Moiseenko V M, Danilova A B, Danilov A O, Turkevich E A, Baldueva I A, Matsko D E
Vopr Onkol. 2008;54(3):303-14.
There is a great variety of histological patterns of skin melanoma and, in particular, that of its metastatic patterns. Malignant melanocytes are capable of influencing tumor-associated antigen expression. As of now, several varieties of melanoma-associated antigens (MAA) have been identified: MART1/melan A, tyrosinase, MITF, gp100, members of MAGE family, S100, CD63 and CD146. Peptides isolated from such molecules can induce MHC-restricted response of cytotoxic T-lymphocytes. It has been shown that level and nature of specific antigen expression caused by melanocytes correlate with tumor stage and a relationship between survival and MAA expression on tumor cells identified. Morphological features, growth pattern and proliferation rate varied in melanoma cell cultures used in our study. Our experiments involved evaluation of changes in the properties of antigens HLA (class 1 and 2), tumor tissue samples and cells isolated from them, which were capable of stable proliferative activity during passages 5, 10, 15, 20, 25, 30 and 35, and assay for MAA content. Levels of the antigens were significantly lower following long-term culturing melanoma cell melanoma cells in vitro. At initial passages (1-5), antigen profile in most cultures was similar to that in tumor tissue samples. Later on each cell population showed greater antigen expression heterogeneity matched by increased number of cells going through mitotic cycle; their nuclei were stained with antibodies to Ki-67. No HLF A/B/C molecule expression took place during tumor cell culturing: stained cells--in 68.9% of cultures (passages 1-5) and 36.3% (passage 35). However, HLA DQ/DP/DR molecule identification showed an inverse relationship: 44.1% (passage 5) while virtually all the cell lines did synthesize those molecules after passage 35. Hence, MAA and MHC (class 1 and 2) antigens expression in tumor cell should be monitored when they are used for preparation of autologuos and allogenic vaccines. In case of allogenic vaccine production, cell lines capable of stable production of MAA should be selected.
皮肤黑色素瘤有多种组织学模式，尤其是其转移模式。恶性黑色素细胞能够影响肿瘤相关抗原的表达。截至目前，已鉴定出多种黑色素瘤相关抗原（MAA）：MART1/黑色素A、酪氨酸酶、MITF、gp100、MAGE家族成员、S100、CD63和CD146。从此类分子中分离出的肽可诱导细胞毒性T淋巴细胞的MHC限制性反应。研究表明，黑色素细胞引起的特异性抗原表达水平和性质与肿瘤分期相关，且已确定肿瘤细胞上MAA表达与生存之间的关系。在我们的研究中，黑色素瘤细胞培养物的形态特征、生长模式和增殖率各不相同。我们的实验涉及评估HLA（1类和2类）抗原、肿瘤组织样本及其分离出的细胞的特性变化，这些细胞在第5、10、15、20、25、30和35代时具有稳定的增殖活性，并检测MAA含量。体外长期培养黑色素瘤细胞后，抗原水平显著降低。在最初几代（1 - 5代）时，大多数培养物中的抗原谱与肿瘤组织样本中的相似。后来，每个细胞群体显示出更大的抗原表达异质性，同时经历有丝分裂周期的细胞数量增加；它们的细胞核用抗Ki - 67抗体染色。在肿瘤细胞培养过程中未发生HLF A/B/C分子表达：染色细胞在68.9%的培养物中（第1 - 5代）和36.3%（第35代）出现。然而，HLA DQ/DP/DR分子的鉴定显示出相反的关系：44.1%（第5代），而实际上所有细胞系在第35代后都合成了这些分子。因此，当肿瘤细胞用于制备自体和异体疫苗时，应监测其MAA和MHC（1类和2类）抗原的表达。在生产异体疫苗的情况下，应选择能够稳定产生MAA的细胞系。
