Immunomanipulation of appetite and body temperature through the functional mimicry of leptin.

Objective: Although current obesity therapies produce some benefits, there is a need for new strategies to treat obesity. A novel proposal is the use of anti-idiotypic antibodies as surrogate ligands or hormones. These anti-idiotypic antibodies carry an internal motif that imitates or mimics an epitope in the antigen (i.e., hormone or ligand). Thus, anti-idiotypic antibodies to several ligands may mimic them in transducing signals when binding to their receptors. Research Methods and Procedures: We developed an anti-idiotypic polyclonal antibody against the region of a leptin monoclonal antibody that competitively binds leptin, mimicking the active site structure of leptin. To test whether our anti-idiotype could also reproduce leptin functions, we examined food intake, body weight, and colonic temperature in male Wistar rats (n = 9) in response to intracerebroventricular administration of the leptin anti-idiotype. Results: Our leptin anti-idiotype induced a significant reduction in food intake coupled with an increase in body temperature comparable to that of leptin. That is, the intracerebroventricular administration of 8.0 microg of leptin anti-idiotype or 5.0 microg leptin significantly increased colonic temperature (Delta 1.9 plusminus 0.11 °C and Delta1.7 plusminus 0.12 °C, respectively). In addition, both decreased 24-hour food intake (-26.4 plusminus 2.4% and -21.9 plusminus 2.2%) compared with the control. The gain in body weight was also decreased by acute administration of the anti-idiotype (-1.4 plusminus 0.28%) and leptin (-1.1 plusminus 0.17%) vs. the phosphate-buffered saline control (1.3 plusminus 0.15%). Discussion: These studies revealed that the leptin anti-idiotype inhibited food intake and enhanced heat production, mimicking leptin's central actions

Immunoneutralization and anti-idiotype production: two-sided applications of leptin

The neuroendocrine and immune systems are linked through a complex bi-directional network, in which hormones modify immune function, and the immune system, through the action of cytokines, affects neuroendocrine responses involved in the maintenance of body homeostasis. The adipocyte-derived, peptide hormone leptin is a pleiotropic molecule belonging to the helical cytokine family. On pp. 182-187, Matarese et al. suggest the possibility of new leptin-based therapeutic strategies for the treatment of both infection and autoimmune disease

Cardiotrophin-1 defends the liver against ischemia-reperfusion injury and mediates the protective effect of ischemic preconditioning

Ischemia-reperfusion (I/R) liver injury occurs when blood flow is restored after prolonged ischemia. A short interruption of blood flow (ischemic preconditioning [IP]) induces tolerance to subsequent prolonged ischemia through ill-defined mechanisms. Cardiotrophin (CT)-1, a cytokine of the interleukin-6 family, exerts hepatoprotective effects and activates key survival pathways like JAK/STAT3. Here we show that administration of CT-1 to rats or mice protects against I/R liver injury and that CT-1-deficient mice are exceedingly sensitive to this type of damage. IP markedly reduced transaminase levels and abrogated caspase-3 and c-Jun-NH2-terminal kinase activation after I/R in normal mice but not in CT-1-null mice. Moreover, the protective effect afforded by IP was reduced by previous administration of neutralizing anti-CT-1 antibody. Prominent STAT3 phosphorylation in liver tissue was observed after IP plus I/R in normal mice but not in CT-1-null mice. Oxidative stress, a process involved in IP-induced hepatoprotection, was found to stimulate CT-1 release from isolated hepatocytes. Interestingly, brief ischemia followed by short reperfusion caused mild serum transaminase elevation and strong STAT3 activation in normal and IL-6-deficient mice, but failed to activate STAT3 and provoked marked hypertransaminasemia in CT-1-null animals. In conclusion, CT-1 is an essential endogenous defense of the liver against I/R and is a key mediator of the protective effect induced by IP

Decreased cardiotrophin-1 levels are associated with a lower risk of developing the metabolic syndrome in overweight/obese children after a weight loss program

Objective: Cardiotrophin-1 (CT-1) shares some similarities with other cytokines, and participates in the control of energy metabolism. Higher circulating levels are observed in obese humans, but little information is gathered in weight loss (WL) programs. Therefore, we aimed to investigate the association of serum CT-1 levels with metabolic variables and the risk of developing metabolic syndrome (MetS) after a WL program in overweight/obese children. Subjects and Methods: Forty-four overweight/obese children (mean age 11.5 yr; 50% males) undergoing a 10-week WL program were enrolled. Subjects were dichotomized at the median of Body Mass Index-Standard Deviation Score (BMI-SDS) change, as high and low responders after intervention. Results: CT-1 levels were significantly reduced (-48 fmol/mL, p=0.043) in the high responder group after the WL program. They had significantly lower body weight (-3.7 kg, p<0.001), body fat mass (-8%, p<0.001), BMI-SDS (-0.78, p<0.001) and waist circumference (-5.4 cm, p<0.001), and a significant improvement in lipid and glucose profiles (p<0.05). Interestingly, decreased CT-1 levels significantly predicted changes in total cholesterol (41%) and LDL-cholesterol (28%). Moreover, in our participants the lower the CT-1 levels, the higher the reduction in MetS risk components, after the 10- week intervention, (p-ANCOVA=0.040, p-trend=0.024). Conclusion: We showed, for the first time, a reduction in serum CT-1 levels after a WL program and this decrease in CT-1 was strongly associated with a reduction in cholesterol levels and in MetS risk factors in overweight/obese children. Our findings may suggest that CT-1 could be an indirect marker for the diagnosis of MetS in this population

Induction of hypothermia, hypoglycemia and hyperinsulinemia after acute leptin immunoneutralization in overnight fasted mice

Acute immunoneutralization of circulating leptin, with an anti-leptin antibody, significantly reduced rectal temperature at 30 min and 75 min post-injection in overnight fasted and at 30 min in overnight fed mice, while no effects in metabolic and ponderal indicators were observed after antibody administration for 22 days. Furthermore, hyperinsulinemia and hypoglycemia were induced by passive immunization against leptin, being both influenced by the post-prandrial status. These experiments confirm through an indirect approach that leptin is involved in energy, but also in glucose homeostasis

Immunohistochemical detection of chloride/bicarbonate anion exchangers in human liver

Sodium-independent Cl-/HCO3- exchange activity has been observed in isolated rat hepatocytes and intrahepatic bile duct epithelial cells, where it is involved in intracellular pH regulation and, possibly, biliary bicarbonate secretion. Monoclonal antibodies to the membrane domain of human chloride/bicarbonate anion exchanger proteins, AE1 and AE2, were prepared so that we might determine by immunohistochemical methods the presence and location of these antiporters in the human liver. To obtain the antibody against AE1, we immunized mice with injections of washed human erythrocytes. The selected monoclonal antibody was found to be specific for the 17-kD proteolytic membrane fragment of AE1 protein. The antibody to AE2 was produced with a 14-mer synthetic peptide, whose sequence corresponds specifically to amino acid residues 871 to 884 in the deduced primary structure of human kidney AE2 protein. When the monoclonal antibody to AE2 peptide was employed for the immunohistochemical study of liver specimens (by both immunofluorescence and immunoperoxidase), a clearly defined staining was present at the canalicular membrane of hepatocytes, as well as the luminal side of the membrane of bile duct epithelial cells from small and medium-sized bile ducts. No staining was observed in the liver parenchyma with the monoclonal antibody to AE1, which instead strongly decorated the erythrocytes in liver blood vessels. We conclude that AE2 immunoreactivity is present in human liver, where it localizes very specifically to the membrane regions, which appear most probably involved in the transport of bicarbonate to bile (i.e., the canalicular membrane of hepatocytes and the apical side of epithelial cells of small and medium bile ducts)

Effect of a beta3-adrenergic agonist on liver glucokinase gene expression in alloxan-diabetic rats

Beta-adrenergic agonists have been shown to elicit powerful hypoglycemic effects when administered to different animal models of diabetes. However, the intimate mechanism involved in this process remains unclear. In this context, treatment of alloxan-induced diabetic rats with the beta3-adrenergic agonist Trecadrine (1 mg x kg(-1) x day(-1), s. c.) for four days, normalized glycemia with no changes in plasma insulin levels. Liver glucokinase, a key enzyme in the regulation of glucose storage in hepatocytes, whose gene expression is significantly decreased in alloxan-diabetic rodents, showed a recovery in its mRNA levels after Trecadrine administration. These data suggest that beta3-adrenergic agonists enhance glucose storage in liver, probably through a non-insulin dependent mechanism of action

Decreased anion exchanger 2 immunoreactivity in the liver of patients with primary biliary cirrhosis

Chloride-bicarbonate anion exchanger 2 (AE2) is expressed in a variety of tissues, including the liver and salivary glands, where it may participate in the generation of hydroionic fluxes into secretions. We have previously reported decreased hepatic levels of AE2 messenger RNA in patients with primary biliary cirrhosis (PBC), a cholestatic condition frequently associated with pluriglandular exocrine failure. Here we investigated the expression of AE2 protein in the liver of PBC patients. Using a monoclonal antibody against an AE2 peptide, immunohistochemistry was performed on liver biopsy specimens from subjects with normal liver (n = 7), patients with PBC (n = 13), and patients with cirrhosis or cholestasis other than PBC (n = 17 and 11, respectively). Immunostaining was graded from 0 to 7, according to its intensity and distribution. AE2 immunoreactivity was observed in normal livers, as previously reported, and in many pathological liver biopsy specimens, being mainly restricted to canaliculi and the luminal membrane of terminal and interlobular bile ducts. Canalicular and ductular scores were significantly reduced in the PBC group compared with each control group (normal liver and cirrhosis or cholestasis other than PBC), whereas no differences in immunoreactivity scores were observed among control groups. When four patients with primary sclerosing cholangitis (PSC) were analyzed, they also differed from those with PBC. These results suggest that PBC is characterized by diminished expression of AE2 in the liver. Reduced levels of this transporter protein might be involved in the pathogenesis of cholestasis in PBC