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Selective Impairment of Cell Antigenicity by Fixation¹

From the Departments of Tumor Biology, and Immunology, Karolinska Institute, School of Medicine, and the Department of Immunology, National Bacteriological Laboratory, Stockholm, Sweden

Abstract

The effects of fixation by formaldehyde, p-formaldehyde, and glutaraldehyde on H-2^a and Moloney leukemia virus (MLV)-associated antigens were evaluated. Two cell types were employed: a) a MLV-induced lymphoma, YAC, which is passaged in ascites form in A/Sn mice or grown in long-term culture, and b) MLV-infected A/Sn and BALB/c mouse fibroblast monolayers. Antigenicity of the suspension cells was assessed by conventional absorbtions of antisera of predetermined titers to H-2^a and MLV, and subsequent titration of these absorbed antisera by the ⁵¹Cr release cytotoxicity test against pertinent target cells. Antigenicity of fibroblast monolayers was assessed directly by a mixed hemadsorption technique. At low concentrations and short fixation periods, p-formaldehyde selectively impaired MLV antigens, preserving H-2^a. At high concentrations and longer fixation periods, p-formaldehyde markedly reduced H-2^a antigenicity while preserving MLV antigens. Fixation of either suspension cells or monolayers with glutaraldehyde (0.25% for 5 min at 20°C) markedly impaired both types of antigenicity.

Footnotes

¹ This work was supported by the Swedish Cancer Society, the Leukemia Society of America, and Contract No 1 CP 33316 within the Virus Cancer Program of the National Cancer Institute.

² R. A. G. is the recipient of a United States Public Health Service Research Career Development Award.

³ Please send reprint requests to: Prof. A. Fagraeus, National Bacteriological Laboratory, 105 21 Stockholm, Sweden.