Changed homing of hemopoietic precursor cells after long-term treatment with parathyroid hormone

Changes in the capacity of hemopoietic stromal microenvironment to promote homing of hemopoietic stem cells from different hierarchical compartments were evaluated in mice treated with parathyroid hormone by determining their 24-h precipitation factor. This parameter did not change for splenic short-living hemopoietic stem cells (splenic CFU) and considerably decreased for the bone marrow of mice treated with parathyroid hormone. For earlier long-living hemopoietic stem cells (cells forming the cobblestone area on day 28) the precipitation factor after injections of parathyroid hormone did not change in the bone marrow and decreased in the spleen. These data suggest that parathyroid hormone decreases the efficiency of homing of short-living hemopoietic stem cells in the bone marrow. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 142, No. 7, pp. 97–100, July, 2006     

Peculiarities of bone marrow hemopoiesis in early aging OXYS rats

Colony-forming activity of bone marrow cells in 3-and 12-month-old Wistar rats does not differ by the number of early and erythroid precursors and by the formation of granulocyte-macrophage colonies. In senescence-accelerated OXYS rats, the number of early and erythroid precursors significantly increases by the age of 12 months and surpasses the corresponding values in Wistar rats. The number of granulocyte-macrophage colonies in OXYS rats does not change with age, but the numbers of these colonies formed at the age of 3 and 12 months in these animals are higher than in Wistar rats. As a result, the total number of hemopoietic colonies in 12-month-old OXYS rats 2-fold surpassed that in 12-month-old Wistar rats. Activation of granulopoiesis and increased numbers of early and erythroid precursors indicate deep changes in the functional status of the hemopoietic stem cell in 1-year-old OXYS rats in the direction characteristic of aging animals. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 7, pp. 96–99, July, 2007     

Effect of amino acids and antibodies against nerve growth factor receptors on the development of organotypic culture of lymphoid tissue

We studied the effects of 20 L-amino acids on organotypic culture of splenic lymphoid tissue from 3-month-old rats were studied in the presence of apoptosis-inducing monoclonal antibodies against low-affinity receptors for nerve growth factor NGFRp75. The influence of amino acids stimulating cell proliferation in explants (lysine, asparagine, and glutamic acid) did not depend on NGFRp75. Hydrophobic amino acids inhibiting the growth zone in isolated application and abolished the inhibition of explant development in the presence of antibodies against NGFRp75. These amino acids can mediate the proapoptotic effect on lymphoid tissue via low-affinity receptors for nerve growth factor. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 2, pp. 218–221, February, 2007     

Generation of free oxygen radicals and clastogenesis in bone marrow cells in MRL/1 mice

The level of chemiluminescence induced by phorbol myristate acetate or opsonized zymosan in the bone marrow cells of 5–6-month-old male MRL/1 mice is significantly higher than that in the same cells of 1–2-month-old mice. The number of chromosome aberrations induced by the prooxidant mutagen dioxydine in bone marrow cellsin vivo is significantly higher in 5–6-month-old MRL/1 mice than in 1–2-month-old mice. It is suggested that these effects are more pronounced in 5–6-month-old animals due to progressive development of rheumatic pathology in this mouse strain. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 123, No. 1, pp. 22–25, January, 1997     

Artificial Aging of Mice

Clinical signs of aging verified by morphometrical analysis of brain tissue were observed in young mice 4 months after administration of brain extract from old mice (5 intraperitoneal injections).     

Opposite effects of antioxidants on anxiety in Wistar and OXYS rats

Passive behavior in the open field test and high anxiety in an elevated plus-maze develop in OXYS rats by the age of 3 months and are regarded as manifestations of early aging. We studied the possibility of preventing these disturbances with vitamin E and whortleberry extract (20 mg/kg for 45 days from the age of 1.5 months). Whortleberry extract alone increased horizontal activity and reduced anxiety of OXYS rats, while anxiety in Wistar rats increased significantly after treatment with both preparations, and especially with vitamin E: the number of entries into open arms of the elevated plus-maze and the time spent there decreased. The results necessitate comprehensive evaluation of the aftereffects of long-term use of antioxidants, acknowledged geroprotectors intended for preventive use. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 6, pp. 685–688, June, 2006     

Toxic effects of d-galactose on thymus and spleen that resemble aging

Continuous low-dose injection of d-galactose induces changes in mice that resemble accelerated aging. As such, these mice have been used as models to study mechanisms of aging. Here, we examined whether repeated (daily, for 60 days) subcutaneous injections (at 50?mg d-galactose/kg) into young adult (i.e., 2-month-old) mice induced changes in key immune system organs that were on par with those associated with aging. The results showed that galactose-treated mice develop histologic changes in their thymic cortical and medullary regions; immunohistochemical analysis revealed unorganized distributions of keratin-5 and keratin-8 proteins in the thymus of these hosts. These histological changes in the thymus of d-galactose-treated mice were also observed in the organs of aged (i.e., 24-month-old control mice); however, in this latter group, these changes were accompanied by a strong infiltration of adipose cells. Galactose-treated mice also evinced alterations within their splenic white and red pulp. Further, ultrastructural analyses of the thymus and spleen of the treated mice revealed increases in irregularly shaped lymphocytes bearing visible pyknosis. It was also seen that levels of autophagy within thymic epithelial cells were greatly decreased in the tissues of the galactose-treated mice, an outcome also seen in aged mice. Lastly, the level of memory T-lymphocytes and percentage of IgM-B220-B-lymphocytes in spleens of the galactose-treated mice were both increased (albeit insignificantly so) relative to values among splenocytes of age-matched control; however, these levels were not clealy as elevated as would be expected in “elderly” mice. Taken together, our results strongly suggest that d-galactose treatment can induce structural changes in the thymus and spleen, and some changes in organ-associated cell phenotypes, that are similar to several effects seen with aging. However, the fact that many endpoints do not appear to be truly reflective of what should be seen in immune system organs/cells of “elderly” mice now calls into question the appropriateness of the use of d-galactose (i.e., is it histologically/immunotoxicologically-proper?) to create age-mimicry in mice.     

Delayed effects of long-term administration of granulocyte colony-stimulating factor to mice

We studied the effects of chronic administration of granulocyte colony-stimulating factor in nonmobilizing doses to mice. Over 18 months of the study, 55% animals of the treatment group died of unknown cause, blood diseases and tumors were found in 20% mice, and in 5% animals pathological changes were absent. Control mice had no diseases (normal values of total and differential leukocyte count). The diagnoses made over the first 7 months mainly included myeloproliferative diseases. Solid tumors were found at later terms. Suppurative inflammation at the site of injection was observed in all mice after 3-month treatment with granulocyte colony-stimulating factor. Our results indicate that chronic administration of granulocyte colony-stimulating factor in low doses leads to the development of etiologically different tumors and sharply reduced animal life span. The use of granulocyte colony-stimulating factor during allogeneic transplantation of hemopoietic stem cells can be hazardous for donors. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 568–573, May, 2008     

Progressive Accumulation of Bacterial Lipopolysaccharide In Vivo During Murine Acute Graft-Versus-Host Disease

In a previous report the authors demonstrated that acute graft-versus-host disease (GVHD) was associated with pathologic amounts of tumour necrosis factor alpha (TNF-α) and the appearance of lipopolysaccharide (LPS) in the blood of GVH reactive mice just prior to death. In this study the authors have investigated the kinetics of LPS accumulation in different organs during the course of acute GVHD using a murine model. Unirradiated C57BL/6×AF1 (B6AF₁) mice were transplanted with C57BL/6 (B6) lymphoid cells and killed at predetermined times after transplantation for LPS analysis. Control animals were injected with either 60×10⁶ B6AF1 lymphoid cells (syngeneic) or 60×10⁶ irradiated (2000 rad) CBA lymphoid cells (allogeneic). Lipopolysaccharide began to appear in the liver and the spleen of GVH reactive mice from day 2 post-transplant and by day 10 all GVH reactive mice tested positive for hepatic and splenic LPS. Low levels of LPS were detected in some control mice from days 2 to 10 post-transplant but LPS was never detected after day 10 in control groups. Total hepatic and splenic LPS in acute GVH reactive mice peaked at a time coincident with the appearance of LPS in the serum and with the onset of mortality. These results demonstrate that tissue levels of LPS increase throughout the course of acute GVHD and are sufficient to trigger the release of pathologic amounts of TNF-α from primed macrophages resulting in the cachexia and mortality associated with acute GVHD in this model.     

Production of lymphocyte-activating factors by mouse macrophages during aging and under the effect of short peptides

Age-specific characteristics of production of lymphocyte-activating factor by mouse peritoneal macrophages and modulation of this production by short synthetic peptides (Vilon, Epithalon, and Cortagen) were studied. The production of lymphocyte-activating factors by macrophages stimulated with lipopolysaccharides in vitro was lower in old animals. The opposite modulating effects of short peptides on the production of lymphocyte-activating factors by resident and lipopolysaccharide-stimulated macrophages in young and old mice were demonstrated for the first time. This is a possible mechanism of immune system dysfunction during aging, which opens new vistas for its correction with short synthetic peptides. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 142, No. 9, pp. 333–336, September, 2006     

Alloreactivity. I. Effects of age and thymic hormone treatment on cell-mediated immunity in C57B1/6NNia mice

C57B1/6NNia mice 1, 12, and 24 months old showed loss of cellular-mediated cytotoxicity with aging. Treatment of the three age groups with different thymic hormone preparations effected their cellular mediated cytotoxicity differently. When cytotoxicity of the thymic hormone treated groups was compared to that of the physiological saline treated group, 1-month-old mice treated with serum thymic factor (FTS) at 1 microgram/mouse and 10 ng/mouse had significantly higher activity, and lower to similar activities at 12 and 24 months; TP5 (active fragment of thymopoietin) at 1 microgram and 10 ng caused significantly higher activity in 1-month-old mice, and lower to higher and significantly lower to similar activity at 12 and 24 months, respectively; TM4 (an analogue of TP5) at 1 ng showed significantly depressed activity in 1-month-old mice, and significantly enhanced activity in 12- and 24-month-old mice; thymosin at 10 micrograms and 1 microgram had slightly lower, but not significant, depression at 1 month, similar activities at 12 months and significantly depressed to higher activity at 24 months. Unimmunized control mice showed significant protection in the 12-month-old mice in comparison to 1- and 24-month-old mice. Different hormone preparations showed age- and dose-dependent effects on the ability of spleen cells to kill P815 mastocytoma. Partial restoration of cytotoxicity was observed in 24-month-old mice treated with FTS, TP5 and thymosin fraction V.     

Telomerase activity in cells of 9-day-old spleen colonies formed by bone marrow of normal and thymectomized mice

Telomerase activity was measured in spleen colonies, in progeny of individual 9-day-old splenic CFU formed by the bone marrow from normal (physiological aging) and thymectomized mice. Cells of spleen colonies expressed telomerase activity. No correlation was found between telomerase activity in spleen colony cells and age of animals. Thymectomy of bone marrow donors had no effect on telomerase activity. Our results suggest that the thymus plays a role in cell aging.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 11, pp. 567–569, November, 2004     

Derivation of a new B-cell line from mouse spleen and its characterization

Prolonged culturing of splenocytes of (CBA×C57B1/6)F₁ hybrid mice in RPMI-1640 medium with 15% fetal calf serum resulted in the derivation of a new B-cell line. After a monolayer of stromal fibroblast-like cells was formed, lymphoid cell growth was observed in 4–5 weeks of splenic cell culturing. All the cells of the splenic line belonged to the B series and expressed IgM on their surface; they did not form lymphoid colonies after injection to lethally irradiated mice, and even after 8-month culturing and several passages they could not be differentiated. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 2, pp. 183–186, February, 1995 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

Emotional state and one-trial learning in OXYS rats with hereditarily elevated production of oxygen radicals

Comparative analysis of unconditioned and conditioned behavior of Wistar and prematurely aging OXYS rats revealed that the latter have significantly reduced locomotor and exploratory activities, increased anxiety in the elevated plus-maze test, spatial disorientation, and abnormal associative learning. OXYS rats can be used as a biological model for studying molecular, neurobiological, and neurochemical mechanisms of brain aging. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 130, No. 8, pp. 155–158, August, 2000     

亚急性衰老小鼠脾脏T细胞CD137表达的研究

目的：探讨CD137分子在衰老小鼠脾脏T细胞表面表达的规律。方法：通过注射D－半乳糖建立亚急性衰老小鼠模型。分离衰老模型组小鼠及正常对照组小鼠的脾脏T细胞，经Con A刺激后进行培养，采用流式细胞术检查Con A刺激后不同培养时间的T细胞上CD137的表达，并进行比较。结果：小鼠脾脏T细胞经过Con A刺激后，正常对照组小鼠T细胞上CD137分子的表达高峰出现于刺激后第6天，平均表达率为48．5％，之后迅速下降。1个月和2个月衰老模型组小鼠T细胞上CD137分子在表达高峰的表达率平均分别为39．1％和32．8％，均显著低于正常对照组小鼠（P＜0．01），其中1个月模型组小鼠T细胞上CD137分子表达高峰的出现时间及下降规律与正常对照组小鼠相同；而2个月衰老模型组小鼠T细胞上CD137分子的表达高峰出现于刺激后第4天，第9天缓慢下降至与正常对照组相同时间的水平。结论：亚急性衰老小鼠T细胞上CD137分子的表达低于正常水平。至衰老过程的后期，T细胞上CD137分子的表达高峰提前出现，表达水平达到峰值后下降速度较正常小鼠缓慢，提示CD137分子在机体抗衰老过程中，具有调节T细胞功能的作用。     

Persistent increase of d-aspartate in d-aspartate oxidase mutant mice induces a precocious hippocampal age-dependent synaptic plasticity and spatial memory decay

The atypical amino acid d-aspartate (d-Asp) occurs at considerable amounts in the developing brain of mammals. However, during postnatal life, d-Asp levels diminish following the expression of d-aspartate oxidase (DDO) enzyme. The strict control of DDO over its substrate d-Asp is particularly evident in the hippocampus, a brain region crucially involved in memory, and highly vulnerable to age-related deterioration processes. Herein, we explored the influence of deregulated higher d-Asp brain content on hippocampus-related functions during aging of mice lacking DDO (Ddo^−/−). Strikingly, we demonstrated that the enhancement of hippocampal synaptic plasticity and cognition in 4/5-month-old Ddo^−/− mice is followed by an accelerated decay of basal glutamatergic transmission, NMDAR-dependent LTP and hippocampus-related reference memory at 13/14 months of age. Therefore, the precocious deterioration of hippocampal functions observed in mutants highlights for the first time a role for DDO enzyme in controlling the rate of brain aging process in mammals.     

Suppressive and antitumor activities of bone marrow cells and splenocytes of AKR mice during aging

In 4-month-old leukemia-prone AKR mice, the ability of bone marrow cells to inhibit proliferation of concanavalin A-stimulated splenocytes and mastocytoma P815 cellsin vitro was sharply increased in the preleukemic period. In 7-month-old mice, differences in natural suppressive activity of bone marrow cells were significant, but less pronounced than in 4-month-old mice. The immunosuppressive activity was not found in the spleen. In 4-month-old AKR mice, in the inhibition of proliferation of mitogen-stimulated splenocytes was increased due to enhanced NO production by bone marrow cells. These findings suggest that the increased antiproliferative activity observed in the bone marrow of AKR mice long before the appearance of clinical manifestations of leukemia is associated with disturbances in differentiation of myeloid progenitor cells and accumulation of natural suppressor cells in the bone marrow. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No. 4, pp. 452–454, April, 1999     

Effect of Transplantation of Splenic Lymphoid Cells on Functional Activity of the Immune and Nervous System in Experimental Animals

We studied the effect of transplantation of splenic lymphoid cells on functional activity of the nervous (orientation and exploratory behavior and expression of genes for interleukin-1β, type 1 interleukin-1 receptor, and erythropoietin in the brain) and immune system (cellular and humoral immune response, proliferative activity of immunocompetent cells, and expression of cytokine genes in splenocytes) in syngeneic animals with different behavioral characteristics. Intravenous injection of the lymphoid fraction of splenocytes from donor mice with a certain behavioral pattern in the open-field test had a modulatory effect on vertical locomotor activity of syngeneic recipient mice. The increase or decrease in vertical locomotor activity due to transplantation of immunocompetent cells was accompanied by specific changes in mRNA level for erythropoietin receptor and type 1 interleukin-1 receptor in brain cells of recipient mice. The regulation of orientation and exploratory behavior was also accompanied by changes in functional activity of the immune system in recipient animals. It was manifested in modulation of proliferative activity of thymic and splenic cells and cytokine gene expression in splenocytes.     

Effect of Thyroxine on the Immune Response of Mice in Vivo and in Vitro

The effect of thyroxine on the immune response of BALB/c mice to sheep erythrocytes was investigated. In mice rendered hypertiyroid by subcutaneous injections of T4 the primary immune response to an injection of SRBC in vivo did not show a consistent increase in splenic anti-SRBC plaque-forming cells. However, the total number of splenic cells in T4-treated mice was generally decreased, and thus, the number of PFC per 10⁶ splenic cells in T4-treated mice was higher than those of saline and buffer controls. In in-vitro primary response to SRBC PFC per culture (3×10⁷ splenic cells) increased significantly in T₄-injected animals as compared with controls. The calculated PFC per spleen also increased significantly. The addition of T₄ to normal splenic cell cultures enhanced the primary immune response to SRBC in vitro. The optimum concentration of T₄ was found to be 10^-8 M. These results indicate a direct enhancing effect of T₄ on the immune response of lymphoid cells. This enhancing effect, however, may be attenuated in vivo by the alteration of the number and/or composition of lymphoid cells brought about directly or indirectly by injections of exogenous thyroxine.     

Effects of long-term treatment of mice with anti-I-J monoclonal antibody and dialyzable leukocyte extract on immune function and lifespan

In 1969 Walford hypothesized that age-related dysfunctions of the immune system may be involved in the pathogenesis of the lesions and disease of aging. Studies were initiated to test whether immunologic interventions intended to maintain the integrity of the immune system would delay the onset of diseases of aging and prolong lifespan. Adult BC3F1 mice were treated with anti-I-J monoclonal antibody, with human dialyzable leukocyte extract, or with saline once a week for one year. Spleen cells from the mice were then assayed for suppressor, T-helper and B-cell activity. Treatment with dialyzable leukocyte extract decreased the elevated nonspecific suppressor activity. Mice treated with anti-I-J antibody had elevated T-helper cell activity. In another experiment, mice were treated weekly with anti-I-J antibody, dialyzable leukocyte extract, or saline from 18 months of age until natural death. The mice were immunized with avian gammaglobulin at 27 and again at 29 months of age. Both types of immunologic intervention resulted in a greater secondary antibody response than that of the saline-treated control mice. Mice treated with anti-I-J antibody survived longer than did mice of the other two groups. There was a correlation between the magnitude of the secondary response of individual mice and their lifespan. The results provide support for the immunologic theory of aging.