Kupiec-Weglinski J W, Heidecke C D, Araujo J L, Abbud-Filho M, Towpik E, Araneda D, Strom T B, Tilney N L
Cell Immunol. 1985 Jun;93(1):168-77. doi: 10.1016/0008-8749(85)90397-1.
(LEW X BN)F1 cardiac allografts are rejected acutely (7 days) in unmodified LEW rats, yet survive indefinitely following cyclosporine (CsA) treatment (15 mg/kg im for 7 days) or in T-cell-deprived (B) recipients. Using these models, the function of T helper cells (Th) in the maintenance phase of CsA-mediated long-term graft survival was examined. With monoclonal antibody immunoaffinity fractionation techniques, Th (W3/25+OX8-) were separated from spleens of CsA-treated hosts 3-4 weeks after grafting (CsA-Th), from specifically sensitized (s-Th), or from normal ungrafted (n-Th) rats. Adoptive transfer of 60 X 10(6) CsA-Th into B recipients produced rejection of donor-specific, but not third-party grafts in 21 +/- 7 days, comparable to s-Th (17 +/- 4 days), but faster than n-Th (4-5 weeks, P less than 0.025). CsA-Th recombined with T cytotoxic/suppressor phenotype (CsA-Tc/s, OX8+W3/25-) in numbers contained in 100 X 10(6) CsA-T cells were ineffectual, even when supplemented with exogenous interleukin 2-rich conditioned medium (IL-2CM); in contrast 100 X 10(6) s-T cells + IL-2CM inevitably caused acute rejection in B hosts (11 +/- 3 days). Increasing numbers of Th incrementally to 100 X 10(6) augmented the effectiveness of s-Th (rejection in 13 +/- 2 days), but did not improve potency of CsA-Th (20 +/- 2 days). Suppressor activity produced by small numbers of contaminating CsA-Tc/s (c. 0.4%, 4-5 X 10(5) cells in 100 X 10(6) CsA-Th) accounted for extended graft survival in B recipients, as this small number of CsA-Tc/s transferred into untreated syngeneic rats increased test graft survival to c. 16 days (P less than 0.001). IL-2 production by spleen cells, depressed during CsA treatment, returned to normal levels 2-3 weeks following drug withdrawal, whereas transfer of CsA-Th into B recipients induced a shift of IL-2 levels from dramatically depressed to normal, findings suggesting normal IL-2 production by CsA-Th. This report demonstrates that an unresponsive state in CsA-treated animals is achieved despite the presence of fully potent donor-specific Th. Active suppressor activity plays a critical role in the maintenance phase of graft survival in rats treated transiently with CsA.
在未接受处理的LEW大鼠中，(LEW×BN)F1心脏异体移植会在7天内被急性排斥，但在接受环孢素（CsA）治疗（15mg/kg，腹腔注射，共7天）后或移植到T细胞缺失（B）受体中时，移植心脏可长期存活。利用这些模型，研究了T辅助细胞（Th）在CsA介导的长期移植存活维持阶段中的作用。采用单克隆抗体免疫亲和分级分离技术，从移植后3 - 4周接受CsA治疗的宿主脾脏中分离出Th（W3/25 + OX8 - ）（CsA - Th），从特异性致敏的（s - Th）或未移植的正常（n - Th）大鼠脾脏中分离出Th。将60×10⁶个CsA - Th过继转移到B受体中，在21±7天内导致供体特异性而非第三方移植被排斥，这与s - Th（17±4天）相当，但比n - Th（4 - 5周，P＜0.025）更快。将100×10⁶个CsA - Th中所含数量的CsA - Th与细胞毒性/抑制性T细胞表型（CsA - Tc/s，OX + 8W3/25 - ）重组，即使补充富含外源性白细胞介素2的条件培养基（IL - 2CM）也无效；相反，100×10⁶个s - Th + IL - 2CM必然会在B宿主中引起急性排斥（11±3天）。将Th数量逐渐增加到100×10⁶可增强s - Th的效力（13±2天发生排斥），但并未提高CsA - Th的效力（20±2天）。少量污染的CsA - Tc/s（约0.4%，100×10⁶个CsA - Th中有4 - 5×10⁵个细胞）产生的抑制活性导致B受体中移植存活时间延长，因为将这少量的CsA - Tc/s转移到未处理的同基因大鼠中会使测试移植的存活时间增加到约16天（P＜0.001）。在CsA治疗期间脾脏细胞产生的IL - 2受到抑制，停药后2 - 3周恢复到正常水平，而将CsA - Th转移到B受体中会使IL - 2水平从显著抑制转变为正常，这些发现表明CsA - Th能正常产生IL - 2。本报告表明，尽管存在完全有活性的供体特异性Th，但CsA处理的动物仍能达到无反应状态。活性抑制活性在CsA短期处理的大鼠移植存活维持阶段起着关键作用。
