Tomohide Hosokawa

Dietary fat alters the fatty acid composition of lymphocyte membranes and the rate at which suppressor capacity is lost

SJL/J mice were fed from conception two nutritionally adequate semi-purified diets that differed only in polyunsaturated to saturated fatty acid content. The effect of diet fat on the fatty acid composition of membranes from spleen and thymus cells was determined. Diet fat was found to significantly alter the fatty acid composition of lymphocyte membrane phosphatidylcholine and phosphatidylethanolamine. Diet also altered the degree of resistance against tolerance-induction.

Effect of food-restriction stress on immune response in mice

Daily 23-h food deprivation for 1-5 days induced gastric ulcers and atrophic changes of the spleen and thymus, accompanied by a rise in plasma cortisol and catecholamine levels in mice. It also modulated several immune cell functions in the spleen including a drop in the B cell population but no change in the mitogen response of the B cells, an increase in T cell population but no change in the L3T4/Lyt2 ratio and an early increase in natural killer activity and O2- production by macrophages. These effects are thought to correlate to the increase in stress-associated humoral factors and this may partly result from stress induced by food restriction.

Age-related enhancement of tumor necrosis factor (TNF) production in mice

We investigated age-related changes in the production of TNF at the cellular level using immunocompetent peritoneal and spleen cells from C3H/He mice of various ages. The density of cultured peritoneal macrophages and spleen cells required for TNF production was at least 5 x 10(5) cells/dish. The optimal concentration of OK-432 for 24-h culture of peritoneal macrophages (1 x 10(6) cells) and spleen cells (1 x 10(7) cells) was 0.5 and 0.1 KE/ml, respectively. Among peritoneal cells, adherent macrophages were the major TNF-producing cells, whilst nonadherent T or B cells alone did not produce TNF after stimulation with OK-432. In the case of spleen cells, T or B cells were involved in the production of TNF when cultured with a few adherent cells in the presence of OK-432. However, T or B cells alone failed to produce TNF. Production of TNF by peritoneal macrophages from both male and female mice increased significantly with aging. In contrast, although TNF production by spleen cells tended to increase with aging, no significant change was noted. The total number of peritoneal and spleen cells, respectively increased up to about 18 months after birth with B cells being principally responsible for this age-related increase. We previously reported that systemic production of TNF increases with aging. The present study of TNF production at the cellular level in mice indicated (1) that TNF production per macrophage increased with aging, and (2) that the number of T and B cells involved in the production of TNF in the presence of macrophages also increased at least up to middle age.

Age-related decline in humoral immunity caused by the selective loss of TH cells and decline in cellular immunity caused by the impaired migration of inflammatory cells without a loss of TDTH cells in SAMP1 mice

We investigated the cellular basis of the age-related decline in antibody (Ab) and delayed-type hypersensitivity (DTH) responses to sheep red blood cells (SRBC) in vivo in short-lived senescence-accelerated mouse (SAM) P1. In SAMP1 mice, age-related decreases in CD4+ T cells in the peripheral blood occurred earlier than in control mice and occurred in parallel with the age-related decline in Ab and DTH responses. In addition, the involution of the thymus was faster. The injection of thymic T cells from young mice before sensitization completely restored the Ab responses in aged SAMP1 mice. These data suggest that the age-related decline in Ab response is due to the age-related early loss of helper-T (TH) cells. On the other hand, the local transfer of spleen cells from sensitized aged donors into the footpads of naive syngeneic recipients evoked strong DTH responses, demonstrating the existence of DTH-mediating T (TDTH) cells in the spleens of aged SAMP1 mice. Moreover, the local injection of naive spleen cells from young donors, together with the antigen, into the footpads caused DTH responses in sensitized aged recipients. These findings indicate that TDTH cells were induced and were able to migrate and function as effector cells in aged mice. When naive spleen cells from aged donors were injected locally into the footpad, they restored the DTH response in aged mice, but this effect did not work if the cells were injected intravenously. This demonstrates that the inflammatory cells of the aged mice were able to work at the local site, but could not migrate there. The intravenous injection of naive spleen cells from young donors restored the DTH response in aged mice, suggesting that the endothelial cells of aged mice were not impaired and permitted the inflammatory cells to migrate into the extravascular tissues. Thus, although the age-related decline of the Ab and DTH responses occur in parallel, we found different effects of aging on TH and TDTH cells in SAMP1 mice. Furthermore, our data suggest that the reason for the low DTH response in aged SAMP1 mice is not the loss of TDTH cells, but rather the impaired migration of inflammatory cells into the local site.

Immune abnormality in relation to nonimmune diseases in sam mice

A series of related strains of senescence-accelerated mouse (SAM) shows strain-unique age-related diseases, such as amyloidosis, deficit in learning and memory, osteoporosis, and brain atrophy, while many of these disease-prone mouse (SAMP) strains have impaired immune activity as young adults, and have a short life span, probably not due directly to the diseases. Because the mean life span was prolonged and the time of the disease onset was delayed by a low-calorie dietary condition or a specific pathogen-free environment, both of which ameliorate the impaired immune activity, the enhancement of immune activity may help decrease the deteriorative process of aging, to that seen in ordinary strains of mice. Studies using the SAMP model may help elucidate the role of immunity in the aging process. Herein, we review the cellular and genetic basis of the immune abnormality in SAMP mice, then discuss the relationship between immune abnormality and development of the age-related disease, senile amyloidosis, findings obtained on SAMP hybrid mice and congenic mice for disease-related genes. Activation of the gene(s) for senile amyloid per se shortened the life span, and the early development of the immune dysfunction primarily seems to be both genetically and physiologically independent of amyloidosis, although the disease may be indirectly modified in the aged with depressed immune activity.

The effect of taurine on age-related immune decline in mice: the effect of taurine on T cell and B cell proliferative response under costimulation with ionomycin and phorbol myristate acetate.

Proliferative responses to the costimulation with phorbol-12-myristate-13-acetate (PMA) and suboptimal doses of ionomycin in the purified T and B cells from old mice were lower than those from young mice. The degree of the age-related decline was more significant in T cells than in B cells. Taurine, a sulfur containing amino acid, augmented the proliferative responses of T cells from both young and old mice. The augmentation of the proliferative response by taurine was more marked in old T cells than in young ones. The concentration of intracellular free calcium ion ([Ca2+]i) was significantly lower in the old T cells under the stimulation with PMA and ionomycin than that in the young ones. In the presence of taurine, the concentration of [Ca2+]i in the old T cell significantly increased under the stimulation. The results indicate that taurine improved the proliferative response of old T cells by the restoration of the increment of the concentration of [Ca2+]i under the stimulation.

Increased Superoxide Anion Production and Glutathione Peroxidase Activity in Peritoneal Macrophages from Autoimmune-Prone MRL/Mp-lpr/lpr Mice

We studied the release of superoxide anion (O-2) in peritoneal macrophages from autoimmuneprone MRL/Mp-lpr/lpr (MRL/l) mice. Compared to resident peritoneal macrophages from control MRL/Mp-+/+ (MRL/n) mice, macrophages from MRL/l mice exhibited an age-related increase of spontaneous and PMA-induced O-2 secretion in association with the development of the autoimmune process. Analysis of the kinetic parameters of NADPH oxidase in macrophages revealed that MRL/l macrophages were in a primed state, as shown by the decreased Km value of the oxidase for NADPH. Furthermore, we studied several key enzymes for their ability to scavenge the oxygen radicals in the macrophages. Among the enzymes studied, only glutathione peroxidase (GSH Px) activity was increased in peritoneal macrophages from MRL/l mice and this change was closely correlated with the increase in O-2 production. Thus, GSH Px activity in macrophages seems to play an important role in macrophage functions under increased oxidative stress.

Neonatal tolerance induction in the thymus to MHC-class II-associated antigens. I. Preferential induction of tolerance to mls antigens and resistance to allo-MHC antigens

Neonatal tolerance inducibility of self-major histocompatibility complex (MHC)-class II-associated antigens was compared with that of allo-class II antigens. BALB/c (H-2d, Mlsb) mice, less than 24 hr after birth, were intravenously injected with bone marrow cells of either (BALB/c X DBA/2)F1 (H-2d, Mlsb/a, semiallogeneic at the Mls locus) or (BALB/c X B10.BR)F1 (H-2d/k, Mlsb; semiallogeneic at the MHC), as antigens. The mice were tested for in vivo immune activity of class II-reactive T cells by means of the popliteal lymph node-swelling assay. They developed tolerance, irrespective of type of antigens, showing profoundly suppressed host-versus-graft reaction, and those tolerized to the allo-MHC antigens accepted skin grafts of the corresponding allogeneic mice. In the thymus and spleen of the Mls-tolerant mice, antigen-specific class II-reactive T-cell activity was completely abolished, without the apparent involvement of suppressor cells. In contrast, the activity in allo-MHC-tolerant mice was not reduced in either thymus or peripheral lymphoid organs, suggesting that systemic hyporesponsiveness is attributable to reversible suppression of immune competent cells. The resistance for cell-level tolerance induction to allo-class II antigens may not be ascribed to the active participation of allo-MHC antigens in prevention of or in escape from tolerance induction or both, since an injection of bone marrow cells of both Mls and H-2-semiallogeneic (DBA/2 X B10.BR)F1 (H-2d/k, Mlsa/b) mice could induce tolerance to Mlsa-H-2d antigens in newborn thymus cells.

The Role of Oxygen Radicals in the Pathogenesis of Gastric Mucosal Lesions Induced in Mice by Feeding-Restriction Stress

To investigate the role of oxygen radicals in the pathogenesis of gastric mucosal lesions induced by psychological stress, we exposed 3-mo-old C3H mice to feeding-restriction stress for 1 to 5 days. Serial changes in the activities of antioxidant enzymes in the gastric mucosa, together with O2- production by macrophages, were measured. The stress increased the plasma cortisol level and started to produce acute gastric lesions (AGL) on the 2nd day. Before the development of AGL, the activity of superoxide dismutase in the gastric mucosa had already decreased and the ability of O2- production by macrophages was enhanced from the 1st day. This suggests that oxygen radicals play some role in the development of AGL induced by the stress.

Ontogeny of macrophage function. I. Phagocytic activity and A-cell activity of newborn and adult mouse peritoneal macrophages.

Phagocytic activity and immune-participating A-cell activity of newborn and adult mouse macrophages were investigated under in vitro conditions. Thioglycollate medium-stimulated newborn (SNB) or adult (SA), and nonstimulated adult (NA) peritoneal macrophages were used. Immune complexes of sheep erythrocytes (SRBC), aldehyde-fixed SRBC, and latex beads were employed in phagocytosis tests. A-cell activity was estimated as the capacity to support primary or secondary responses of macrophage-deprived adult spleen cells to SRBC. Results obtained were: 1) phagocytic activity of NA macrophages was highest and that of SNB macrophages was higher than SA macrophages, 2) no difference was observed in A-cell activity for secondary response among SNB, SA, and NA macrophages, and 3) SNB macrophages were lacking in A-cell activity for primary response. These results suggest that the substantial A-cell function should be evaluated in primary responses, and that the A-cell and pahgocytic functions do not necessarily accompany each other.