Research Staff: Albert Sylvester, B.S., Research Technician
Students: (1) Noshin Kathuria, 2nd year Ph.D. student in Microbiology and Immunology, (2) Kush Modi, M.D., 2nd year Masters in Public Health student, (3) Shikata Mudhaka, M.D., 2nd year Infectious Disease Fellow and (4) Amanda Michael, M.D., 1st year Internal Medicine Resident.
Research Interests: Our laboratory carries out highly translational research in the area of vaccinology, immunology and infectious disease. The laboratory aims to develop next generation optimized DNA vaccine based strategies that target antigen-specific cellular and humoral immune responses to mucosal sites.
Research
DNA Vaccines
In the past decade and a half, the DNA vaccine concept has been tested and applied against various pathogens and tumor antigens. In theory, this conceptually safe, non-live vaccine approach represents a unique and technically simple means to induce immune responses. Importantly, DNA vaccines drive not only humoral immunity but also the holy grail of live infection, cellular immunity — in particular, the elusive induction of killer cytotoxic T cells (CTL). The mechanisms by which DNA vaccines produce antigen-specific immunity in vivo are under intense investigation, with an idealized model presented in Figure 1. The optimized gene sequence of interest (antigenic or immune adjuvant genes) is generated synthetically or by PCR, enzymatically inserted into the multiple cloning region of the plasmid backbone, purified, and delivered to the inoculation site by several delivery methods to the skin, subcutaneum or muscle. Using host cellular machinery, the plasmid enters the nucleus of transfected myocytes (1) and resident antigen presenting cells (APCs) (2); here the plasmid components initiate gene transcription, protein production in the cytoplasm, followed by formation of foreign antigens as proteins as peptide strings. The cell provides natural post-translational modifications to the antigen so that native protein conformations are maintained and customize the antigens in a similar manner to that induced by live infection with recombinant vectors.
These host-synthesized antigens then can become the subject of immune surveillance in the context of both MHC class-I and class-II of the vaccinated individual. APCs play a dominant role in the induction of immunity of DNA vaccines by presenting vaccine-derived endogenous peptides on major histocompatibility complex (MHC) I molecules following either their direct transfection by the plasmid vaccine (2) or by cross-presentation of cell associated exogenous antigens, for example due to APC engulfment of apoptotic transfected cells (3). In addition, APCs mediate the display of peptides on MHC II molecules after secreted protein antigens that have been shed from transfected cells are captured and processed within the endocytic pathway (4). Antigen-loaded APCs traffic to the draining lymph node (DLN) via the afferent lymphatic vessel (5) where they ‘present’ peptide antigens in combination with costimulatory molecules to naïve T cells, providing the necessary secondary signals to initiate an immune response and expansion of T cells (6). In response to peptide-bound MHC molecules and costimulatory second signals, activated CD4 T helper cells secrete cytokines during cell-to-cell interaction with B cells and bind to costimulatory molecules that are required for B cell activation (7). In addition, shed antigen can be captured by specific high affinity immunoglobulins expressed on the surface of B cells in the DLN; these then present processed antigen to T helper cells, thereby facilitating the induction of an effective B cell response. In theory, once migrating T cells have been primed in the DLN they could be restimulated and further expanded at the site of immunization via presentation of peptide–MHC complexes displayed by transfected muscle cells. These processes coordinately elicit specific immunity against plasmid-encoded antigen by activating both T and B cells which, now “armed”, can traffic through efferent lymphatic system (8) and provide a surveillance system. Together, the two arms of the immune system induced specifically following DNA vaccination can create a powerful defense against most infectious diseases.
Figure 1. Induction of Cellular and Humoral Immunity by DNA Vaccination
Image: Kutzler and Weiner (2008). DNA Vaccines: Ready for Prime Time? Nature Reviews Genetics Vol.9:1-13.
DNA vaccines against HIV-1 that target mucosal immune responses
A large and highly specialized innate and adaptive mucosal immune system protects the mucosal surface against potential pathogens, including HIV, and contributes almost 80% of all immunocytes that are accumulated in the mucosa-associated lymphoid tissue (MALT). The mucosal tissues are also one of the main interfaces between environmental antigens and the immune system. Based on a multitude of data there are several features desired in an HIV vaccine immunogen. Such an immunogen should induce strong and broad humoral and cellular immunity. Furthermore, as HIV is in general a sexually transmitted disease and the cells of the gut are preferentially targeted for viral destruction, an immunogen should be capable of inducing in particular mucosal as well as systemic immune responses. An effective vaccination modality that could elicit mucosal-specific and long-lasting immune response against mucosal invading pathogens has been an area of intense investigation.
In collaboration with the University of Pennsylvania, a major ongoing effort for the laboratory’s HIV-1 vaccine development includes an adjuvant strategy that encodes novel mucosal targeting chemokines, CCL25, -27 and -28, used to elicit HIV-1-specific T cells and immunoglobulin A at mucosal issues. A focus of the laboratory is to quantitatively measure the immunological function and phenotype of responding optimized DNA vaccine-induced antigen-specific CD8+ T cells (CTL), CD4+ T cells (T helper) as well as immunoglobulin isotypes (including sIgA) at mucosal sites, through the use of luminex, ELISpot, flow cytometric analysis, immunohistochemistry and ELISA assays. Moreover, new DNA vaccine delivery techniques including electroporation/ electrostimulation are underway and have dramatically improved potency.
Novel immune therapeutics to treat Clostridium difficile
The importance of studying Clostridium difficile (C. diff) lies in the fact that this bacterium is no longer considered a nuisance disease. In fact, public and scientific concern in the U.S. precipitated a joint CDC/FDA/HHS workshop May 2006 to discuss the problem. In addition, hospitals are experiencing marked increased rates of disease in the US, Canada, and Europe, while physicians are documenting markedly increased relapse rates following treatment with metronidazole. Mortality rates due to C. diff are increasing, and new strains produce higher levels of toxin and are resistant to antibiotics. C. diff. is a gram-positive, anaerobic, spore-forming bacterium recognized as the major cause of hospital-acquired diarrhea. While as many as 3% of healthy outpatient adults may be colonized with the organism, this rate increases dramatically following hospitalization. Disease results from the production of one of two major cytotoxins, toxins A and B, whose clinical manifestations range from mild diarrhea to severe colitis with potentially life-threatening complications, such as toxic megacolon/perforation. Since 2000, extensive publications have described prevalence rate resurgence, elevated clinical severity and increased patient reoccurrence rates of C. diff disease in North America and Europe. These events have renewed scientific interest in the development of novel approaches to disease treatment and prevention, including toxin-specific vaccines. Our laboratory is currently testing a highly optimized mucosal DNA vaccine strategy to treat this debilitating disease.
Examination of immune responses in Intravenous drug users
Substantial evidence from both clinical observations and animal experiments has shown that acute and chronic opioid administration exerts inhibitory effects on peripheral humoral and cellular immune responses including antibody production, natural killer cell activity, cytokine expression, and phagocytic activity, as well as increased sensitivity to sepsis during withdraw. Few studies have examined whether the opioid-induced peripheral immune suppression observed is also seen in the gastrointestinal lymphoid tissue, where a large and highly specialized innate and adaptive mucosal immune system protects the mucosal surface against potential pathogens and contributes to almost 80% of total lymphocytes in the body. In addition, immunosuppressed populations (transplantation, pregnancy, chemotherapy, drug abuse) present a unique challenge for potential receipt of vaccines due to decreased CTL and antibody recall responses. In light of this, the laboratory is working to further understand the role opioid administration plays in the regulation of host mucosal immune responses.
Selected Publications (See all Michele Kutzler's publications in PubMed.)
(Note: Wetzel is Michele Kutzler's maiden name - see pubmed articles)
"DNA Vaccines: Ready for Prime Time?" (review article)
M.A. Kutzler and D.B. Weiner
Nature Reviews Genetics (in press) Vol.9:1-13, 2008
"Sustained suppression of SHIV89.6P replication in macaques by vaccine-induced CD8+ memory T cells"
J. Yin, A. Dai, M.A. Kutzler, A. Shen, J. LeCureux, M.G. Lewis, T. Waldmann, D.B. Weiner, and J.D. Boyer
AIDS, Vol. 22(14):1739-48, 2008
"Heterosubtypic protection against pathogenic human and avian influenza viruses via in vivo electroporation of synthetic consensus DNA antigens"
D.J. Laddy, J. Yan, M. Kutzler, D. Kobasa, G.P. Kobinger, A.S. Khan, J. Greenhouse, N.Y. Sardesai, R. Draghia-Akli, and D.B. Weiner
PLoS ONE, 3:e2517, 2008
"Treatment Interruption as a tool to measure changes in immunologic response to HIV-1" (review article)
M.A. Kutzler and J.M. Jacobson
Current Opinion in HIV and AIDS, Vol.3 (2):131-135, 2008
"DAMGO-induced expression of chemokines and chemokine receptors: the role of TGF-{beta}1"
C. Happel, A.D. Steele, M.J. Finley, M.A. Kutzler, and T.J. Rogers
Journal of Leukocyte Biology, 83:956-963, 2008
"SIV DNA Vaccine Co-Administered with an Engineered Adjuvant Plasmids Enhances CD8 SIV Specific Cellular Immune Responses in Cynomolgus Macaques"
J.D. Boyer, T.M. Robinson, M.A. Kutzler, S.A. Calarota, D. Choo, S. Sackiley, V. Roopchand, M.K. Sidhu, K. Muthumani, M. Lewis, G. Pavlakis, T. Waldmann, D. Weiner
Proceedings of the National Academy of Sciences, USA, 104:18648-18653, 2007
"Epitope-driven TB vaccine development: a streamlined approach using immuno-informatics, ELISpot assays, and HLA transgenic mice"
J.A. McMurry, S. Kimball, J.H. Lee, D. Rivera, W. Martin, D.B. Weiner, M. Kutzler, D.R. Sherman, H. Kornfeld, and A.S. De Groot
Current Molecular Medicine, 7:351-368, 2007
"Enhanced cellular immune responses elicited by an engineered HIV-1 subtype B consensus-based envelope DNA vaccine"
J. Yan, H. Yoon, S. Kumar, M.P. Ramanathan, N. Corbitt, M. Kutzler, A. Dai, J.D. Boyer, and D.B. Weiner
Molecular Therapy, 15:411-421, 2007
"Comparative ability of plasmid IL-12 and IL-15 to enhance cellular and humoral immune responses elicited by a SIVgag plasmid DNA vaccine and alter disease progression following SHIV(89.6P) challenge in rhesus macaques"
S.Y. Chong, M.A. Egan, M. Kutzler, S. Megati, A. Masood, V. Roopchard, D. Garci-Hand, D.C. Montefiori, J. Quiroz, M. Rosati, E.B. Schadeck, J.D. Boyer, G.N. Pavlakis, D.B. Weiner, M. Sidhu, J.H. Eldridge, and Z.R. Israel
Vaccine, Vol.25(26):4967-82, 2007
"Smallpox: Pathogenesis and Host Immune Responses Relevant to Vaccine and Therapeutic Strategies. Microorganisms and Bioterrorism" (book chapter)
M.A. Kutzler, K.E. Ugen, and D.B. Weiner
Series: Infectious Agents and Pathogenesis. Friedman, Herman; Anderson, Burt; Bendinelli, Mauro (Eds.) p. 63-81. Illus. ISBN: 978-0-387-28156-8 (print) 978-0-387-28159-9 (online), 2006
"Regression of subcutaneous B16 melanoma tumors after intratumoral delivery of an IL-15-expressing plasmid followed by in vivo electroporation"
K.E. Ugen, M.A. Kutzler, B. Marrero, J. Westover, D. Coppola, D.B. Weiner, and R. Heller
Cancer Gene Therapy, 13:969-974, 2006
"Novel strategy for generation of mucosal immune responses against HIV-1 following systemic vaccination" (abstract)
M.A. Kutzler, K.A. Schoenly, K. Muthumani, R.M. Parkinson, H. Maquire, K. Ugen, and D. Weiner
Retrovirology, 3 Suppl 1:S30, 2006
"Mapping of immune responses following wild-type and mutant ABeta42 plasmid or peptide vaccination in different mouse haplotypes and HLA Class II transgenic mice"
M.A. Kutzler, C. Cao, Y. Bai, H. Dong, P.Y. Choe, V. Saulino, L. McLaughlin, A. Whelan, A.Y. Choo, D.B. Weiner, and K.E. Ugen
Vaccine, 24:4630-4639, 2006
"Coimmunization with an optimized IL-15 plasmid results in enhanced function and longevity of CD8 T cells that are partially independent of CD4 T cell help"
M.A. Kutzler, T.M. Robinson, M.A. Chattergoon, D.K. Choo, A.Y. Choo, P.Y. Choe, M.P. Ramanathan, R. Parkinson, S. Kudchodkar, Y. Tamura, M. Sidhu, V. Roopchand, J.J. Kim, G.N. Pavlakis, B.K. Felber, T.A. Waldmann, J.D. Boyer, and D.B. Weiner
Journal of Immunology, 175:112-123, 2005
"Developing an epitope-driven tuberculosis (TB) vaccine"
A.S. De Groot, J. McMurry, L. Marcon, J. Franco, D. Rivera, M. Kutzler, D. Weiner, and B. Martin
Vaccine, 23:2121-2131, 2005
"HIV vaccine development by computer assisted design: the GAIA vaccine"
A.S. De Groot, L. Marcon, E.A. Bishop, D. Rivera, M. Kutzler, D.B. Weiner, and W. Martin
Vaccine, 23:2136-2148, 2005
"SIV DNA vaccine co-administered with IL-12 expression plasmid enhances CD8 SIV cellular immune responses in cynomolgus macaques"
J.D. Boyer, T. M. Robinson, M.A. Kutzler, R. Parkinson, S. A. Calarota, M. K. Sidhu, K. Muthumani, M. Lewis, G. Pavlakis, B. Felber, and D. Weiner
Journal of Medical Primatology, 34:262-270, 2005
"Developing DNA vaccines that call to dendritic cells" (editorial)
M.A. Kutzler and D. B. Weiner
The Journal of Clinical Investigation, 114:1241-1244, 2004
"Engineering DNA vaccination as an approach to HIV immune therapy" (review article)
J.D. Boyer, M.A. Kutzler, M.A. Chattergoon, S.A. Calarota, G. Pavlakis, R.P. Sekaly, R.R. MacGregor, and D.B. Weiner
Clinical and Applied Immunology Reviews, 3:183-197, 2003
"Selective inactivation of CCR5 and decreased infectivity of R5 HIV-1 strains mediated by opioid-induced heterologous desensitization"
I. Szabo, M.A. Wetzel, N. Zhang, A.D. Steele, D. E. Kaminsky, C. Chen, L. Y. Liu-Chen, F. Bednar, E.E. Henderson, O. M. Howard, J. J. Oppenheim, and T.J. Rogers
Journal of Leukocyte Biology, 74:1074-1082, 2003
"Low power millimeter wave irradiation exerts no harmful effect on human keratinocytes in vitro"
I. Szabo, M. R. Manning, A. A. Radzievsky, M.A. Wetzel, T.J. Rogers, and M. C. Ziskin
Bioelectromagnetics, 24:165-173, 2003
"The effect of X4 and R5 HIV-1 on C, C-C, and C-X-C chemokines during the early stages of infection in human PBMCs"
M.A. Wetzel, A.D. Steele, E.E. Henderson, and T.J. Rogers
Virology, 292:6-15, 2002
"Interactions of opioid receptors, chemokines, and chemokine receptors" (review article)
I. Szabo, M. Wetzel, L. McCarthy, A. Steele, E.E. Henderson, M.Z. Howard, J.J. Oppenheim, and T.J. Rogers
Advances in Experimental Medicine and Biology 493:69-74, 2001
"Opioids, opioid receptors, and the immune response" (review article)
L. McCarthy, M. Wetzel, J.K. Sliker, T.K. Eisenstein, and T.J. Rogers
Drug and Alcohol Dependence, 62:111-123, 2001
"Cell-density-regulated chemotactic responsiveness of keratinocytes in vitro"
I. Szabo, M.A. Wetzel, and T.J. Rogers
The Journal of Investigative Dermatology 117:1083-1090, 2001
"The synthetic peptide WKYMVm attenuates the function of the chemokine receptors CCR5 and CXCR4 through activation of formyl peptide receptor-like 1"
B.Q. Li, M.A. Wetzel, J.A. Mikovits, E.E. Henderson, T.J. Rogers, W. Gong, Y. Le, F.W. Ruscetti, and J.M. Wang
Blood, 97:2941-2947, 2001
"Desensitization of chemokine receptor CCR5 in dendritic cells at the early stage of differentiation by activation of formyl peptide receptors"
Y. Le, M.A. Wetzel, W.W. Shen Gong, T.J. Rogers, E.E. Henderson, and J.M. Wang
Clinical Immunology, 99:365-372, 2001
"Down-regulation of the chemokine receptor CCR5 by activation of chemotactic formyl peptide receptor in human monocytes"
W. Shen, B. Li, M.A. Wetzel, T.J. Rogers, E.E. Henderson, S.B. Su, W. Gong, Y. Le, R. Sargeant, D.S. Dimitrov, J.J. Oppenheim, and J.M. Wang
Blood, 96:2887-2894, 2000
"Mu-opioid induction of monocyte chemoattractant protein-1, RANTES, and IFN-gamma-inducible protein-10 expression in human peripheral blood mononuclear cells"
M.A. Wetzel, A.D. Steele, T.K. Eisenstein, M.W. Adler, E.E. Henderson, and T.J. Rogers
Journal of Immunology, 165:6519-6524, 2000
In The News
Inovio Invited to Present at International Vaccine Conferences
September 24, 2010
Yahoo! Finance, Forbes, PharmaLive
Michele Kutzler was quoted in the press release “Inovio Invited to Present at International Vaccine Conferences” published by several media outlets including Forbes, PharmaLive, and Yahoo! Canada.
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