Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition.
Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition.
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DOI：
10.1084/jem.153.5.1198
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发表时间：
1981-05-01
期刊：
影响因子：
--
通讯作者：
Marrack P
中科院分区：
文献类型：
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作者：
Kappler JW;Skidmore B;White J;Marrack P
我们开发了一种生产抗原特异性、H - 2限制性T细胞杂交瘤的方法。融合中的肿瘤细胞伴侣本身是一个T细胞杂交瘤，FS6 - 14.13.AG2（或其衍生物），它在受到伴刀豆球蛋白A刺激时可被诱导产生生长因子白细胞介素 - 2（IL - 2），但没有已知的抗原特异性。融合中的正常T细胞伴侣是一群淋巴结T细胞母细胞，它们通过体内免疫，接着在体外用抗原刺激以及在含IL - 2的培养基中进行克隆扩增，从而高度富集了抗原特异性、H - 2限制性T细胞。这些融合产生了在培养中持续生长的杂交瘤。相当大比例的杂交瘤表现出在受到适当H - 2类型的照射脾细胞呈递的特异性抗原刺激时产生IL - 2的能力。产生了四个克隆的抗原/H - 2特异性杂交瘤系。AO - 40.10在由携带I - A(k)的细胞呈递鸡卵白蛋白（OVA）时产生反应。DC1.18.3在由I - A(d)呈递牛肉细胞色素c的脱辅基形式时产生反应。AODK - 10.4在由I - A(d)呈递钥孔血蓝蛋白（KLH）时产生反应。AODK - 1.16也对由H - 2(d)的I区产物呈递的KLH产生反应，但数据与I - J - I - E(d)区的产物或者由I - A(d)和I - E(d)/I - C(d)两者成分组成的组合分子都相符。巧合的是，AO - 40.10被证明与定位于K - I - A区的H - 2(b)产物具有意外的同种异体反应性。这些杂交瘤作为单克隆材料的来源，对于研究具有H - 2限制性抗原特异性的T细胞上的受体应该是极有价值的。我们还通过将AO - 40.10的一个抗氮鸟嘌呤克隆与具有不同抗原/H - 2特异性的正常T细胞融合，产生了具有两种抗原/H - 2特异性的T细胞杂交瘤。许多杂交瘤保留了对OVA加H - 2(a)以及对第二种抗原/H - 2组合的反应性。没有一个对OVA加第二种H - 2类型或者对第二种抗原加H - 2(a)产生反应。其中一个杂交瘤被成功克隆，产生了AOFK - 11.11.1系。它保留了识别从一个亲本遗传的OVA加I - A(k)以及从另一个亲本遗传的KLH加I - A(f)的能力。它不识别OVA加I - A(f)或者KLH加I - A(k)。这些结果对描述与H - 2产物相关联识别的抗原的T细胞受体性质的模型有一定影响。它们不支持抗原和H - 2由两个独立的T细胞受体分别识别的模型。
We developed a method for production of antigen-specific, H-2-restricted T cell hybrids. The tumor cell partner in the fusions was itself a T cell hybrid, FS6-14.13.AG2 (or its derivatives), which could be induced to produce the growth factor, interleukin-2 (IL-2), in response to a challenge with concanavalin A, but had no known antigen specificity. The normal T cell partner in the fusions was a population of lymph node T cell blasts that had been highly enriched in antigen-specific, H-2-restricted T cells by in vivo immunization, followed by in vitro challenge with antigen and clonal expansion in IL-2-containing medium. These fusions produced hybrids that grew constitutively in culture. A sizable proportion of the hybrids demonstrated the ability to produce IL-2 in response to a challenge with specific antigen presented by irradiated spleen cells of the appropriate H-2 type. Four cloned antigen/H-2-specific hybrid lines were produced. AO-40.10 responded to chicken ovalbumin (OVA) when presented by I-A(k)-bearing cells. DC1.18.3 responded to the apo form of beef cytochrome c when presented with I-A(d). AODK-10.4 responded to keyhole limpet hemocyanin (KLH) presented with I-A (d). AODK-1.16 also responded to KLH presented by a product of the I region of H-2(d), but the data were consistent with either a product of the I-J-I-E(d) region or a combinatorial molecule with elements from both I-A(d) and I-E(d)/I-C(d). Coincidentally, AO-40.10 was shown to have an unexpected alloreactivity with a product of H-2(b) mapping to the K-I-A region. These hybrids should prove invaluable as sources of monoclonal material for the study of the receptor(s) on T cells with H-2-restricted antigen specificities. We also generated T cell hybrids with two antigen/H-2 specificities by fusing an azaguanine-resistant clone of AO-40.10 to normal T cells with a different antigen/H-2 specificity. Many of the hybrids retained reactivity to OVA plus H-2(a) and to the second antigen/H-2 combination. None reacted to either OVA plus the second H-2 type or to the second antigen plus H-2(a). One of these hybrids was successfully cloned to produce the line AOFK- 11.11.1. It retained the ability to recognize OVA plus I-A(k) inherited from one parent, and KLH plus IA(f) inherited from the other. It did not recognize OVA plus IA(f) or KLH plus I-A(k). These results have some bearing on models describing the nature of T cell receptors for antigen recognized in association with H-2 products. They do not support models in which antigen and H-2 are recognized separately by two independent T cell receptors.