T-Cell Populations and Cytokine Expression Are Impaired in Thymus and Spleen of Protein Malnourished BALB/c Mice Infected with Leishmania infantum

Visceral leishmaniasis (VL) is a parasitic infectious disease that causes significant morbidity and mortality in the tropical and subtropical regions of the world. Although infections with visceralizing Leishmania may be asymptomatic, factors such as undernutrition increase the likelihood of progressing to clinical disease. Protein malnutrition, the most deleterious cause of malnutrition in developing countries, has been considered as a primary risk factor for the development of clinical VL. However, data regarding the immunological basis of this association are scarce. With the aim to analyze the effects of protein malnutrition on Leishmania infantum infection, we used BALB/c mice subjected to control or low protein isocaloric diets. Each animal group was divided into two subgroups and one was infected with L. infantum resulting in four study groups: animals fed 14% protein diet (CP), animals fed 4% protein diet (LP), animals fed 14% protein diet and infected (CPi), and animals fed 4% protein diet and infected (LPi).The susceptibility to L. infantum infection and immune responses were assessed in terms of body and lymphoid organ weight, parasite load, lymphocyte subpopulations, and cytokine expression. LPi mice had a significant reduction of body and lymphoid organ weight and exhibited a severe decrease of lymphoid follicles in the spleen. Moreover, LPi animals showed a significant decrease in CD4⁺CD8⁺ T cells in the thymus, whereas there was an increase of CD4⁺ and CD8⁺ T cells percentages in the spleen. Notably, the cytokine mRNA levels in the thymus and spleen of protein malnourished-infected animals were altered compared to the CP mice. Protein malnutrition results in a drastic dysregulation of T cells and cytokine expression in the thymus and spleen of L. infantum-infected BALB/c mice, which may lead to defective regulation of the thymocyte population and an impaired splenic immune response, accelerating the events of a normal course of infection.     

Effects of Dietary Selenium on Selenoprotein W Gene Expression in the Chicken Immune Organs

Selenoprotein W (SelW) is expressed in the immune systems of mammals. However, its pattern of expression in the immune organs of birds is still unclear. To investigate the distribution of SelW and effects of dietary Se levels on the SelW mRNA expression in the immune organs of birds, 1-day-old male chickens were fed either a commercial diet or an Se-supplemented diet containing 0.601, 1.058, 1.514, or 2.427?mg Se per kilogram, and 1.0, 2.0, 3.0 or 5.0?mg sodium selenite per kilogram for 90?days. The immune organs (spleen, thymus, and bursa of Fabricius) were collected and examined for Se content and SelW mRNA levels. The mRNA expression of SelW was detected in all the tissues. Although Se content was the highest in the spleen, the remarkable stability of the SelW mRNA level was observed in this organ during different times of dietary Se supplementation. Se-supplemented diet can make the SelW expression levels higher within a certain range in thymus and bursa of Fabricius. The present study demonstrates that SelW is widely expressed in immune organs of birds and that Se-supplementation of the feed increases SelW expression in the thymus and the bursa of Fabricius.     

Dietary supplementation of organic or inorganic chromium modulates the immune responses of broilers vaccinated with Avian Influenza virus vaccine

Dietary supplementation with the organic chromium (Cr) has been shown to positively affect the immune function of poultry. However, to our knowledge, no experiment has been done to directly compare the impacts of Cr chloride and chromium picolinate (CrPic) on the immune responses of broilers vaccinated with Avian Influenza (AI) virus vaccine. Therefore, the present experiment was conducted to investigate the effects of supplemental Cr sources (Cr chloride and CrPic) and levels on the growth performance and immune responses of broilers vaccinated with AI virus vaccine so as to provide an effective nutritional strategy for improving immune function of broilers. A total of 432 1-day (d)-old male broiler chicks were used in a 1 plus 2×4 design. Chickens were given either a diet without Cr supplementation (control) or diets supplemented with 0.4, 0.8, 1.6, or 3.2 mg Cr/kg as either Cr chloride or CrPic for 42 d. Compared to the control, dietary Cr supplementation had no effect (P>0.05) on average daily gain, average daily feed intake and gain : feed of broilers during the starter and grower phases, but increased (P<0.05) the relative weights of bursa of fabricius on d 21 and thymus, spleen, or bursa of fabricius on d 42, serum antibody titers against AI virus on d 21, 28, 35 and 42, blood T-lymphocyte transformation rate on d 28 and 42, blood T-lymphocyte percentage on d 42, and serum interleukin-2 contents on d 28. Broilers fed the diets supplemented with the inorganic Cr chloride had higher (P<0.05) weights of thymus, spleen and bursa of fabricius than those fed the diets supplemented with the CrPic on d 42. In addition, broilers fed the diets supplemented with the CrPic had higher (P<0.05) antibody titers against AI virus than those fed the diets supplemented with the inorganic Cr chloride on d 21 and 35. These results indicate that dietary Cr supplementation improved immune responses of broilers vaccinated with AI virus, and the inorganic Cr chloride was more effective than the CrPic in increasing the relative weights of lymphoid organs, however, the CrPic was more effective than the inorganic Cr chloride in enhancing the serum antibody titer against AI virus.     

Effect of hypokinesia on nucleic acid and protein metabolism in the lymphoid organs of rats

Metabolism of nucleic acids and protein by lymphoid cells of the rat spleen and thymus was studied under conditions of 22-day hypokinesia. It was shown that in the course of hypokinesia the loss of cellular mass by the spleen and thymus was associated with varied biochemical changes in the remaining lymphoid cells. The thymocytes showed a significant activation of nucleic acid and protein biosynthesis. Meanwhile in spleen lymphocytes, DNA and RNA metabolism was inhibited with no appreciable changes in protein metabolism. Potential mechanisms of changes in metabolism of thymus and spleen lymphocytes under long-term hypokinesia are discussed.     

The influence of dietary protein deficiency on haemopoietic cells in the mouse.

These experiments examined the effect of a diet limited only in protein (4% by weight) on haemopoietic stem cells in mice. This diet places severe restrictions on growth and cell proliferation and this was reflected in lower numbers of colony forming units (CFUs) and in vitro colony forming cells (CFCs). Differences were apparent in the response of different organs to this stress; for instance, the incidence of spleen CFUs fell sharply from around 40/mg spleen tissue to 1-4/mg spleen tissue after 3 weeks on a low protein diet. This selective loss did not occur in bone marrow where total CFUs remained proportional to cellular content. Yet a third pattern was shown by thymus CFUs--although the numbers were low these increased from 16/thymus in normal mice to 132/thymus in deprived mice. This was the only organ examined which showed an increase. The effects of a return to a high protein (18%) diet showed that the spleen was the most responsive organ. By day 5 after the return to 18% protein the spleen contained as many CFUs per million cells as the bone marrow. During this time the content of CFU in the spleen had increased some 50-fold whereas bone marrow CFUs only doubled. The spleen assumes the major reconstructive role during the refeeding process.     

免疫应答期间脑和淋巴器官中去甲肾上腺素含量的变化

应用高效液相色谱－电化学检测法（ＨＰＬＣ－ＥＣＤ）测定大鼠在用绵羊红细胞（ＳＲＢＣ）免疫后第２－７天期间,下丘脑、海马、脑干、胸腺和脾脏中去甲肾上腺素（ＮＡ）含量的变化。实验结果表明,下丘脑内ＮＡ含量在免疫后第４－７天明显增加,其中第７天有回降趋势。海马内ＮＡ含量在第４－５天显著增多。而胸腺和脾脏中ＮＡ水平在第４－５天均明显降低。脑干在免疫应答期间无明显改变。以上结果说明,体液免疫应答可影响脑和淋     

Effects of selenium supplementation on virus-induced inflammatory heart disease

The effects of 10 wk of selenium (Se) supplementation (5 ppm) in drinking water on immune responses and resistance to a myocarditic Coxsackie virus B3 (CB3) infection were studied in female Balb/c mice. Se supplementation reduced CB3-induced mortality: at day 14 postinoculation, survival was 58% in the Se-treated group as compared to 25% in the untreated group. Whole-blood glutathione peroxidase (GSH-Px) activity was elevated by 68% (p < 0.001) and Se content in the liver by 24% (p < 0.001). Red (RBC) and white blood cell (WBC) counts, as well as the number of cells in the spleen and thymus, were unaffected. The cellular counts of T-lymphocytes (CD4⁺, CD8⁺) and natural killer (NK⁺) cells in the blood were not affected. However, the CD4⁺/CD8⁺ ratio (5.2) tended to increase after Se supplementation (5.9). The spleen lymphoproliferative response to T and B-cell mitogens were increased by 9 and 43%, respectively (ns), in the Se-supplemented group. The total NK cell activity in blood and spleen showed minor increases, but when the activity in the blood was expressed per cell, the increase amounted to 35% (ns) with Se supplementation. The inflammatory and necrotic lesions in the ventricular myocardium at 7 and 14 d postinoculation were not significantly reduced by Se treatment, probably owing to the increased survival with Se even of mice with the most pronounced heart damage; comparable untreated mice were estimated to have died at day 14. Results indicate that modest doses of Se can improve immune function, which may increase the general resistance to this viral infection.     

THE INFLUENCE OF DIETARY PROTEIN DEFICIENCY ON HAEMOPOIETIC CELLS IN THE MOUSE

These experiments examined the effect of a diet limited only in protein (4% by weight) on haemopoietic stem cells in mice. This diet places severe restrictions on growth and cell proliferation and this was reflected in lower numbers of colony forming units (CFUs) and in vitro colony forming cells (CFCs). Differences were apparent in the response of different organs to this stress; for instance, the incidence of spleen CFUs fell sharply from around 40/mg spleen tissue to 1 -4/mg spleen tissue after 3 weeks on a low protein diet. This selective loss did not occur in bone marrow where total CFUs remained proportional to cellular content. Yet a third pattern was shown by thymus CFUs–although the numbers were low these increased from 16/thymus in normal mice to 132/thymus in deprived mice. This was the only organ examined which showed an increase. The effects of a return to a high protein (18 %) diet showed that the spleen was the most responsive organ. By day 5 after the return to 18% protein the spleen contained as many CFUs per million cells as the bone marrow. During this time the content of CFU in the spleen had increased some 50-fold whereas bone marrow CFUs only doubled. The spleen assumes the major reconstitutive role during the refeeding process.     

海藻酸钠寡糖对小鼠免疫及 抗氧化功能的影响

目的研究海藻酸钠寡糖对小鼠免疫及抗氧化活性的影响。方法采用不同浓度的海藻酸钠寡糖分别对小鼠连续灌胃15d后,测量小鼠生长性能、小鼠胸腺和脾脏指数以及小鼠血浆中SOD和GSH的活性。结果海藻酸钠寡糖能促进小鼠的生长性能,提高小鼠的胸腺和脾脏指数。其中给小鼠灌注高浓度的海藻酸钠寡糖15d后,与对照组相比,小鼠的特定生长率和增重率分别提高了37.0%和48.5%(P0.01),小鼠胸腺和脾脏指数分别提高了18.8%和21.7%(P0.01)。海藻酸钠寡糖还能增加小鼠的抗氧化活性,与对照组相比,灌注高浓度海藻酸钠寡糖组小鼠的SOD和GSH-Px活性分别提高了92.6%和35.9%(P0.01)。结论海藻酸钠寡糖能促进小鼠的生长性能,增强小鼠的免疫和抗氧化功能。     

Effects of a protein-free diet on the DNA-synthesizing and dividing cells in the lymphoid organs of rats. Changes induced by phytohemagglutinin and cortisone invivo

Levels of ³H-thymidine-labeled lymphocytes on autoradiographs of cell smears and mitotic indices (M.I.) were determined in the popliteal lymph nodes (PLN), spleen and thymus of rats fed with a balanced or a protein deprived (PD) diet for 7 weeks. The latter diet reduced the number of labeled cells per 10³ in the spleen but not in the PLN's and the thymus. The M.I.'s were reduced 50% in the spleen and dropped drastically in the thymus. Subplantar injection of phytohemagglutinin (PHA) provoked a sharp increase in the M.I.'s of the PLN's and the spleen, and also moderately increased the labeling index in the PLN's in normal rats. In PD rats PHA no longer influenced the mitoses in the PLN's and the spleen while it considerably increased the proportion of labeled cells in the latter. These discrepancies between DNA-synthesis and mitoses in the thymus and the spleen suggest a premitotic block of the cell cycle after protein deprivation. The effects of a 5 day-cortisone treatment resembled in a large measure the changes induced by a protein-free diet including the arrest of the cell cycle. The thymic lymphocytes were more sensitive to the hormone in PD than in normal rats. On the contrary, in the PLN's and the spleen mainly cortisone-resistant cells remained alive among the DNA-synthesizing lymphocytes after protein deprivation. All these data agree with the hypothesis of an intervention of endogenous glucocorticoids in the protein deficiency-induced lymphoid involution.     

B-cell suppression of antibody response to sheep red blood cells in mice of high- and low-responding genotypes.

CBA and C57B1 mice (high and low responders to sheep red blood cells, respectively) were injected intravenously with syngeneic lymph node, marrow, spleen, or thymus cells together with sheep red blood cells (SRBC), and the production of antibody-forming cells (AFC) was assayed in the spleen. Transfer of lymph node, marrow, spleen, or thymus cells led to a significant enhancement of immune responsiveness in low-responding C57B1 mice. In contrast, transfer of marrow, lymph node, or spleen cells to high-responding CBA mice was accompanied by a decline in AFC production. These effects were magnified if syngeneic cell donors had been primed with SRBC; suppression in CBA mice and stimulation in C57B1 mice were especially pronounced after transfer of SRBC-primed lymphoid cells. Pretreatment of CBA donors with cyclophosphamide in a dose causing selective B-cell depletion completely abrogated the suppression of immune responsiveness. A large dose (10⁷) of syngeneic B cells injected together with SRBC suppressed the accumulation of AFC in both CBA and C57B1 mice. No suppression of immune responsiveness was observed after transfer of intact thymus cells, hydrocortisone-resistant thymocytes, or activated T cells. We conclude that suppression of the immune response to SRBC is induced by B cells. At the same time, there is a possibility that the addition of “excess” B cells acts as a signal, triggering suppressor T cells.     

Changes in proliferation activity and relative distributions of lymphoid cell subpopulations in congenitally athymic nu/nu mice and lurcher mice with spontaneous olivopontocerebellar degeneration

Changes observed in mice with congenital damage of some part of the CNS-neuroendocrine-immune regulatory system are described. nu/nu mice with congenital absence of thymus and Lurcher mice with spontaneous olivopontocerebellar degeneration displayed changes in the histoarchitecture of adrenal gland, immune organs (thymus, spleen, axillar lymph nodes) and intestine. Changes were also observed in IgM+, IgG+, CD4+ and CD8+ lymphoid cell subpopulations in the main lymphoid organs--the spleen and axillar lymph nodes and in the proliferative ability of whole lymphoid cell populations. The extreme decrease of lymphoid T-cell subpopulations in athymic nu/nu mice is the consequence of the absence of thymus, the organ of their maturation. On the other hand, a relative increase of B-cell subpopulations was found in this mouse strain. A relative decrease of CD4+ lymphocytes and a different influence of immunization on B-cell subpopulations were found in the spleen in neurodeficient Lurcher mice. The high percentage of apoptotic cells, cells in the S-phase of cell cycle and increased proliferation index in nu/nu mice suggest that the turnover and renewal of lymphoid cells in the spleen in nu/nu mice is more rapid than in control immunocompetent BALB/c mice.     

Immunocytochemical detection of Ig-positive cells in blood, lymphoid organs and the gut associated lymphoid tissue of the turbot (Scophthalmus maximus)

The present study was designed to search for the sites of the B-cell lineage in the different lymphoid organs of turbot (Scophthalmus maximus) by immunoperoxidase staining with a rabbit polyclonal antiserum against deglycosylated turbot IgM (TUDG-6). A turbot immunoglobulin (Ig) fraction, isolated by protein A, was checked for purity by gel filtration and SDS-PAGE under reducing conditions. The turbot IgM was deglycosylated and used to raise an antiserum. The antiserum titre was evaluated in ELISA. It was then used to analyse turbot peripheral blood leucocytes for membrane and cytoplasmic Ig and for immunohistochemistry with turbot lymphoid tissues. Very low numbers of Ig+ cells were found in thymus sections. In sections of spleen, Ig+ cells were observed in white pulp, around ellipsoids but were mostly concentrated and associated with melanomacrophage centers (MMCs). The lymphoid Ig+ cells in the kidney tended to be dispersed among haematopoietic and granulopoietic cell populations and were in intimate association with the MMCs and blood vessels. This association between MMCs and Ig+ cells in the spleen and the kidney, is discussed with respect to the role played by these organs in the immune system of fish. Last, the lymphoid population in the gut associated lymphoid tissue (GALT) of turbot was characterised with respect to staining for Ig. Immunoreactive cells were rarely detected in the epithelial layer although many lymphocytes were present, but they were frequently observed in the lamina propria, presumably as part of the GALT and involved in mucosal immune responses.     

Effect of Dietary Supplementation of Organic Chromium on Performance, Carcass Traits, Oxidative Parameters, and Immune Responses in Commercial Broiler Chickens

An experiment was conducted to study effect of supplementing graded concentrations (0, 100, 200, 300, or 400 μg/kg diet) of organic chromium (Cr) on performance, carcass traits, oxidative stress, and immune responses in commercial broiler chickens reared in open-sided poultry house under tropical climatic conditions. Each diet was fed ad libitum to eight replicates consisting of six birds in each pen from 1 to 42 days of age. Body mass gain and feed efficiency at 21 and 42 days of age and relative mass of liver, abdominal fat, and ready to cook yields at 42 days of age were not affected (P > 0.05) by supplementing organic Cr in broiler diet. Body mass loss during pre-slaughter holding period (12 h) reduced and relative breast mass increased nonlinearly (P < 0.01) with concentration of Cr in diet. Lipid peroxidation decreased, while activities of glutathione peroxidase and glutathione reductase in plasma increased nonlinearly with Cr supplementation. The ratio between heterophyl and lymphocyte was not affected (P > 0.05) with Cr supplementation in broiler diet. Relative mass of lymphoid organs (bursa, spleen, and thymus) and antibody production to Newcastle disease vaccination were not affected (P > 0.05) by the treatments employed. The cell-mediated immunity (lymphocyte proliferation ratio) increased nonlinearly with dietary Cr concentration. The results of the present study indicated that supplementation of organic Cr did not influence body mass and feed efficiency. However, supplementation of the Cr reduced pre-slaughter holding losses, increased the relative mass of breast, and increased antioxidant status and lymphocyte proliferation in broilers reared in tropics.     

Quantitative study of regression and regeneration of the thymic cell population after X-irradiation in the newtPleurodeles waltlii Michah

Summary The changes in cell numbers of different thymic cell populations and the conditions governing the regeneration of these populations and the thymus itself were examined after X-irradiation (700 rads) of different parts of the body. The general effects of the irradiation were studied in each experimental group in terms of mortality and growth rate. The particular effects on each thymic cell population were studied by the measurement of mitotic activity and of evaluation of the changes in numbers among these populations in the thymus itself, and were compared with the effects in the granulopoietic layer of the liver and in the spleen. The great reduction in the number of lymphocytes after irradiation demonstrates that they are more radiosensitive than other cell types; this reduction can be compensated for by the arrival of new lymphoid cells originating from other lymphoid organs (if they have been protected from irradiation) and by allowing thymic regeneration. Thus, irradiation has indirect effects on non-irradiated areas, and demonstrates that the lymphoid cell population has a high potential for multidirectional migration.     

微生态制剂对断奶仔兔生产性能及免疫的影响

目的研究微生态制剂对断奶仔兔生产性能、免疫功能的影响,为开发应用新型添加剂提供依据。方法选择64只35日龄、体重相近的断奶仔兔,随机分为4组,每组设2个重复,每个重复8只。实验期为40 d。对照组Ⅰ饲喂基础饲粮,实验组Ⅱ、Ⅲ、Ⅳ在基础日粮中分别添加0.1%、0.3%、0.5%的微生态制剂。测定日增重、饲料报酬、发病率、死亡率、胸腺指数、脾脏指数及T淋巴细胞ANAE阳性细胞百分率等指标。结果实验组Ⅱ、Ⅲ、Ⅳ日增重分别比对照组I提高了10.6%（P〈0.05）、23.967%、27.0%（P〈0.01）,饲料报酬分别提高了6.8%、14.3%、15.1%,发病率分别降低了12.5%、18.7%、18.7%,死亡率分别降低了12.5%、12.5%、18.7%,脾脏指数分别提高了11.5%（P〈0.05）、29.5%、34.4%（P〈0.01）,胸腺指数分别提高了2.3%、15.9%、17.9%,T淋巴细胞阳性率分别提高了17.7%（P〈0.05）、26.9%、28.4%（P〈0.01）。结论在断奶应激的情况下,饲料中添加微生态制剂可提高仔兔的生产性能和免疫功能。     

Post-radiation disorders in the cyclic AMP system of mouse spleen and thymus lymphoid cells]

Early alterations in the enzyme activities controlling cyclic adenosine-3',5'-monophosphate (cAMP) metabolism after the irradiation (800 rad) of mice were found in the lymphoid cells of the spleen and thymus. Postradiation disturbances in these activities' ration induced alterations in cAMP steady-state concentrations in the cell. It was also demonstrated that irradiation reduced lymphoid cell ability to accumulate cAMP in response to isoproterenol administration.