The Journal of Immunology, Vol 143, Issue 12 3917-3924, Copyright © 1989 by American Association of Immunologists

ARTICLES

Relationship between T cell receptors and antigen-binding factors. II. Common antigenic determinants and epitope recognition shared by T cell receptors and antigen-binding factors

M Iwata, K Katamura, RT Kubo, HM Grey and K Ishizaka
Johns Hopkins University School of Medicine, Good Samaritan Hospital, Baltimore, MD 21239.

The T cell hybridomas 231F1 and 12H5 constitutively secrete glycosylation-inhibiting factor (GIF) and glycosylation-enhancing factor (GEF), respectively, which lack affinity for OVA-coupled Sepharose. When the 231F1 and 12H5 cells were stimulated by OVA-pulsed syngeneic macrophages, however, GIF and GEF produced by the cells had affinity for OVA. Both the OVA-binding GIF from the 231F1 cells and OVA- binding GEF from the 12H5 cells bound to a mAb against TCR-alpha beta and a mAb against TCR-alpha, suggesting a serologic relationship between TCR and OVA-binding factors. However, the OVA-binding GIF and GEF bound to mAb 14-12 and 14-30, respectively. Because these mAb do not bind TcR alpha beta-chains, it appears that the Ag-binding factors are different from TCR itself. The OVA-binding factors from both 12H5 cells and 231F1 cells do not bind to urea-denatured OVA. The binding of the factors to OVA Sepharose was inhibited by a peptide corresponding to residues 307-317 (P307-317) in the native OVA, but not by the peptide corresponding to residues 323-339 (P323-339). Furthermore, the OVA-binding factors bound to P306-319-coupled Sepharose but not to P323- 339-coupled Sepharose, and were recovered by elution of the former Sepharose at acid pH. The binding of OVA to anti-OVA antibodies was not inhibited by either peptide. Inasmuch as the 231F1 cells and 12H5 cells can be stimulated by P307-317 in the context of a MHC product, it appears that the Ag-binding factors and TCR-alpha beta on the cell sources of the factors may recognize the same epitope in the OVA molecules. The results also showed that Ag-binding factors and antibodies recognize distinct epitopes in the Ag molecules.