Fairchild R L, Moorhead J W
Cell Immunol. 1987 Mar;105(1):147-60. doi: 10.1016/0008-8749(87)90064-5.
To study further soluble factors which regulate contact sensitivity (CS) to 2,4-dinitrofluorobenzene (DNFB), hapten-primed spleen cells from BALB/c mice were used to make T-cell hybridomas. A hybrid constitutively producing a suppressor factor was identified and cloned (clone 3-10). Incubation of BALB/c DNFB immune lymph node cells (LNC) in the 3-10 supernatant suppressed the ability of the immune cells to transfer CS to DNFB. The passive transfer of CS to oxazalone or to 2,4,6-trinitrochlorobenzene (TNCB) was not suppressed by the 3-10 factor. The hapten specificity of the 3-10 factor further was demonstrated by the ability of DNFB immune LNC but not LNC from unsensitized or from TNCB-sensitized mice to absorb the factor. The 3-10 factor also was adsorbed by DNFB-immune LNC from mice that were syngeneic with BALB/c mice at the K locus of the MHC (e.g., B10.D2 and D2.GD). Pretreatment of DNFB-immune LNC with monoclonal anti-Kd antibody or with anti-DNP antibodies blocked the ability to adsorb the factor. These results indicated that the 3-10 suppressor factor binds to DNP/H-2Kd complexes on immune LNC. Nylon wool-purified T cells (83% Thy-1.2+) from DNFB-immune LNC were able to adsorb the factor as well as unseparated immune LNC. Furthermore, treatment of immune LNC with anti-Thy-1.2 plus C' abrogated the ability of the cells to adsorb the factor, indicating that the cellular target of the 3-10 factor is a T cell. In addition, treatment of the immune LNC with an autoantiidiotypic antiserum (CS 231) plus C', which depletes DNP-specific delayed-type hypersensitivity effector T (TDH) cells, also abrogated the ability of the cells to adsorb the factor. Finally, the suppressor factor was adsorbed and eluted from DNP affinity columns but was not adsorbed by TNP affinity columns. Collectively, these results indicate that although the monoclonal 3-10 suppressor factor has affinity for DNP, focusing of the factor on the TDH cells requires recognition of DNP in the context of the appropriate MHC determinant, Kd.
为了进一步研究调节对2,4 - 二硝基氟苯（DNFB）接触敏感性（CS）的可溶性因子，使用来自BALB/c小鼠的经半抗原致敏的脾细胞制备T细胞杂交瘤。鉴定并克隆了一株组成性产生抑制因子的杂交瘤（克隆3 - 10）。将BALB/c DNFB免疫淋巴结细胞（LNC）在3 - 10上清液中孵育，可抑制免疫细胞将CS转移至DNFB的能力。3 - 10因子未抑制CS向恶唑酮或2,4,6 - 三硝基氯苯（TNCB）的被动转移。DNFB免疫LNC而非未致敏或TNCB致敏小鼠的LNC吸收该因子的能力，进一步证明了3 - 10因子的半抗原特异性。3 - 10因子也被来自在主要组织相容性复合体（MHC）K位点与BALB/c小鼠同基因的小鼠（如B10.D2和D2.GD）的DNFB免疫LNC吸附。用单克隆抗Kd抗体或抗DNP抗体预处理DNFB免疫LNC可阻断其吸附该因子的能力。这些结果表明，3 - 10抑制因子与免疫LNC上的DNP/H - 2Kd复合物结合。来自DNFB免疫LNC的尼龙毛纯化T细胞（83% Thy - 1.2 +）与未分离的免疫LNC一样能够吸附该因子。此外，用抗Thy - 1.2加补体（C'）处理免疫LNC可消除细胞吸附该因子的能力，表明3 - 10因子的细胞靶点是T细胞。另外，用自身抗独特型抗血清（CS 231）加补体处理免疫LNC，可耗尽DNP特异性迟发型超敏反应效应T（TDH）细胞，这也消除了细胞吸附该因子的能力。最后，抑制因子可从DNP亲和柱上吸附并洗脱，但不能被TNP亲和柱吸附。总体而言，这些结果表明，尽管单克隆3 - 10抑制因子对DNP有亲和力，但该因子聚焦于TDH细胞需要在合适的MHC决定簇Kd的背景下识别DNP。
