By Henry G. Kunkel (ed.), Frank J. Dixon (ed.)
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Additional resources for Advances in Immunology, Vol. 31
First, T cells were cultured for 4 days with a protein antigen, resulting in the production of helper cells; the function of the helper T cell was assayed in a second culture by mixing the T cells with unprimed B cells 30 EXIIL R . UNANUE and the appropriate hapten-protein conjugate, later measuring the anti-hapten response. The anti-hapten response-usually to DNPwas measured by a plaque-forming assay. Carrier-specific helper T cells were generated, inducing a small to modest number of plaque-forming cells with rather large experimental variations.
Our studies indicated that serum treated with 2-ME and then alkylated with iodoacetamide generated a 65,000-MW molecule that migrated electrophoretically like albumin and promoted growth of B cells and T cells (Sidman and Unanue, 1978; also in Braun and Unanue, 1980). The active moiety appeared, therefore, to be closely related to albumin. The later studies of Opitz et al. , 1978, 1980): Using affinity chromatography with Affi-Gel Blue columns, the 2-MEgenerated moiety behaved exactly like albumin; furthermore, purified bovine albumin treated with 2-ME also generated a growth-promoting activity.
Nevertheless, the results in the various publications appeared to be highly consistent. The generation of helper T cells to hemocyanin or to (TG)-AL required the presence of accessory phagocytic cells in the culture. The 2-ME did not replace the need for macrophages. Macrophages pulsed with the antigen were also highly efficient in generating helper T cells. McDougal and Gordon (1977) developed a similar system. In their experiments, cortisone-resistant thymocytes developed into helper T cells upon culture with hemocyanin or fowl y-globulin.
Advances in Immunology, Vol. 31 by Henry G. Kunkel (ed.), Frank J. Dixon (ed.)