Mark Wooten, Ph.D.
Professor
Chair, Institutional Biosafety Committee
Director, BSL3 Laboratory
Host/Pathogen Interactions in Lyme Disease and Melioidosis
Office: HEB 241
笔丑辞苍别:听419.383.6818
Fax: 419.383.3002
E-mail Address: R.Mark.Wooten@utoledo.edu
Media Release:
Dr. Wooten's laboratory is interested in the host/pathogen interactions that lead to the development of two different infectious diseases: Lyme disease and melioidosis.
Lyme disease听(i.e. Lyme borreliosis) is a particularly interesting and complicated malady that involves two major events: 1) persistent infection of the host by the spirochetal bacterium听Borrelia burgdorferi听and 2) the response of the host's innate immune defenses to the organism, which produces the inflammation that leads to the symptoms of Lyme disease while attempting to clear the persistent infection.听B. burgdorferi听is a highly infectious tick-borne bacterial obligate parasite that is especially adept at evading host defenses, disseminating widely via spirochetal motility through dense tissues, and persisting long-term within almost any tissue of the body. The infected host mounts a vigorous immune response to these spirochetes, as evidenced by the production of inflammatory soluble mediators and large quantities of听B. burgdorferi-specific antibodies. Although passive transfer of听B. burgdorferi-specific antiserum can prevent naive mice from subsequent infection, the immune response elicited during natural infection is usually unable to clear the infection, resulting in a persisting bacterial reservoir that can re-emerge under various conditions. This persistence in target tissues promotes prolonged stimulation of the host's innate defenses via interaction with endogenous bacterial lipoproteins, resulting in activation of immune pathways that appear to mediate much of the inflammatory pathology indicative of Lyme disease. Based on these dynamics, we are interested in utilizing the well-established murine model of Lyme disease to address the following areas:
- Identification of host immune cell types and mediators that are important in controlling the abilities of听B. burgdorferi听to infect, invade, and persist in mammalian host tissues.
- Identification of host signaling pathways that modulate the inflammatory pathology that is characteristic for Lyme disease.
- Usage of intravital microscopy techniques that allow direct visualization of the interactions between听B. burgdorferi听and different immune cell populations within the intact skin of living, infected mice over time.
- Assessment of the importance of听B. burgdorferi听motility and chemotaxis mechanisms for establishing acute and persistent infection in vertebrate and invertebrate hosts.
- Using our findings from the above experiments to identify targets for vaccines or other therapies that restore effective immune clearance.
Phagocytosis of B. burgdorferi by a dermal professional antigen-presenting cell.听 Transgenic mice (I-Ab-GFP expressing) possessing green MHC class II-expression were infected with DsRed-expressing B. burgdorferi intradermally into the ear.听 Intact ear tissues were imaged using intravital confocal microscopy on living mice at 24h post-infection. The four images were taken at 6-minute intervals (left-to-right).
Melioidosis听is a human and animal disease that is caused by infection with听Burkholderia pseudomallei, which is endemic within different tropical and subtropical regions worldwide. Acute disease can lead to fulminant septicemia with mortality rates of 40-90%, even with vigorous antibiotic and supportive therapy. Chronic disease can also develop, with recrudescence occurring months to years after initial exposure. While there are some predisposing factors that make certain human populations more susceptible to developing this disease (e.g. diabetes, alcohol abuse, renal disease, etc.), exposure to听B. pseudomallei-containing aerosols is reported to have an LD50听鈮�100 organisms in mice. Based on these properties, B. pseudomallei is considered a Tier 1 select agent with the highest potential for misuse as a biological weapon. This organism is also quite resistant to many classes of antibiotics and there is currently no vaccine. Thus, there is great interest in identifying targets for preventative and/or curative treatments for these infections.
Persistence within innate immune and other cell types appears to be central to the development of melioidosis, providing them with an environment where they can proliferate and spread cell-to-cell via actin polymerization, thus enabling the bacteria to spread and evade many humoral immune mediators. While relatively little is known about the molecular basis for听B. pseudomallei听惫颈谤耻濒别苍肠别 in vivo, our research suggests that these bacteria quickly escape the phagosome and evade intracellular killing, preventing bacterial clearance and generation of an effective adaptive immune response. We believe a better understanding of the basic biology of phagocyte subversion by this bacterium would greatly facilitate the development of preventative and curative treatments. We are utilizing the murine model of melioidosis, as well as human in vitro models, to address the following areas:
- Identification of mechanisms that virulent听B. pseudomallei听strains utilize to circumvent efficient clearance by macrophages/neutrophils.
- Identification of outer membrane proteins expressed by听B. pseudomallei听that might serve as virulence factors/vaccine candidates.
- Identification of bacterial mechanisms for evading complement-mediated killing
- Testing vaccines/therapeutics in animal models to assess effectiveness and identify immune correlates of clearance.
| Global distribution of Burkholderia pseudomallei. The map represents the global distribution of B. pseudomallei based on consensus evidence gathered from January 1910 to September 2022. Green color represents complete consensus on the absence of B. pseudomallei and red represents complete consensus on the presence of B. pseudomallei. To obtain updated global consensus evidence and perform this analysis, we used a weighted scoring system with a method modified from a previous modelling study (Supplementary Information).
B. pseudomallei polymerizes and attaches to actin filaments and uses it to disseminate from infected cells. In this micrograph, bacteria are stained green and actin within murine macrophages is stained red.
Dr. Wooten received his Bachelors (Zoology/Chemistry; 1985) and Masters (Microbiology/Immunology; 1990) from the University of Arkansas (Fayetteville) under the supervision of Dr. Jim Saunders. He received his Ph.D. (Microbiology/Immunology; 1995) from the University of Mississippi Medical Center under the supervision of Dr. Jan Bly. He completed post-doctoral training at the University of Utah College of Medicine in the laboratory of Dr. Janis Weis. Dr. Wooten joined the Department of Medical Microbiology and Immunology at the 淫妻社 College of Medicine in May 2001.
Current Funding:
Identification of immunoprotective antigens for preventing Lyme disease
NIH/NIAID (R01)
Period of support: 09/01/2026 - 08/31/2031
Principal Investigator: R. Mark Wooten
Factor H-Fc fusions as novel therapeutics for Burkholderia pseudomallei infections
National Institute of Allergy and Infectious Diseases (R41)
Period of Support: 08/01/23 - 07/31/25
Principal Investigator:听 R. Mark Wooten
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Representative Publications:
Breidenbach, J.D., B.W. French, T.T. Gordon, A.L Kleinhenz, F.K. Khalaf, J.C. Willey, J.R. Hammersley, R. M. Wooten, E. Crawford, N.N. Modyanov, D. Malhotra, S.T. Haller and D.J. Kennedy.听 Short-term, Low Dose Microcystin-LR Aerosol Induces Inflammatory Responses in Healthy Human Primary Airway Epithelium. In press
(2021). Microcystin-LR (MC-LR) triggers inflammatory responses in macrophages. Int J Mol Sci. 2021 Sep 14;22(18):9939. doi: 10.3390/ijms22189939. PMID:听34576099.
(2021). Interactions between the pathogenic Burkholderia and the complement system: A review of potential immune evasion mechanisms.听 Front Cell Infect Microbiol. 听(2021) Sep 30;11:701362. doi: 10.3389/fcimb.2021.701362. PMID:听34660335
(2020).听 Immune Response to Borrelia: Lessons from Lyme Disease Spirochetes. Curr Issues Mol Biol. DOI: 10.21775/cimb.042.145. PMID: 33289684.
. (2016).听 Borrelia burgdorferi CheY2 is dispensable for chemotaxis or motility but crucial for the enzootic life cycle of the spirochete.听 Infect Immun. 2016 Dec 29;85(1):e00264-16.doi:10.1128/IAI.00264-16
. (2016) The听Borrelia burgdorferi听CheY3 response regulator is essential for chemotaxis and completion of its natural infection cycle.听听May 20. doi: 10.1111/cmi.12617
听(2015) Spirochetal motility and chemotaxis in the natural enzootic cycle and development of Lyme disease. Curr Opin Microbiol 28: 106-113.
听听(2015) 听Motor rotation is essential for the formation of the periplasmic flagellar ribbon, cellular morphology, and Borrelia burgdorferi persistence within Ixodes tick and murine hosts. 听Infect Immun. 2015 May;83(5):1765-77. doi: 10.1128/IAI.03097-14.
听听(2014) 听Delineating the importance of serum opsonins and the bacterial capsule in affecting the uptake and killing of Burkholderia pseudomallei by murine neutrophils and macrohages. 听PLoS Negl Trop Dis. 2014 Aug 21;8(8):e2988. doi: 10.1371/journal.pntd.0002988.
听听(2013) 听Borrelia burgdorferi elicited-IL-10 suppresses the production of inflammatory mediators, phagocytosis, and expression of co-stimulatory receptors by murine macrophages and/or dendritic cells. 听PLoS One. 2013 Dec 19;8(12):e84980. doi: 10.1371/journal.pone.0084980
听 (2013) 听Secretion of growth factors from macrophages when culture with microparticles. J. Biomed. Mater. Res., Part A. Nov; 101(11):3170-80.
听 (2012) 听Capsule influences the deposition of critical complement C3 levels required for the killing of听Burkholderia pseudomallei听via NADPH-oxidase induction by human neutrophils. PLoS One. 2012;7(12):e52276. doi: 10.1371/journal.pone.0052276.
, RG.听 (2012) 听Human platelets efficiently kill IgG-opsonized听E. coli. FEMS Immunol Med Microbiol. 65:78-83.
听 (2012) 听ELISA-based measurement of antibody responses and PCR-based detection profiles can distinguish between active infection and early clearance of听Borrelia burgdorferi. Clin Dev Immunol. 2012:138069.
, ER.听 (2010) Identification and characterization of听Burkholderia mallei听and听B. pseudomallei听adhesions for human respiratory epithelial cells. BMC Microbiol. 10:250-269.
.听(2009) Roles for phagocytic cells and complement in controlling relapsing fever infection.听 J Leukoc Biol. 2009 Sep; 86(3):727-736.
Lazarus JJ, Kay MA, McCarter AL, Wooten RM.听(2008) Viable Borrelia burgdorferi enhances interleukin-10 production and suppresses activation of murine macrophages.听Infect Immun.听Mar;76:1153-1162.
*听 (2007)听 Borrelia burgdorferi binding of host complement regulator factor H is not required for efficient mammalian infection.听 Infect. Immun. 75:3131-3139.听 *These labs contributed equally to this manuscript.
听(2007)听 The Moraxella catarrhalis autotransporter McaP is a conserved surface protein that mediates adherence to human epithelial cells through its N-terminal passenger domain.听 Infect. Immun.听75:314-324.
听(2006)听 Gene expression profiling reveals unique pathways associated with differential severity of lyme arthritis. 听J. Immunol. 177:7930-7942.
听(2006)听 IL-10 Deficiency promotes increased Borrelia burgdorferi clearance predominantly through enhanced innate immune responses.听 J Immunol. 177: 7076-7085.
(2006) Identification of aFrancisella tularensis LVS outer membrane protein that confers adherence to A549 human lung cells.听 FEMS Microbiol. Lett. 263:102-108.
听(2004) Effects of vLsE Complementation on the Infectivity of Borrelia burgdorferi Lacking the Linear Plasmid Ip28-1. Infect. Immun., 72:6577-6585.
听(2003) Effect of Complement Component C3 Deficiency on Experimental Lyme Borreliosis in Mice. Infect. Immun.,听71:4432-4440.
听(2003) Tripalmitoyl-S-Glyceryl-Cysteine-Dependent OspA Vaccination of Toll-Like Receptor 2-Deficient Mice Results in Effective Protection from听Borrelia burgdorferi听Challenge. Infect. Immun.,听71:3894-3900.
听(2002) Toll-like receptor 2 plays a pivotal role in host defense and inflammatory response to听Borrelia burgdorferi. Vector Borne Zoonotic Dis,听2:275-278.
. (2002) Toll-like receptor 2 is required for innate, but not acquired, host defense to听Borrelia burgdorferi. J. Immunol.,听168:348-355.听
听(2001) Host-pathogen interactions promoting inflammatory Lyme arthritis: use of mouse models for dissection of disease processes. Curr. Opin. Micro.,听4:274-279.听
听(2001) Resistance to Lyme disease in decorin-deficient mice. J. Clin. Invest.,听107:845-852.听
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