Iwakami N, Kikuchi A, Kunishita T, Yamamoto H, Nonaka I, Kamo I
National Institute of Neuroscience, Tokyo, Japan.
Immunology. 1996 Jan;87(1):108-12.
Lymphoproliferative activities produced by cloned thymic myoid cell 871207B were analysed by immunological and biochemical methods. The lymphoproliferative activities were separated into two fractions by DEAE-Sepharose CL-6B chromatography: one is in the fraction passed through the column and the other in the fraction eluated from the column with a low concentration of NaCl. The eluated fraction induced the proliferation of interleukin-1 (IL-1)-dependent D10N4 M cells. This activity was abrogated by an anti-IL-1 alpha antibody, but not an anti-IL-1 beta antibody. Expression of IL-1 alpha mRNA was also detected in 871207B cells. The thymocyte proliferative activity found in the fraction passed through the DEAE-Sepharose column was further separated into three fractions by heparin-Sepharose column chromatography: (1) the fraction passed through the column, (2) the fraction weakly bound to the column, and (3) the fraction firmly bound to the heparin column. The fraction passed through the heparin column sustained the growth of IL-6-dependent MH60.BSF-2 cells. IL-6-specific mRNA was found in 871207B cells. The thymocyte proliferative activity of the fraction firmly bound to the heparin column was neutralized with an anti-IL-7 antibody. The biological activity of the fraction weakly bound to the column remained to be elucidated. These results suggest that thymic myoid cells produce IL-1 alpha, IL-6, IL-7 and unidentified lympho-stimulatory factors, all of which play significant roles in many steps of T-cell development in the thymus.
采用免疫学和生物化学方法分析了克隆的胸腺肌样细胞871207B产生的淋巴细胞增殖活性。通过DEAE-琼脂糖CL-6B层析将淋巴细胞增殖活性分离为两个组分:一个在通过柱的组分中,另一个在用低浓度NaCl从柱上洗脱的组分中。洗脱组分诱导白细胞介素-1(IL-1)依赖性D10N4 M细胞增殖。该活性被抗IL-1α抗体消除,但不被抗IL-1β抗体消除。在871207B细胞中也检测到IL-1α mRNA的表达。通过DEAE-琼脂糖柱的组分中发现的胸腺细胞增殖活性通过肝素-琼脂糖柱层析进一步分离为三个组分:(1)通过柱的组分,(2)与柱弱结合的组分,和(3)与肝素柱牢固结合的组分。通过肝素柱的组分维持IL-6依赖性MH60.BSF-2细胞的生长。在871207B细胞中发现了IL-6特异性mRNA。与肝素柱牢固结合的组分的胸腺细胞增殖活性用抗IL-7抗体中和。与柱弱结合的组分的生物学活性有待阐明。这些结果表明,胸腺肌样细胞产生IL-1α、IL-6、IL-7和未鉴定的淋巴细胞刺激因子,所有这些因子在胸腺中T细胞发育的许多步骤中都发挥着重要作用。