Production of functional human T-T hybridomas in selection medium lacking aminopterin and thymidine.
Production of functional human T-T hybridomas in selection medium lacking aminopterin and thymidine.
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在缺乏氨基蝶呤和胸苷的选择培养基中生产功能性人 T-T 杂交瘤。
DOI：
10.1073/pnas.79.23.7484
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发表时间：
1982
影响因子：
11.1
通讯作者：
Engleman,EG
中科院分区：
文献类型：
--
作者：
Foung,SK;Sasaki,DT;Grumet,FC;Engleman,EG
免疫活化T细胞和恶性T细胞系之间杂交瘤的产生提供了可溶性T细胞衍生产物的潜在无限来源。最近，已经描述了人T-T杂交体;然而，它们的使用受到生长缓慢和染色体不稳定性的阻碍，这至少部分地是由于在传统的次黄嘌呤/氨基蝶呤/胸苷（HAT）选择培养基中存在胸苷。在这份报告中，我们描述了一个快速增长的次黄嘌呤磷酸核糖基转移酶缺陷的人类T细胞系命名为J3 R7的发展，使用阿扎胞苷/次黄嘌呤（AH）培养基作为HAT培养基的替代选择培养基，并通过使用J3 R7系和AH选择技术生产功能性T-T杂交瘤。在AH培养基中选择的杂交种比在HAT培养基中生长的杂交种在数量上大4倍，在生长速度上快3倍。从HAT培养物中没有获得稳定的克隆，而AH衍生的杂交体可以通过有限稀释法容易地克隆。杂交的证据包括：（i）杂交细胞中染色体数目约为J3 R7细胞的两倍;（ii）Leu-3a表面抗原在杂交细胞上的存在，Leu-3a表面抗原存在于正常辅助T细胞上，但不存在于J3 R7细胞上;（iii）正常T细胞伴侣和J3 R7细胞系的HLA抗原的表达;和（iv）来自多个杂交克隆但不是来自J3 R7细胞系的白细胞介素2的组成型分泌。到目前为止，这些克隆已保持其快速生长，染色体数目，表面表型，和白细胞介素2的组成性分泌4个月。
The production of hybridomas between immunologically activated T cells and malignant T-cell lines offers a potentially unlimited source of soluble T-cell-derived products. Recently, human T-T hybrids have been described; however, their use has been hampered by slow growth and chromosomal instability due at least in part to the presence of thymidine in the traditional hypoxanthine/aminopterin/thymidine (HAT) selection medium. In this report, we describe the development of a rapidly growing hypoxanthine phosphoribosyltransferase-deficient human T-cell line designated J3R7, the use of azaserine/hypoxanthine (AH) medium as an alternative selection medium to HAT medium, and the production of functional T-T hybrids by using the J3R7 line and the AH selection technique. Hybrids selected in AH medium were 4-fold greater in number and 3-fold faster in growth rate than hybrids grown in HAT medium. No stable clones were obtained from HAT cultures whereas AH-derived hybrids could be readily cloned by the method of limiting dilution. Evidence for hybridization included (i) the presence of approximately twice the number of chromosomes in hybrids than in J3R7 cells; (ii) the presence on hybrid cells of the Leu-3a surface antigen, present on normal helper T cells but not on J3R7 cells; (iii) the expression of HLA antigens of both the normal T-cell partner and the J3R7 line; and (iv) the constitutive secretion of interleukin 2 from multiple hybrid clones but not from the J3R7 cell line. Thus far, these clones have maintained their rapid growth, chromosome number, surface phenotype, and constitutive secretion of interleukin 2 for 4 months.