Schreurs M W, de Boer A J, Schmidt A, Figdor C G, Adema G J
Department of Tumour Immunology, University Hospital Nijmegen St Radboud, The Netherlands.
Melanoma Res. 1997 Dec;7(6):463-70. doi: 10.1097/00008390-199712000-00004.
Melanocyte lineage-specific antigens, like gp100, have been shown to be recognized by tumour-infiltrating lymphocytes isolated from melanoma patients. Therefore these antigens might be used as targets for specific anti-melanoma immunotherapy. To investigate this potential in syngeneic mouse models, we cloned and characterized the murine homologue of gp100 from a B16 murine melanoma cDNA library. The isolated cDNA clone encodes a protein of 626 amino acids, sharing 79.7% sequence homology with human gp100. Expression of murine gp100 was restricted to cells of the melanocyte lineage, as determined by Northern and Western analysis. However, like human gp100, reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed low levels of transcription of the murine gp100 gene in all the cell lines and normal tissues tested. In contrast, murine tyrosinase, another melanocyte differentiation antigen, mimics human tyrosinase expression and did show absolute melanocyte lineage-specific expression. The characterization of murine gp100 will allow investigation of anti-gp100 immune responses in C57BL/6 mice using the syngeneic B16 tumour model and the possible adverse effects of such immunity on normal melanocytes.
黑色素细胞谱系特异性抗原，如gp100，已被证明可被从黑色素瘤患者体内分离出的肿瘤浸润淋巴细胞识别。因此，这些抗原可能用作特异性抗黑色素瘤免疫疗法的靶点。为了在同基因小鼠模型中研究这种潜力，我们从B16小鼠黑色素瘤cDNA文库中克隆并鉴定了gp100的小鼠同源物。分离出的cDNA克隆编码一个由626个氨基酸组成的蛋白质，与人类gp100的序列同源性为79.7%。通过Northern和Western分析确定，小鼠gp100的表达仅限于黑色素细胞谱系的细胞。然而，与人类gp100一样，逆转录-聚合酶链反应（RT-PCR）分析显示，在所测试的所有细胞系和正常组织中，小鼠gp100基因的转录水平较低。相比之下，另一种黑色素细胞分化抗原小鼠酪氨酸酶，模仿人类酪氨酸酶的表达，并且确实表现出绝对的黑色素细胞谱系特异性表达。小鼠gp100的特性鉴定将有助于利用同基因B16肿瘤模型研究C57BL/6小鼠中抗gp100免疫反应以及这种免疫对正常黑色素细胞可能产生的不利影响。
