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MONOCLONAL ANTIBODIES TO FETAL EPIDERMIS

胎儿表皮单克隆抗体

基本信息

批准号：
3079011
负责人：
ALFRED T LANE
金额：
$ 7.06万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-08-01 至 1986-07-31
项目状态：
已结题

项目摘要

The structural embryology of fetal skin has been well studied and characterized; little is known about its function or biochemical characteristics. For this reason, fetal, neonatal and infant skin will be used to develop monoclonal antibodies to epidermal componentS of periderm, stratum intermedium, basal cells, basement membrane zone and congenital melanocytic nevi. Monoclonal antibodies will be generated by fusion of P3/NS1 (HGPRT-) mouse myeloma cells with the spleens of immunized mice. The mice will be immunized with human epidermal and dermal cellular components from tissues obtained from abortions, still births, neonatal deaths, neonatal foreskins and infants with congenital melanocytic nevi. Fused cells will be cultured in a media containing hypoxanthine, methotrexate and thymidine to exclude unfused HGPRT- P3/NS1 cells. Hybrid cells producing antibodies to the immunizing tissues will be cloned by dilution in individual culture wells. Those single colonies producing antibodies to the immunizing tissues will be evaluated for tissue specificity by radiommunoprecipitation and immuofluorescence against fetal, neonatal and adult skin and against human tissue culture cell lines. Monoclonal antibodies that are not against common cell surface antigens (Beta2 microglobulin, HLA) but are present in fetal and absent in more mature epidermis and those present in mature epidermis and absent in fetal skin will be studied further. The molecular weight and isoelectric point of the antigenic component reacting with the monoclonal antibody will be identified by SDS gel electrophoresis, electroblotting to nitrocelloluse paper and immunoperoxidase staining. Ultrastructural subcellular localization will be done by immunoelectron microscopy. Once developed, the specific monoclonal antibodies will be used to evaluate cutaneous neoplasms (squamous cell carcinoma, basal cell carcinoma, melanoma) non-neoplastic proliferations (keratoacanthoma, epidermal appendageal tumors, seborrheic keratosis, warts) and a broad range of other skin diseases (epidemolysis bullosa, psortasis, congenital melanocytic nevi). Correlation will be attempted between antigenic presence in fetal, neonatal or adult skin with function of epidermal cells. Current monoclonal and human antibodies will be used to evaluate fetal and neonatal skin for the presence of known antigens in order to establish sequence and progression of fetal development.

胎儿皮肤的结构胚胎学已经得到了很好的研究，特征;对其功能或生物化学知之甚少特色因此，胎儿、新生儿和婴儿的皮肤将用于开发针对牙周炎表皮成分的单克隆抗体，中间层、基底细胞、基底膜带和先天性黑素细胞痣单克隆抗体将通过以下融合产生： P3/NS 1（HGPRT-）小鼠骨髓瘤细胞与免疫小鼠的脾脏。小鼠将用人表皮和真皮细胞免疫从流产、死产、新生儿死亡、新生儿包皮和患有先天性黑色素细胞痣的婴儿。融合细胞将在含有次黄嘌呤的培养基中培养，甲氨蝶呤和胸苷以排除未融合的HGPRT-P3/NS 1细胞。混合产生针对免疫组织的抗体的细胞将被克隆，在单个培养威尔斯孔中稀释。那些产生将评估免疫组织的抗体通过放射免疫沉淀和免疫荧光对胎儿特异性，新生儿和成人皮肤以及抗人组织培养细胞系。不针对常见细胞表面抗原的单克隆抗体（β 2微球蛋白，HLA），但存在于胎儿和缺乏在更多的成熟表皮和那些存在于成熟表皮和缺乏在胎儿皮肤将进一步研究。分子量和等电点与单克隆抗体反应的抗原组分的量将是 SDS凝胶电泳、硝酸纤维素电印迹鉴定纸和免疫过氧化物酶染色。超微结构亚细胞通过免疫电子显微镜进行定位。一旦开发出来，特异性单克隆抗体将用于评估皮肤肿瘤（鳞状细胞癌、基底细胞癌、黑色素瘤）非肿瘤性增生（角化棘皮瘤、表皮附件肿瘤，脂溢性角化病，疣）和广泛的其他皮肤疾病（大疱性痣、银屑病、先天性黑色素细胞痣）。将尝试将胎儿、新生儿和新生儿中的抗原存在或具有表皮细胞功能的成人皮肤。目前的单克隆和人类抗体将用于评估胎儿和新生儿皮肤的存在已知抗原，以确定序列和进展胎儿的发育。