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Isolation and Localization of RNA Fractions Able to Transfer Tumor-specific Delayed Hypersensitivity in Vitro¹

Department of Microbiology, The University of Texas Health Science Center, San Antonio, Texas 78284

RNA fractions were prepared by sucrose density gradient centrifugation from hot-cold phenol, RNA-rich extracts of lymphoid tissues from strain 2 guinea pigs hyperimmunized to line 1 or line 10 tumors. Each RNA fraction was assessed for its ability to convert nonsensitized strain 2 peritoneal exudate cells to a state of specific sensitivity for line 1- or line 10-solubilized tumor antigens. An RNA fraction residing between the 4 S and 18 S peaks, designated as Fraction "B," transferred line 10 or line 1 sensitivity in 12 experiments. Twelve additional RNA extracts containing 2 subfractions prepared from RNA fraction B, designated as B₁ and B₂, also transferred line 1 or line 10 sensitivity in 14 experiments. Except for 3 experiments where the 4 S or 18 S material transferred tumor-specific sensitivity, RNA fractions corresponding to approximately 4 S, 18 S, 22 S, and 28 S were unable to transfer tumor-specific sensitivity to nonsensitized peritoneal exudate cells. Treatment of fraction B with RNase results in complete loss of ability to transfer immunobiological activity.

¹ This investigation was supported by NIH Contract 72-3205 and in part by Grant 1 ROI CA 17958-01A1 from the National Cancer Institute. Presented in part at the Annual Meeting of the American Association of Cancer Research in Houston, Texas, 1974 (14).

Received 5/21/76. Accepted 8/27/76.