concanavalin-a and Obesity

Concanavalin A-induced hepatotoxicity was compared in lipodystrophic aP2-nSREBP-1c transgenic mice (LD mice) lacking adipose tissue, obese leptin-deficient ob/ob mice, and lean wild-type (WT) mice. Serum leptin and adiponectin were low in LD mice, whereas ob/ob mice had undetectable leptin, but high adiponectin. Protection from hepatotoxicity was observed in ob/ob, but not in LD mice, despite low cytokine levels and reduced T cell activation and hepatic natural killer T cells in both groups. Administration of adiponectin protected LD mice from hepatotoxicity without altering cytokine levels. In contrast, administration of leptin heightened disease susceptibility by restoring cytokine production. Neutralization of TNF alpha protected LD mice from liver damage. Increased in vivo susceptibility to the hepatotoxic effect of TNF alpha was observed in LD mice. In vitro, adiponectin protected primary hepatocytes from TNF alpha-induced death, whereas leptin had no protective effect. In conclusion, although leptin increases susceptibility to hepatotoxicity by regulating cytokine production and T cell activation, adiponectin protects hepatocytes from TNF alpha-induced death.

Topics: Adiponectin; Animals; Apoptosis; Autoimmune Diseases; CCAAT-Enhancer-Binding Proteins; Concanavalin A; Cytokines; DNA-Binding Proteins; Hepatitis; In Situ Nick-End Labeling; Intercellular Signaling Peptides and Proteins; Killer Cells, Natural; Leptin; Lipodystrophy; Lymphocyte Activation; Mice; Mice, Obese; Mice, Transgenic; Obesity; Sterol Regulatory Element Binding Protein 1; T-Lymphocytes; Transcription Factors; Tumor Necrosis Factor-alpha

Impaired immune function linked to obesity has been shown in both human and animal studies. The purpose of this work was to evaluate the effects of a 4-week energy restriction (50% of total energy intake) on immune function in previously diet-induced (cafeteria) overweight rats. Flow cytometric analysis revealed that the number of spleen T helper cells were significantly (P < 0.05) elevated in control and overweight energy-restricted rats as compared with groups fed ad libitum groups. The proliferative response of splenocytes to phytohaemaglutinin and concanavalin A from overweight rats after energy restriction was significantly (P < 0.05) higher compared to overweight nonrestricted rats. The cytotoxic activity of natural killer cells tended to be lower in overweight rats compared to controls. Finally, control rats under the dietary deprivation period presented higher levels of uncoupling protein 2 mRNA and lower levels of leptin receptor mRNA compared with the reference control group. These results suggest that energy restriction is able to restore, at least in part, the impaired immune response commonly observed in overweight animals.

Topics: Animals; Blood Glucose; Body Weight; Cell Division; Concanavalin A; Cytotoxicity, Immunologic; Diet, Reducing; Energy Intake; Flow Cytometry; Immunity; Ion Channels; Killer Cells, Natural; Male; Membrane Transport Proteins; Mitochondrial Proteins; Obesity; Phytohemagglutinins; Rats; Rats, Wistar; Receptors, Cell Surface; Receptors, Leptin; RNA, Messenger; Spleen; T-Lymphocytes, Helper-Inducer; Triglycerides; Uncoupling Protein 2

We investigated the effects of a diet containing EPAX-7010, rich in PUFAs such as eicosapentaenoic acid [20:5(n-3)] and docosahexaenoic acid [22:6(n-3)], i.e., a PUFA/EPAX regimen, on T-cell activation in diabetic pregnant rats and their obese pups.. Mild hyperglycemia in pregnant rats was induced by intraperitoneal injection of streptozotocin on Day 5 of gestation. T-cell blastogenesis was assayed by using (3)H-thymidine, whereas intracellular free calcium concentrations ([Ca(2+)]i) were measured by using Fura-2 in diabetic pregnant rats and their obese offspring.. Concavalin-A-stimulated T-cell proliferation was decreased in both pregnant diabetic rats and their obese pups as compared with control animals. Feeding the PUFA/EPAX diet restored T-cell proliferation in both groups of animals. We also employed ionomycin, which at 50 nM opens calcium channels, and thapsigargin (TG), which recruits [Ca(2+)]i from endoplasmic reticulum pool. We observed that ionomycin-induced increases in [Ca(2+)]i in T-cells of diabetic mothers and obese offspring were greater than in those of control rats. Furthermore, feeding PUFA/EPAX diet diminished significantly the ionomycin-evoked rise in [Ca(2+)]i in diabetic and obese animals. TG-induced increases in [Ca(2+)]i in T-cells of diabetic pregnant rats and their obese offspring were greater than in those of control rats. The feeding of the experimental diet significantly curtailed the TG-evoked increases in [Ca(2+)]i in both diabetic and obese rats.. Together, these observations provide evidence that T-cell activation and T-cell calcium signaling are altered during gestational diabetes and macrosomia. Hence, dietary fish oils, particularly eicosapentaenoic acid and docosahexaenoic acid, may restore these T-cell abnormalities.

Topics: Animals; Calcium; Concanavalin A; Diabetes Mellitus, Experimental; Dietary Fats, Unsaturated; Fatty Acids, Omega-3; Female; Fetal Macrosomia; Gestational Age; Ionomycin; Lymphocyte Activation; Obesity; Pregnancy; Pregnancy in Diabetics; Rats; Rats, Wistar; Signal Transduction; Spleen; T-Lymphocytes; Thapsigargin

Leptin-deficient ob/ob mice are protected from Con A-induced hepatitis. However, it is unclear whether leptin deficiency or obesity itself is responsible for this protection. To address this question, wild-type (WT) obese mice with high serum leptin levels were generated by injection of gold thioglucose (WT GTG). Both Con A-injected WT and WT GTG mice developed hepatitis, whereas no hepatic damage was observed in ob/ob mice. Moreover, TNF-alpha and IFN-gamma levels as well as expression of the activation marker CD69 were elevated in liver mononuclear cells of WT and WT GTG mice, but not in ob/ob mice following administration of Con A. The liver of WT and WT GTG mice had the same percentage of NK T cells, a lymphocyte population involved in Con A-induced hepatitis. This population decreased equally in both WT and WT GTG mice after Con A injection. In contrast, the liver of ob/ob mice contained 50% less NK T cells compared to WT and WT GTG mice. Furthermore, no decrease in NK T cells was observed in Con A-injected ob/ob mice. We conclude that leptin-deficiency, not obesity, is responsible for protection from Con A-induced hepatitis.

Topics: Animals; Aurothioglucose; Body Weight; Chemical and Drug Induced Liver Injury; Concanavalin A; Female; Interferon-gamma; Killer Cells, Natural; Leptin; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Obese; Obesity; T-Lymphocyte Subsets; Tumor Necrosis Factor-alpha

Leptin is an adipocyte-secreted hormone that centrally regulates weight control. However, leptin receptor is expressed not only in the central nervous system, but also in other systems such as reproductive and hematopoietic tissues. Human leptin has previously been shown to enhance cytokine production by murine peritoneal macrophages and human circulating monocytes. In this paper we have assessed the presence of leptin receptors in peripheral human T lymphocytes and we have studied their functional role. Both CD4(+) and CD8(+) T lymphocytes express leptin receptors. Moreover, we show that human leptin dose-dependently enhances proliferation and activation of human circulating T lymphocytes when they are costimulated by PHA or Con A. Leptin alone was not able to activate T lymphocytes. To confirm a direct effect of leptin on T lymphocytes, monocytes were extracted by adhesion to culture flasks. The early activation surface marker CD69 was then induced in both CD4(+) and CD8(+) T lymphocytes after 8 h stimulation with PHA or Con A. Leptin dose-dependently enhanced stimulated CD69 expression. Moreover, leptin dose-dependently enhanced the expression of the late activation markers CD25 and CD71 in both CD4(+) and CD8(+) T lymphocytes after 48 h stimulation with PHA or Con A. Finally, we have found that leptin modulates CD4(+) T lymphocyte activation toward Th1 phenotype by stimulating the synthesis of IL-2 and IFN-gamma. These results demonstrate the presence of the leptin receptor in human circulating CD4(+) and CD8(+) T lymphocytes and a functional role of leptin as a modulator (enhancer) of lymphocyte stimulation with a shift toward Th1 cytokine-production profile. This function of leptin may have some relevance in the pathophysiology of immunologic alterations related to obesity.

Topics: Antigens, CD; Carrier Proteins; Cell Division; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Flow Cytometry; Gene Expression; Humans; Interferon-gamma; Interleukin-2; Leptin; Lymphocyte Activation; Molecular Weight; Monocytes; Obesity; Phytohemagglutinins; Receptors, Cell Surface; Receptors, Leptin; T-Lymphocytes; Th1 Cells

We reported previously that obesity is a risk factor for deteriorating cellular immune functions in aging. However, the mechanism by which obesity decreases cellular immunity remains to be elucidated. To determine the mechanism of the decrease in cellular immunity with obesity, lean (Fa/?) and obese (fa/fa) 12-mo-old Zucker rats were used. The mitogen response of splenic lymphocytes in obese Zucker rats was significantly lower than that of lean Zucker rats, which was not restored by in vitro treatment with indomethacin (10 micromol/L), an inhibitor of prostaglandin E2 (PGE2). In addition, PGE2 production by splenic lymphocytes was not greater in obese than in lean Zucker rats. Glucose consumption by splenic lymphocytes after in vitro incubation with concanavalin A (conA) for 48 h was also significantly lower in obese Zucker rats. Expression of glucose transporter 1 (GLUT-1), analyzed by Western blot analysis, was lower in splenic lymphocytes of obese than in lean Zucker rats. However, the expression of the conA receptor in splenic lymphocytes, analyzed by flow cytometry with fluorescein isothiocyanate-conjugated conA, was not significantly different between lean and obese Zucker rats. In conclusion, the decreased mitogen response of splenic lymphocytes in obese Zucker rats may be due in part to the decreased uptake of glucose as the main energy source for lymphocytes at the stage of proliferation and may be associated with the decreased expression of GLUT-1.

Topics: Animals; Blood Glucose; Concanavalin A; Cyclooxygenase Inhibitors; Dinoprostone; Female; Glucose; Immunity, Cellular; Indomethacin; Insulin; Lymphocytes; Mitogens; Monosaccharide Transport Proteins; Obesity; Rats; Rats, Zucker; Spleen; Triglycerides

Although obese people have been reported to have a higher incidence of infections and some types of cancer, the immunocompetence of obese subjects remains poorly understood. To investigate whether obesity affects immunity, we studied obese subjects (BMI > 30 kg/m2) whose health was uncomplicated by any other disorder, including hyperglycemia. We compared mitogen-induced blastogenic response of peripheral blood lymphocytes in 34 obese subjects (mean +/- s.e. BMI: 38.4 +/- 2.0 kg/m2) and 35 non-obese controls (BMI: 21.3 +/- 0.4 kg/m2) who were matched for age and sex. The effects of weight reduction were also evaluated in 19 obese persons (BMI: 36.4 +/- 1.8 kg/m2) on a very low calorie diet. Mean (+/- s.e.) intracellular incorporation of [3H]-thymidine, on stimulation of T lymphocytes with either phytohaemagglutinin (PHA) or concanavalin A (Con A), and B lymphocytes with pokeweed mitogen, was significantly diminished in obese subjects compared to non-obese controls (47552 +/- 6917 vs. 83720 +/- 6252 cpm, P < 0.001; 30301 +/- 6018 vs. 45942 +/- 3723 cpm, P < 0.05; 13669 +/- 2971 vs. 23735 +/- 2048 cpm, P < 0.01, respectively). After weight reduction (BMI: 27.8 +/- 1.2 kg/m2), the mean T lymphocyte responses to PHA and Con A were increased significantly vs. baseline (98404 +/- 2444 vs. 50337 +/- 9516 cpm, P < 0.05 and 69523 +/- 15480 vs. 36695 +/- 8006 cpm, P < 0.05, respectively). Depressed blastogenesis of B lymphocytes was also augmented but was not statistically significant. The results suggest that obese subjects have underlying immune impairment in responsiveness of lymphocytes and that these impairments are reversible with adequate weight reduction.

Topics: Adult; Blood Glucose; Body Mass Index; Concanavalin A; Diet, Reducing; DNA; Female; Humans; Insulin; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Obesity; Phytohemagglutinins; Pokeweed Mitogens; Regression Analysis; Weight Loss

The effect of hyperlipaemic serum on mitogen-induced T lymphocyte proliferation was investigated with cynomolgus monkeys. The mitogen-induced blastogenesis was remarkably inhibited when either hyperlipaemic or normal monkey lymphocytes were incubated with hyperlipaemic sera. Hyperlipaemic serum also inhibited ConA-induced interleukin 2 (IL-2) production as well as IL-2 receptor (IL-2R) expression of normal monkey lymphocytes. On the other hand, it showed slight inhibition of T-cell proliferation induced by adding recombinant human IL-2 to IL-2R-positive normal monkey lymphocytes. These results indicate that hyperlipaemic serum inhibited an early stage of T-cell autocrine activation pathway including IL-2 production and IL-2R expression.

Topics: Animals; Concanavalin A; Female; Hyperlipidemias; Interleukin-2; Lymphocyte Activation; Macaca fascicularis; Obesity; Receptors, Interleukin-2; Recombinant Proteins; T-Lymphocytes

In contrast to systemic autoimmunity, spontaneous autoimmune thyroiditis of Obese strain (OS) chickens is associated with a marked T cell hyperreactivity in vitro, i.e. an increased proliferation and interleukin 2 (IL 2) secretion in response to Concanavalin A (ConA). In the present study we report an enhanced capacity of OS peripheral lymphoid cells (splenocytes and peripheral blood lymphocytes, PBL) to adsorb fluorescein isothiocyante (FITC) labelled ConA, but not phytohemagglutinin (PHA). However, the elevated ConA binding cannot be a prerequisite for in vitro ConA hyperreactivity as OS thymocytes are normal with respect to ConA binding but nonetheless exhibit elevated responses to this mitogen. Moreover, ConA binding does not correlate with the frequency of cells able to express IL 2 receptors upon short term ConA stimulation. The percentage of ConA activatable cells was found to be increased in OS- PBL as compared to normal control PBL, but was unaltered in OS splenocytes. This finding points to a further mechanism of T cell hyperreactivity in OS chicks in addition to the previously reported defects in nonspecific immunosuppression. Finally, enumeration of cells in the S phase revealed that enhanced proliferation of OS T lymphocytes was not restricted to the in vitro response to ConA and phytohemagglutinin (PHA) but also occurs in vivo.

Topics: Animals; Cell Division; Chickens; Concanavalin A; Disease Models, Animal; Lymphocyte Activation; Lymphoid Tissue; Obesity; Receptors, Concanavalin A; T-Lymphocytes; Thyroiditis, Autoimmune

The enhanced T cell reactivity (ConA hyperresponsiveness and IL 2 hypersecretion) of spleen lymphocytes of Obese strain (OS) chickens with spontaneous autoimmune thyroiditis has recently been shown to be due to a defect in macrophage-derived non-specific suppressor factors that regulate IL 2 secretion and IL 2-promoted T lymphoblast proliferation in normal healthy animals. In the present study, we present several lines of evidence that the increased T cell response of peripheral blood lymphocytes (PBL) of OS chickens is due to mechanisms entirely different from the described dysregulation of splenic T cells: 1) In contrast to the splenic macrophages, peripheral blood monocytes of OS animals are not deficient in the production of IL 2 antagonistic activity (IAA); 2) therefore, cocultivation of PBL from OS and Normal White Leghorn (NWL) chickens in communicating culture chambers did not abrogate the difference in Con A response as previously observed with spleen lymphocytes. 3) Immunofluorescence with a monoclonal antibody (INN CH 16) against the chicken IL 2 receptor revealed enhanced numbers of mitogen activatable T cells in OS PBL but not OS spleen lymphocytes. 4) After prolonged Con A stimulation of PBL, OS and NWL lymphoblasts did not differ from each other in functional aspects. In contrast to this, Con A lymphoblasts from OS spleens exhibited enhanced staining with INN CH 16 in parallel with an increased proliferative response to IL 2. Thus, the primary T cell dysfunction involved in the development of autoimmune disease in OS chickens is the result of at least two separate regulatory defects.

Topics: Animals; Cell Communication; Chickens; Concanavalin A; In Vitro Techniques; Interleukin-2; Lymphocyte Activation; Obesity; Spleen; T-Lymphocytes; Thyroiditis, Autoimmune

Topics: Animals; Concanavalin A; Culture Techniques; Cyclic AMP; Diazoxide; Homeostasis; Humans; Hyperinsulinism; Insulin; Insulin Resistance; Liver; Lymphocytes; Obesity; Rats; Receptor, Insulin