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Fractionation of Lymphocytes Involved in the Generation of Cell-Mediated Cytotoxicity over Insolubilized Conjugated Histamine Columns¹

From the Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, and the Division of Clinical Pharmacology, Department of Medicine and Pharmacology, the Cardiovascular Research Institute, University of California Medical Center, San Francisco, California 94143

Abstract

Spleen cells from normal BALB/c mice were cultured in vitro with irradiated C57BL/6 stimulating cells. Five days later the T cell-mediated cytotoxic activity of the effector cells was assessed with a ⁵¹Cr-release assay that used H-2^bEL-4 tumor cells as targets. Before the BALB/c responding lymphocytes were sensitized they were fractionated by passing the spleen cells over insolubilized histamine rabbit serum albumin Sepharose columns (H-RSA-S) or over rabbit serum albumin Sepharose (RSA-S) control columns. Fractionation of cells over the H-RSA-S columns depleted or significantly reduced the cytotoxic potential of the unretained cells. All cytotoxic potential was recovered when the cells that adhered to the H-RSA-S were eluted from the columns. In contrast, no effect on responsiveness was detected after the cells had been fractionated over the control column. The loss of response potential by the cells that did not adhere to H-RSA-S could not be accounted for by removal of macrophages nor by the concentration of cells with suppressor activity in the effluent. These cell fractionation studies raise the possibility but do not prove that cytotoxic precursor cells may express amine receptors that could be responsible for their retention by insolubilized histamine columns.

Footnotes

¹ This work was supported in part by National Institutes of Health Grants HL-15851 and GM-16496.

² Present address: Department of Surgical Sciences, Clinical Research Center, Watford Road, Harrow, Middlesex, England.

³ Present address: Department of Hormone Research, the Weizmann Institute of Science, Rehovot, Israel.