Functional analysis of the immunosenescence of the human B cell system: dissociation of normal activation and proliferation from impaired terminal differentiation into IgM immunoglobulin-secreting cells.

The abilities of B cells from 24 young (mean 26 yr) and 24 elderly (mean 86 yr) humans to proliferate and differentiate into immunoglobulin-secreting cells (ISC) were investigated. Initial studies in young subjects demonstrated that a Staph protein A (SpA)-driven system could simultaneously assess the proliferative and differentiative capabilities of B cells resulting in IgM production. B cell proliferative responses were found to be partially T cell-dependent, whereas differentiation was absolutely T cell-dependent. Also, no significant differences could be detected in the abilities of nonproliferating allogeneic and autologous T cells to support B cell responsiveness. Although B cells from elderly subjects continuously exposed to SpA displayed proliferative responses equal to young subjects, the differentiation of B cells from elderly subjects into IgM ISC was markedly reduced as compared to young subjects. Analyses of results from co-culture experiments showed that the differentiation impairments of B cells from some elderly subjects could be partially corrected by allogeneic T cells from young subjects, whereas the impairments of others were more refractory. Moreover, T cells from elderly subjects were able to promote the differentiation of B cells from young subjects. Other experiments in elderly subjects showed that significant impairments of B and T cell functions rarely coexisted and that compensatory increases in B or T cell function were not evident. Thus, B cells from certain elderly humans have intrinsic impairments of differentiation required for optimal IgM production even though activation and proliferation remain normal in the presence of SpA. These impairments in differentiation are sometimes improved by T cells from young subjects, although in some elderly individuals, the differentiative impairments fail to be reversed.

Analysis of cellular heterogeneity in mouse thymus cultures.

Analysis of 5 to 6 d primary cultures of cells derived from murine thymus glands revealed a heterogeneous population of cells rather than "pure" reticuloepithelial cell cultures as was assumed previously by other investigators. The monolayer cultures consisted of at least three cell types: thymus epithelial cells, macrophagelike epithelioid cells, and fibroblasts. Surprisingly, about 50% of the cells had positive cytochemical staining reactions for acid phosphatase and nonspecific esterase. The sme cells phagocytized carbon particles, latex beads, and yeast. Furthermore, these cells could be removed from the initial cell suspension by phagocytosis of carbonyl iron, followed by magnetic separation, but once they had adhered to the substratum they were resistant to trypsin removal. All of these findings supported the conclusion that about 50% of the cells in the monolayers were macrophages. The other cells present were thymus epithelial cells and a small number of fibroblasts. Both of the latter types of cell were cytochemically negative, did not phagocytize particulate material, and were not removed by carbonyl iron treatment, but were removed by treating the monolayer with trypsin. The findings in this report indicated that epithelioid morphology alone was inadequate to identify correctly the cell types found in thymus cultures and that the use of such cultures as a model to study in vitro the maturation of certain immunological functions has been based on assumptions here shown to be incorrect.

Effects of vitamin B6 deficiency on thymic epithelial cells and T lymphocyte differentiation.

Dietary vitamin B6 (pyridoxine) deficiency in young Lewis rats results in a reduction of T lymphocyte numbers and defects of cellular immunocompetence. In vitro studies of thymic epithelial (TE) cells, responsible for inducing T lymphocyte differentiation, revealed that maintenance on a vitamin B6 deficient diet for 2 weeks resulted in a severe defect in TE cell function. When the deficient animals were returned to a normal diet, TE cell function was restored. Exposure of lymphoid precursors from neonatally thymectomized or vitamin B6-deficient donors to normal TE monolayers resulted in their conversion to functional T lymphocytes, as measured by their response in MLR and to mitogens. However, TE monolayers from vitamin B6-deficient animals were unable to effect such a maturation of T lymphocytes. Therefore, it is suggested that the defect in cellular immunocompetence following this dietary deficiency is due, at least in part, to the inability of TE cells to effect the differentiation of T lymphocyte precursors to functional T lymphocytes. The dietary deficiency does not, however, impair lymphoid precursors, which can be stimulated to further differentiation by exposure to normal TE cell monolayers.

In vivo effects of rabbit anti-mouse brain serum on theta bearing lymphocytes of AKR mice.

The in vivo effect of rabbit anti-AKR mouse brain-associated serum (RAMB) was determined on theta bearing lymphocytes present in the spleens and thymuses of mature C3H mice and AKR mice staged into preleukemic, leukaemic and overtly leukaemic states. Following seven daily injections of RAMB serum, the splenic plaque-forming cell (PFC) response to sheep erythrocytes (SRBC) and the percentage of theta-bearing lymphocytes in the spleen were significantly decreased in the C3H and the preleukaemic AKR mice. Decreases in thymic weight and thymocyte numbers were also apparent. Determinations of theta antigen density using in vitro cytoxicity tests indicated that splenic and thymic T lymphocytes (thymus-derived) remaining in the RAMB-treated C3H and preleukemic AKR mice consisted primarily of cells bearing less of the theta surface antigen. Histopathological studies of tissues from these two treated groups revealed cortical lymphocyte depletion in the thymus, and marginal and periarteriolar depletion in the spleen. Leukaemic AKR mice, administered seven injections of RAMB serum, demonstrated less dramatic changes in thymus weight, histopathology and theta-bearing cell percentages when compared with the data from the preleukaemic AKR or matureC3H mice. The results from testing overtly luekaemic AKR mice administered RAMB serum for the 7 or 9 days did not demonstrate differences from findings from groupsof overtly leukaemic control mice. These data indicate that the in vivo activity of RAMB serum in C3H mice and preleukaemic AKR mice is directed primarily toward the less mature T-lymphocyte population. This influence of RAMB serum is lesspronounced in leukaemic and the more overtly leukaemic mice, suggesting that a decreasedpopulation of RAMB-susceptible lymphocytes are present in these animals.

Induction of T-cells differentiation in vitro by thymus epithelial cells.

Thymus-reticular epithelial cells (TE-cells) were grown in a cell culture devoid of any lymphocytic elements. These cells were able to induce T-cell differentiation in spleen cells from T-dificient mice as expressed by con-A responsiveness and GvH reactivity. It was also shown that xenogeneic rat TE cells were as effective in the induction of T-cell differentiation in vitro as syngeneic TE cells. This system is therefore ideal for the study of T-cell development.