Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.
Detection of cellular antibodies in SIV immunized monkeys.
Symp Nonhum Primate Models AIDS. 1992 Nov 17-20;10:abstract no. 132. Unique Identifier : AIDSLINE PRIM10/93200972 Yee JL; Jennings MB; Acevedo R; Gardner MB; Department of Medical Pathology, University of California, Davis.
Abstract:
Understanding the specificity and function of antibodies to cells and/or cellular components in immunized monkeys is important in vaccine development. In a retrospective study using frozen pre- and post-immunization sera from monkeys immunized with various inactivated SIVmac (grown in human cell lines)/adjuvant combinations and controls, we have set up assays to detect anti-cell reactivities: Anti-CD4: Diluted monkey sera were reacted in an enzyme immunoassay using recombinant CD4 coated plates (E.I. DuPont, Inc.) followed by horseradish peroxidase conjugated goat anti-monkey immunoglobulin and O-phenylene-diamine as the detection system. Reactivity was detected in all but one monkey immunized with inactivated SIV plus adjuvant. (The non-reactive serum was from a monkey which was not protected in subsequent challenge experiments.) A much lower signal was detected in sera from monkeys receiving no immunization, adjuvant alone, and inactivated SIV alone. Anti-Hut-78: Hut-78 cells were incubated with serum plus FITC conjugated goat anti-human immunoglobulin and analyzed by FACS. Results paralleled those seen in the CD4 assay. The sera were also tested by Western immunoblot against a lysate of Hut-78 cells. Cells were washed, lysed by freeze-thaw cycles, and pelleted for use as antigen. Post-immunization sera from all monkeys reacted to more bands than their pre-immunization sera did. One of the reactive bands was also identified by HLA-DR antisera. Neither anti-CD4 nor anti-DR, in dual staining studies, significantly blocks the reactivity of sera from immunized monkeys against Hut-78 cells.
Keywords: Animal Antigens, CD4/*IMMUNOLOGY Blotting, Western Cell Line Haplorhini Human Immunity, Cellular Immunoenzyme Techniques Immunoglobulins/IMMUNOLOGY SIV/GROWTH & DEVELOPMENT/*IMMUNOLOGY Vaccines, Inactivated/*IMMUNOLOGY Viral Vaccines/*IMMUNOLOGY ABSTRACT 930630
M9361048
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Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.
