Bigby M, Vargas R, Sy M S
Department of Pathology, Massachusetts General Hospital, Boston.
J Immunol. 1989 Dec 15;143(12):3867-72.
We produced a series of T cell hybridomas that produce IL-2 when cultured with syngeneic APC coupled to FITC or TNP. These hybridomas are hapten specific and Ia restricted. The hybridomas were used to detect hapten-bearing APC in draining lymph nodes of mice sensitized with trinitrochlorobenzene or FITC in vivo. Hapten-bearing APC capable of stimulating the hybridomas were detectable in draining lymph nodes of hapten-painted mice within 3 h after sensitization. The ability of lymph node APC to stimulate the hybridomas peaked at 24 h and declined by 48 h. The dendritic cell subpopulation was the subpopulation of cells that were found in the regional lymph nodes of hapten-painted animals that were capable of stimulating the hybridomas to produce IL-2. Prior treatment of the skin with low dose UVB irradiation before epicutaneous application of contact sensitizers significantly reduced the capacity of hapten-bearing APC to stimulate the hybridomas. This observation was corroborated by results obtained from flow microfluorometry analysis of lymph node cells from FITC-sensitized mice. Lymph node dendritic cells obtained from FITC-painted mice contain a brightly staining group of cells by flow microfluorometry analysis. Lymph node dendritic cells from FITC-painted, UVB-irradiated mice did not contain this brightly staining population. These results indicate that low dose, local UVB irradiation may affect APC migration and/or function. We believe that these hybridomas will prove to be useful tools in the study of the development and regulation of contact hypersensitivity.
我们制备了一系列T细胞杂交瘤,当与偶联异硫氰酸荧光素(FITC)或三硝基苯(TNP)的同基因抗原呈递细胞(APC)一起培养时,这些杂交瘤可产生白细胞介素-2(IL-2)。这些杂交瘤具有半抗原特异性且受Ia限制。这些杂交瘤被用于检测在体内用三硝基氯苯或FITC致敏的小鼠引流淋巴结中携带半抗原的APC。致敏后3小时内,在涂抹半抗原的小鼠引流淋巴结中可检测到能够刺激杂交瘤的携带半抗原的APC。淋巴结APC刺激杂交瘤的能力在24小时达到峰值,并在48小时下降。树突状细胞亚群是在涂抹半抗原动物的局部淋巴结中发现的能够刺激杂交瘤产生IL-2的细胞亚群。在经皮应用接触性致敏剂之前,先用低剂量紫外线B(UVB)照射皮肤,可显著降低携带半抗原的APC刺激杂交瘤的能力。对FITC致敏小鼠的淋巴结细胞进行流式微量荧光测定分析所获得的结果证实了这一观察结果。通过流式微量荧光测定分析,从涂抹FITC的小鼠获得的淋巴结树突状细胞含有一组染色明亮的细胞。从涂抹FITC且经UVB照射的小鼠获得的淋巴结树突状细胞不含有这种染色明亮的群体。这些结果表明,低剂量局部UVB照射可能影响APC的迁移和/或功能。我们相信,这些杂交瘤将被证明是研究接触性超敏反应的发生和调节的有用工具。