The goal of this research proposal is to identify and characterize the molecular mechanisms of oral streptococci that contribute to the pathogenesis of streptococcus-associated nephritides and to develop specific prophylactic and therapeutic measures for these diseases. We have shown that S. mutans, S. pyogenes and S. sanguis produce a heparin-- binding protein (HBP) that also binds to heparan proteoglycans in basal laminae of animal tissues, particularly in kidney, heart and liver. This cationic protein is loosely associated with the bacterial cell wall and is released along with other components, such as lipoteichoic acid (LTA), into the cell environment during growth.
One specific aim i s to define the pathogenic properties of purified HBP and LTA in rabbits particularly their abilities to mediate deposition in kidneys of circulating immune complexes and/or formation of immune complexes in situ. Kidney tissue will be analyzed for pathology using histologic and immunohistologic stains.
A second aim i s to evaluate antibody production to HBP and LTA by rabbits during immunization with nephritis-associated strains of streptococci or with other streptococcal strains. The relative quantities and properties of HBP on bacterial surfaces and in culture medium will be determined using monoclonal antibodies. Ultrastructural location of the antigens on streptococci will be determined using immunoelectron microscopy.
The third aim i s to isolate the structural genes of HBP from these bacteria and determine the nucleotide sequences to further characterize this virulence factor.
The fourth aim i s to generate HBP-deficient strains of streptococci, using insertion mutagenesis, and to compare them with isogeneic parental strains to prove the pathogenic properties of this protein. The information derived from the experiments proposed in this application will further our understanding of all infection-associated nephritides in humans and may ultimately permit the development of safe streptococcal vaccines to protect against infection without risk of tissue injury.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE005696-12
Application #
3219588
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1981-02-01
Project End
1997-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
12
Fiscal Year
1993
Total Cost
Indirect Cost
Name
State University of New York at Buffalo
Department
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Stinson, Murray W; Alder, Susan; Kumar, Sarmishtha (2003) Invasion and killing of human endothelial cells by viridans group streptococci. Infect Immun 71:2365-72
Zhang, L; Ignatowski, T A; Spengler, R N et al. (1999) Streptococcal histone induces murine macrophages To produce interleukin-1 and tumor necrosis factor alpha. Infect Immun 67:6473-7
Barnard, J P; Stinson, M W (1999) Influence of environmental conditions on hydrogen peroxide formation by Streptococcus gordonii. Infect Immun 67:6558-64
Juarez, Z E; Stinson, M W (1999) An extracellular protease of Streptococcus gordonii hydrolyzes type IV collagen and collagen analogues. Infect Immun 67:271-8
Stinson, M W; McLaughlin, R; Choi, S H et al. (1998) Streptococcal histone-like protein: primary structure of hlpA and protein binding to lipoteichoic acid and epithelial cells. Infect Immun 66:259-65
Barnard, J P; Stinson, M W (1996) The alpha-hemolysin of Streptococcus gordonii is hydrogen peroxide. Infect Immun 64:3853-7
Choi, S H; Zhang, X; Stinson, M W (1995) Dynamics of streptococcus histone retention by mouse kidneys. Clin Immunol Immunopathol 76:68-74
Winters, B D; Ramasubbu, N; Stinson, M W (1993) Isolation and characterization of a Streptococcus pyogenes protein that binds to basal laminae of human cardiac muscle. Infect Immun 61:3259-64
Hyzy, J; Sciotti, V; Albini, B et al. (1992) Deposition of circulating streptococcal lipoteichoic acid in mouse tissues. Microb Pathog 13:123-32
Glurich, I; Winters, B; Albini, B et al. (1991) Identification of Streptococcus pyogenes proteins that bind to rabbit kidney in vitro and in vivo. Microb Pathog 10:209-20

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