PMID: 6169779Oct 1, 1981Paper

In vitro generation of antigen-specific hemolytic plaque-forming cells from human peripheral blood mononuclear cells

The Journal of Experimental Medicine
J Misiti, T A Waldmann

Abstract

We have described a culture and assay system for the sensitization of human peripheral blood mononuclear cells with a T cell-dependent antigen, sheep erythrocytes, in the absence of nonspecific stimulatory agents and with the subsequent generation of macroscopic hemolytic plaques. We have shown that the antibody produced by the plaque-forming cells generated in this culture system is specific for the sensitizing antigen, and that the plaques created are not false plaques because their formation is inhibited by cycloheximide. The success of this system can be attributed to several critical factors including large numbers of peripheral blood mononuclear cells (5 x 10(6) culture), a prolonged period of incubation (10-11 d), continuous rocking during the entire period of incubation, culturing in large (35-mm) flat-bottomed culture dishes in the presence of human plasma, and the appropriate antigen concentration (5 x 10(6) sheep erythrocytes/culture). Furthermore, the generation of macroscopic hemolytic plaques requires plaquing sensitized peripheral blood mononuclear cells in target cell monolayers fixed in an agarose matrix with an incubation period of 2-3 h. We have further shown that the antigen-specific response measured by thi...Continue Reading

References

Oct 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·C A JanewayH Wigzell
Dec 1, 1976·The Journal of Clinical Investigation·S BroderT A Waldmann
Oct 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·A S Fauci, K R Pratt
Aug 26, 1966·Science·R I Mishell, R W Dutton
Sep 1, 1967·The Journal of Experimental Medicine·R I Mishell, R W Dutton

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Citations

Oct 10, 1985·Journal of Immunological Methods·J M WeilerR Yarchoan
Jan 1, 1983·Developmental and Comparative Immunology·F M McCorkle, G A Leslie
Jan 1, 1986·International Journal of Immunopharmacology·M L VillaE Clerici
Jan 1, 1988·International Journal of Immunopharmacology·M L VillaE Clerici
Oct 1, 1990·Toxicology Letters·R M Pillai, R R Watson
Oct 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·K InabaS Muramatsu
Aug 1, 1983·The Journal of Clinical Investigation·H C LaneA S Fauci
Jan 1, 1991·Immunopharmacology and Immunotoxicology·A MisefariD Venza Teti
Jan 1, 1984·Scandinavian Journal of Immunology·E Beckmann, D Levitt
Dec 1, 1983·European Journal of Immunology·J Kagan, Y S Choi
Sep 30, 1982·Journal of Immunological Methods·C L Reading

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