Thomas J. Inzana

BIOGRAPHICAL SKETCH Provide the following information for the key personnel in the order listed for Form Page 2. Follow this format for each person. DO NOT EXCEED FOUR PAGES.

NAME Thomas Joseph Inzana		POSITION TITLE Tyler J. and Frances F. Young Professor of Bacteriology
EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, and include postdoctoral training.)
INSTITUTION AND LOCATION	DEGREE (if applicable)		YEAR(s)	FIELD OF STUDY
University of Georgia, Athens, GA	B.S.		1975	Microbiology
University of Georgia, Athens, GA	M.S.		1978	Medical Microbiology
University of Rochester, School of Medicine, Rochester, NY	Ph.D.		1982	Microbiology
Baylor College of Medicine, Houston, TX	Postdoc.		1984	Fellowship in Med. Lab. Microbiol. Public Health.

A. Positions and Honors. List in chronological order previous positions, concluding with your present position. List any honors. Include present membership on any Federal Government public advisory committee.

Assistant Professor Coll/. Vet. Med. Dept. Vet. Microbiol.Pathol.

Washington State University 7/84-6/87

Assistant Professor and Director VA-MD Reg. Coll. Vet. Med., Depart.

Clinical Microbiology Biomed. Sci. Pathobiol. Virginia Tech 7/87-4/91

Visiting Associate Professor SUNY at Buffalo School of Medicine 6/92-8/92

Associate Professor and Director VA-MD Reg. Coll.Vet. Med.

Clinical Microbiology Dept. Biomed. Sci. Pathobiol. 4/91-4/96

Visiting Professor Univ. Penn. School of Medicine 1/00-5/00

Coordinator, Cent. Mol. Med. Infectious. Dis. VA-MD Reg. Coll. Vet. Med. 1/98-7/02

Professor and Director, Clinical Microbiology VA-MD Reg. Coll. Vet. Med., Depart. 4/96-present

Tyler J. and Frances F. Young Prof. Bacteriol. VA-MD Reg. Coll. Vet. Med., Depart.

Biomed. Sci. Pathobiol. Virginia Tech 4/03-present

Diplomate: American Board of Medical Microbiology

Fellow: American Academy of Microbiology

Member: American Society for Microbiology, American Academy of Microbiology, American Association of Veterinary Laboratory Diagnosticians, American Association for the Advancement of Science, Conference of Research Workers in Animal Diseases (CRWAD), International Endotoxin Society, Phi Zeta National Veterinary Medicine Honor Society

Pfizer Award for Research Excellence: 2003

Patents:

Inzana, T. J. 1995. Application of noncapsulated mutants of Actinobacillus pleuropneumoniae for use in live vaccines against swine pleuropneumonia. Patent Number 5,429,818 (In commercial use by Boehringer-Ingelheim as APP-LAC).

Inzana, T.J. and C. Ward. 7/2000. Defined, recombinant mutants of Actinobacillus pleuropneumoniae for use as vaccine candidates against swine pleuropneumonia. Patent number 6,086,894.

Inzana, T.J. 12/4/2001. Recombinant vaccine for diseases caused by encapsulated organisms. Patent Number 6,326, 001.

B. Selected peer-reviewed publications (in chronological order from a total of 78).

Ward, C.M., and T.J. Inzana. 1994. Resistance to bactericidal antibody and complement in Actinobacillus pleuropneumoniae is mediated by capsular polysaccharide and blocking antibody. J. Immunol. 153:2110-2121

Moxon, E.R., B.E. Gewurz, J.C. Richards, T. Inzana, M.P. Jennings, and D.W. Hood. 1996. Phenotypic switching of Haemophilus influenzae. Mol. Microbiol. 19:1149-1150.

Ward, C. K., and T. J. Inzana. 1997. Identification and characterization of a DNA region involved in the export of capsular polysaccharide by Actinobacillus pleuropneumoniae serotype 5a. Infect. Immun. 65:2491-2496.

Inzana, T.J., J. Hensley, J. McQuiston, A. J. Lesse, A. A. Campagnari, S. M. Boyle, and M. A. Apicella. 1997. Phase variation and conservation of lipooligosaccharide epitopes in Haemophilus somnus. Infect. Immun. 65:4675-4681.

Cox, A.D., M.D. Howard, J.-R. Brisson, M. van der Zwan, P. Thibault, M.B. Perry, and T.J. Inzana. 1998. Structural analysis of the phase variable lipooligosaccharide from Haemophilus somnus strain 738. Eur. J. Biochem. 253:507-516.

Ward, C.K., M.L. Lawrence, and T.J. Inzana. 1998. Cloning and mutagenesis of a serotype-specific DNA region involved in encapsulation and virulence of Actinobacillus pleuropneumoniae serotype 5a: concomitant expression of serotype 5a and 1 capsular polysaccharides in recombinant A. pleuropneumoniae serotype 1. Infect. Immun. 66:3326-3336.

Wu, Y., J.H. McQuiston, A. Cox, T.D. Pack, and T.J. Inzana. 2000. Molecular cloning and mutagenesis of a DNA locus involved in lipooligosaccharide biosynthesis in Haemophilus somnus. Infect. Immun. 68:310-319.

McQuiston, J.H., J.R. McQuiston, A.D. Cox, Y, Wu, S. M. Boyle, and T.J. Inzana. 1999. Characterization of a DNA region containing 5’-(CAAT)_n-3’ DNA sequences involved in lipooligosaccharide biosynthesis in Haemophilus somnus. Microb. Pathogen. 28:301-312.

Howard, M.D., A.D. Cox, J.N. Weiser, G.G. Schurig, and T.J. Inzana. 2000. Antigenic diversity in Haemophilus somnus lipooligosaccharide and investigation of the phosphorylcholine epitope. J. Clin. Microbiol. 38:4412-4419.

Inzana, T.J. and B. Fenwick. 2001. Serologic diagnosis of Actinobacillus pleuropneumoniae in swine by capsular polysaccharide-biotin/streptavidin-enzyme-linked immunosorbent assay. J. Clin. Microbiol. 39:1279-1282.

Inzana, T.J., G. Glindemann, A.D. Cox, W. Wakarchuk, and M.D. Howard. 2002. Incorporation of N-acetylneuraminic acid into Haemophilus somnus lipooligosaccharide (LOS): Enhancement of resistance to serum and reduction of LOS antibody binding. Infect. Immun. 70:4870-4879.

Bandara, A.B., M.L. Lawrence, H.P. Veit, and T.J. Inzana. 2003. Association of Actinobacillus pleuropneumoniae capsular polysaccharide with Virulence in Pigs. Infect. Immun. 71:3320-3328.

Cox, A.D., M.D. Howard, and T.J. Inzana. 2003. Structural analysis of the lipooligosaccharide from the commensal Haemophilus somnus strain 1P. Carbohydr. Res. 338:1223-1228.

Jessing, S.G., Ø. Angen, and T.J. Inzana. 2003. Evaluation of a multiplex PCR test for simultaneous identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6. J. Clin. Microbiol. 41:4095-4100.

Sylte, M.J., T.J. Inzana, C.J. Czuprynski. 2004. Reactive oxygen and nitrogen intermediates contribute to Haemophilus somnus lipooligosaccharide-mediated apoptosis of bovine endothelial cells. Vet. Immunol. Immunopathol. 97:207-217.

Hüssy, D, Y Schlatter, R Miserez, T Inzana, and J Frey. 2004. PCR-based identification of serotype 2 strains of Actinobacillus pleuropneumoniae biovars I and II. Vet. Microbiol. 99:307-310.

St Michael, F. MD Howard, J Li, AJ Duncan, TJ Inzana, and AD Cox. 2004. Structural analysis of the lipooligosaccharide from the commensal Haemophilus somnus genome strain 129Pt. Carbohydr Res. 339:529-535.

Inzana, T.J., G. Glindemann, and G. Snider. 2004. Characterization of a wildtype strain of Francisella tularensis isolated from a cat and review of the literature. J. Vet. Diagn. Invest. 16:374-381.

Mohd-Zain, Z., S.L. Turner, A.M. Cerdeño-Tárraga. A.K. Lilley, T.J. Inzana, A.J. Duncan, R.M. Harding, D.W. Hood, T.E. Peto, and D.W Crook. Transferable Antibiotic Resistance Elements in Haemophilus influenzae Share a Common Evolutionary Origin with a Diverse Family of Coherent Genomic Islands. J. Bacteriol. 186: 8114-8122.

Howard, MD, JH Boone, V Buechner-Maxwell., GG Schurig, and TJ Inzana. 2004. Inhibition of Bovine Macrophage and Polymorphonuclear Leukocyte Superoxide Anion Production by Haemophilus somnus. Microbial Pathogen. 37:263-271.

Inzana TJ, G. Glindemann, B Fenwick, J Longstreth, D Ward. 2004. Risk assessment of transmission of capsule-deficient, recombinant Actinobacillus pleuropneumoniae. Vet. Microbiol.104:63-71.

C. Research Support. List selected ongoing or completed (during the last three years) research projects (federal and non-federal support). Begin with the projects that are most relevant to the research proposed in this application. Briefly indicate the overall goals of the projects and responsibilities of principal investigator identified above.

ONGOING Dates of Project and level of support Percent Effort

1. DAMD17-03-1-0008 Inzana (PI) 3/03-3/07 $1,068,274 30%

US Dept of Defense

Investigation of the capsule of Francisella tularensis LVS.

The major goals of this project are to purify and characterize the capsule from F. tularensis; To isolate the genes responsible

for capsule formation; To identify outer membrane proteins capable of illiciting a cellular immune response, and to develop biosensors using the above information for rapid detection of F. tularensis.

2. NIH 501722 Inzana (PI) 10/03-10/08 $910,148 15%

University of Maryland subcontract as part of Mid-Atlantic RCE(subcontract from NIH)

Vaccines and immunomodulatory therapies for tularemia

The major goals of this project are to conjugate the capsule of F. tularensis to various types of proteins and in the presence

of different adjuvants evaluate the capability of the conjugate to induce a cellular immune response and determine if

protection can be provided against lethal challenge in mice and primates.

3. 2001-52100-11314 Inzana (PI) 9/01-9/05 $650,000 10%

NSF-USDA/CSREES

Complete genome sequencing of the Bovine Pathogen Haemophilus somnus

The major goal of this project is to sequence and annotate the genome of Haemophilus somnus strain 2336.

4. 99-35204-7670 and 2003-35204-13637 Inzana (PI) 8/99-9/06 $520,000 20%

USDA/CREES

Molecular basis of LOS phase variation, phosphocholine expression, and sialylation in Haemophilus somnus virulence and colonization

The major goals of this project are to investigate the genetic basis of LOS phase variation and sialylation, and determine how phase variation and sialylation affects virulence and pathogenesis in animal model systems.

COMPLETED

1. 98-35204-6811 Inzana (PI) 12/98-11/01 $180,000 15%

USDA/CREES

The molecular basis for Actinobacillus pleuropneumoniae capsule in virulence

The major goals of this project were to determine if the type or quantity of capsular polysaccharide affected virulence in the swine pathogen Actinobacillus (Haemophilus) pleuropneumoniae. This was done by knocking out capsule biosynthesis genes and complementing the mutation to alter the quantity of the original capsule type or cloning

capsule genes from another serotype in order to change the type of capsule expressed.

2. 97-39210-5006 Inzana (PI) 10/97-10/00 $182,000 15%

USDA/CREES

Risk assessment of release of recombinant Actinobacillus pleuropneumoniae

The goals of this research were to determine the risk of a recombinant, attenuated strain of A. pleuropneumoniae to transfer a kanamycin resistance gene from its genome to other bacteria in the animal or for the strain to persist in the animal.