Construction of antigen-specific suppressor T cell hybridomas from spleen cells of mice primed for the persistent IgE antibody formation.
Attempts were made to generate Ag-specific suppressor T cells from Ag-primed spleen cells by using glycosylation inhibiting factor (GIF). BDF1 mice were primed with alum-absorbed OVA and their spleen cells were stimulated with OVA. Ag-activated T cells were then propagated in IL-2-containing conditioned medium. Incubation of the T cells with OVA-pulsed syngeneic macrophages resulted in the formation of IgE-potentiating factor and glycosylation-enhancing factor that has affinity for OVA, i.e., OVA-specific glycosylation-enhancing factor. However, if the same Ag-activated splenic T cells were propagated in the IL-2-containing medium in the presence of GIF T cells obtained in the cultures formed IgE-suppressive factors and OVA-specific GIF on antigenic stimulation. Thus we constructed T cell hybridomas from the Ag-activated T cells propagated by IL-2 in the presence of GIF. A representative hybridoma, 71B4, formed OVA-specific GIF on incubation with OVA-pulsed macrophages of BDF1 mice or C57B1/6 mice. However, if the same hybridoma cells were incubated with OVA alone or with OVA-pulsed macrophages of H-2k or H-2d strains, they produced GIF that had no affinity for OVA. The OVA-specific GIF bound to OVA-Sepharose but did not bind to BSA-Sepharose or KLH Sepharose. Intravenous injections of the OVA-specific GIF from the hybridoma suppressed the IgE and IgG1 anti-DNP antibody response of BDF1 mice to DNP-OVA, but failed to suppress the anti-hapten antibody responses of the strain to DNP-keyhole limpet hemocyanin, indicating that the factors suppressed the antibody response in a carrier-specific manner. However, the same OVA-specific GIF failed to suppress the anti-hapten antibody response of DBA/1 mice to DNP-OVA, suggesting that the immunosuppressive effects of the factors is MHC restricted.
Iwata
M
M
Subdepartment of Immunology, Johns Hopkins University School of Medicine, Baltimore, MD 21239.
Ishizaka
K
K
D000818:Animals; D002352:Carrier Proteins / Q000276:immunology; D002459:Cell Fusion; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006031:Glycosylation; D006825:Hybridomas / Q000276:immunology* / Q000378:metabolism; D007073:Immunoglobulin E / Q000096:biosynthesis*; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000276:immunology; D051379:Mice; D008806:Mice, Inbred AKR; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008811:Mice, Inbred DBA; D010047:Ovalbumin / Q000276:immunology; D029584:Prostatic Secretory Proteins; D051381:Rats; D011917:Rats, Inbred Lew; D013154:Spleen / Q000276:immunology* / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism
D002352:Carrier Proteins; D000939:Epitopes; D008222:Lymphokines; D029584:Prostatic Secretory Proteins; C043076:beta-microseminoprotein; C013059:immunoglobulin-binding factors; D007073:Immunoglobulin E; D010047:Ovalbumin