In Vitro Immune Response to the 2,4,6-Trinitrophenyl Determinant in Aged C57BL/6J Mice: Changes in the Humoral Immune Response to, Avidity for the TNP Determinant and Responsiveness to LPS Effect with Aging

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Abstract
An in vitro anti-TNP response of the spleen cells from aged C57BL/6J mice showed approximately 4-fold less PFC than did that from young adult mice. Anti-θ serum-treated young spleen cells gave an anti-TNP response that was definitely greater than the response of the anti-θ serum-treated aged spleen cells in the presence of the exogenous activated thymus cells as helper cells. These results suggested that the deficits in B cells may be partly responsible for the imparied anti-TNP response of the aged spleen cells. To examine further the capacity of stem cells in the bone marrow to generate B cells responsible for anti-TNP response in the spleen, we injected i.v. 1.5 to 2.0 × 107 bone marrow cells from young or aged mice into lethally irradiated syngeneic recipients that had previously been thymectomized. Four to 6 weeks later, 107 spleen cells from the two groups of these recipient mice were immunized with TNP-SRBC in the presence of the exogenous activated thymus cells and assayed for anti-TNP PFC. The response of the aged marrow-derived B cells was approximately one-half of that of the young marrow-derived B cells. The avidity for TNP determinant of the antibodies produced by the PFC was determined by the plaque-inhibition technique. The avidity of the antibodies produced by the aged mice was approximately 33 times lower than that by the young mice. Anti-TNP response of the young spleen cells were markedly enhanced by the addition of LPS to the cultures, whereas no or little enhancement of the response was induced in the aged spleen cells even in the presence of high concentration of LPS. In contrast, DNA synthesis of both the young and aged spleen cells was comparably stimulated by 1 µg/ml and 10 µg/ml of LPS, however, it was rather less in the aged spleen cells at a concentration of 100 µg/ml. Mechanisms responsible for the changes in avidity and responsiveness to LPS with aging are discussed.

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