Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.
Analysis of cytotoxic T lymphocyte responses to SIV proteins in SIV-infected macaques using antigen-specific stimulation with recombinant vaccinia and fowl poxviruses.
AIDS Res Hum Retroviruses. 1994 May;10(5):551-60. Unique Identifier : AIDSLINE MED/95000927 Kent SJ; Stallard V; Corey L; Hu SL; Morton WR; Gritz L; Panicali DL; Greenberg PD; Department of Medicine, University of Washington, Seattle 98195.
Abstract:
Methods to analyze CD8+ CTL responses to simian immunodeficiency virus (SIV)-encoded proteins are essential to understand lentivirus immunopathogenesis and protective immune responses. Recombinant infectious shuttle vectors are useful for analyzing CTL responses to many viruses, including HIV. Therefore, CTL responses in SIV-infected Macaca fascicularis to SIV env and SIV gag/pol were evaluated using specific antigen stimulation with recombinant vaccinia (rVV) and fowl poxviruses (rFPV) containing SIV genes. Peripheral blood mononuclear cells from SIV-infected animals were stimulated with autologous cells infected with rVV expressing SIV env/gag/pol, and CTLs specific for SIV env and for SIV gag/pol were detected by testing for lytic activity in target cells expressing these genes separately. Lymphocyte subset purifications from the effector population demonstrated that the CTL response was mediated by CD8+ cells, and the use of brefeldin A to selectively block antigen presentation in association with MHC class I products affirmed this cytolytic activity was class I restricted. The use of rVV to analyze responses to SIV genes is potentially problematic in hosts immunized to vaccinia. Fowl poxvirus is an alternative virus that has many of the molecular advantages of vaccinia virus but is genomically distinct. Therefore, the ability of rFPV to expand and detect SIV-specific CTLs was evaluated. Although there was no cytopathic effect following infection with rFPV, macaque cells infected with this vector did express rFPV gene products, and could be used as stimulator and target cells to detect SIV-specific CD8+ CTLs. The results suggest that these recombinant viral vectors can be used to specifically stimulate CD8+, MHC class I-restricted CTLs reactive to SIV proteins, and should facilitate evaluating CTL responses in both SIV-infected animals and animals vaccinated against SIV.
Keywords: Animal Antigens, Viral/GENETICS Cell Line Comparative Study Concanavalin A/PHARMACOLOGY Fowlpox Virus/GENETICS Fusion Proteins, gag-pol/GENETICS/IMMUNOLOGY Gene Products, env/GENETICS/IMMUNOLOGY Genetic Vectors Macaca fascicularis Phenotype Recombination, Genetic Retroviridae Proteins/GENETICS/*IMMUNOLOGY Retroviridae Proteins, Oncogenic/GENETICS/IMMUNOLOGY Simian Acquired Immunodeficiency Syndrome/IMMUNOLOGY Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. SIV/GENETICS/*IMMUNOLOGY T-Lymphocytes, Cytotoxic/*IMMUNOLOGY Vaccinia Virus/GENETICS Viral Fusion Proteins/GENETICS/IMMUNOLOGY JOURNAL ARTICLE
950130
M9510842
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Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.
