The long-term goal of this study is to understand the role in pathogenesis and immunity of an environmentally regulated surface polysaccharide of Staphylococcus aureus chemically characterized as poly-N-succinyl-B-1-6 glucosamine (PNSG). PNSG has previously been determined to be the protective capsular polysaccharide/adhesin (PS/A) antigen of Staphylococcus epidermidis, raising the possibility that PNSG could be used as a """"""""pan-staphylococcal"""""""" vaccine. To define the role of PNSG in pathogenesis of S. aureus infection 5 different PNSG-deficient S. aureus strains representative of major lineages will be constructed by genetic means via interruption of the genes in the intracellular adhesin (ica) locus that encodes proteins needed for synthesis of PNSG. Isogenic parental, mutant and ica complemented strains will be evaluated in vitro to determine the role of PNSG in promoting S. aureus adherence to catheters and in providing resistance of bacterial cells to phagocytic killing by leukocytes and complement. The same strains will also be tested for infectious capability in several animal systems of S. aureus infection, including animals actively and passively immunized with ica-deleted S. aureus and normal human serum to reflect the immunologic status of humans, who have high levels of natural antibody to S. aureus surface antigens. Because PNSG isolated from some strains of staphylococci have up to 30 percent of the succinate substituents on the polyglucosamine backbone replaced by acetate, purified PNSG, with differing ratios of succinate and acetate substituents on the polyglucosamine backbone, will be produced for immunologic studies. Rabbits will be immunized with the variants and sera assessed for antibody titer and opsonic killing ability. The PNSG variant structures will be used to immunize mice to evaluate their ability to generate protective immunity in the same systems used for the study of the role of PNSG in S. aureus virulence. In addition, passive protection by the rabbit sera raised to the variant PNSG constructs will be evaluated in the animal systems. All the above mentioned studies will provide new and useful information regarding pathogenesis and immunity of staphylococcal infections, stressing the use of animal systems that reflect naturally acquired immunity in humans to S. aureus. By the end of these studies we expect to have a clear understanding of the role of PNSG in virulence, as determined in a variety of staphylococcal infection systems, the immunochemical properties of PNSG that can engender protective immunity, and the types of S. aureus infections wherein PNSG-specific immunotherapies show the most potential for success.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI046706-02
Application #
6497308
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
2001-02-01
Project End
2006-01-31
Budget Start
2002-02-01
Budget End
2003-01-31
Support Year
2
Fiscal Year
2002
Total Cost
$373,476
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02115
Skurnik, David; Cywes-Bentley, Colette; Pier, Gerald B (2016) The exceptionally broad-based potential of active and passive vaccination targeting the conserved microbial surface polysaccharide PNAG. Expert Rev Vaccines 15:1041-53
Gauguet, Stefanie; D'Ortona, Samantha; Ahnger-Pier, Kathryn et al. (2015) Intestinal Microbiota of Mice Influences Resistance to Staphylococcus aureus Pneumonia. Infect Immun 83:4003-14
Cerca, Filipe; França, Angela; Pérez-Cabezas, Begoña et al. (2014) Dormant bacteria within Staphylococcus epidermidis biofilms have low inflammatory properties and maintain tolerance to vancomycin and penicillin after entering planktonic growth. J Med Microbiol 63:1274-83
Lu, Xi; Skurnik, David; Pozzi, Clarissa et al. (2014) A Poly-N-acetylglucosamine-Shiga toxin broad-spectrum conjugate vaccine for Shiga toxin-producing Escherichia coli. MBio 5:e00974-14
Babra, Charlene; Tiwari, Jully G; Pier, Gerald et al. (2013) The persistence of biofilm-associated antibiotic resistance of Staphylococcus aureus isolated from clinical bovine mastitis cases in Australia. Folia Microbiol (Praha) 58:469-74
Pier, Gerald B (2013) Will there ever be a universal Staphylococcus aureus vaccine? Hum Vaccin Immunother 9:1865-76
Cywes-Bentley, Colette; Skurnik, David; Zaidi, Tanweer et al. (2013) Antibody to a conserved antigenic target is protective against diverse prokaryotic and eukaryotic pathogens. Proc Natl Acad Sci U S A 110:E2209-18
Bentancor, Leticia V; Routray, Abhisek; Bozkurt-Guzel, Cagla et al. (2012) Evaluation of the trimeric autotransporter Ata as a vaccine candidate against Acinetobacter baumannii infections. Infect Immun 80:3381-8
Yoong, Pauline; Cywes-Bentley, Colette; Pier, Gerald B (2012) Poly-N-acetylglucosamine expression by wild-type Yersinia pestis is maximal at mammalian, not flea, temperatures. MBio 3:e00217-12
Komarova, Bozhena S; Tsvetkov, Yury E; Pier, Gerald B et al. (2012) Synthesis of pentasaccharides corresponding to the glycoform II of the outer core region of the Pseudomonas aeruginosa lipopolysaccharide. Carbohydr Res 360:56-68

Showing the most recent 10 out of 38 publications