Progress 08/01/01 to 06/30/06
Outputs
We have determined the molecular mechanisms of antibody-mediated clearance of B. bronchiseptica and B. pertussis. These mechanisms require antibodies, complement activation, FcR binding,and neutrophils recruitment/activation. The two closely related subspecies differ in some very interesting aspects of this process. B. bronchiseptica is rapidly cleared by antibodies, and efficiently stimulates TLR4 via its lipopolysaccharide (LPS) to recruit/activate neutrophils, and rapidly overcomes animals lacking TLR4. B. pertussis does not stimulate TLR4 as efficiently, and expresses a toxin, pertussis toxin, that prevents the efficient recruitment of neutrophils. B. pertussis is therefore not efficiently cleared from the lower respiratory tract by antibodies. In collaboration with Reka Albert and Ottar Bjornstad we have examined the effects of avoiding antibody mediated clearance on the within-host and between-host dynamics of this organism to show that this characteristic is
critical to the biology of the organism that persists in immune populations.
Impacts
The major challenge to the control of B. pertussis disease is the ability of this organism to remain endemic even in highly vaccinated populations. Our data show that Pertussis Toxin gives B. pertussis the ability to infect immunized hosts, which would allow B. pertussis to circulate through vaccinated individuals, where it causes variable disease that usually goes undocumented. Understanding how these organisms can be controlled by immunity, and the specific mechanisms they use to avoid clearance, can allow us to target these mechanisms specifically, allowing us to block this pathway and thereby let the immune system more efficiently clear the infection.
Publications
- Pilione, M. R. and Harvill, E. 2006. The Bordetella type three secretion system inhibits IFN delta production that is required for efficient antibody-mediated bacterial clearance. Infection and Immunity. 74(2):1043-9.
- Pilione M.R. and Harvill E. T. 2006. CD11b is required for the resolution of inflammation induced by Bordetella bronchiseptica respiratory infection. Cellular Microbiology. 8(5):758-68.
- Joo, J., Gunney, M., Cases, M., Hudson, P., Albert, R., and Harvill, E.T. 2006. Bacteriophage-mediated intraspecific competition: Experiment and theory. Proceedings of Royal Society. 273(1595):1843-8.
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Progress 01/01/05 to 12/31/05
Outputs
Our work so far has determined the molecular and mechanistic details of antibody-mediated clearance of B. bronchiseptica and B. pertussis. Specifically, these mechanisms require antibodies and two important antibody effector functions, complement activation and FcR binding. Since we also determined that neutrophils are required, and must bear both complement receptors and FcRs on their surface, and that macrophages are required to recruit those neutrophils, we have compiled a complete model for the molecular mechanisms of bacterial clearance of the bordetellae. Importantly, we have identified relevant differences between B. bronchiseptica and B. pertussis that appear to relate to their different interactions with their respective hosts. In a significant expansion beyond the stated goals of this grant, we have determined that Pertussis Toxin, only expressed by B. pertussis, prevents rapid antibody-mediated bacterial clearance.
Impacts
Our work so far has determined the molecular and mechanistic details of antibody-mediated clearance of B. bronchiseptica and B. pertussis. Specifically, these mechanisms require antibodies and two important antibody effector functions, complement activation and FcR binding. Since we also determined that neutrophils are required, and must bear both complement receptors and FcRs on their surface, and that macrophages are required to recruit those neutrophils, we have compiled a complete model for the molecular mechanisms of bacterial clearance of the bordetellae. Importantly, we have identified relevant differences between B. bronchiseptica and B. pertussis that appear to relate to their different interactions with their respective hosts. In a significant expansion beyond the stated goals of this grant, we have determined that Pertussis Toxin, only expressed by B. pertussis, prevents rapid antibody-mediated bacterial clearance.
Publications
- Harvill, E.T. Lee, G., Grippe, V.K., and Merkel, T.J. 2005. Complement Depletion Renders C57BL/6 Mice Sensitive to the Bacillus anthracis Sterne Strain. Infection and Immunity. 737:4420-2.
- Bjornstad, O.N. and Harvill, E.T. 2005. Evolution and Emergence of Bordetella in Humans. Trends in Microbiology. 138:355-9.
- Skinner, J.A., Pilione, M.R., Shen, H., Harvill, E.T. and Yuk, M.H. 2005. Bordetella type III secretion modulates dendritic cell function resulting in immunosuppression and bacterial persistence. J. Immunol. 175:4647-4652.
- Mann, P.B., Preston, A., Kennett, M.J., and Harvill, E.T. 2005. Comparative TLR4 Mediated Innate Host Defense to Bordetella Infection. Infection and Immunity. 7312:8144-52.
- Kirimanjeswara, G.S., Mann, P.B., Pilione, M.R., Kennett, M.J. and Harvill, E.T. 2005. The complex mechanism of antibody-mediated clearance of Bordetella from the lungs requires TLR4. J. Immunol. 17511:7504-11.
- Kirimanjeswara, G.S., Agosto, L.M., Kennett, M.J., Bjornstad, O.N. and Harvill, E.T 2005. Pertussis toxin inhibits neutrophil recruitment to delay antibody-mediated clearance of Bordetella pertussis. Journal of Clinical Investigations. 11512:3594-601.
- Wolfe, D., Kirimanjeswara, G.S., and Harvill, E.T. 2005. Clearance of Bordetella parapertussis from the lower respiratory tract requires humoral and cellular immunity. Infection and Immunity. 73:6508-6513.
- Hughes, M.A., Green, C.S., Lowchyj, L., Lee, G., Grippe, V.K., Smith, Jr., M.F., Huang, C.L., Harvill, E.T and Merkel, T.J. 2005. MyD88-dependent signaling confers protection to Bacillus anthracis spore challenge in mice: Implications for Toll-Like Receptor Signaling. Infection and Immunity. 7311:7535-40.
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Progress 01/01/04 to 12/31/04
Outputs
The work in our lab over the past year has substantially advanced our understanding of the interactions between the respiratory pathogens of the genus Bordetella and its natural hosts. In using the mouse model we have been able to use many molecular tools to manipulate host immunity to infection, thereby determining the immune functions involved in efficient control of bacterial numbers. In addition, we have begun to characterize the factors involved in pathogenesis, which is often independent of bacterial numbers. We have found that it is, however, dependent on certain bacterial genes. The combination of genetic manipulation of bacterial factors, comparative analysis of human and animal pathogens and manipulation of host immunity continue to generate insight into the complex interactions between the bordetellae and their mammalian hosts.
Impacts
We want to determine which immune functions are important in the control/clearance of the bordetellae. We have used the combination of adoptive transfer of antibodies and knockout mice lacking antibody effector functions to identify antibodies as critical to the immune response and have now determine the mechanism of antibody-mediated clearance of B. bronchiseptica and are currently investigating what appear to be substantial differences to that of B. pertussis.
Publications
- Mann, P.B., Kennett, M.J., and Harvill, E.T. 2004. Toll-Like Receptor 4 Is Critical to Innate Host Defense in a Murine Model of Bordetellosis. J Infect Dis. 2004 Mar 1;189(5):833-836.
- Pishko, E.J., Kirimanjeswara, G.S, Gopinathan, L., Pilione, M.R., Kennett, M.J. and Harvill, E.T. 2004 Antibody-mediated Bacterial Clearance from the Lower Respiratory Tract of Mice Requires Complement Component C3. Eur J Immunol. 34(1):184-93.
- Liu, M., Liu, Y., Doulatov, S.R., Gingery, M., Eiserling, F.A., Baker, S., Davis, P., Preston, A., Maskell, D.J., Harvill, E.T., Parkhill, J. and Miller, J.F. 2004 Genomic and genetic analysis of Bordetella bacteriophages encoding reverse transcriptase-mediated tropism-switching cassettes. J Bacteriol. 186(5):1503-17.
- Mann, P.B., Elder, K.D., Kennett, M.J., and Harvill, E.T. 2004, TLR4 Dependent Early Elicited TNFa Expression is Critical for Innate Host Defense Against Bordetella bronchiseptica. Infection and Immunity. 72(11):6650-8.
- Elder, K.D. and Harvill, E.T. 2004. Strain-Dependent Role of BrkA during Bordetella pertussis Infection of the Murine Respiratory Tract. Infection and Immunity. 72(10):5919-24.
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Progress 01/01/03 to 12/31/03
Outputs
We have developed a mouse model of bordetellosis in which both bacterium and host immunity can be manipulated genetically to characterize their interactions at the molecular level. We will now use this model to dissect the host immune response factors that are important in controlling and/or clearing B. bronchiseptica and B. pertussis infections. Mice with specific immunodeficiencies will be used to determine the role of individual immune functions. We have used this approach to identify antibodies as one important factor and have now determine the mechanism of antibody-mediated clearance. We have compared these two bordetellae in the mouse model to dissect the host immune response factors that are important in controlling and/or clearing infection. Mice with specific immunodeficiencies have been used to determine the role of individual immune functions. We have identified antibodies as necessary and sufficient to clear all three bordetellae from the lower respiratory
tract and have determined the mechanism of antibody-mediated clearance. Antibodies require both complement and Fc receptors to clear B. bronchiseptica from the lungs, but only complement in the trachea. In addition, we have shown that complement acts by opsonizing B. bronchiseptica for phagocytosis via CR3. However, different mechanisms are at work in antibody-mediated clearance of B. pertussis. Although antibodies are required for clearance, adoptively transferred antibodies alone have no effect on B. pertussis in naive mice until after endogenous immunity is generated, and they have no effect in mice that lack adaptive immunity. In addition, we have examined the contributions of antibody effector functions to B. pertussis clearance and have determined that complement is not required, but FcRs are, although the spatial and temporal parameters and magnitude of their contribution remains to be determined.
Impacts
We want to determine which immune functions are important in the control/clearance of the bordetellae. We have used the combination of adoptive transfer of antibodies and knockout mice lacking antibody effector functions to identify antibodies as critical to the immune response and have now determine the mechanism of antibody-mediated clearance of B. bronchiseptica and are currently investigating what appear to be substantial differences to that of B. pertussis.
Publications
- Burns, V.C., Pishko, E.J., Preston, A., Maskell, D.J. and Harvill, E.T. 2003. The Role of Bordetella O-antigen in Respiratory Tract Infection. Infection and Immunity. 71(1):86-94.
- Kirimanjeswara, G.S., Mann, P.B. and Harvill, E.T. 2003. The Role of Antibodies in Immunity to Bordetella Infections. Infection and Immunity. 71(4):1719-1724.
- Preston, A., Maxim, E., Toland, E., Pishko, E.J., Harvill, E.T., Caroff, M. and Maskell, D.J. 2003. Bordetella bronchiseptica PagP is a Bvg-regulated lipid A palmitoyl transferase that is required for persistent colonisation of the mouse respiratory tract. Molecular Microbiology. 48(3):725-736.
- Pishko, E.J., Betting, D.J., Hutter, C.S. and Harvill, E.T. 2003. Bordetella pertussis Acquires Resistance to Complement Mediated Killing in vivo. Infection and Immunity. 71(9):4936-42.
- Pilione, M.R., Pishko, E.J., Preston, A. and Harvill, E.T. 2003 pagP is Required for Resistance to Antibody-Mediated Complement Lysis During Bordetella bronchiseptica Respiratory Infection. Accepted for Publication.
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Progress 01/01/02 to 12/31/02
Outputs
B. pertussis and B. parapertussis cause whooping cough in humans and B. bronchiseptica causes Atrophic Rhinitis and other respiratory tract diseases that result in millions of dollars of damage to US agriculture yearly. B. bronchiseptica also infects a very broad range of mammals and causes many other diseases including kennel cough in dogs and snuffles in rabbits. Current vaccines do not protect from infection, resulting in the persistent colonization of a large proportion (ninety percent) of herds. We have developed a mouse model of this disease in which both bacterium and host immunity can be manipulated genetically to characterize their interactions at the molecular level. We will now use this model to dissect the host immune response factors that are important in controlling and/or clearing B. bronchiseptica infection in comparison to other bordetellae. Mice with specific immunodeficiencies will be used to determine the role of individual immune functions. We
have used this approach to identify antibodies as one important factor and will now determine the mechanism of antibody-mediated clearance. Together these experiments will reveal which immune functions are important in the control/clearance of B. bronchiseptica. The goal of this research project is to understand how the immune response controls Bordetella infections.
Impacts
We have compared the three bordetellae in the mouse model to dissect the host immune response factors that are important in controlling and/or clearing infection. Mice with specific immunodeficiencies have been used to determine the role of individual immune functions. We have identified antibodies as necessary and sufficient to clear all three bordetellae from the lower respiratory tract and have determined the mechanism of antibody-mediated clearance. Antibodies require both complement and Fc receptors to clear B. bronchiseptica from the lungs, but only complement in the trachea. However, different mechanisms are at work in antibody-mediated clearance of B. pertussis and B. parapertussis. Based on the results it will be possible to rationally design new vaccines that maximally induce the appropriate immune response, allowing for more effective control and possibly eradication of this important pathogen
Publications
- Heininger, U., Cotter, P.A., Fescemyer, H, Martinez de Tejada, G., Yuk, M.H., Miller, J.F. and Harvill, E.T. 2002 Comparative Phenotypic Analysis of the Bordetella parapertussis Strain Selected for Genomic Sequencing, Infection and Immunity, 70(7): 3777-3784.
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Progress 01/01/01 to 12/31/01
Outputs
Bordetella bronchiseptica causes or contributes to Atrophic Rhinitis in swine, Kennel Cough in dogs, Snuffles in rabbits, and multiple other diseases in domestic and wild animals. Vaccines for B. bronchiseptica have not been shown to be effective in preventing disease, and no vaccine for any Bordetella subspecies has been shown to affect colonization rates. Consequently B. bronchiseptica is endemic in many herds and kennels. We have investigated the mechanisms of protective immunity to B. bronchiseptica infection to enable the design of improved vaccines and therapies to eliminate this important pathogen. We have recently determined that antibodies are necessary and sufficient to rapidly clear B. bronchiseptica from the lower respiratory tract of mice, one of its natural hosts. Subsequently, we used mice lacking specific antibody effector functions to determine their respective contributions to antibody-mediated effects. Using this approach we have identified specific
antibody effector functions that are involved in antibody-mediated bacterial clearance and shown that different effector functions are involved in different regions of the respiratory tract.
Impacts
This work is defining the role of various host immune functions in control and clearance of bacterial respiratory disease. This information will allow for the rational design of improved vaccines and therapeutics to control and/or eliminate respiratory pathogens from domestic animal herds.
Publications
- Le, L.Q., Kabarowski, J.H.S., Weng, Z., Satterthwaite, A.B., Harvill, E.T., Jensen, E.R., Miller, J.F., Witte, O.N. 2001. Mice Lacking the Orphan G Protein-Coupled Receptor G2A Develop a Late-Onset Autoimmune Syndrome. Immunity 14:561-571.
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