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• Research & Scholarship Committee, Group on Faculty Affairs, Association of American Medical Colleges

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 staff: Albert Sylvester, BS, Research Technician

Students

• Kush Modi, MD, second-year master's in public health student
• Shikata Mudhaka, MD, second-year infectious disease fellow
• Amanda Michael, MD, first-year internal medicine resident

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.

Philadelphia Inquirer (November 20, 2020)

Mobilized: Drexel Researchers Face Down The Coronavirus
Pulse Fall 2020

This Week in Virology (September 27, 2015)

Exel (2015)

The Bulletin (November 10, 2014)

Fierce Biotech Research (August 5, 2014)

"New Research Tests C. Difficile Vaccine Efficacy"
(August 4, 2014)

highlights of our C. diff vaccine outcome
(August 1, 2014)

WHYY's radio program "The Pulse" (July 31, 2014)

Science Daily (July 31, 2014)

Philly.com (January 22, 2013)

Selected Publications

(See all . Note: Wetzel is Michele Kutzler's maiden name - .)

Gary EN, Kathuria N, Makurumidze G, Curatola A, Ramamurthi A, Bernui ME, Myles D, Yan J, Pankhong P, Muthumani K, Haddad E, Humeau L, Weiner DB, Kutzler MA
Vaccine, 38(11):2626-2635, Epub 2020 Feb 10, Mar 4, 2020

"Developments towards a prophylactic hepatitis C virus vaccine"
BP Latimer, EN Gary and MA Kutzler
Current Immunology Reviews, 12:2, 2016

"Chemokine Adjuvanted Electroporated-DNA Vaccine Induces Substantial Protection from Simian Immunodeficiency Virus Vaginal Challenge"
M A Kutzler, M C Wise, N A Hutnick, Z Moldoveanu, M Hunter, M Reuter, S Yuan, J Yan, A Ginsberg, A Sylvester, B Pahar, D Carnathan, N Kathuria, A S Khan, D Montefiori, N Y Sardesai, M R Betts, J Mestecky, P Marx and D B Weiner.
Nature Mucosal Immunology, advance online publication, May 6, 2015; doi:10.1038/mi.2015.31

"An Optimized, Synthetic DNA Vaccine Encoding the Toxin A/Toxin B RBDs of Clostridium difficile Induces Protective Antibody Responses In Vivo"
Baliban, SM, Michael, A, Shammassian B, Mudakha S, Khan A S, Cocklin S, Zetner I, Latimer BP, Bouillaut L, Hunter, M, Marx P, Sardesai NY, Welles SL, Jacobson JM, Weiner DB, Kutzler MA.
Infect Immun; 82(10):4080-91, Oct 2014

"Neuronal ferritin heavy chain and drug abuse affect HIV-associated cognitive dysfunction"
Pitcher J, Abt A, Myers J, Han R, Snyder M, Graziano A, Festa L, Kutzler M, Garcia F, Gao WJ, Fischer-Smith T, Rappaport J, Meucci O.
The Journal of clinical investigation 124, 656-669, 2014

"Strong HCV NS3/4a, NS4b, NS5a, NS5b-specific cellular immune responses induced in Rhesus macaques by a novel HCV genotype 1a/1b consensus DNA vaccine"
Latimer, B., R. Toporovski, J. Yan, P. Pankhong, M. P. Morrow, A. S. Khan, N. Y. Sardesai, S. L. Welles, J. M. Jacobson, D. B. Weiner, and M.A. Kutzler
Hum Vaccin Immunother;10(8), June 20, 2014

"Generation of Antigen-Specific Immunity Following Systemic Immunization with DNA vaccine encoding CCL25 Chemokine Immunoadjuvant"
Kathuria N., Kraynyak K.K., Carnathan, D., Betts, M., Weiner, D.B., and Kutzler, M.A.
Human Vaccine & Immunotherapeutics. Nov 1;8(11), 2012

"DNA-based HIV vaccines do not induce generalized activation in mucosal tissue T cells"
Reuter M.A., Yuan, S., Marx, P.A., Kutzler, M.A., Weiner, D.B., Betts, M. R.
Human Vaccine & Immunotherapeutics 2012 Oct 30;8(11). [Epub ahead of print], 2012

"Non-contact helium-based plasma for delivery of DNA vaccines: Enhancement of humoral and cellular immune responses"
Connolly RJ, Chapman T, Hoff AM, Kutzler MA, Jaroszeski MJ, Ugen KE
Hum Vaccin Immunother. 2012 Aug 16;8(11). [Epub ahead of print], 2012

"High antibody and cellular responses induced to HIV-1 clade C envelope following DNA vaccines delivered by electroporation"
Yin, J., A. Dai, J. Lecureux, T. Arango, M. A. Kutzler, J. Yan, M. G. Lewis, A. Khan, N. Y. Sardesai, D. Montefiore, R. Ruprecht, D. B. Weiner, and J. D. Boyer
Vaccine 29: 6763-6770, 2011

"Long-term programming of antigen-specific immunity from gene expression signatures in the PBMC of rhesus macaques immunized with an SIV DNA vaccine"
Belisle, S. E., J. Yin, D. J. Shedlock, A. Dai, J. Yan, L. Hirao, M. A. Kutzler, M. G. Lewis, H. Andersen, S. M. Lank, J. A. Karl, D. H. O'Connor, A. Khan, N. Sardesai, J. Chang, L. Aicher, R. E. Palermo, D. B. Weiner, M. G. Katze, and J. Boyer
PLoS One 6: e19681, 2011

"Novel peptides based on HIV-1 gp120 sequence with homology to chemokines inhibit HIV infection in cell culture"
Chertov, O., N. Zhang, X. Chen, J. J. Oppenheim, J. Lubkowski, C. McGrath, R. C. Sowder, 2nd, B. J. Crise, A. Malyguine, M. A. Kutzler, A. D. Steele, E. E. Henderson, and T. J. Rogers
PLoS One 6: e14474, 2011

"Opioid-induced chemokine expression requires NF-kappaB activity: the role of PKCzeta"
Happel C, Kutzler M, Rogers TJ
J Leukoc Biol; 89:301-309, 2011

"Systemic immunization with CCL27/CTACK modulates immune responses at mucosal sites in mice and macaques"
Kraynyak KA, Kutzler MA, Cisper NJ, Khan AS, Draghia-Akli R, Sardesal NY, Lewis M G,Yan J, Weiner DB
Vaccine; 28:1942-1951, 2010

"Plasmids encoding the mucosal chemokines CCL27 and CCL28 are effective adjuvants in eliciting antigen-specific immunity in vivo"
Kutzler MA, Kraynyak KA, Nagle SJ, Parkinson RM, Zharikova, Chattergoon DM, Maguire H, Muthumani K, Ugen K, Weiner DB
Gene Ther; 17;72-82, 2010

"Plasmid-encoded IL-15Rα adjuvants specific immune responses induced by a DNA vaccine in vivo"
Kraynyak KA, Kutzler MA, Cisper NJ, Laddy DJ, Morrow MP, Waldmann TA, Weiner DB
Hum Gene Ther; 20:1143-1156, 2009

"Coimmunization with an optimized IL15 plasmid adjuvant enhances humoral immunity via stimulating B cells induced by genetically engineered DNA vaccines expressing consensus JEV and WNV E DIII"
Ramanathan MP, Kutzler MA, Kuo YC, Yan J, Liu H, Shah V, Bawa A, Selling B, Sardesai NY, Kim JJ, Weiner DB
Vaccine; 27:4370-4380, 2009

"High dose of plasmid IL-15 inhibits immune responses in an influenza non-human primates immunogenicity model"
Yin, J., A. Dai, D. J. Laddy, J. Yan, T. Arango, A. S. Khan, M. G. Lewis, H. Andersen, M. A. Kutzler, R. Draghia-Akli, D. B. Weiner, and J. D. Boyer
Virology 393: 49-55, 2009

"Ultra-Fast Low Concentration Detection of Candida Pathogens Utilizing High Resolution Micropore Chip"
R. Mulero, D.H.Lee, M.A.Kutzler, J.M. Jacobson and M.J. Kim.
Sensors. Vol. 8;1-X, 2008.

(review article)
M.A. Kutzler and D.B. Weiner
Nature Reviews Genetics (in press) Vol.9:1-13, 2008

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

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

(review article)
M.A. Kutzler and J.M. Jacobson
Current Opinion in HIV and AIDS, Vol.3 (2):131-135, 2008

C. Happel, A.D. Steele, M.J. Finley, M.A. Kutzler, and T.J. Rogers
Journal of Leukocyte Biology, 83:956-963, 2008

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

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

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

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

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

(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

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

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

A.S. De Groot, J. McMurry, L. Marcon, J. Franco, D. Rivera, M. Kutzler, D. Weiner, and B. Martin
Vaccine, 23:2121-2131, 2005

A.S. De Groot, L. Marcon, E.A. Bishop, D. Rivera, M. Kutzler, D.B. Weiner, and W. Martin
Vaccine, 23:2136-2148, 2005

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