Detrick-Hooks B, Smith H G, Bast R C, Dunkel V C, Borsos T
J Immunol. 1976 May;116(5):1324-31.
Naturally soluble tumor antigens were detected in the ascites fluid of guinea pigs bearing an ascites tumor and from exhausted tissue culture media of cultured tumor cells. Two antigenically distinct cell lines of diethylnitrosamine-induced strain-2 guinea pig hepatomas (line-10 and line-1) served as the source of tumor antigens. Tumor antigen activity was detected by four different techniques: immunodiffusion, inhibition of complement-mediated cytotoxicity, inhibition of membrane immunofluorescence, and delayed cutaneous hypersensitivity. With syngeneic tumor-specific antiserum, line-10 guinea pig tumor antigens were detected by immunofluorescence in the concentrated ascites and tissue culture fluids. With a xenogenic antiserum, demonstrated to be tumor specific, line-10 tumor antigens were detected not only in the concentrated ascites and tissue culture fluids but also in two of the partially purified fractions of these fluids. When the line-10 concentrated ascites and its fraction I were subjected to ultracentrifugation at 300,000 x G for 1 hr, the antigen activity was retained in the supernatant and thus by this criterion the tumor antigens detected in these samples are soluble. Immunodiffusion data indicate that more than one antigen is present in the line-10 system since three lines of precipitation were detected when line-10 concentrated ascites was reacted with the line-10 tumor-specific antiserum. In contrast to this, the line-10-concentrated tissue culture fluid displayed only one line of precipitation. Although tumor antigens could not be demonstrated in the other antigenically distinct tumor cell line, line-1, by immunodiffusion or inhibition of membrane immunofluorescence, inhibition of complement-mediated cytotoxicity was able to detect tumor antigens in the line-1 concentrated ascites and tissue culture fluids.
在携带腹水瘤的豚鼠腹水中以及培养的肿瘤细胞耗尽的组织培养基中检测到天然可溶性肿瘤抗原。二乙基亚硝胺诱导的2型豚鼠肝癌的两种抗原性不同的细胞系(10号线和1号线)用作肿瘤抗原的来源。通过四种不同技术检测肿瘤抗原活性:免疫扩散、补体介导的细胞毒性抑制、膜免疫荧光抑制和迟发型皮肤超敏反应。用同基因肿瘤特异性抗血清,通过免疫荧光在浓缩腹水和组织培养液中检测到10号线豚鼠肿瘤抗原。用一种已证明为肿瘤特异性的异种抗血清,不仅在浓缩腹水和组织培养液中检测到10号线肿瘤抗原,而且在这些液体的两个部分纯化级分中也检测到。当10号线浓缩腹水及其I级分在300,000×G下超速离心1小时时,抗原活性保留在上清液中,因此根据这一标准,在这些样品中检测到的肿瘤抗原是可溶的。免疫扩散数据表明,10号线系统中存在不止一种抗原,因为当10号线浓缩腹水与10号线肿瘤特异性抗血清反应时,检测到三条沉淀线。与此形成对比的是,10号线浓缩组织培养液仅显示一条沉淀线。尽管通过免疫扩散或膜免疫荧光抑制在另一种抗原性不同的肿瘤细胞系1号线中未证明有肿瘤抗原,但补体介导的细胞毒性抑制能够在1号线浓缩腹水和组织培养液中检测到肿瘤抗原。