The principal goal of the Virology Core is to provide virological support for Projects 1`through 4 and the Immunology Core. This will be accomplished in 3 specific aims.
Aim 1. To provide standardized viral stocks for in vitro assays and macaque challenge studies. Titered viral stocks for challenge will include SHIVs, provided by Dr. Yichen Lu, Virus Research Institute., Cambridge, MA., for Projects 1 through 3 and HIV- 2SBL6669, provided by Dr. G. Biberfeld, Stockholm, Sweden, for Project 4. In addition, strains of HIV-1, vaccinia constructs, and transforming herpes viruses will be grown and provided as needed to the projects and Immunology Core. Genetically, monotypic viruses obtained by transfection of infectious molecular clones will be provided as needed to ascertain the fine specificities of immune responses. The IHV has a library of infectious molecular clones and appropriate vector systems available for this purpose.
Aim 2. To evaluate challenged rhesus macaques for infectious. Infection will be monitored by plasma p24/27 antigen, virus co-cultivation, and by quantitative plasma viral RNA measurements. For Projects 1 through 3, a NASBA method that detects SIV gag sequences will be used. Project 4 uses HIV-2SBL6669 as the challenge virus and a HIV-2 specific NASBA is under development by our collaborators. Alternatively, QC-PCR will be used to determine viral loads for this virus. In addition to these methods, it may be of interest to determine the phenotype or genotype of viruses that break through vaccination (c.f., Projects 1 and 3). The Virology Core will isolate these viruses and characterize relevant phenotypes and genotypes.
Aim 3. TO carry out assays for neutralizing antibodies and to quantify VSFs. Neutralizing antibody assays will be carried out by the Virology Core. Several assays are online that detect antibodies specific for primary isolates and laboratory strains of virus. The Virology Core will also quantify VSF activity in supernatants provided by the Immunology Core. These assays will be overseen by Mr. Mike Merges, who has extensive experience in the performance and interpretation of such assays.
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2006) Cholera toxin indirectly activates human monocyte-derived dendritic cells in vitro through the production of soluble factors, including prostaglandin E(2) and nitric oxide. Clin Vaccine Immunol 13:106-15|
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2005) Pasteurella multocida toxin activates human monocyte-derived and murine bone marrow-derived dendritic cells in vitro but suppresses antibody production in vivo. Infect Immun 73:413-21|
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2004) Calcium signaling through phospholipase C activates dendritic cells to mature and is necessary for the activation and maturation of dendritic cells induced by diverse agonists. Clin Diagn Lab Immunol 11:77-82|
|Abdelwahab, Sayed F; Cocchi, Fiorenza; Bagley, Kenneth C et al. (2003) HIV-1-suppressive factors are secreted by CD4+ T cells during primary immune responses. Proc Natl Acad Sci U S A 100:15006-10|
|Bagley, K C; Shata, M T; Onyabe, D Y et al. (2003) Immunogenicity of DNA vaccines that direct the coincident expression of the 120 kDa glycoprotein of human immunodeficiency virus and the catalytic domain of cholera toxin. Vaccine 21:3335-41|
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2003) An enzymatically active a domain is required for cholera-like enterotoxins to induce a long-lived blockade on the induction of oral tolerance: new method for screening mucosal adjuvants. Infect Immun 71:6850-6|
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2002) Pertussis toxin and the adenylate cyclase toxin from Bordetella pertussis activate human monocyte-derived dendritic cells and dominantly inhibit cytokine production through a cAMP-dependent pathway. J Leukoc Biol 72:962-9|
|Bagley, Kenneth C; Abdelwahab, Sayed F; Tuskan, Robert G et al. (2002) Cholera toxin and heat-labile enterotoxin activate human monocyte-derived dendritic cells and dominantly inhibit cytokine production through a cyclic AMP-dependent pathway. Infect Immun 70:5533-9|
|Devico, Anthony L; Fouts, Timothy R; Shata, Mohamed T et al. (2002) Development of an oral prime-boost strategy to elicit broadly neutralizing antibodies against HIV-1. Vaccine 20:1968-74|
|Kamin-Lewis, R; Abdelwahab, S F; Trang, C et al. (2001) Perforin-low memory CD8+ cells are the predominant T cells in normal humans that synthesize the beta -chemokine macrophage inflammatory protein-1beta. Proc Natl Acad Sci U S A 98:9283-8|