Department of Veterinary Pathology - Jeffrey J. Adamovicz

Building Address: University of Missouri, College of Veterinary Medicine, Department of Veterinary Pathobiology, 209D Connaway Hall, Columbia, MO 65211-5130

Phone Number: 573 882-3261

Email: adamoviczj@missouri.edu

Lab Website: http://rbl.missouri.edu/

Our lab has broad interests in zoonotic infectious disease and vaccinology. I continue to be fascinated and drawn to both newly emerging diseases such as Zika virus and canine dysautonomia as well as better understood older pathogens such as the causative agents of plague, anthrax, meliodosis, brucellosis and tularemia. My current research interests are in the development of vaccines and animal models for zoonotic diseases and the pathogenesis of facultative intracellular bacteria.

Our lab is currently working on a human vaccine for Burkholderia pseudomallei, the causative agent of meliodosis. Human disease caused by Burkholderia spp. is a serious problem in many parts of the world including infections acquired by immunocompromised patients and those with cystic fibrosis. Traditional antimicrobial therapy is protracted and problematic. There is an urgent need to develop vaccines for Burkholderia infections, particularly those caused by highly pathogenic B. pseudomallei and B. mallei as well as less pathogenic B. multivorans and B. cenocepacia. This project is a multi-disciplinary effort with collaborating laboratory groups.

We have also embarked on collaborative studies for Zika Virus, a recently re-emerged flavivirus. In order to better support testing of both novel vaccines and therapeutic studies we have worked to develop a small animal model for ZIKV. This new model will allow us to study the host immune response to ZIKV and vector (mosquito) transmission studies.

Our lab also has a continuing interest in immunity to Y. pestis, B. abortus and other facultative intracellular bacteriaparticularly the role of DCs as vaccine targets or as targets to alter the pathogenesis of disease. There is a paucity of data on how bacteria interact with dendritic cells and thwart both innate and adaptive immune functions. We are interested in defining methodologies to select and test important T cell epitopes as part of rationally designed vaccines which will induce more effective T cell responses to facultative intracellular bacteria.

This project leverages structural vaccinology employing an in silico approach followed by in vitro and in vivo assessments of efficacy. Antibodies with antibacterial effects against Burkholderia have been identified and found to be protective in a mouse model of infection. T cell immunity has also been shown to play an important role in protection. However, no one has developed a protective vaccine for Burkholderia. Because of these findings, the nature of protective immunity to Burkholderia spp. remains elusive with some advocating the role of antibody while others argue for T cell-based protective mechanisms. Researchers have worked to identify protective vaccines against Burkholderia with some conserved candidates having been identified including its lipopolysaccharide (LPS) and its flagellar protein FliC. Unfortunately, Burkholderia spp. can phase shift and modify its LPS, escaping anti-LPS antibody based protection. Therefore, we will focus our epitope discovery efforts on the flagellar protein FliC. Putative protective T cell epitopes have been proposed for FliC, but have been found to be non-protective. Immunogenic, bactericidal B cell epitopes have also been identified for FliC, but have also have not been shown to be non-protective in vivo. We propose to include these observations and data into our search for FliC linked B and T cell epitopes. These proof of concept studies may support our search for additional vaccine target antigens.

Zika Virus Animal Model Development and Pathogenesis.
Zika virus (Flaviviridae, Flavivirus, [ZIKV]) is an arthropod-borne virus (arbovirus) transmitted by mosquitoes of the genus Aedes. Currently, there are no FDA-approved antivirals or vaccines available to treat or prevent ZIKV infections. Major efforts are being undertaken to develop vaccines and therapeutics against the virus. An important tool needed for these efforts is an appropriate animal model that mimics ZIKV pathogenicity as observed in humans and also supports the viral transmission cycle between the vertebrate host and the invertebrate mosquito vector. Immune deficient mice A129 (INFR-/-) and AG129 (INFR-/-) but not wild-type mice have been shown to be susceptible to needle delivered ZIKV infection indicating that these immune-deficient mice can be useful for studying ZIKV pathogenesis and Type I interferon response. However, it has also been shown that there are important differences in viral targets of the Type I interferon response between mice and humans, specifically regarding the inhibition of STAT2 by ZIKV and dengue virus (DENV) NS5, which is known to occur in humans but not in mice . We recently demonstrated that genetically engineered STAT2 knockout (KO) golden Syrian hamsters (Mesocricetus auratus) are highly susceptible to ZIKV infection via mosquito bite. We also showed that timed-pregnant STAT2 KO hamsters injected with ZIKV at 8 days post-coitus led to the infection of the placenta and fetal brains. Regarding implantation, the reproductive system of hamsters is more similar to that of humans in comparison to that of mice as in the latter, the placenta does not achieve its final structure until halfway through gestation. We hypothesize that genetically-engineered, immune-compromised STAT2 KO hamsters are an appropriate model for mimicking the ZIKV transmission cycle and for investigating ZIKV pathogenesis, fetal infection and sexual transmission to better support the development of vaccines and/or antiviral compounds.

The development of a brucellosis vaccine for cattle is a strategic goal of the USDA and CABs. Towards this end we first choose to test the current vaccine RB51 in multiple doses. RB51 vaccination is currently a single dose calf-hood vaccine. This effort began in 2012 in and continues. Our initial studies were to evaluate of the role of CMI in efficacy of Experimental Alternate Schedule of Live Attenuated RB51 Vaccine against Brucellosis in Cattle. We have completed this project recently and our data show that multi-dose RB51 vaccination prevents vertical transmission and significantly lowers bacterial burden in the multi-dose treated cows. In particular our data show that vaccine boosting during pregnancy does not induce abortion as was previously thought. We seek to extend these studies to better define the bovine immune response to RB51 and or develop more defined sub-unit vaccines for brucellosis.

Dendritic Cell (DC) Biology and Immunity to B. abortus.
Understanding the role of DCs in infection and in particular infection with B. abortus is an understudied area but critical to develop an understanding of immunity to this facultative intracellular pathogen. Our efforts are focused on in vitro studies of monocyte derived or stem cell derived DC precursors. Our hypothesis is that RB51 infected DCs can stimulatecognate T cells and improve CMI. This stimulation may be enhanced with TLR agonists. This past year we have worked to develop immortalized DC subsets, established an MOI value for monocyte derived DCs exposed to RB51 vaccine strain, established an improved bovine DC yield from bovine stem cells and demonstrated activation of T cells derived from RB51 vaccinated cattle by bovine derived DCs.

Selected Publications: My primary contributions to science have focused on the development of vaccines for zoonotic bacterial diseases. Most important in the development and testing of next generation vaccines for plague and anthrax. I had begun this work in the 1995, at this time there were no effective vaccines available against inhalation plague and not much was understood about immune mechanisms of protection. An effective vaccine against inhalation anthrax was available but was criticized for it’s reactegenicity and extensive and lengthy dose regimen. We undertook basic and applied research to develop a de novo vaccine for plague and an improved vaccine for anthrax. The focus on both vaccines was preliminary efficacy data followed by robust product development studies to create IND products and then to assist advanced development of these medical products through full FDA licensure. Both vaccine candidates were submitted as investigational new drugs and have completed phase three clinical trials.

Earliest publications were on development and testing of novel recombinant plague vaccine:

Heath, D.G., G.W. Anderson, Jr., J.M. Mauro, S.L. Welkos, G.P. Andrews, J. Adamovicz, A.M. Friedlander. (1998). Protection against experimental bubonic and pneumonic plague by a recombinant capsular F1-V antigen fusion protein vaccine. Vaccine 16:1131-1137.

Andrews, G.P., S. Strachan, G.E. Benner, A.K. Sample, G.W. Anderson Jr., J.J. Adamovicz, S.L. Welkos, J.K. Pullen, A.M. Friedlander. (1999). Protective efficacy of recombinant Yersinia outer proteins against bubonic plague caused by encapsulated and non-encapsulated Yersinia pestis. Infect Imm 67:1533-1537.

Jarrett, C.O., F. Sebbane, J.J. Adamovicz, G.P. Andrews, B.J. Hinnebusch. (2004). A flea-borne transmission model to evaluate vaccine efficacy against naturally acquired bubonic plague. Infect Imm 72:2052-2056.

Publications on scalable improvements for manufacture of plague vaccine and vaccine regimen:

Powell, B.S., G.P. Andrews, J.T. Enama, S. Jendrek, C. Bolt, P. Worsham, J.K. Pullen, W. Ribot, H. Hines, L. Smith, D.G. Heath, J.J. Adamovicz. (2005). Design and testing for a non-tagged F1-V fusion protein as vaccine antigen against bubonic and pneumonic plague. Biotech Prog 21:1490-1510.

A. Glynn, C.J. Roy, B.S. Powell, J.J. Adamovicz, L.C. Freytag, J.D. Clements. (2005). Protection against aerosolized Yersinia pestis challenge following homologous and heterologous prime-boost with plague antigens. Infect Imm 73:5256-5261.

J.L. Goodin, D.F. Nellis, B.S. Powell, V.V. Vyas, J.T. Enama, L.C. Wang, P.K. Clark, S.L. Giardina, J.J. Adamovicz, D.F. Michiel. (2007). Purification and protective efficacy of monomeric and modified Yersinia pestis Capsular F1-V Fusion Proteins for Vaccination against Plague. Protein Expression and Purification 53:63-79.

P.A. Arlen, M. Singleton, J.J. Adamovicz, Y. Ding, A. Davoodi-Semiromi, H. Daniell. (2008). Effective plague vaccination via oral delivery of plant cells expressing F1-V antigens in chloroplasts. Infect Immun. 76:3640-50.

Publications on mechanisms of protective immunity and immune correlates development:

Parent, M.A., K.N. Berggren, I.K. Mullarky, F.M. Szaba, L.W. Kummer, J.J. Adamovicz, S.T. Smiley. (2005). Yersinia pestis V protein epitopes recognized by CD4 T cells. Infect Imm 73:2197-2204.

T. Jones, J.J. Adamovicz, S.L. Cyr, C.R. Bolt, N. Bellerose, L.M. Pitt, G.H. Lowell, D.S. Burt (2006). Intranasal proteosomeTM-based F1-V vaccine elicits respiratory and serum antibody responses and protects mice against lethal aerosolized plague infection. Vaccine 24:1625-1632.

J. Bashaw, S. Norris, S. Weeks, S. Trevino, J.J. Adamovicz, S. Welkos. (2007). Development of in vitro correlate assays of immunity to infection with Yersinia pestis. Clin. Vaccine Immunol. 14:605-16.

S. Welkos, S. Norris, J. Adamovicz. (2008). A modified caspase-3 assay correlates with immunity of nonhuman primates to infection by Yersinia pestis. Clin Vaccine Immunol. 15:1134-37.

S.F. Little, W.M. Webster, H. Wilhelm, B. Powell, J. Enama, J.J. Adamovicz. (2008). Evaluation of quantitative anti-F1 IgG and anti-V IgG ELISAs for use as an in-vitro based potency assay of plague vaccine in mice. Biologicals. 36:287-95.

P. Fellows, J. Adamovicz, J. Hartings, R. Sherwood, W. Mega, T. Brasel, E. Barr, L. Holland, W. Lin, A. Rom, W. Blackwelder, J. Price, S. Morris, D. Snow, M.K. Hart (2010). Protection in mice passively immunized with serum from cynomolgus macaques and humans vaccinated with recombinant plague vaccine (rF1V). Vaccine. Nov16;28(49):7748-56

S. Lin, S. Park, J.J. Adamovicz, J. Hill, J.B. Bliska, C.K. Cote, D.S. Perlin, K. Amemiya, S.T. Smiley. (2010). TNF- and IFN- contribute to F1/LcrV-targeted immune defense in mouse models of fully virulent pneumonic plague. Vaccine. Dec16;29(2):357-62

C.Y. Lindsey, S.F. Little, S. Norris, B. Powell, J.J. Adamovicz*. (2012). Validation of quantititative ELISA for the measurement of anti-Yersinia pestis F1 and V antibody concentration in non-human primate sera. J. Immuno Assays & Immunochem. Jan;33(1):91-113

B. Powell, W. Ribot, J. Adamovicz, G. Andrews. (2004). Structural Characterizations of Protein Antigens for the Next Generation Vaccines against Anthrax and Plague. U.S. Government Technical Report

W. J. Ribot, B. S. Powell, B. E. Ivins, , S. Little, W. M. Johnson, T. A. Hoover, S.L. Norris, J. Adamovicz, A.M. Friedlander, and G. P. Andrews. (2006). Comparative efficacy of protective antigen isoform vaccines against Bacillus anthracis spore challenge in rabbits. Vaccine 24:3469-3476.

B.S. Powell, J. T. Enama, W.J. Ribot, W.M. Webster, S. Little, T. Hoover, J.J. Adamovicz, G.P. Andrews. (2007). Multiple Asparagine Deamidation of Bacillus anthracis Protective Antigen Causes Charge Isoforms whose Complexity Correlates with Reduced Biological Activity. Proteins: Structure, Function and Bioinformatics. 68:458-79.

PATENTS:
U.S. 10/987,533 and PCT/US2004/38480 filed 11/12/2004. Awarded 18 May 2010 #7,718,779; Prophylactic, and therapeutic monoclonal antibodies (specific for the virulence (V) antigen of Yersinia pestis). Tran C. Chanh, Gerald P. Andrews, Jeffrey J. Adamovicz and Bradford S. Powell.