Aiello F B, Musiani P, Maggiano N, Larocca L M, Piantelli M
Clin Exp Immunol. 1985 Aug;61(2):433-9.
By panning procedures employing T6 and T3 monoclonal antibody, human thymocytes were fractionated into two subpopulations depleted of T6- or T3-positive (T6+, T3+) cells. Unfractionated thymocytes and T6- and T3-depleted subpopulations were separately cultured for 48 h in RPMI 1640 medium with 10% FCS or in HB 101 serum-free medium. Determining the phenotype of unfractionated thymocytes at various time intervals, a time-dependent increase of T3+ cells was observed. An inverse relationship was found between the percentage of T3+ cells and the T6 and peanut agglutinin (PNA) reactive thymocytes. When the surface antigen expression in the T3-depleted population (greater than 95% T6+ and PNA+ cells) was analysed, a strong increase of T3+ cells and a complementary reduction of T6+ and PNA+ cells was evidenced. During that time the surface phenotype of the T6-depleted population (greater than 80% T3+ cells) showed the same trend of differentiation, as the other thymocyte preparations. These results indicate that a conspicuous fraction of human thymocytes and particularly of those characterized by a cortical phenotype (PNA+ and T6+ cells), are able to express mature T-cell antigens when cultured in vitro in the absence of the thymic microenvironment influence. However, the in vitro acquisition of a mature phenotype is not accompanied by a parallel achievement of the capacity to respond to mitogens such as PHA or T3 monoclonal antibody.
通过使用T6和T3单克隆抗体的淘选程序,人胸腺细胞被分离成两个亚群,分别去除了T6阳性(T6+)或T3阳性(T3+)细胞。未分离的胸腺细胞以及去除T6和T3的亚群分别在含有10%胎牛血清的RPMI 1640培养基或HB 101无血清培养基中培养48小时。在不同时间间隔测定未分离胸腺细胞的表型,观察到T3+细胞呈时间依赖性增加。发现T3+细胞的百分比与T6和花生凝集素(PNA)反应性胸腺细胞之间呈负相关。当分析去除T3的群体(超过95%为T6+和PNA+细胞)的表面抗原表达时,发现T3+细胞显著增加,而T6+和PNA+细胞则相应减少。在此期间,去除T6的群体(超过80%为T3+细胞)的表面表型与其他胸腺细胞制剂表现出相同的分化趋势。这些结果表明,相当一部分人胸腺细胞,特别是那些具有皮质表型(PNA+和T6+细胞)的胸腺细胞,在体外培养且不受胸腺微环境影响时能够表达成熟T细胞抗原。然而,体外获得成熟表型并不伴随着对诸如PHA或T3单克隆抗体等有丝分裂原反应能力的相应获得。