Genetic mapping of human chromosome 11: use of human cell surface antigens.
Genetic mapping of human chromosome 11: use of human cell surface antigens.
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人类 11 号染色体的遗传图谱：使用人类细胞表面抗原。
DOI：
10.1007/bf01534934
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Jones,C
中科院分区：
文献类型：
--
作者：
Jones,C
构建人类染色体精细结构图谱的能力具有重要的现实意义和理论意义。此前，我们分离并鉴定了一个人-中国仓鼠体细胞杂种，其唯一的人类染色体为人的11号染色体(1)。这种名为J1的杂交种表达许多人类细胞表面抗原，这些标记已被证明在制定区域地图策略方面有用。Oda和Puck博士表明，用人类细胞免疫的实验动物产生的抗血清在补体存在的情况下对人类细胞有毒性，但不会杀死中国仓鼠细胞。这项技术被用来鉴定由含有人11号染色体(3，4)的杂交细胞表达的人细胞表面抗原。最近生产的识别这些抗原的克隆抗体使我们能够扩展这一分析(5-7)。我们通过对这些抗原标记的选择，获得了一系列J1的亚克隆，其中11号染色体的不同部分已被删除(8，9)。这个杂交小组在11号染色体(9-11)的区域定位中很有用。A~基因是我们研究的细胞表面抗原之一，位于11号染色体(11，12)的p13区，该区域与Wilms肿瘤无虹膜综合征(13)密切相关。虽然被选择丢失该区域的缺失杂交体在确定该区域方面是有用的，但它们不能用于进一步研究该区域的基因。为此，我们利用Goss和Harris开发的技术改进，开发了一组包含部分11号染色体的新杂交种(14)。亚克隆JL-11只含有11号染色体短臂，经高剂量X射线照射(8000rad)后与中国仓鼠卵巢细胞融合。由于辐射会导致广泛的染色体碎裂，新的杂交细胞将包含来自捐赠者细胞的不同遗传物质片段。为了分离出含有a基因的11个片段的杂交种，用抗A1单抗处理融合群体，并在包被山羊抗鼠免疫球蛋白(15)的细菌培养板上“淘洗”。表达a~的细胞与鼠单抗结合并附着在包被的平板上。用这种方法恢复了一系列表达Al的杂交种。利用间接免疫荧光和同工酶分析(7，9)，克隆并分析了另外两种人类细胞表面抗原(A3和mer-2)a~，以及乳酸脱氢酶A(LDHA)和酸性磷酸酶2(ACP2)两种同工酶的表达。对三个具有代表性的辐射杂交种及其亲本细胞JL-11的分析结果如表1所示。
The ability to construct a fine structure map of the human chromosomes has both practical and theoretical importance. Previously we isolated and characterized a human-Chinese hamster somatic cell hybrid which contains human chromosome 11 as its only human chromosome (1). This hybrid, J1, expresses a number of human cell surface antigens, and these markers have proved to be useful in developing regional mapping strategies. Drs. Oda and Puck showed that antisera elicited in experimental animals immunized with human cells are toxic to human cells in the presence of complement but do not kill Chinese hamster cells (2). This technique was used to identify human cell surface antigens expressed by the hybrid cell containing human chromosome 11 (3, 4). The recent production of monoclonal antibodies which recognize these antigens has allowed us to extend this analysis (5-7). We have derived a series of subclones of J1 by selection against these antigen markers in which varying portions of chromosome 11 have been deleted (8, 9). This hybrid panel has been useful in the regional mapping of chromosome 11 (9-11). The gene encoding a~, one of the cell surface antigens we have studied, is located in the p13 region of chromosome 11 (11, 12) which is of particular interest because of the association of this region with the Wilms tumor anirdia syndrome (13). Although the deletion hybrids which have been selected to lose this region have been useful in defining the region, they cannot be used to study further the genes in this region. For this reason we have developed a new set of hybrids containing portions of chromosome 11 using a modification of techniques developed by Goss and Harris (14). The subclone Jl-ll which contains only the short arm of chromosome 11, was subjected to a high dose of X-irradiation (8000 rads) and was then fused with Chinese hamster ovary cells. Because the irradiation causes extensive chromosome fragmentation, the new hybrids will contain different fragments of genetic material from the donor cell. To isolate the hybrids which have retained a fragment of 11 containing the a~ gene, the fused population was treated with anti-a1 monoclonal antibody and" panned" on bacterial culture plates coated with goat antimouse immunoglobulin (15). Cells which expressed a~ bound the mouse monoclonal antibody and attached to the coated plates. A series of hybrids which expressed al were recovered in this manner. They have been cloned and analyzed for the expression of a~, two other human cell surface antigens (a3 and MER-2), and the two isozymes lactic dehydrogenase A (LDHA) and acid phosphatase 2 (ACP2), using indirect immunofluorescence and isozyme analysis (7, 9). Results of this analysis on three representative irradiation hybrids and their parental cell Jl-ll are shown in Table 1.