LYME BORRELIOSIS & CO-INFECTIONS Last Chance for Patients with Unknown Causes

Demonstration: Borrelia burgdorferi s.l., Babesia, Bartonella, Anaplasma/Ehrlichia

Béla P. Bózsik, M.D.

Lyme Borreliosis Foundation, 1991

Sinaia 2011

Last chance, but

WHY?

Borrelia-binding antibodies were detected with disparate results by parallel serological investigations with different reagents and ELISA+WB methods.

CW Ang et al 2011. August.

After ~30 years of detecting Borrelia burgdorferi sensu lato.

What does it mean?  Parallel investigations were happened & revealed problems  The sero-response to Borrelia burgdorferi sensu lato is especially unique

‒ IgM-response can be existing in chronic stage ‒ IgM&IgG-responses can be separated each other: switching inhibited ‒ Independent immune responses exist to the different bacterial component with the characters abovementioned ‒ Specific antibodies can be exhausted producing even seronegativity ‒ The two-tier method has failed the 15% of patients at least

 The evaluation practice has been failed even more seronegatives  The present MC-antigens gave a good chance to get more informations about the causative agents & pathogenetic processes, but it were lost by the present practice.

The suggested flow-chart info

Special connection were in every time among the Microscopy-Germs-Physicians

1. Microscope helps to discover the micro-Cosmos−

~ 1783. letters of the uneducated Dutch merchant(~200) to the Royal Academy ~

2. Then the facts were hidden(?) for decades.. 3. 1905. The Microscopy was re-discovered & applied in the diagnosis of Syphilis up to nowadays. 4. 1909. NewSpirochetewasdiscoveredinblood ofthyroidectomized Chambers, H. − Royal Free Hospital; BMJ, 1909 & Lancet 913,1:1728.

5. 1981. Lyme Spirochete could be discovered with microscope, but ithasnotbeensuggestedasdiagnosticmethod − Manual of Microbiology 6. Foransweringlet us see What has happened during tick-biting

Diagnostic Definitive Symptoms Indicate promt treatment

Lyme borreliosis

Caused by Circulating Spirochetes Since Setout • Early (I.) − Days-Weeks, • Late (II.) − Months(6-12),

• Chronic (III.) Stages − Years.

It is chronic damage of the whole body. The symptoms deeply depend on the fitness LBC of the body, the co-infections and other bystander diseases. Paresis seu Dysfunctio Nervi cranialis I.—XII.

Special Morphology gives Special Way for Spirochetes

Through flagellar tube & central shedding can be connected the protoplasmic space with the extrabacterial material & BLEBS.

Three Easy Solutions 1. Artificial products & „filaments” disturbing and cooking the results. Answer: new reagent that – Prevent the development of artificial products – Save extremely vulnerable Spirochetes – Support their movement, – That help their identification.

Three Easy Solutions 2. Few Lyme Spirochetes are circulating for detecting them Answer: – The volume of blood samples should be fair enough for both the patients and the method – That is centrifuged & concentrated by ~ 1000 times – The sediment of different layer were investigated without fixing and staining − detecting nearly normal states of the causative agents.

Three Easy Solutions 3. The intraerythrocyter & intraleukocyter germs in TBDs need the analysis of Plasma-sediment − only, in the first phase Buffy coat − in the present method − in the present method RBC WBC − in the present method altogether gave the best result. In these layers there were proved the blood-cells with causative agents.

– – – –

I. Basic phenomenon Dividing in Human blood sample

II. Basic phenomenon Nativity of GEMMA

You can see on video

GEMMA the discoid structure AFM picture

Miklóssy personal gift

Brørson personal gift

III.Basic phenomenon Bleb & Shedding

BLEB is the macromoleculecular groups of Osp-s and

other components of Borrelia. − It can be developing during cultivation − it is a protecting mechanism of Borrelia. SHEDDING helps Borrelias to be free from harming molecules − in this way macromolecular IC goes into the body: − it solves the paradoxon of few germ wide symptoms

IMMUNOCYTOLOGY (1990-2002): Indirect immuno-fluorescence with monoclonals binding to OspA & flagellin (donated by prof.Barbour)

Two types of blebbing are PERIFERIAL and CENTRAL ones. •Periferial type big immunocomplex has OspA antigen among others, it does not influences either movement or division. •Central type is exclusively for flagellin based immunocomplexes and Borrelia burgdorferi sensu lato were immobilized. anti-flagellaris

B. garinii−the Short Spirocheta+IC

B.Afzelii

the Long one

BLEBS, the regular & the irregular

The so called „Filaments” and their

Extrabacterialis Protein of Borrelia burgdorferi .s.l.

Patients’ Blood sample (27/72/105; 72/14/14) Phosphorwolframic-acid (1%) negativ staining, 1000x

140 131

WB-reactivity of 137 consecutive Lyme patients 120

%

100

IgM IgG

p100 VlsE p41s p39 OspA OspC p41g p41a p18 18,3 9,5 19,0 30,7

40,0 95,6

2,2 9,5

8,0 8,0

28,5 10,2

27,0 15,3

28,5 17,5

0,7 5,1

80

60

55 42

39

40 25

39

37

26 21

20

13

13

11

11

24

14 7

3

1

0 p100

VlsE

p41s

p39

IgM

OspA

OspC

p41g

IgG

p41a

p18

25

WB-reactivity patterns in Lyme borreliosis

non-WEIGHTED Summarized reactivity

6/137 has both FLAGELLIN-patterns prove the change in sero-response

20

MS 8/15 patients 15

MS 4/10 patient

10

5

0 p100

VlsE

p41

p39

OspA OspC p41-G

p41-A

p18

p100

VlsE

p41

p39

OspA OspC

p41-G

p41-A

recombinant-Antigen palettes for Borrelia burgdorferi sensu lato IgM IgG Group Red(IgM): MS 8/15 patients-21 determinations Group Blue(IgG): 10 patients 12 determinations

p18

FLAGELLIN-patterns in MS – Flagellin-pattern „I” with reactivity to: • • • •

p41-common with IgG molecules p41-common with IgM molecules p41-B.garinii, p41-b.afzelii

– Flagellin-pattern „II” with only IgG Reactivity • p41-common • p41-B.garinii • p41-B.afzelii

WB-reactivity & Copaxone Patient 1.

Patient 2.

Daily change

37oC

IgM-IgG switching after 4 years of Copaxone treatment

WB-reactivity & Copaxone Patient 3.

WB-reactivity & Copaxone Patient 4.

MULTIPLEX PARESIS. ESPECIALLY AFTER A TRANSITORY IMPROVEMENT CAUSED BY STEROID

HOMOLOGY Sequence homology

cross-reaction has been detected between − Spirochaete(Borrelia) Flagellin & − Human Myelin Basic Protein + Human Axons Weigelt W et al. 1993, Fikrig E et al. 1993.

Sigal LH wgroup BBA 1993

Copaxone − glatiramer acetat (6,4 kD) is a RANDOM polymer of four amino acids Found in myelin basic protein: glutamic acid, lysine, alanine, tyrosine, and may work as a decoy for the immune system.

That can work contrary to BLEBS.

Borrelia burgdorferi sensu lato fakultatív intracellularis parazita ~ 5x2,5+25x1,3

Hasonlóan sikertelenek a TIGECYCLINE-nel végzett kísérletek, de monotherápiás próbálkozások is; hiába károsítja a GEMMÁKat. Alapítványunk 1994 óta vitatja a monotherápia gyógyító hatását és helyette kidolgozta a KOMBINÁLT KEZELÉST.

Babesiosis • Intraerythrocyter parazita • Számos altörzs

– There are over 100 species of Babesia identified – B.microti, USA – B.divergens, EU

• Változatos klinikai megjelenés – – – –

Cephalalgia, neuropathia hyperhidrosis nocturnalis Cardiomyopathia, dyspnoe Anaemia − akutan veseelégtelenség, halál

• Növekvő transfúziós fertőzésekről írnak

Bartonellosis • Intraerythrocyter parazita • Számos altörzs két(?) humán pathogen – B.henselae – B.quintana

• Változatos klinikai megjelenés – – – – –

Dermatitis CSD, dyspigmentatio Angiopathia, vasoproliferatio Lymphadenopathia, lymphadenitis Cardiomyopathia, Endocarditis, dyspnoe Cephalalgia, neuropathia….

• Transfúziós fertőzés-veszély fennáll

Albert Edward BARTON THOMPSON

Dehio+al; Vet.Res.(2009)40:29

Rolain et al; Lancet 2002

Chronic & Persistent Stage

ANAPLASMA/EHRLICHIA • Intracellular/Intraleukocyter parazita • Számos altörzs két humán pathogen(?) – Anaplasma phagocytophylum – A.monocytotica/E.chaffeensis

• Változatos klinikai megjelenés – – – –

cephalalgia hyperhidrosis nocturnalis Neuropathia−neutropenia Cardiomyopathia, dyspnoe

• Növekvő transfúziós fertőzésekről írnak

Rikihisa, 2010;2011

Anaplasma Humán vérmintában

Chronicity & Autoimmunity Erythrophagocytosis

Apoptosis & Morules

Would it be permitted/advised to be treating in subclinical state

PLEASE keep the HOPE

Nil nocere Nil admirari Salus aegroti suprema lex, esto