Bb BBK32-dependent vascular adhesion in vivo

Topics with information and discussion about published studies related to Lyme disease and other tick-borne diseases.
RitaA
Posts: 2766
Joined: Thu 1 Jul 2010 8:33

Bb BBK32-dependent vascular adhesion in vivo

Postby RitaA » Fri 26 Oct 2012 2:52

http://onlinelibrary.wiley.com/doi/10.1 ... 5/abstract

Vascular binding of a pathogen under shear force through mechanistically distinct sequential interactions with host macromolecules

Tara J. Moriarty1,2,*,
Meiqing Shi2,†,
Yi-Pin Lin4,
Rhodaba Ebady1,
Hong Zhou3,‡,
Tanya Odisho1,
Pierre-Olivier Hardy2,
Aydan Salman-Dilgimen2,
Jing Wu5,§,
Eric H. Weening5,¶,
Jon T. Skare5,
Paul Kubes3,
John Leong4,
George Chaconas2,*

Author Information

1 Matrix Dynamics Group, Faculty of Dentistry, and Department of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, ON, Canada
2 Snyder Institute for Chronic Diseases, Departments of Biochemistry & Molecular Biology and Microbiology and Infectious Diseases, University of Calgary, Calgary, AB, Canada
3 Snyder Institute for Chronic Diseases, Department of Physiology and Pharmacology, University of Calgary, Calgary, AB, Canada
4 Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA, USA
5 Department of Microbial and Molecular Pathogenesis, Texas A&M Health Science Center, Texas A&M University, Bryan, TX, USA

Article first published online: 24 OCT 2012

DOI: 10.1111/mmi.12045

© 2012 Blackwell Publishing Ltd

Summary

Systemic dissemination of microbial pathogens permits microbes to spread from the initial site of infection to secondary target tissues and is responsible for most mortality due to bacterial infections. Dissemination is a critical stage of disease progression by the Lyme spirochaete, Borrelia burgdorferi. However, many mechanistic features of the process are not yet understood. A key step is adhesion of circulating microbes to vascular surfaces in the face of the shear forces present in flowing blood. Using real-time microscopic imaging of the Lyme spirochaete in living mice we previously identified the first bacterial protein (B. burgdorferi BBK32) shown to mediate vascular adhesion in vivo. Vascular adhesion is also dependent on host fibronectin (Fn) and glycosaminoglycans (GAGs). In the present study, we investigated the mechanisms of BBK32-dependent vascular adhesion in vivo. We determined that BBK32–Fn interactions (tethering) function as a molecular braking mechanism that permits the formation of more stable BBK32–GAG interactions (dragging) between circulating bacteria and vascular surfaces. Since BBK32-like proteins are expressed in a variety of pathogens we believe that the vascular adhesion mechanisms we have deciphered here may be critical for understanding the dissemination mechanisms of other bacterial pathogens.

RitaA
Posts: 2766
Joined: Thu 1 Jul 2010 8:33

Re: Bb BBK32-dependent vascular adhesion in vivo

Postby RitaA » Fri 26 Oct 2012 6:14

I just noticed that supplementary information (although not the complete article) is available here:

http://onlinelibrary.wiley.com/doi/10.1 ... 5/suppinfo

Filename / Format / Size / Description

mmi12045-sup-0001-si.pdf ; 2341K ; Supporting Information

mmi12045-sup-0002-si.mov ; 7310K ; Supporting Information

mmi12045-sup-0003-si.mov ; 6413K ; Supporting Information

RitaA
Posts: 2766
Joined: Thu 1 Jul 2010 8:33

Re: Bb BBK32-dependent vascular adhesion in vivo

Postby RitaA » Fri 26 Oct 2012 7:12

From Dr. Moriarty's Lab:

http://piil.matrixdynamics.ca/lab_abstracts.html

The posters are especially interesting, but need to be magnified (by clicking on + ) to read.

Here are a few snippets from just one particularly interesting example:

1. Andrew Kao, Anil Bansal, Kyung-Phil Kim, Nupur Gupta, Rhodaba Ebady, Samir Patel and Tara Moriarty. 2012. "Lyme disease: Developing an ELISA-based diagnostic assay" . Research poster presented at the University of Toronto Infectious Diseases/Microbiology Research Day, University of Toronto, June 26, 2012.


http://piil.matrixdynamics.ca/abstracts ... poster.pdf

Lyme Disease: Developing an ELISA Based Diagnostic Assay

Abstract

Lyme disease (LD) is the most common vector-borne disease in temperate climates. The CDC reports a >350% increase in U.S. incidence over the last 15 years. LD is caused by the transmitted bacterium Borrelia burgdorferi (Bb), and is readily treated by antibiotics if infection is diagnosed early. CDC-recommended methods for serological testing for LD have several limitations, including: 1) low sensitivity of the assay in early stages of Bbinfection; 2) poor diagnostic range, resulting in the inability to distinguish between active and prior infections; 3) most current diagnostic tests use Bblysates or peptides from two infection-associated antigens (VlsE and OspC) to detect serum antibodies against Bb; however, the expression patterns and functions of these antigens during infection are not well understood. Recently, we determined that the Bb early antigen BBK32 mediates bacterial adhesion to vascular surfaces during bloodborne pathogen dissemination. We are producing a library of purified BBK32 sequence variants, domains and peptides for investigation of the BBK32-mediated vascular adhesion mechanism and epitope-mapping of dissemination-blocking BBK32 antibodies. We will use these BBK32 variants to develop an ELISA assay for identification of BBK32 peptides which are most highly immunogenic early in infection (IgM antibodies) and which exhibit a diagnostic range sensitive enough to distinguish between current and previous Bb infections. Development and validation of ELISA assays will be carried out using serum samples from control patients and clinically- and laboratory- diagnosed LD patients obtained from the OAHPP Public Health Laboratory. The sensitivity and specificity of the BBK32-based ELISA assay will be compared to the sensitivity and specificity of commercial kits used for diagnosis of LD. Earlier diagnosis of LD using BBK32-based ELISA assays could lead to more rapid introduction of antibiotic therapy, and more sensitive methods of determining the efficacy of antibiotic treatment.


[snip]

CURRENT LYME DISEASE DIAGNOSTIC ASSAYS:

•Most current diagnostic tests use whole Borrelia lysates or selected antigens to detect serum antibodies against B. burgdorferi.
•ELISA with C6 peptide (26 mer synthetic peptide analogue of the invariable region 6 (IR6) of the VIsE variable major protein like sequences has been shown to be highly sensitive and specific for the detection of B. burgdorferi infection. The C6 assay has a limited diagnostic range.
•More recent assay developments: Luciferase immuno-precipitation system (LIPS) developed for diagnosing B. burgdorferi, infections uses a synthetic antigen called VOVO (consists of repeated antigenic peptide sequences, VIsE-OspC-VIsE-OspC).
•Problem: Genetic variation in antigens in different bacterial strains


[snip]

WHY BBK32:

BBK32 is an early antigen, conserved, less variation between strains, critical for blood-borne dissemination stage of disease and likely a target of host antibody response

RitaA
Posts: 2766
Joined: Thu 1 Jul 2010 8:33

Re: Bb BBK32-dependent vascular adhesion in vivo

Postby RitaA » Fri 26 Oct 2012 7:15

For anyone interested in learning more about Dr. Moriarty or the current level of funding for the research her team is conducting, I'm including this link and information:

http://www.utoronto.ca/dentistry/facult ... iarty.html

Current academic / hospital appointments

2010 to present - Assistant Professor, Faculty of Dentistry, University of Toronto
2011 to present - Assistant Professor, Faculty of Medicine, Department of Laboratory Medicine & Pathobiology

Education

07/2005 - 07/2010 Postdoctorate - Biochemistry & Molecular Biology, Microbiology & Infectious Diseases, The University of Calgary, Canada
09/1999 - 05/2005 Doctorate (PhD), Doctor of Philosophy, Anatomy and Cell Biology, McGill University, Canada
09/1997 - 11/1999 Bachelor's, Bachelor of Science, Anatomy and Cell Biology, McGill University, Canada
09/1990 - 05/1997 Bachelor of Arts, Anthropology, McGill University, Canada

[snip]

Research interests

Pathogen hematogenous dissemination mechanisms
Spirochete pathogenesis mechanisms (especially Borrelia species)
Lyme disease/Lyme arthritis

Shear force-regulated vascular adhesion mechanisms
Real-time 3D single cell resolution imaging of pathogen infection & dissemination in living hosts
Host-pathogen interactions
Mouse models of periodontal disease

Recent publications

1. Lee, W.-Y., Moriarty, T.J., Wong, C.H., Zhou, H., Strieter, R.M., van Rooijen, N., Chaconas, G. and Kubes, P. 2010. “An intravascular immune response to Borrelia burgdorferi involves Kupffer cells and iNKT cells.” Nature Immunology 11 (4): 295-302.

2. Moriarty, T.J. and Chaconas, G. 2009. “Identification of the determinant conferring permissive substrate usage in the telomere resolvase, ResT.” Journal of Biological Chemistry 284 (35): 23293-301.

3. Tourand, Y., Deneke, J., Moriarty, T.J. and Chaconas, G. 2009. “Characterization and in vitro reaction properties of 19 unique hairpin telomeres from the linear plasmids of the Lyme disease spirochete.” Journal of Biological Chemistry. 284 (11): 7264-72.

4. Norman, M.U., Moriarty, T.J. (co-first author), Dresser, A.R., Millen, B., Kubes, P. and Chaconas, G. 2008. “Molecular mechanisms involved in vascular interactions of the Lyme disease pathogen in a living host.” PLoS Pathogens. 4(10): e1000169.

5. Moriarty, T.J. (co-first author), Norman, M.U., Colarusso, P., Bankhead, T., Kubes, P. and Chaconas, G. 2008. “Real-time high resolution 3D imaging of the Lyme disease spirochete adhering to and escaping from the vasculature of a living host.” PLoS Pathogens 4(6): e1000090.

Awards and Honours

2012 CIHR Bhagirath Singh Early Career Award in Infection and Immunity
2010 Postdoctoral Research Award; University of Calgary Department of Biochemistry and Molecular Biology (research achievements 2008-2010); Calgary, Canada
2010 Textbook cover; Borrelia: Molecular Biology, Host Interaction and Pathogenesis; Eds. D. Scott Samuels and Justin D. Radolf. 2010.
2009 Paper of the Week/Cover of publication issue/Author Spotlight: Journal of Biological Chemistry; Paper: Moriarty & Chaconas, 2009; top 1% of reviewed manuscripts
2008 Poster Prize; Imaging Host Pathogen Interactions symposium, Heidelberg, Germany
2006 – 2009 Postdoctoral Fellowship, National fellowship; Canadian Institutes of Health Research, Canada
2005 – 2008 Postdoctoral Fellowship; Provincial fellowship: Alberta Heritage Foundation for Medical Research, Canada; in years with CIHR fellowship overlap, received Incentive Award from AHFMR instead of full fellowship
2005 Dean’s Honour List, Ph.D. graduating class of McGill University
2005 Postdoctoral Fellowship; Provincial fellowship: Fonds de la recherche en santé Québec, Canada; declined
2005 Nomination for the Alice Wilson award, Royal Society of Canada; CIHR nominated its most highly rated female post-doctoral fellows pursuing their research in Canada during 2004 and 2005 for this award.
2003 McGill Graduate Studies Fellowship; McGill University Department of Anatomy & Cell Biology (research achievements 2002-2003)
2003 McGill Graduate Studies Fellowship; Institutional award: McGill University, Montréal, Canada
2003 Poster Prize; 3rd Annual McGill Experimental Medicine Graduate Students’ Society poster competition (1st prize, Cancer category), Montréal, Canada
2002 – 2005 Doctoral Research Scholarship; National scholarship: Canadian Institutes of Health Research, Canada
2002 Poster Prize; 2nd Annual McGill Experimental Medicine Graduate Students’ Society poster competition (1st prize, Cancer category), Montréal, Canada
1999 Challenge Program Bursary; Institutional award: Lady Davis Institute of Medical Research/ Human Resources Development Canada, Montréal, Canada
1997 Challenge Program Bursary; Institutional award: Regroupement des services aux aînés de centre-ville (Downtown Seniors’ Service Network)/ Human Resources Development Canada, Montréal, Canada

[snip]

Current research grants [I'm only including those that pertain to Lyme disease]

04/2012 - 03/2013 CIHR Bhagirath Singh Early Career Award in Infection and Immunity
Title: Mechanisms of vascular dissemination in the Lyme disease spirochete
Principal Applicant: Moriarty, T.J.
Amount: $25,000
Duration: lump sum prize;
Awarded for highest ranked new investigator operating grant in the fields of infection and immunity in the 2011 CIHR operating grant competitions

04/2012 - 03/2017 Canadian Institutes of Health Research (Operating Grant)
Title: Mechanisms of vascular dissemination in the Lyme disease spirochete
Principal Applicant: Moriarty, T.J.
Amount: $713,435
Duration: 5 years

01/2012-12/2012 National Research Fund for Tick-borne Diseases
Title: Mechanisms of metabolic syndrome-enhanced susceptibility to disseminated Lyme disease
Principal Applicant: Moriarty, T.J.
Amount: $60,000 USD
Duration: 1 year

05/2011 - 04/2016 Natural Sciences and Engineering Research Council of Canada (Discovery Grant)
Title: Spirochete coordination of adhesion and motility in the presence of fluid shear forces
Principal Applicant: Moriarty, T.J.
Amount: $150,000 ($30,000/year)
Duration: 5 years + 1 year automatic extension

03/2011 Canada Foundation for Innovation (Leaders Opportunity Fund/Ontario Research Fund (ORF) Small Infrastructure Fund)
Title: High resolution real-time 3D imaging of bacterial infection in living hosts: Investigation of blood-borne dissemination mechanisms of the Lyme disease pathogen
Principal Applicant: Moriarty, T.J
Amount: $637,024;
Duration: lump sum

07/2011—06/2012 The Banting Research Foundation (New Investigator competition)
Title: Vascular Adhesion Mechanisms of the Lyme Disease Spirochete
Principal Applicant: Moriarty, T.J.
Amount: $20,000;
Duration: 1 year

ChuckG
Posts: 99
Joined: Thu 17 Mar 2011 23:45
Location: Berkeley

Re: Bb BBK32-dependent vascular adhesion in vivo

Postby ChuckG » Sat 27 Oct 2012 1:05

Steere identified BBK32 as p35. Barbour et al 2008 found BBK32 to be strongly immunogenic (IgG) in both early and late lyme. Calculated molecular mass is 40.8 kDa so it is in the shadow of FlaB and BmpA,B,&D.

RitaA
Posts: 2766
Joined: Thu 1 Jul 2010 8:33

Re: Bb BBK32-dependent vascular adhesion in vivo

Postby RitaA » Sat 27 Oct 2012 1:21

Thanks very much for this useful information, Chuck.

I've been wondering a lot about BBK32 lately, and it helps to know that it's the same thing as Allen Steere's p35.

It does seem that Moriarty and Barbour are in agreement when it comes to BBK32/p35 being found in both early and late Lyme disease.

Thanks again.


Return to “Published Studies”

Who is online

Users browsing this forum: Brandwatch [Bot] and 4 guests