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Enhancement of Immune Response Potential of Mouse Lymphoid Cells Fractionated over Insolubilized Conjugated Histamine Columns¹

Department of Chemical Immunology, The Weizmann Institute of Science, Rehovot, Israel, Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, and Division of Clinical Pharmacology, Departments of Medicine and Pharmacology, University of California Medical Center, San Francisco, California 94122

Abstract

Spleen cells from (BALB/c C57BL/6)F₁ mice which had been immunized 7 days earlier with SRBC were either fractionated over columns of histamine-rabbit serum albumin-Sepharose beads or were untreated. Equal numbers of filtered and unfiltered cells were injected into irradiated, syngeneic recipients with SRBC. The kinetics of direct and indirect PFC generated by the two groups of transferred cells were similar. However, 4.5 more direct and 5.8 more indirect PFC were detected per unit number of spleen cells injected in the recipients of the filtered than in those of the unfiltered cells. In addition to enhanced PFC responses, spleen cell fractionation over the histamine-coated beads also resulted in slight but significant increases in the numbers of nucleated cells per recipient spleen. These findings indicate that removal of cells adherent to histamine-coated beads enhances the humoral immune response potential of mouse spleen cells. A similar result was obtained when histamine column-fractionated thymocytes were injected together with unfractionated bone marrow cells. In contrast, no enhancing effect was observed when fractionated marrow cells were mixed with unseparated thymocytes. Fractionation of spleen cells over poly-L-lysine-Sepharose columns, which bind mouse spleen cells and are similar in charge to histamine-rabbit serum albumin-Sepharose, had no detectable immunologic effect on transferred spleen cells. The possibility that the columns of insolubilized conjugated histamine retain thymus-derived lymphocytes which express amine receptors and function to regulate humoral immunity is considered.

Footnotes

¹ This research was supported by Agreement 06-035 with the National Institutes of Health, United States Public Health Service, and United States Public Health Service Grants HL 09964 and GM 16496.

² Present address: Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014.

³ Present address: Division of Clinical Pharmacology, Departments of Medicine and Pharmacology, University of California Medical Center, San Francisco, California 94122.

⁴ Part of this work was performed during tenure of a special fellowship of the U. S. Public Health Service 1 FO, 3HL50244-01, while KLM was on sabbatical leave from the Division of Clinical Pharmacology, Departments of Medicine and Pharmacology, University of California Medical Center, San Francisco, California 94122.