The Journal of Immunology, Vol 141, Issue 10 3342-3348, Copyright © 1988 by American Association of Immunologists

ARTICLES

Soluble factors in tolerance and contact sensitivity to 2,4-dinitro- fluorobenzene in mice. IX. A monoclonal T cell suppressor molecule is structurally and serologically related to the alpha/beta T cell receptor

RL Fairchild, RT Kubo and JW Moorhead
Department of Microbiology/Immunology, University of Colorado School of Medicine, Denver 80262.

Ts cells from mice tolerized with dinitrobenzene sulfonate produce a DNP-specific, MHC-restricted soluble suppressor factor (SSF) which regulates contact sensitivity to 2,4-dinitro-fluorobenzene. Previous studies have shown that the SSF-producing T cells and the soluble factor have the same hapten/MHC specificity suggesting that SSF may represent a secreted form of the Ts membrane receptor. The relationship between TCR proteins and SSF was investigated by examining the structural and serologic properties of a monoclonal DNP/H-2Kd-specific suppressor molecule produced by a Ts hybridoma. Reduction followed by alkylation abrogated the ability of the 3-10 molecule to inhibit transfer of contact sensitivity to 2,4-dinitro-fluorobenzene, indicating that intact disulfide bonds were a required structural property for suppression. Reduction of the 3-10 molecule followed by affinity chromatography on DNP-coupled Sepharose beads indicated that the 3-10 suppressor molecule is a dimer and that one of its chains binds to cell-free DNP. Serologic properties of the 3-10 molecule were examined by determining the ability of pan-reactive rabbit anti-TCR antibodies and anti-V beta 8 mAb KJ16.133 and F23.1 to adsorb suppressor activity from 3-10 culture supernatant and affinity purified 3-10 ascites material. All three reagents adsorbed the suppressor activity whereas control antibodies had no effect. When 3-10 material was passed through a F23.1-conjugated Sepharose affinity column, suppressor activity was recovered in the column eluate but not in the effluent fraction. When the 3-10 molecule was reduced and separated into its two chains (i.e., DNP-binding and non-DNP-binding chains), it was found that the anti-V beta 8 antibody F23.1-bound to the non-DNP- binding chain of the suppressor molecule. Collectively, these results indicate that the monoclonal 3-10 suppressor molecule is structurally similar to the alpha/beta TCR and suggest that the 3-10 molecule expresses a determinant encoded by the V beta 8 family of TCR genes. These results are consistent with our hypothesis that these suppressor molecules represent a secreted form of the TCR expressed on the surface of the DNP-specific Ts.

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