Detection and characterization of cytotoxic T lymphocyte precursors in the murine intestinal intraepithelial leukocyte population.
Detection and characterization of cytotoxic T lymphocyte precursors in the murine intestinal intraepithelial leukocyte population.
复制标题
小鼠肠上皮内白细胞群中细胞毒性 T 淋巴细胞前体的检测和表征。
作者：
P. Ernst;D. Clark;K. Rosenthal;A. Befus;J. Bienenstock
从小肠粘膜获得高度纯化的上皮内白细胞（IEL）制剂。分离固有层（LPL）的白细胞，并与IEL进行表型比较，以验证IEL受LPL的污染最小。因为大约80%的IEL表达通常与细胞毒性/抑制性T淋巴细胞相关的Lyt-2抗原，所以我们希望确定该群体中是否存在细胞毒性T细胞的前体。为了产生细胞毒性细胞，将CBA/J小鼠（H-2k）的IEL和脾细胞与经辐照的同种异体脾细胞（H-2d或H-2b）在单向混合白细胞反应（MLR）中共培养。4 ~ 6天后，检测培养细胞对51 Cr标记的H-2d或H-2b肿瘤或ConA刺激的淋巴母细胞靶细胞的反应，并比较同种异体抗原刺激的IEL和脾细胞的特异性。来自两种组织的细胞毒性细胞显示同种异体靶的抗原特异性裂解。用针对Thy-1.2、Lyt-1.1或Lyt-2.1抗原的单克隆抗体加上补体处理由上皮内或脾脏前体产生的效应细胞，可使细胞毒性降低85%至100%，尽管只有20%至50%的细胞被溶解。同种异体抗原特异性和表面抗原表型的培养IEL细胞是相同的脾细胞，并允许我们得出结论，IEL含有细胞毒性T淋巴细胞前体（CTLp）。进一步表征显示，与脾脏一样，上皮内CTLp为Thy-1+和Lyt-1+，它们的沉降速度相同，但不同于上皮内自然杀伤细胞。尽管80%的IEL是Lyt-2+，但IEL群体中CTLp的频率估计比脾脏中低三倍至五倍，并且Lyt-2+细胞显示不是CTLp的富集来源。因此，大多数IEL的功能仍然未知。然而，在该群体中存在CTLp，其可能能够被管腔抗原刺激。
Highly purified preparations of intraepithelial leukocytes (IEL) were obtained from the small intestinal mucosa. Leukocytes from the lamina propria (LPL) were isolated and phenotypically compared with IEL to verify that IEL were minimally contaminated by LPL. Because approximately 80% of IEL expressed the Lyt-2 antigen usually associated with cytotoxic/suppressor T lymphocytes, we wished to determine if precursors for cytotoxic T cells were present in this population. In order to generate cytotoxic cells, IEL and spleen cells from CBA/J mice (H-2k) were co-cultured with irradiated allogeneic spleen cells (H-2d or H-2b) in a one-way mixed leukocyte reaction (MLR). Four to six days later, the cultured cells were assayed against 51Cr-labeled H-2d or H-2b tumor or Con A-stimulated lymphoblast target cells, and the specificity of alloantigen-stimulated IEL and spleen cells was compared. The cytotoxic cells derived from both tissues displayed antigen-specific lysis of the allogeneic targets. Treatment of effector cells, generated from intraepithelial or splenic precursors, with monoclonal antibodies against Thy-1.2, Lyt-1.1, or Lyt-2.1 antigens plus complement, decreased cytotoxicity 85 to 100%, even though only 20 to 50% of the cells were lysed. The alloantigen specificity and surface antigen phenotype of the cultured IEL cells were identical to those of spleen cells and allowed us to conclude that IEL contained a cytotoxic T lymphocyte precursor (CTLp). Further characterization showed that, like spleen, the intraepithelial CTLp was Thy-1+ and Lyt-1+ and their sedimentation velocity was the same but differed from intraepithelial natural killer cells. Although 80% of IEL were Lyt-2+, the frequency of CTLp in the IEL population was estimated to be threefold to fivefold lower than in spleen, and the Lyt-2+ cells were shown not to be an enriched source of CTLp. Thus, the function of the majority of the IEL is still not known. However, there exists within this population CTLp, which may be capable of being stimulated with luminal antigens.