Studies of the induction and expression of T cell mediated immunity. VI. Heterogeneity of lytic efficiency exhibited by isolated cytotoxic T lymphocytes prepared from highly enriched populations of effector-target conjugates.
Studies of the induction and expression of T cell mediated immunity. VI. Heterogeneity of lytic efficiency exhibited by isolated cytotoxic T lymphocytes prepared from highly enriched populations of effector-target conjugates.
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T 细胞介导的免疫的诱导和表达的研究。
DOI：
10.4049/jimmunol.126.1.208
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发表时间：
1977
影响因子：
4.4
通讯作者：
B. Bonavida
中科院分区：
文献类型：
--
作者：
E. Grimm;B. Bonavida
在开发了通过特异性结合同种异体靶细胞来高度富集细胞毒性T淋巴细胞（CTL）的方法之后，已经检查了分离的同种异体免疫细胞毒性淋巴细胞（CTL）的裂解效率。通过将免疫淋巴细胞（体外和体内产生）与同种异体肿瘤靶细胞混合制备的小鼠效应细胞-靶细胞缀合物通过不连续胎牛血清（FCS）梯度的速率沉降进行富集。底层含有80 - 90%的结合淋巴细胞，或3- 6倍的富集和约50%的回收率。通过微量稀释技术分离，然后通过单细胞，细胞介导的细胞毒性（CMC）试验证明，平均70%的结合淋巴细胞从初级和次级反应裂解其附着的靶细胞。任何免疫T细胞群中CTL的绝对频率通过用在单细胞CMC测定中观察到杀伤的那些缀合淋巴细胞的因子校正缀合物的百分比来确定。使用这些技术的CTL富集和单细胞测定，已经获得的结果提供了证据的CTL裂解潜力的异质性。单个CTL的内在裂解效率通过定量1373缀合的淋巴细胞裂解靶所需的时间间隔来确定。在37°C下，对于初级CTL需要长达5小时，对于次级CTL需要长达4小时，以实现最大的单靶裂解。此外，平均1° CTL显示出比平均2° CTL更慢的杀伤速率。发现50%的1°淋巴细胞裂解所需的时间为2.5小时，2°淋巴细胞裂解所需的时间为1.2小时，表明细胞毒性淋巴细胞的裂解潜力是异质的。CTL的裂解效率的异源性也通过实验显示，表明结合淋巴细胞的频率与通过51 Cr释放测量的细胞毒性之间的相关性较差。
The lytic efficiency of isolated alloimmune cytotoxic lymphocytes (CTL) has been examined, following the development of a method to highly enrich for cytotoxic T lymphocytes (CTL) by specific binding to allogeneic target cells. Murine effector-target cell conjugates, prepared by mixing populations of immune lymphocytes (generated in vitro and in vivo ) with allogeneic tumor target cells, were enriched by rate sedimentation through a discontinuous fetal calf serum (FCS) gradient. The bottom layer contained 80 to 90% conjugated lymphocytes, or an enrichment of 3- to 6-fold and a recovery of approximately 50%. Isolation by a microdilution technique, followed by single-cell, cell-mediated cytotoxicity (CMC) assay proved that an average of 70% of the conjugating lymphocytes from both primary and secondary responses lysed their attached target cell. The absolute frequency of CTL in any immune T cell population was determined by correcting the percentage of conjugates by the factor of those conjugating lymphocytes observed to kill in the single-cell CMC assay. Using these techniques of CTL enrichment and single-cell assay, results have been obtained that provide evidence for heterogeneity of CTL lytic potential. The intrinsic lytic efficiency of individual CTL was determined by quantitation of the time intervals required for target lysis by 1373 conjugated lymphocytes. Up to 5 hr for primary and 4 hr for secondary CTL was necessary to achieve maximum single-target lysis at 37°C. In addition, the average 1° CTL displayed a slower rate of killing than the average 2° CTL. It was found that the time required for lysis by 50% of the 1° lymphocytes was 2.5 hr and for 2° lymphocytes was 1.2 hr, demonstrating that cytotoxic lymphocytes are heterogeneous in their lytic potential. Heterogeneity of lytic efficiency by CTL was also shown by experiments demonstrating a poor correlation between the frequency of conjugating lymphocytes and the cytotoxicity measured by 51 Cr release.