In vivo activity of affinity-purified helper factor from antigen-specific helper clone.
Supernatant from culture of a virally transformed OVA-specific helper T clone (C-41) was examined for the presence of soluble helper factor. Inoculation of helper clone supernatant into DNP-KLH-primed mice enhanced the IgG anti-DNP response when given with DNP-OVA. The C-41 supernatant did not trigger the DNP-primed B cells in mice when injected with hapten (DNP) coupled to an unrelated carrier (BSA). The carrier-dependent helper activity of C-41 supernatant in vivo demonstrates the presence of an antigen-specific T helper factor in the media of the cultured helper clone. Extensive immunization of F1(C57BL X BALB/c) mice with the helper clone resulted in the production of anti C-41 antibodies. Monoclonal antibodies prepared from the immunized mice were screened for specificity of binding to other transformed T lines and clones, some specific to OVA. Monoclonal antibodies that stained the C-41 cells exclusively were considered clone-specific. Supernatants of the helper clone were passed over columns of anti-clone-specific antibodies. The eluates from three antibodies were active as antigen-specific helper factor, i.e., they elevated the IgG anti-DNP response in vivo in a linked recognition fashion in the presence of DNP-OVA. The affinity-purified factor was inactive when injected with DNP-BSA or DNP-BSA + OVA. Thus, we describe the antigen-specific immune function of a clone-produced helper factor in normal mice.
Azar
Y
Y
Falek
P R
PR
Kagan
J
J
Ben-Sasson
S Z
SZ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D002457:Cell Extracts / Q000276:immunology; D002846:Chromatography, Affinity; D002999:Clone Cells / Q000276:immunology / Q000378:metabolism; D004136:Dinitrobenzenes / Q000276:immunology; D000939:Epitopes; D005260:Female; D007376:Interleukin-2 / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010047:Ovalbumin / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D002457:Cell Extracts; D004136:Dinitrobenzenes; D000939:Epitopes; D007376:Interleukin-2; D010047:Ovalbumin