Borrelia Miyamotoi

[Roxy]

@VerlorengezondheidM, aanvullende informatie bij Bron viewtopic.php?f=6&t=1492&start=50#p26570

BMJ Case Reports
14 februari 2013
Case study - 'Seronegative lyme neuroborreliosis in a patient using rituximab' by Willemijn Alexandra van Dop, Marie-José Kersten, Bob de Wever, and Joppe Willem Hovius; Bron https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3604215/

Abstract
A 66-year-old woman presented with severe shooting pains throughout her back and legs, followed by progressive deafness, weight loss and headache. She had a history of marginal zone B-cell lymphoma stage IV-B, for which she was successfully treated with immunochemotherapy and rituximab maintenance therapy. A relapse was suspected, but chemotherapy was not administered, since, despite elaborate investigations, malignancy could not be proven. Because of a history of tick bites she was tested for antibodies against Borrelia burgdorferi in serum and cerebrospinal fluid (CSF), which were negative. However, a B burgdorferi PCR on CSF came back positive. The patient was treated for seronegative Lyme neuroborreliosis with ceftriaxone intravenously and dramatically improved. This case presentation demonstrates that, in immunocompromised patients, it is important not to solely rely on antibody testing and to use additional diagnostic tests to avoid missing or delaying the diagnosis.

Outcome and follow-up
During these 3 weeks she improved dramatically, which further strengthened the diagnosis. She gained weight, the complaints of headache and vertigo disappeared and her hearing loss slowly improved. Paired serology on blood and CSF 7 weeks later still revealed no antibodies against B burgdorferi.

One-and-a-half years later, a routine MRI of the myelum showed no evidence for relapse of the lymphoma and the hearing ability had completely returned. During neurological follow-up the patient reported a slight unsteadiness of gait, which was attributed to residual postinfectious complications. The rituximab maintenance therapy had been discontinued during her hospital admission, but despite the slow repopulation of B-lymphocyte subsets, 1.5 years after her initial presentation, there were still no antibodies against B burgdorferi (figure 3).

Discussion
In conclusion, this case report is an important reminder of the fact that in patients treated with drugs such as rituximab, which interfere with the immune response, serological diagnostics are not always reliable. In immunocompromised patients with a high suspicion of infectious diseases, such as infection with B burgdorferi, it is therefore important not to rely solely on antibody testing but to use several additional diagnostic tests (e.g, PCR and culture) to avoid missing or delaying the diagnosis. Lyme neuroborreliosis, although sometimes easy to diagnose, can be difficult to diagnose, and when the clinical suspicion is high, like in our case, additional diagnostic tests or sometimes even empiric treatment should be considered.

De Lyme (neuroborreliose) patiënte is succesvol behandeld met ceftriaxone.

[VerlorengezondheidM]

Is It Lyme Disease or Tick-Borne Relapsing Fever (TBRF)?

Bron: YouTube (IGeneX Inc.), kijk en luister meer

[VerlorengezondheidM]

LGLC Living Lands Series 2020: Lyme and tick-borne diseases

Professor Holly Ahern, Associate Professor of Microbiology, SUNY Adirondack

Bron: YouTube, kijk en luister meer

P.S.; hoor ik dat nou goed dat een Borrelia Miyamotoi besmetting door de Ixodes teek niet wordt geassocieerd met een rash of ben ik soms abuis?

[VerlorengezondheidM]

The Complement Binding and Inhibitory Protein CbiA of Borrelia miyamotoi Degrades Extracellular Matrix Components by Interacting with Plasmin(ogen)

Ngoc T. T. Nguyen1, Florian Röttgerding1, Gayatri Devraj1, Yi-Pin Lin2, Arno Koenigs1,3 and Peter Kraiczy1*

Bron: Frontiersin, lees meer

[Roxy]

'The Truth About Lyme online conference broadcasting on February 6 - 10, 2021'; Bron https://aspire.care/featured/the-truth- ... onference/ en Bron https://truthaboutlyme.com/?ref=gabriella%20

The Importance of Testing for Borellia miyamotoi
ASPIRE talks with Shannon L. Delaney, MD, on the latest study of Borellia miyamotoi in those with persistent symptoms and suspected tick-borne illness; Bron https://aspire.care/featured/borellia-m ... e-illness/

..Shannon: For this study, we looked at antibodies to B. miyamotoi. The lab we used in the study was Imugen, which was a tick-borne specialty lab that Quest Diagnostics recently acquired. Now you can get the test through Quest. I don’t know of any other lab doing this testing for TBRF, specifically B. miyamotoi other than Igenix. Their test is a little different; they do a Western Blot for TBRF, after that lab, they can do the speciation of what type of Relapsing Fever is positive. I don’t believe they can do that speciation in New York State. Quest Diagnostics is the only lab I know of that does this particular antibody test. Any doctor can order it through Quest.

The PCR test for B. miyamotoi is more widely available at different labs, but again we focused on antibodies for this study. Any time a patient is out of the acute early phase of the disease, a PCR test will not be reliable, so you really need to test for the antibodies.

At Columbia, in the second opinion Lyme consult practice, patients are not typically in the acute phase of the disease course, so in the setting of chronic disease, you look at the antibodies. I have not seen a positive PCR test yet, even in the patients who were antibody positive for B. miyamotoi. My best guess is that the PCR test will be positive when the patient is having early symptoms such as fever/flu-like illness, but after that phase, you should really look at antibodies because that gives you a reflection of what the immune system has seen in the past..

Frontiers in Medicine
27 oktober 2020
'Borrelia miyamotoi Serology in a Clinical Population With Persistent Symptoms and Suspected Tick-Borne Illness'; Bron https://www.frontiersin.org/articles/10 ... 67350/full
Shannon L. Delaney 1,2*, Lilly A. Murray 1,2, Claire E. Aasen 1, Clair E. Bennett 1,2, Ellen Brown 1,2 and Brian A. Fallon 1,2
1 Lyme & Tick-Borne Diseases Research Center, Columbia University Irving Medical Center, New York, NY, United States
2 New York State Psychiatric Institute, Columbia University Irving Medical Center, New York, NY, United States

Kort onderzoeksrapport artikel (case study)

Methods
This study included 82 patients consecutively screened for B. miyamotoi and B. burgdorferi antibodies as part of a clinical workup for tick-borne illness at the Columbia University Irving Medical Center from June 2017 to October 2018. Chart review was approved by the New York State Psychiatric Institute IRB. Clinic patients were seeking a second opinion to determine whether their persistent polymorphic symptoms were attributable to tick-borne infection. Patients frequently endorsed chronic symptoms of fatigue, pain, neurocognitive, and psychiatric problems.

Evaluations consisted of comprehensive physician assessments, serologic testing, and questionnaires. The GLP-Q assay was an indirect EIA for detection of antibody to B. miyamotoi, performed at Imugen, Inc. in Norwood, MA. As previously reported, they use the GlpQ gene sequence from B. miyamotoi (GenBank accession number AY368276) as the basis for cloning and expression as a 38-kDa recombinant protein (rGlpQ) (6). B. burgdorferi antibodies were assessed using the C6 peptide ELISA and IgM and IgG western immunoblot. Patients were designated B. miyamotoi positive based on IgG or IgM GlpQ seroreactivity (defined as >1 to the value calculated for the highest result on the standard curve). Patients were designated Lyme-positive if they met 2017 CDC surveillance criteria for definite or probable Lyme disease, having an EM skin lesion or multisystem clinical symptoms with at least 5 positive IgG bands on the Western blot.

Patients rated their symptoms using the General Symptom Questionnaire-30 (GSQ-30), a measure specifically developed to assess multisystem symptom burden in patients with early Lyme disease and post-treatment Lyme disease syndrome (12). Patients also completed the Beck Depression Inventory-II, Cognitive Failures Questionnaire, Fatigue Severity Scale, Epworth Sleepiness Scale, McGill VAS Pain Scale, and Zung Anxiety Scale. Mann-Whitney U tests were conducted to compare B. miyamotoi-positive and B. burgdorferi-positive patients on these measures.

Results
Of the 82 patients, 21 (26%) tested positive for B. miyamotoi by anti-GlpQ ELISA; all were IgG positive and IgM negative. Of these 21 patients, five also met CDC surveillance criteria for definite or probable Lyme disease. Of the 61 patients who tested negative for B. miyamotoi, 22 met criteria for definite or probable Lyme disease. The remaining 39 patients tested negative for both B. miyamotoi and B. burgdorferi antibodies.

The B. miyamotoi-positive group reported significantly more sleepiness on the Epworth Sleepiness Scale (Md = 9 vs. 4, U = 42.00, z = −2.51, p = 0.01) and significantly more pain on the VAS (Md = 5.80 vs. 2.70, U = 58.00, z = −2.02, p = 0.04) than the group with probable or definite Lyme disease. On the total score and individual item level, there were no significant between-groups differences on the GSQ-30, but the B. miyamotoi-positive group did endorse being bothered more by headaches than the B. burgdorferi-positive group (Md = 3 vs. 2 at a trend level, U = 52.50, z = −1.79, p = 0.07).

Data for B. miyamotoi-positive patients is presented in Supplementary Table 1. Mean age was 34 years. Eight of 21 (38%) reported hospitalization (seven medical and one psychiatric) since symptom onset, three for cardiac and two for neurologic abnormalities. All 21 received prior antibiotic treatment, of whom 20 received at least 2 weeks of doxycycline or amoxicillin. Sixteen of the 21 patients lived in the Northeast/Mid-Atlantic USA. Of the remaining five patients, two lived in California, two lived in Florida, and one lived in Illinois. Of the 82 patients in the study, 80 (98%) had not been previously tested for B. miyamotoi infection. Among the 21 patients positive on the GlpQ ELISA, 18 were also tested using the Lyme C6 ELISA and none were positive.

Discussion
..Given that B. miyamotoi disease is an emerging tick-borne illness, further basic science research and in-vitro models are needed to clarify the mechanisms and optimal treatment of B. miyamotoi disease.

Funding
The Lyme and Tick-borne Diseases Research Center at Columbia University Irving Medical Center, established by the Lyme Disease Association and the Global Lyme Alliance, provided financial support for this study.

Quest Diagnostics - Licenses and Accreditation; Bron https://www.questdiagnostics.com/home/a ... ditations/

De 'Phelix Phage Borrelia test' is een niet-gevalideerde test, in een lopend (wetenschappelijk) onderzoek en wordt uitgevoerd door Red Laboratories dat geen ISO 15189 certificaat heeft van het BELAC België (de Belgische Accreditatie-instelling - Medische laboratoria (MED); Bron https://economie.fgov.be/nl/themas/kwal ... atoria-med hoe betrouwbaar is de PCR test waar mee wordt gewerkt en hoe betrouwbaar is een positieve uitslag op B. miyamotoi enkel op basis van de test?

[VerlorengezondheidM]

microorganisms-09-00154.pdf

Borrelia miyamotoi—An Emerging Human Tick-Borne
Pathogen in Europe

(1.09 MiB) 448 keer gedownload

Risk of Tick-Borne Disease May Be "Drastically Underestimated" in Western US

Kristi Rosa

Bron: Contagion live.com, lees meer

[Roxy]

Borrelia miyamotoi en Relapsing Fever in verschillende landen

MDPI
12 januari 2021
Review: 'Borrelia miyamotoi - An Emerging Human Tick-Borne Pathogen in Europe' by Katarzyna Kubiak, Magdalena Szczotko, and Małgorzata Dmitryjuk; Bron https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827671/
Borrelia miyamotoi in Europa; Bron 'Table 1'

..Worldwide, B. miyamotoi prevalence in questing Ixodes ticks ranges from 0.2 to 10% [42,43,44,77,82]. This pathogen has been detected in all three tick life stages (larvae, nymphs and adults) [68,83,84]. In European populations of I. ricinus,
B. miyamotoi was identified in 0.1-2% larvae [78,85], 0.4-2.8% nymphs [12,36,78,79] and 3.0-4.3% of adults [78,79]

Spanje (2018):
'Borrelia miyamotoi: Should this pathogen be considered for the diagnosis of tick-borne infectious diseases in Spain?' by Ana M Palomar, Aránzazu Portillo, Paula Santibáñez, Sonia Santibáñez, José A Oteo; Bron https://pubmed.ncbi.nlm.nih.gov/29187292/

Resultaten: 12,9% van de met Borrelia geïnfecteerde teken droegen B. miyamotoi en 87,1% Borrelia burgdorferi sensu lato.

Zweden (2010):
'Prevalence and diversity of Borrelia species in ticks that have bitten humans in Sweden' by Peter Wilhelmsson, Linda Fryland, Stefan Börjesson, Johan Nordgren, Sven Bergström, Jan Ernerudh, Pia Forsberg, Per-Eric Lindgren; Bron https://pubmed.ncbi.nlm.nih.gov/20844223/

Resultaten: 1% van de met Borrelia geïnfecteerde teken droegen B. miyamotoi en 99% Borrelia burgdorferi sensu lato.

Duitsland (2020):
'Borrelia miyamotoi and Borrelia burgdorferi (sensu lato) identification and survey of tick-borne encephalitis virus in ticks from north-eastern Germany' by Cristian Răileanu, Oliver Tauchmann, Ana Vasić, Elisabeth Wöhnke, Cornelia Silaghi; Bron https://pubmed.ncbi.nlm.nih.gov/32106890/

Resultaten: ongeveer 25% van de met Borrelia geïnfecteerde teken droegen B. miyamotoi en 75% Borrelia burgdorferi sensu lato.

Nederland (2017):
Publicatie - 'Borrelia miyamotoi in vectors and hosts in The Netherlands'; Bron https://pubmed.ncbi.nlm.nih.gov/28065617/

Out of 3360 unfed (or questing) nymphs, 313 (9.3%) were infected with B. burgdorferi s.l., 70 (2.1%) were infected with B. miyamotoi, and 14 (0.4%) were co-infected with B. burgdorferi s.l. and B. miyamotoi. Co-infection of B. burgdorferi s.l. with B. miyamotoi occurred more often than expected from single infection prevalences (p=0.03).

Artikel Biowetenschap en maatschappij (2020) - 'Koorts na een tekenbeet – soms iets anders dan lymeziekte?' over Borrelia miyamotoi; Bron https://www.biomaatschappij.nl/koorts-n ... ymeziekte/

België en Nederland (2014):
Publicatie - 'The presence of Borrelia miyamotoi, a relapsing fever spirochaete, in questing Ixodes ricinus in Belgium and in The Netherlands'; Bron https://www.researchgate.net/publicatio ... etherlands

In this study, we show the results of PCR screenings for B. miyamotoi in flagged Ixodes ricinus from Belgium and the Netherlands. B. miyamotoi was detected in nine of thirteen, and three of five locations from the Netherlands and Belgium, respectively. These outcomes indicate that B. miyamotoi is more spread than previously thought. The mean infection rate
B. miyamotoi was 1.14% for Belgium and 3.84% for the Netherlands.

V.S
Joseph J. Burrascano Jr - Making Rational Choices when Testing for Tick-Borne Diseases - Ilads 2020; Bron https://www.ilads.org/ilads-conference/ ... g-rational

DIVERSITY OF TICK-BORNE PATHOGENS- Need to test for co-infections
In a 2018 study that tested ticks submitted by patients to Igenex, 11.3% contained Lyme Borrelia, 3.4% contained tick-borne relapsing fever (TBRF) Borrelia, 3.3% contained Rickettsia, 2.4% contained Babesia, 2.1% contained Ehrlichia and 0.4% contained Bartonella.
A concurrent study in which over 10,000 patient samples tested by Igenex were reported, 37.3% were positive for Babesia species, 32.1% for Lyme Borrelia, 27.7% for TBRF, 19.1% for Bartonella, 16.7% for Anaplasma, 12.8% for Rickettsia and 6.9% for Ehrlichia. Positives were found in patient specimens from nearly every state.

LYME BORRELIOSIS
Igenex immunoblots for Lyme Borrelia are able to detect antibodies to eight distinct strains and species, and when tested using validation samples from the CDC, New York, California, the CAP and NY Biologicals, sensitivity was 100% when interpreted with Igenex interpretative criteria, and 89% with CDC criteria. Specificity was 97% when using Igenex criteria and 99.3% when using CDC criteria. Because not every patient generates detectable levels of antibody, adding PCR testing in this cohort can increase disease detection by 12% (294 specimens tested).

TBRF
Igenex immunoblots for TBRF are able to detect antibodies to six distinct species of TBRF. In well-characterized, PCR-positive patient samples, sensitivity was 100% (convalescent serum) and testing using a wide variety of specimen types revealed 97% specificity.

Ierland; Bron https://www.iddoctor.eu/docs/Crypto-Inf ... ambert.pdf
Plos One
15 april 2019
'Metagenomic 16S rRNA gene sequencing survey of Borrelia species in Irish samples of Ixodes ricinus ticks' by John S. Lambert, Michael John Cook, John Eoin Healy, Ross Murtagh, Gordana Avramovic, Sin Hang Lee; Bron https://journals.plos.org/plosone/artic ... ne.0209881

Questing Ixodes ricinus nymph samples were taken at six localities within Ireland.

The overall rate of borrelial infection in the Irish tick population was 5%, with a range from 2% to 12% depending on the locations of tick collection. The most prevalent species detected was B. garinii (70%) followed by B. valaisiana (20%) and B. miyamotoi (10%).

Red Laboratories - 'Are we looking at the wrong culprit'?: Bron https://www.ilads.org/ilads-conference/ ... se-culprit en Bron https://www.iddoctor.eu/docs/Crypto-Inf ... atovic.pdf en Bron https://www.youtube.com/watch?v=Mr2hKGOjCZM er is in 2020 gerapporteerd dat 60% van de teken die zij hebben getest positief waren voor Borrelia miyamotoi-fagen terwijl slechts 15% positief was voor Borrelia burgdorferi sensu lato-fagen en de percentages overeen kwamen met de resultaten van de geteste samples (bloed, urine, speeksel) van mensen uit verschillende landen in Europa en de VS.

Pathogens found in positive ticks:
Borrelia burgdorferi sl: 17% pos
Borrelia miyamotoi: 60% pos
Borrelia Relapsing Fever Group 23% pos
25% of positive ticks were for 2 pathogens

Unexpected high rate of B. miyamotoiand Relapsing Fever Borrelia Phages in tested ticks.

This result is in line with those obtained on human samples:
Since July2019, over 2100 results from patients originating various countries have been obtained.
Testing included mainly late stage/chronic patients and the aggregated data are showing 30% negative results and 70% positive among which over 60% indicated the presence of specific Borrelia miyamotoi phages.

With respect to the obtained results, a question raised: are we searching for the wrong culprit with Lyme-disease testing?

Interessant is wat het antwoord op de gestelde vraag zal gaan worden. Is het juist of onjuist?

[VerlorengezondheidM]

Transovarial transmission of Borrelia spp., Rickettsia spp. and Anaplasma phagocytophilum in Ixodes ricinus under field conditions extrapolated from DNA detection in questing larvae

Daniela Hauck, Daniela Jordan, Andrea Springer, Bettina Schunack, Stefan Pachnicke, Volker Fingerle & Christina Strube

Bron: parasitesandvectors.biomedcentral.com, lees meer

[Roxy]

@VerlorengezondheidM,
BMC
Parasites & Vectors
Publicatie 7 april 2020
Open access studie - 'Transovarial transmission of Borrelia spp., Rickettsia spp. and Anaplasma phagocytophilum in Ixodes ricinus under field conditions extrapolated from DNA detection in questing larvae' by Daniela Hauck, Daniela Jordan, Andrea Springer, Bettina Schunack, Stefan Pachnicke, Volker Fingerle & Christina Strube; Bron https://parasitesandvectors.biomedcentr ... 20-04049-7

Conclusions
Detection of pathogen DNA in questing I. ricinus larvae provides evidence for transovarial transmission of Rickettsia spp. and Borrelia spp. under field conditions. In consequence, I. ricinus larvae might serve as a source of human or animal infection with these pathogens. However, further studies investigating the percentage of viable transovarially transmitted Rickettsia and Borrelia species are necessary. As expected, transovarial transmission of A. phagocytophilum was rare.

Table 1 'Detection of pathogenic microorganisms in I. ricinus larvae nests'; Bron https://parasitesandvectors.biomedcentr ... 7/tables/1
Locatie (Samplelocatie): op verschillende locaties in Duitsland in een tijdsperiode van 2011 t/m 2018 in verschillende maanden (Bemonsteringsmaand) met in totaal 1.500 teken larven (Aantal onderzochte larven (totaal aantal larven)). Percentages van
B. miyamotoi worden in deze tabel en in de studie van de onderzoekers niet genoemd.

Amsterdam AMC - wetenschappelijk onderzoek 'Ticking on Pandora's Box'; https://www.amc.nl/web/specialismen/exp ... as-box.htm en ZonMw - wetenschappelijk onderzoek 'Ticking on Pandora's Box' looptijd 2017 - 2022; Bron https://www.zonmw.nl/nl/onderzoek-resul ... doras-box/
Vervolg onderzoek - 'Pathogenese en het ziektebeeld na infectie met Borrelia Miyamotoi'; Bron https://www.amc.nl/web/specialismen/exp ... amotoi.htm

..Huidig onderzoek
In Nederlandse Ixodes teken komen behalve de Borrelia burgdorferi sensu lato bacterie – de veroorzaker van de ziekte van Lyme – en teken encefalitis virus (TBEV), ook andere teken overdraagbare ziekteverwekkers voor. In de studie ‘Ticking on Pandora’s Box’ wordt gekeken hoe vaak Borrelia miyamotoi, Anaplasma phagocytophilum, Neoehrlichia mikurensis, Rickettsia species of Babesia species een acuut koortsend ziektebeeld na een tekenbeet kunnen veroorzaken. Daarnaast wordt er onderzoek gedaan naar de aangeboren immuunrespons tegen Borrelia miyamotoi. Deze bacterie kan koorts en griepachtige symptomen veroorzaken en in immuungecompromitteerde patiënten leiden tot meningoencephalitis. Kennis over de interactie van Borrelia miyamotoi met het immuunsysteem is van belang voor het begrijpen van de pathogenese en het ziektebeeld dat na infectie met deze bacterie kan ontstaan..

[Roxy]

Scientific Reports; Bron https://www.nature.com/srep/
Publicatie 2 maart 2021
Open access studie - 'Interaction between Borrelia miyamotoi variable major proteins Vlp15/16 and Vlp18 with plasminogen and complement' by Frederik L. Schmidt, Valerie Sürth, Tim K. Berg, Yi-Pin Lin, Joppe W. Hovius & Peter Kraiczy; Bron https://www.nature.com/articles/s41598-021-84533-x

Abstract
Borrelia miyamotoi, a relapsing fever spirochete transmitted by Ixodid ticks causes B. miyamotoi disease (BMD). To evade the human host´s immune response, relapsing fever borreliae, including B. miyamotoi, produce distinct variable major proteins. Here, we investigated Vsp1, Vlp15/16, and Vlp18 all of which are currently being evaluated as antigens for the serodiagnosis of BMD. Comparative analyses identified Vlp15/16 but not Vsp1 and Vlp18 as a plasminogen-interacting protein of B. miyamotoi. Furthermore, Vlp15/16 bound plasminogen in a dose-dependent fashion with high affinity. Binding of plasminogen to Vlp15/16 was significantly inhibited by the lysine analog tranexamic acid suggesting that the protein–protein interaction is mediated by lysine residues. By contrast, ionic strength did not have an effect on binding of plasminogen to Vlp15/16. Of relevance, plasminogen bound to the borrelial protein cleaved the chromogenic substrate S-2251 upon conversion by urokinase-type plasminogen activator (uPa), demonstrating it retained its physiological activity. Interestingly, further analyses revealed a complement inhibitory activity of Vlp15/16 and Vlp18 on the alternative pathway by a Factor H-independent mechanism. More importantly, both borrelial proteins protect serum sensitive Borrelia garinii cells from complement-mediated lysis suggesting multiple roles of these two variable major proteins in immune evasion of B. miyamotoi.

Material and methods
Bacterial strains and culture conditions
B. miyamotoi HT31 was cultivated at 33 ºC in a modified Kelly-Pettenkofer medium supplemented with 40% fetal calf serum as previously described86. B. garinii G1 was cultured until mid-exponential phase (5 × 107 cells per ml) at 33 °C in Barbour-Stoenner-Kelly (BSK-H) medium (Bio&SELL, Feucht, Germany). For production of hexahistidine (His6)-tagged proteins, Escherichia (E.) coli BL21 (DE3) cells (New England Biolabs, Frankfurt, Germany) grown in yeast tryptone (YT) broth at 37 °C were utilized.

Human serum, proteins, and antibodies
Human serum (NHS) was collected from healthy blood donors as described previously50. Human glu-plasminogen was purchased from Haematologic Technologies (Essex Junction, VT, USA) and urokinase plasminogen activator (uPA) (Merck, Darmstadt, Germany) were used for the activation of plasminogen to plasmin. The chromogenic substrate S-2251 (D-Val-Leu-Lys p-nitroanilide dihydrochloride) were from Sigma-Aldrich (Steinheim, Germany). Factor H, Factor B, C3b, and C5 were purchased from Complement Technology (Tyler, TX, USA). Polyclonal anti-plasminogen antibody was purchased from Acris Antibodies (Herford, Germany), and the monoclonal anti-plasminogen antibody (clone 10-V-1) was from Calbiochem, Merck, Darmstadt, Germany). The polyclonal anti-FH and anti-C3 antibody were obtained from Merck Biosciences (Bad Soden, Germany) and the polyclonal anti-C5, anti-Factor B antibody as well as the neoepitope-specific monoclonal anti-C5b-9 antibody was from Quidel (San Diego, CA, USA). The mouse anti-His antiserum was obtained from Novagen (Merck Darmstadt, Germany) and Qiagen (Hilden, Germany) and the horseradish peroxidase (HRP)-conjugated immunoglobulins were purchased from Dako (Hamburg, Germany).

Discussion
..In summary, we here describe additional roles of variable major proteins Vlp15/16 and Vlp18 involved in immune evasion. Overall, elucidating the complex interplay of these multifactorial proteins in vivo will certainly proof to be an important step towards understanding the pathogenic processes triggered by the relapsing fever spirochete B. miyamotoi.