Sensi M, Orosz C G, Bach F H
J Immunol. 1984 Jun;132(6):3218-25.
It has been shown that peritoneal exudate cells (PEC) from BALB/c mice immunized with minor histocompatibility antigens presented by DBA/2 or B10.D2 spleen cells are capable of lysing syngeneic YC8 tumor cells in a 4-hr 51Cr-release assay. In this study, we employed limiting dilution analysis to determine the frequency of CTL precursors (CTL-P) reactive against both the specific DBA/2 (or P815) target and the syngeneic tumor YC8. The mean frequency of anti-DBA/2 CTL-P in PEC from BALB/c mice immunized with DBA/2 was 1/302. Between one-third and one-fifth of limiting dilution microcultures that exhibited lytic activity against DBA/2 lymphoblasts (or P815) were also able to lyse YC8. No lysis of YC8 was observed in the absence of a parallel lysis on DBA/2 lymphoblasts or P815 target cells. T cell clones, derived by micromanipulation from microcultures selected for cytotoxic activity against YC8 and/or P815, maintained either the specific anti-allogeneic or the doubly reactive ( antiallogeneic plus anti-syngeneic tumor) phenotype. Fourteen clones (six specific and eight doubly reactive) were tested for cytotoxic activity on a panel of target cells with different haplotypes. All showed H-2-restricted specificity for minor histocompatibility antigens shared by DBA/2 and B10.D2. The restriction element for some of the clones mapped in the K region of the H-2 complex, whereas for other clones the restriction element mapped in the D region; both K- and D-restricted clones were able to lyse YC8. When the clones that exhibited lysis on YC8 were tested on two other BALB/c tumor targets, LSTRA, a Moloney virus induced lymphoma, and RL male-1, a radiation induced lymphoma, two of seven were found to lyse all three syngeneic tumor targets equally well, but not syngeneic BALB/c blasts. These clones were functionally categorized as conventional CTL because they were unable to proliferate when cultured with antigen in the absence of exogenous lymphokines, and were unable to produce lymphokine with IL 2 activity when stimulated by the appropriate splenocytes. When tested in vivo in a Winn assay, a strong anti-tumor activity against YC8 was exerted by the anti-DBA/2 clones DY4 -3 and DY16 -3. These clones lysed both YC8 and the immunizing target cells in vitro. No in vivo effect in neutralizing YC8 tumor growth was observed with clone D2-1, a clone that lysed DBA/2 targets but not YC8 in vitro.
已经表明，用DBA/2或B10.D2脾细胞呈递的次要组织相容性抗原免疫的BALB/c小鼠的腹腔渗出细胞（PEC），在4小时的51Cr释放试验中能够裂解同基因的YC8肿瘤细胞。在本研究中，我们采用有限稀释分析来确定针对特异性DBA/2（或P815）靶标和同基因肿瘤YC8具有反应性的CTL前体（CTL-P）的频率。用DBA/2免疫的BALB/c小鼠的PEC中抗DBA/2 CTL-P的平均频率为1/302。在对DBA/2淋巴母细胞（或P815）表现出裂解活性的有限稀释微培养物中，三分之一到五分之一也能够裂解YC8。在没有对DBA/2淋巴母细胞或P815靶细胞进行平行裂解的情况下，未观察到YC8的裂解。通过显微操作从选择用于针对YC8和/或P815具有细胞毒性活性的微培养物中获得的T细胞克隆，维持特异性抗异基因或双重反应性（抗异基因加抗同基因肿瘤）表型。对14个克隆（6个特异性和8个双重反应性）在一组具有不同单倍型的靶细胞上测试细胞毒性活性。所有克隆均显示对DBA/2和B10.D2共有的次要组织相容性抗原具有H-2限制性特异性。一些克隆的限制元件定位于H-2复合体的K区域，而其他克隆的限制元件定位于D区域；K-和D-限制性克隆均能够裂解YC8。当对在YC8上表现出裂解的克隆在另外两个BALB/c肿瘤靶标上进行测试时，一个莫洛尼病毒诱导的淋巴瘤LSTRA和一个辐射诱导的淋巴瘤RL male-1，发现7个中有2个能够同样良好地裂解所有三个同基因肿瘤靶标，但不能裂解同基因的BALB/c胚细胞。这些克隆在功能上被归类为传统CTL，因为它们在没有外源性淋巴因子的情况下与抗原一起培养时不能增殖，并且在受到适当的脾细胞刺激时不能产生具有IL-2活性的淋巴因子。在Winn试验中进行体内测试时，抗DBA/2克隆DY4 -3和DY16 -3对YC8发挥了强大的抗肿瘤活性。这些克隆在体外裂解YC8和免疫靶细胞。克隆D2-1在体外裂解DBA/2靶标但不裂解YC8，在体内未观察到中和YC8肿瘤生长的作用。
