: J Clin Microbiol, 2011
high
NA
Brettschneider: J Clin Microbiol, 1998
30%
89%
NA
Harris & Stephens: J Spiro Tick-Borne Dis,
1995
Cell culture
Bacteria
PCR
DNA
LDA
Borrelia antigens
in urine
ELISA/IFA
IgM / IgG
0 - high
low
poor
Western Blot
IgM / IgG
0 - higher
low
poor
C6 LPE
Antibody to C6 of VlsE
0 - high
low
NA
ELiSpot
Cytokines
NA
NA
NA
CXCL13 in CSF
Chemokines
99%
96%
NA
Aguero-Rosenfeld: Clin Microbiol Rev,
2005
Aguero-Rosenfeld: Clin Microbiol Rev,
2005
Aguero-Rosenfeld: Clin Microbiol Rev,
2005
AID; Schwarzbach: Borreliosis Center
Augsburg
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
Improved detection of Lyme disease
• The lymphocyte transformation test used – MELISA –
is standardized, validated and widely published
• MELISA Lyme testing uses well defined Borrelia100%
specific antigens
© Copyright MELISA Medica Foundation
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 lymphocytes
2) Morphology - evaluation under microscope
• Results are produced as a value on a Stimulation Index
Lymphoblast
Dividing lymphoblast
Macrophage
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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
MELISA Lyme testing in Europe
• 4 recombinant Borrelia antigens tested on 244 patients with
suspected Lyme disease
o
o
o
o
Osp C (afzelii)
p41-internal fragment-1 (garinii)
p42-internal fragment-2 (afzelii)
100%
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
improvement
Valentine-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.
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:
100%
o 4 standard recombinant antigens
listed previously from B.afzelii
and garinii
o 4 new recombinant antigens derived from B.b.sensu stricto
o 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.garinii
o 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)
12
06/2005 before DOX, symptomatic
08/2005 after DOX, asymptomatic
10
Stimulation index
• 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 test
8
6
4
2
0
OspC
Valentine-Thon et al, Diagn Microbiol Infect Dis, 2007
p41-1
p41-2
p100
Lyme Case 2 (F 44 yrs old)
• European patient
• Bitten by a tick, developed
erythema migrans
• Serologically positive
• Doxycycline (DOX) treatment
and symptoms disappeared
• Negative Follow-up MELISA
Lyme test
Valentine-Thon et al, Diagn Microbiol Infect Dis, 2007
12/03 after AMX, symptomatic
03/04 still symptomatic
04/04 after DOX, asymptomatic
20
Stimulation index
• Amoxycillin (AMX) treatment
but symptoms persisted
• Doctor ordered test with
MELISA Lyme
• MELISA Lyme test was
positive
24
16
12
8
4
0
OspC
p41-1
p41-2
p100
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.
© Copyright MELISA Medica Foundation
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
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Swedish patients with suspected
Lyme disease
90
% weakly positive
80
Positive (%)
70
% positive
60
50
40
30
20
10
0
Lyme
Metals
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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
© Copyright MELISA Medica Foundation
Courtesy of Dr Dagmar Magnusson, DDS
Silver (E174)
Titanium
dioxide
(E171)
Gold (E175)
Lead,
Cadmium,
Beryllium
Metals in
food and
drink
Inorganic and
Methylmercury
Nickel
Arsenic &
Molybdenum
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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
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Effects of mercury in the cardiovascular
system (apart from possible allergy)
1. Oxidative stress
2. Inflammation
3. Thrombosis
4. Vascular smooth muscle proliferation and migration
5. Endothelial dysfunction
6. Dyslipidemia (oxidation of high-density lipoprotein and
paraxonase)
7. Immune dysfunction
8. Mitochondrial dysfunction
Ref: Houston, Role of mercury toxicity in hypertension, cardiovascular disease and stroke.
J Clinical Hypertension, v 13, p.621-627
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Deposition of radioactive Hg in various organs of a mouse
after injection
Deposited
Hg
Courtesy of Dr Seo
White areas: deposited mercury
Accumulation of metals in breast cancer
and healthy breast tissue biopsies
400
53,174
10,937 995
816
17,075
3,741
350
Blue: Breast cancer
Red: Healthy tissue
300
250
200
150
100
50
21
42
39
6.92.1
0
(µg/kg)
Fe
Ni
Cr
Zn
Hg
16
Cd
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
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Genetics determine metal susceptibility
Healthy
Skin allergy (dermatitis, oral lichen)
Skin allergy, cardiovascular disease,
thyroiditis
Autoimmune diseases;
MS, RA, psoriasis
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Yellow canaries
How to diagnose susceptible groups?
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Diagnostic tests
•
•
•
•
Blood mineral analysis (serum or whole blood)
Urine (pre- or post-chelation)
Hair mineral analysis screening
MELISA testing for hypersensitivity
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Metal allergy (cellular hypersensitivity)
Ni
Hg
Positive response:
Cell growth and division
Negative response :
No cell growth or division
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How metals cause autoimmunity
Metals bind to SH groups – changing enzyme and protein structure
SH
Hg, Ag, Au, Ni, Ti
Own cells
Tolerated by immune
system
S-Hg, S-Ag, S-Au,
S-Ni, S-Ti
“Foreign” cells
Attacked by immune
system
→ Allergy
→ Autoimmunity
Zzz…
007… With
license to kill!
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Fatigue is frequent in chronic diseases
Hippocampus
Stimulation of
hypothalamuspituitary-adrenal
(HPA) axis can be
caused by
inflammation
products, so called
cytokines.
STRESS
Pituitary
Hypothalamus
Glucocorticoids
Pituitary
Hormones
ACTH
Adrenals
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Most frequent metal allergens
Positive tests %
3,162 patients with CFS, 116 healthy subjects
Metal-specific lymphocytes: biomarkers of sensitivity in man
Stejskal, V, et al. Neuroendo Lett 1999; 20:289-298
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Reactivity to metals, CFS group
and control subjects
MELISA in 111 patients and 116 controls
2% worsened
20% unchanged
78% improved
Health after dental metal
replacement in metal-allergic
patients with CFS
Metal-specific lymphocytes: biomarkers of sensitivity in man
Stejskal, V, et al. Neuroendo Lett 1999; 20:289-298
• 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)
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Cellular hypersensitivity to
metals might trigger FM
Lymphocyte responses in patients
with FM and in healthy controls
80%
Healthy Controls SI ≥ 3
70%
Healthy Controls SI ≥ 5
60%
Patients SI ≥3
50%
Patients SI ≥ 5
40%
30%
20%
10%
0%
Case 1: MBM
53 yr F with fibromyalgia
16
Stimulation index
14
1994
12
1997 After removal
2005 After removal
10
8
6
4
2
0
Gold
Palladium
Nickel
Case 2: IS
69 yr F with fibromyalgia
12
10
1994
1997 Post removal
Stimulation index
8
6
4
2
0
Inorganic mercury
Phenyl mercury
Cadmium
Nickel
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
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Case 3: SC
• After additional crowns and bridgework
experienced muscle pain and weakness –
diagnosis of fibromyalgia
• Aged 53: MELISA testing
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Case 3: MELISA test
Stimulation
Index
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Case 3: MELISA test
Stimulation
Index
Crowns made up of 75% nickel
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Case 3: MELISA treatment
• 2009/2010: Tubal clips removed, all dental
work replaced with composite and zirconium
• August 2010: Dramatic health improvement,
no IBS symptoms, no fibromyalgia symptoms
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Patient questionnaire
www.melisa.org/patient-questionnaire.php
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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
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Case 4: MELISA test
Metal
Molybdenum
Nickel
Manganese
MELISA SI
36.0
10.1
10.1
Result
Strongly positive
Strongly positive
Strongly positive
Surgical steel wires contained:
Nickel
18%
Molybendum
3%
Manganese
2%
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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
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Multiple sclerosis affects 2.5 million
people worldwide
Neuron
Plaques
(demyelination)
Oligodendrocyte
Myelin
Role of oligodendrocytes
in the demyelination
process
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Multiple sclerosis study
• 106 patients with multiple sclerosis
o 82 F and 24 M
• 144 healthy controls
o 80 F and 64 M
• Results:
o Highly significant difference in lymphocyte reactivity to
following metals: MeHg, Au, Pd, Pb, Ti, Ni
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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
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Case 5
• MELISA test strongly positive to nickel
• Removed nickel-containing metal crown
MRI 2011
MRI 2012
Lesions are gone
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Case 6: BB
47 yr F with MS
• 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)
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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,
•
• 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)
Stejskal unpublished)
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Thank you for your attention!
[email protected]
Skype: verastejskal
www.melisa.org