Anichini A, Mortarini R, Fossati G, Parmiani G
Int J Cancer. 1986 Oct 15;38(4):505-11. doi: 10.1002/ijc.2910380409.
Fifty-six tumor clones isolated by cloning in soft agar from early cultures (before the 10th in vitro passage) of two different human metastatic melanomas (Me9229 and Me28) were characterized by FACS analysis for surface expression of class-I and class-II HLA antigens and of melanoma-associated antigens (MAA) with a panel of 15 monoclonal antibodies (MAbs). A marked phenotypic heterogeneity involving MAA and/or HLA markers was observed among the clones derived from both tumors. The differences among the tumor clones and between them and the uncloned melanoma were qualitative and quantitative for each antigen considered. Clones derived from Me9229 expressed the same HLA profile as the parental culture (class I+, class II-) while strong heterogeneity was observed for MAA expression. Clones from Me28 presented a marked heterogeneity for class-I and class-II HLA antigens but were more homogeneous for MAA. The phenotype of the clones was repeatedly checked over the first month in culture and found to remain generally unchanged and not linked to the cell cycle. However, major changes in antigenic expression of the clones could be observed upon treatment with recombinant interferon-gamma (rIFN-gamma): class-I and -II HLA antigens could be induced or augmented while a moderate inhibition was seen on MAA expression. Furthermore, an apparent hierarchy in expression and/or induction of class-II antigens by rIFN-gamma was observed among the tumor clones. DR antigens were more frequently expressed (Me28 clones) and upon treatment with rIFN-gamma reached higher levels than DP and DQ products. Taken together these results indicate that antigenic heterogeneity for MAA and HLA antigens can be detected in cells isolated from early cultures of human metastatic melanomas and suggest that the original uncloned tumor might be considered as a complex mixed population made up of a number of neoplastic cells each expressing a distinct phenotype which can be modulated by lymphokines such as IFN-gamma.
通过软琼脂克隆从两种不同的人类转移性黑色素瘤（Me9229和Me28）的早期培养物（体外传代第10代之前）中分离出56个肿瘤克隆，用一组15种单克隆抗体（MAb）通过流式细胞术分析其I类和II类HLA抗原以及黑色素瘤相关抗原（MAA）的表面表达。在源自这两种肿瘤的克隆中观察到涉及MAA和/或HLA标志物的明显表型异质性。对于所考虑的每种抗原，肿瘤克隆之间以及它们与未克隆的黑色素瘤之间的差异在定性和定量上都是存在的。源自Me9229的克隆表达与亲代培养物相同的HLA谱（I类阳性，II类阴性），而MAA表达则观察到强烈的异质性。来自Me28的克隆在I类和II类HLA抗原方面表现出明显的异质性，但在MAA方面更为均一。在培养的第一个月内对克隆的表型进行了反复检查，发现其总体上保持不变，且与细胞周期无关。然而，在用重组干扰素-γ（rIFN-γ）处理后，可以观察到克隆抗原表达的主要变化：I类和II类HLA抗原可以被诱导或增强，而MAA表达则受到适度抑制。此外，在肿瘤克隆中观察到rIFN-γ对II类抗原表达和/或诱导存在明显的等级差异。DR抗原更频繁地表达（Me28克隆），在用rIFN-γ处理后达到的水平高于DP和DQ产物。综上所述，这些结果表明，在从人类转移性黑色素瘤早期培养物中分离的细胞中可以检测到MAA和HLA抗原的抗原异质性，并表明原始的未克隆肿瘤可能被视为由许多肿瘤细胞组成的复杂混合群体，每个肿瘤细胞都表达一种独特的表型，这种表型可以被诸如IFN-γ等淋巴因子调节。
