Forty-six cultured cell lines of diverse human tumor origins, including 25 melanoma cell lines, were HLA allotyped with the use of a modified eosin complement-dependent cytotoxicity test in combination with absorption and two-color fluorescence techniques. In 10 cases (1 renal cell carcinoma line and 9 melanoma cell lines), the cell line donors had been HLA typed a few years before the cell line-typing project had started and in 13 cases (1 renal cell carcinoma line and 12 melanoma cell lines), the cell line donors were currently available for comparative typing of lymphocytes. HLA-typing results suggested that most cell lines expressed genetically appropriate HLA antigens, although 1 cell line had more than two HLA antigens for one HLA locus and 2 cell lines lacked expression of one or more HLA antigens in comparison with donor typing. One hepatoma cell line, 1 of 2 of the bladder carcinoma cell lines tested, and 17 of the 25 melanoma cell lines expressed DR alloantigens in addition to their HLA-A, B, and C locus antigens. For 9 of the melanoma cell lines, comparisons with donor DR alloantigens could be made, and all these cell lines had exactly the same DR allospecificities as those found on donor B-lymphocytes. HLA typing of cell lines can be used as an adjunct to polymorphic isoenzyme marker tests to verify their patient source and lack of contamination by another cell line, and HLA typing should be used to determine the antigen composition of cells used in the preparation of reagents for immunotherapy or in studies of tumor-specific immunity.
HLA typing of amniotic fluid cells has been used for the prenatal diagnosis of the HLA linked diseases congenital adrenal hyperplasia (21-OH-deficiency (21-OH-def) type) and complement C4 deficiency and it has also been used for the prenatal determination of paternity. There are, however, technical difficulties in this test associated with the weak expression of some B locus antigens on amniotic fluid cells, and theoretical difficulties related to associations between particular HLA antigens and the 21-OH-def allele. Since certain HLA-B locus antigens are found in significantly increased frequencies among patients with 21-OH-def, there is a relatively high incidence of HLA-B homozygosity among the patients and over 40 per cent of the parents of these patients share one or more HLA-B locus antigens. Results of some prenatal HLA typing tests may thus be difficult to interpret, and supplementary tests should be used whenever possible. HLA typing of amniotic cells is, however, the only available procedure for prenatal diagnosis of C4 deficiency and it is the best available procedure for prenatal determination of paternity. A modification of our original procedure allows HLA typing to be performed with increased numbers of HLA typing sera, and sera with optimum reactivity for amniotic fluid cells have now been selected for the definition of most of the more commonly expressed HLA antigens. Although amniotic fluid cells do not express DR Antigens, amniotic fluid cells can be typed for the HLA-linked marker glyoxalase I (GLO) and this may be the informative for prenatal diagnosis in some cases.
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