The H-2K('b) major histocompatibility complex glycoprotein antigens from EL4 and from a carbohydrate-variant subline, WD1, were purified on an anti-H-2K('b) (B8-24-3) monoclonal antibody affinity column. Previous studies have demonstrated that the cell-surface forms of the H-2 molecules from the WD1 variant line are complex but lack the terminal sialic acid residues. Each purified antigen was reconstituted along with purified cytoskeletal proteins into membrane vesicles of defined lipid composition, protein composition, and lipid/protein ratio. / A sensitive microassay was refined to detect the purified H-2K('b) antigens in the liposomes. The assay utilizes the liposomes to inhibit specific conjugate formation between allogeneically primed (H-2('d)) anti-H-2('b)) cytotoxic T cells and H-2('b) target cells. As little as 5 ng of purified H-2K('b) plus 23 ng cytoskeletal proteins in vesicles inhibited conjugate formation to 50% of the maximum inhibition observed. This inhibition was shown to be specific in two ways: (1) control vesicles containing purified H-2('k) proteins inhibited conjugate formation but only at 5- to 7-fold higher H-2('k) concentrations, and (2) the same H-2K('b)-containing vesicles did not inhibit nonspecific conjugate formation. / Liposomes containing H-2K('b) from either EL4 or WD1 similarly inhibited specific conjugate formation between EL4 target cells and the allogeneically primed cytotoxic T cells. These results suggest that the presence or absence of the terminal sialic acid residues on the two N-asparagine-linked complex carbohydrates of H-2K('b) did not affect the recognition step between the CTL and vesicles. / Source: Dissertation Abstracts International, Volume: 45-09, Section: B, page: 2902. / Thesis (Ph.D.)--The Florida State University, 1984.
| Identifer | oai:union.ndltd.org:fsu.edu/oai:fsu.digital.flvc.org:fsu_75415 |
| Contributors | FIGARD, STEVE DAVID., Florida State University |
| Source Sets | Florida State University |
| Detected Language | English |
| Type | Text |
| Format | 122 p. |
| Rights | On campus use only. |
| Relation | Dissertation Abstracts International |
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