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In Vitro Studies of the Genetically Determined Unresponsiveness to Thymus-Independent Antigens in CBA/N Mice¹

From the Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20014 and the Department of Clinical and Experimental Immunology, Naval Medical Research Institute, National Naval Medical Center, Bethesda, Maryland 20014

Abstract

The X-chromosome-linked B lymphocyte defect of CBA/N mice has been studied in vitro by comparing the ability of (CBA/N x DBA/2)F₁ (X^-/X^- x X⁺/Y) male (X^-/Y) and female (X^-/X⁺) spleen cells to respond to the thymus-independent antigen DNP (or TNP)-AECM-Ficoll. (CBA/N x DBA/2)F₁ male spleen cells failed to generate significant in vitro anti-TNP antibody responses to DNP- or TNP-AECM-Ficoll, in contrast to spleen cells from F₁ female (X^-/X⁺) mice which responded normally to these T-independent antigens. Spleen cells from male F₁ mice responded almost as well as F₁ female cells to the thymus-dependent antigen, TNP-sheep red blood cells (TNP-SRBC) in vitro. Adding F₁ male cells to F₁ female cells failed to reduce the response of the latter to DNP-AECM-Ficoll, suggesting that the inability of F₁ male cells to respond was not due to active suppression. The response of F₁ male spleen cells to TNP-SRBC was not impaired by adding high concentrations of TNP-AECM-Ficoll indicating that the mechanism of unresponsiveness was not tolerance induction in all TNP-specific precursors. Lymphocytes from F₁ male mice were capable of forming anti-TNP antibody after stimulation with lipopolysaccharide (LPS) in high concentrations; DNP-AECM-Ficoll had no effect no this polyclonal response.

B lymphocytes from mice bearing only the X-chromosome of the CBA/N strain thus display a profound defect in B cell activation. This functional defect may represent either an inability of the defective B cells to be activated by thymus-independent antigens or the absence of a sub-class of B cells which respond to thymus-independent antigens.

Footnotes

¹ This work was supported in part by the Bureau of Medicine and Surgery Work Unit No. MR 041.02.01.0020B2GI and CICC 3-06-132. The opinions or assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large. The animals used in this study were handled in accordance with the provisions of Public Law 89-54 as amended by Public Law 91-579, the "Animal Welfare Act of 1970" and the principles outlined in the "Guide for the Care and Use of Laboratory Animals," U. S. Department of Health, Education and Welfare Publication No. (NIH) 73-23.

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