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Critical evaluation of the anti-complement immunofluorescence (ACIF) test for Epstein-Barr virus-associated nuclear antigen (EBNA) in known EBV-associated cell lines was carried out. The ACIF procedure, in which the cell smears were thoroughly air-dried and fixed with carbon tetrachloride (for 15 min at room termperature), provided reproducible staining results with minimum variations. The fixed smears coulb be stored for at least 5 weeks at minus 80 degrees C without loss of reactivity. On the basis of the above evaluation, a total of 59 human cell lines, which had unknown or uncertain association with EBV, were examined for the presence or absence of EBNA by the ACIF procedure. All of the cell lines tested could be judged positive or negative for EBNA. All of 21 lines growing as monolayer or in a mixed state of monolayer and suspension, were negative. Of 38 lines grown in suspension, 36 lymphoblastoid lines were positive but the remaining 2 lines (seemingly not lymphoblastoid) were negative.

By means of a semiquantitative antigenic analysis in the reaction of precipitation in agar with the use of a monospecific serum, obtained by the authors, dynamical changes have been studied in the content of the tissue-specific antigen in cells of subcutaneous connective tissue in rats: in the original and tripsinzed tissues, in 3, 6, 16-day and 1--2--3 month and malignized cultures, as well. It was found that at the 3, 11--13 day and then in 2--3 months of explantation there was a decrease in the content of the tissue-specific antigen, that is positively correlated with the degree of cell differentiation.