IMMUNOBIOLOGY OF CUTANEOUS T CELL LYMPHOMA

皮肤T细胞淋巴瘤(CTCL)是一种嗜表皮性的恶性肿瘤 辅助性/诱导性T淋巴细胞随后系统性地 转移性的。在每个单独的患者中，CTCL细胞表达一个 抗原的独特T细胞受体(TCR) 同时是克隆性的标志，肿瘤特异性抗原和 与正常T细胞的历史联系，它们是从正常T细胞进化而来的。 在这个项目中，CTCL TCR的特点将被开发出来 实现四个相互关联的目标。第一，提高敏感度 诊断技术，并获得CTCL细胞用于深入研究 在研究中，将采取两步走的方法。外周血和 CTCL的淋巴准备将大大丰富 用单克隆法进行荧光激活细胞分选 识别肿瘤相关CTCL的抗体(MAb)BE2 抗原。恶性细胞克隆性群体的存在 得到的细胞悬浮液将通过Southern印迹建立 对它们重排的TCR基因进行分析。第二，这些BE2阳性 CTCL细胞将用于产生克隆型(抗TCR)单抗(mAb- L)，对于更敏感的身份识别和 相关患者的CTCL细胞分析。第三，另一个 将产生一组针对独特型决定簇的单抗(mAb-2 单抗-L。一些单抗-2将在结构上模拟CTCL TCR抗原- 结合部位并将用于冰冻的直接检测 皮肤切片以识别CTCL细胞和 鉴定正常“防御性”AN：L-CTCL淋巴细胞与之反应 CTCL TCR。这种自然或诱导的抗CTCL免疫是 将针对CTCL TCR的独特决定因素进行审查 抗CTCL抗体对靶CTCL细胞TCRs的调节作用 CD3，一种与TCR共调节的膜分子。第四, 将对人类和老鼠细胞进行研究，以确定 CTCL细胞体外暴露于紫外线(UVA)-- 照射后的8-甲氧补骨脂素(8-MOP)增强免疫原性 CTCL患者的这些细胞通过改变膜的抗原性。 由于CTCL细胞优先受到8-MOP/UVA的损伤，其 修复总可提取核DNA和In损伤的能力 将评估特定基因(TCR、IL2受体和IL2)，如 将这些基因产物的膜表达。这个 8-MOP/UVA对小鼠免疫功能的影响 细胞色素C小鼠致瘤T细胞杂交瘤细胞疫苗的研究 在体内的生长情况将通过使用 可与这些杂交瘤的TCR选择性结合的可用单抗 细胞。总而言之，这些研究应该显著增强 对CTCL免疫生物学的认识。

Cutaneous T cell lymphoma (CTCL)is a malignancy of epidermotropic helper/inducer T lymphocytes which subsequently become systemically metastatic. In each individual patient, CTCL cells express a distinctive T cell receptor (TCR) for antigen which is simultaneously a marker of clonality, a tumor specific antigen and an historical link with the normal T cells from which they evolved. In this project, the features of CTCL TCRs will be exploited to achieve four interrelated goals. First, to increase sensitivity of diagnostic techniques and to obtain CTCL cells for in-depth studies, a two step approach will be taken. Peripheral blood and lymph node preparations will be substantially enriched for CTCL cells by fluorescence-activated cell sorting with monoclonal antibody (mAb) BE2 which recognizes a CTCL tumor-associated antigen. The presence of clonal populations of malignant cells in the resulting cell suspensions will be established by Southern blot analysis of their rearranged TCR genes. Second, these BE2-positive CTCL cells will be used to produce clonotypic (anti-TCR) mAbs (mAb- l), Important for the even more sensitive identification and analysis of CTCL cells in the relevant patients. Third, another set of mAbs (mAb-2) will be produced against idiotypic determinants of mAb-l. Some mAb-2 will structurally mimic the CTCL TCR antigen- binding sites and will be used in direct examination of frozen sections of skin to recognize putative targets for CTCL cells and to identify normal "defensive" an:l-CTCL lymphocytes reactive with the CTCL TCR. That such natural or induced anti-CTCL immunity is directed at unique determinants of the CTCL TCRs will be examined by modulation of the TCRs on target CTCL cells by antibody against CD3, a membrane molecule which comodulates with the TCR. Fourth, human and murine cells will be studied to determine whether extracorporeal exposure of CTCL cells to ultraviolet A (UVA)-- irradiated 8-methoxypsoralen (8-MOP) increases the immunogenicity of these cells in CTCL patients by altering membrane antigenicity. Since CTCL cells are preferentially injured by 8-MOP/UVA, their capacity to repair damage in total extractable nuclear DNA and in specific genes (TCR, IL2 receptor and IL2) will be assessed, as will the membrane expression of the products of these genes. The effects of 8-MOP/UVA on the capacity of an established anti- cytochrome C murine tumorigenic T cell hybridoma to vaccinate against its own in vivo growth will be assessed through use of an available mAb which binds selectively to the TCR of those hybridoma cells. Together, these studies should markedly enhance understanding of the immunobiology of CTCL.