Interleukin 10 protects mice against staphylococcal enterotoxin B-induced lethal shock.

We investigated the ability of interleukin 10 (IL-10) to protect mice against lethal shock induced by staphylococcal enterotoxin B (SEB). Treatment of mice with IL-10 prevented the death of mice injected with SEB in a dose-dependent manner. IL-10-mediated protection was apparent when administered either prior to or concurrent with SEB but was less effective when administered following SEB injection. This observation indicates that IL-10 is capable of regulating T-cell activation in vivo.     

Protection against TNF-induced lethal shock by soluble guanylate cyclase inhibition requires functional inducible nitric oxide synthase

Hypotension and shock observed in sepsis, SIRS, and tumor necrosis factor (TNF) or cytokine-based cancer treatment are the consequence of excessive nitric oxide (NO) production and subsequent soluble guanylate cyclase (sGC)-mediated vascular smooth muscle relaxation. We demonstrate here that, while NO synthase (NOS) inhibitors exacerbated toxicity, inhibitors of sGC activation protected against TNF-induced lethality, bradycardia, and hypotension. Importantly, sGC inhibition did not interfere with the antitumor activity of TNF. Using NOS inhibitors or iNOS-deficient animals, we furthermore observed that no protection against TNF toxicity could be obtained in the absence of NO. These data imply that iNOS- (and not eNOS-) derived NO is an endogenous protective molecule indispensable to survive a TNF challenge and exerting this beneficial effect via sGC-independent mechanisms.     

Immunostimulatory properties of the Leishmania infantum heat shock proteins HSP70 and HSP83

Emerging evidence indicates that the heat shock proteins (HSPs), a set of highly evolutionary conserved proteins, are playing essential roles in both normal processes of the immune system and specific immune responses. In a previous work, we demonstrated that the Leishmania infantum HSP70 possesses remarkable immunostimulatory properties. In the present work, we have extended the study to another HSP from this parasite, the HSP83. We show that this protein also has an adjuvant effect to an accompanying protein by stimulation of the humoral response when both proteins are fused and co-administered to BALBjc mice. The analysis of the IgG isotypes, IgG1 and IgG2a, indicated that the immunisations with the Leishmania HSPs, mainly the HSP70, potentiate a Thl-type response. It was found that the amino-terminal domain of the HSP70, the most evolutionary conserved region of the molecule, maintains the ability to stimulate the humoral response, whereas the carboxyl-terminal domain does not have a similar effect. Unexpectedly, we found that the L. infantum HSP70 and HSP83 recombinant proteins stimulated the proliferation of spleen cells from unprimed BALB/c mice. Remarkably, this proliferation was abolished either by thermal denaturing of the proteins or by using specific antibodies. The use of the T-cell inhibitor cyclosporin A in the splenocytes proliferation assays suggested that both T- and non-T-cells are stimulated by the Leishmania HSPs. These findings may be relevant for therapeutic and prophylactic applications.     

Gelatinase B deficiency protects against endotoxin shock

Gelatinase B or matrix metalloproteinase-9 (MMP-9) is stored in the tertiary granules of polymorphonuclear leukocytes. These cells are key effectors in acute inflammatory diseases such as sepsis. Endotoxin leads to rapid release of gelatinase B from these granules in vitro and in vivo, but the role of this enzyme in bacterial sepsis and endotoxin shock remains unclear. We studied the clinical course of endotoxinemia and its relation with the expression of gelatinase B from the pool of circulating leukocytes in adult as well as in young mice in a model of endotoxin-induced shock and compared wild-type with gelatinase B-deficient mice. The gelatinase B-deficient mice were resistant to endotoxin shock, which implies that specific MMP-9 inhibition constitutes anapproach for the treatment of septic shock syndromes.     

Gene delivery of SOCS3 protects mice from lethal endotoxic shock

Suppressor of cytokine signaling 3 (SOCS3) was reported as a feedback inhibitor of cytokine receptor signaling byinhibiting the JAK-STAT signal transduction pathway.We sought to test the anti-endotoxic septic shock effect ofliposome mediated gene delivery of SOCS3 in a lethal endotoxic shock mouse model.BALB/c mice were injectedintraperitoneally with 200 μg pcDNA3.1-SOCS3 cationic liposomes,while pcDNA3.1-IL-10 and empty vector aspositive and negative control respectively.Forty-eight hours after gene delivery,mice were challenged with 4 μg ofE.coli 0127:B8 LPS and 18 mg D-GaIN administered i.p.90 min later,serum TNF-α level was determined.Survivalover the next 48h was evaluated.Peritoneal macrophages from survival mice were stimulated in vitro with 1 μg/mlLPS for 18h,and the supernatants were harvested for determination of the amount of TNF-α.We found that genedelivery of SOCS3 significantly increase the mouse survival rate from 27.8±9.6% of control group to 61.1±9.6%(p<0.01).In comparison with control group (218±13pg/ml) and sham delivery group (219±22pg/ml),genedelivery of SOCS3 reduced the level of serum TNF-α(68±9pg/ml) significantly(p<0.01).Furthermore,genedelivery of SOCS3 displayed the capacity of prevention of tolerance of peritoneal macrophages to LPS.Thesefindings suggest that gene delivery of SOCS3 mediated by liposome is a promising approach for endotoxic septicshock treatment.Cellular & Molecular Immunology.2005;2(5):373-377.     

榄香烯复合瘤苗HSP70与HSP70BCG对巨噬细胞功能影响的比较

目的 :比较Hca F榄香烯复合瘤苗HSP70 (HSP70 HTCV)和卡介苗HSP70 (HSP70 BCG)对小鼠腹腔或脾脏巨噬细胞功能的影响 ,分析HSP70 HTCV诱导抗瘤免疫作用的机制。方法 :给正常BALB C小鼠腹腔注射HSP70 HTCV 或HSP70 BCG ,共 3次 ,并用多聚甲醛固定的Hca F细胞体内冲击。收集腹腔和脾脏的巨噬细胞 ,并用HSP70 HTCV或HSP70 BCG 体外再致敏 ,用MTT法测细胞毒活性、细胞的增殖和分泌TNF的能力 ,用中性红吞噬试验测巨噬细胞的吞噬能力。结果 :用HSP70 HTCV 免疫小鼠的腹腔巨噬细胞对Hca F的细胞毒活性强于用HSP70 BCG 免疫小鼠的腹腔巨噬细胞 (4 2 7%VS 31 0 % ,P <0 0 5 ) ;用HPS70 HTCV 免疫小鼠的脾脏巨噬细胞分泌TNF的能力高于用HSP70 BCG免疫小鼠的脾脏巨噬细胞 ,细胞培养上清对L92 9的细胞毒活性分别为 5 7 4 %和 35 9% (P <0 0 5 ) ,脾脏巨噬细胞吞噬中性红的能力的变化两组之间无明显差别 ;免疫巨噬细胞的上述功能均高于未经免疫的对照小鼠的巨噬细胞 (P <0 0 1) ,而三组巨噬细胞的增殖改变无明显差别 (P >0 0 5 )。结论 :HSP70 HTCV 或HSP70 BCG 免疫都能增强巨噬细胞功能 ,但HSP70 HTCV免疫诱导的巨噬细胞对肿瘤细胞有更强的杀伤活性     

Nitrosonisoldipine is a selective inhibitor of inflammatory caspases and protects against pyroptosis and related septic shock

Pyroptosis is a type of acute cell death that mainly occurs in immune cells. It is characterized with robust release of inflammatory cytokines and has emerged to play a critical role in the pathogenesis of sepsis-associated immune disorders. In this study, we screened for pyroptotic inhibitors with the ultimate goal to benefit sepsis treatments. Accidentally, we identified that nitrosonisoldipine (NTS), a photodegradation product of calcium channel inhibitor nisoldipine, inhibits noncanonical pyroptosis. Using murine immortalized BM-derived macrophage and human THP-1 cell line, we further discovered that NTS not only inhibits noncanonical pyroptosis mediated by caspase-11 or caspase-4 but also canonical pyroptosis mediated by caspase-1. Mechanistically, NTS directly inhibits the enzyme activities of these inflammatory caspases, and these inhibitory effects persist despite extensive washout of the drug. By contrast, apoptosis mediated by caspase-3/-7 was not affected by NTS. Mice pretreated with NTS intraperitoneally displayed improved survival rate and extended survival time in LPS- and polymicrobe-induced septic models, respectively. In conclusion, NTS is a selective inhibitor of inflammatory caspases that blocks both the noncanonical and canonical pyroptotic pathways. It is safe for intraperitoneal administration and might be used as a prototype to develop drugs for sepsis treatments.     

Heat shock proteins HSP90, HSP70 and GRP78 expression in medullary thyroid carcinoma

BackgroundMedullary thyroid carcinoma management consists mainly of surgical resection and is largely chemoresistant. There is ongoing effort to discover novel therapies for medullary thyroid carcinoma. Increased levels of heat shock proteins have been associated with multiple cancers and are being studied as potential therapeutic targets. The purpose of this study was to determine the expression levels of heat shock proteins 90 and 70 and of glucose related protein 78 in medullary thyroid carcinoma tissues compared with normal thyroid tissues.Methods20 tissue specimens of medullary thyroid carcinoma and 10 specimens of thyroids without malignancy were analyzed by immunohistochemistry.ResultsMedullary thyroid carcinoma specimens showed 27% higher expression level of heat shock protein 90 immunostaining, and a 43% higher expression level of heat shock protein 70 immunostaining versus normal controls. These differences, however, were not statistically significant. A significantly higher expression level was noted for glucose related protein 78 in the medullary thyroid carcinoma specimens than in the controls.ConclusionThis study indicates increased expression levels of heat shock proteins 90 and 70 and glucose related protein 78 levels in medullary thyroid carcinoma. These findings, though preliminary imply that these proteins may have a role in medullary thyroid carcinoma's tumor biology and may have and future therapeutic options. Larger cohorts are needed to corroborate these results.     

Pyrrolidine dithiocarbamate protects mice from lethal shock induced by LPS or TNF-alpha.

Although important advances have been made in the development of antibiotics and medical intensive care technology in recent years, systemic response to infection remains a major health problem, with growing incidence and high mortality rates. Here we demonstrate the ability of the antioxidant agent pyrrolidine dithiocarbamate (PDTC) to inhibit the in vivo activation of NF-kappaB in lung and liver tissues, as well as the systemic release of TNF-alpha in lipopolysaccharide (LPS)-treated mice. The in vivo effect of PDTC on NF-kappaB activation in liver tissues involved the inhibition of both LPS-induced I kappaB-alpha degradation and the translocation of the p50 and p65 NF-kappaB subunits to the nucleus. In addition to protecting mice against lethal LPS doses, PDTC curtailed TNF-alpha-induced lethal shock. This effect was observed even after LPS injection, and when PDTC was administered at a time when TNF-alpha was already at maximum levels in serum. PDTC-treated mice survived despite high IL-1beta and IL-6 levels, induction of VCAM-1 and ICAM-1 expression or leukocyte infiltration in tissues known to be associated with LPS-induced shock, indicating that PDTC does not act by modifying these responses. Taken together, these results indicate that PDTC interferes with the production as well as the action of TNF-alpha, and points to a possible approach toward the treatment of septic shock.     

Thermal adaptation activates HSP70 synthesis, inhibits overproduction of nitric oxide, and protects the body from acute hypotension during heat shock

The role of HSP70 and nitric oxide in antihypotensive effects of thermal adaptation was studied. Western blot analysis and electron paramagnetic resonance were used to determine the contents of HSP70 and nitric oxide. Protective effect of adaptation was evaluated by the limitation of blood pressure drop after heat shock. The formation of protective effects, accumulation of HSP70, and development of the ability to decrease nitric oxide overproduction had similar dynamic patterns and appeared at the same period. Quercetin, an inhibitor of HSP70 synthesis, prevented the development of protective effects. The data suggest that HSP70 accumulated during adaptation prevents heat shock-induced hypotension by restricting NO over-production and interfering with its cytotoxic effects. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 11, pp. 507–510, November, 1999     

Detection of heat shock protein 70 (HSP70) and anti-HSP70 antibodies in the serum of normal individuals

Heat shock or stress proteins (Hsp) are typically regarded as being intracellular proteins that have a range of functions including the maintenance of cellular integrity. Members of the Hsp70 family of molecules have been implicated in the processing and presentation of antigen and the cross reactivity of lymphocytes specific for pathogen-derived heat shock proteins with self Hsp70 has been suggested to be an underlying cause of certain autoimmune diseases. This study reports the presence of soluble Hsp70 in the peripheral circulation of normal individuals. Concentrations of soluble Hsp70 in females were approximately twice those in males. Circulating anti-Hsp70 antibodies were detected in all individuals assessed, but there were no differences between males and females. However, there was a significant correlation between soluble Hsp70 concentration and antibody levels in males, but not females. The physiological role for circulating heat shock proteins is intriguing, but currently unknown. These findings extend our previous observations that Hsp60 is present in the peripheral circulation and support the proposition that soluble heat shock proteins may play a regulatory role in either the prevention or protection of pathophysiological processes involving inadvertent immunorecognition or cross-recognition of heat shock proteins.     

Detection of heat shock protein 70 (HSP70) and anti-HSP70 antibodies in the serum of normal individuals

Heat shock or stress proteins (Hsp) are typically regarded as being intracellular proteins that have a range of functions including the maintenance of cellular integrity. Members of the Hsp70 family of molecules have been implicated in the processing and presentation of antigen and the cross reactivity of lymphocytes specific for pathogen-derived heat shock proteins with self Hsp70 has been suggested to be an underlying cause of certain autoimmune diseases. This study reports the presence of soluble Hsp70 in the peripheral circulation of normal individuals. Concentrations of soluble Hsp70 in females were approximately twice those in males. Circulating anti-Hsp70 antibodies were detected in all individuals assessed, but there were no differences between males and females. However, there was a significant correlation between soluble Hsp70 concentration and antibody levels in males, but not females. The physiological role for circulating heat shock proteins is intriguing, but currently unknown. These findings extend our previous observations that Hsp60 is present in the peripheral circulation and support the proposition that soluble heat shock proteins may play a regulatory role in either the prevention or protection of pathophysiological processes involving inadvertent immunorecognition or cross-recognition of heat shock proteins.     

A rabies-specific human monoclonal antibody that protects mice against lethal rabies.

According to a recommendation from WHO (World Health Organisation) for prevention of a possible rabies infection, active vaccination has to be combined with application of immunoglobulin to get a fast protective effect. At present, preparations of purified human or equine rabies-specific immunoglobulin are used. We have generated a human rabies-specific monoclonal antibody (huMAb) by immortalization of human B-cells with Epstein Barr Virus (EBV), followed by fusion with a mouse myeloma cell. The resulting clone TW-1 secrets an IgG1 lambda huMAb which specifically reacts in ELISA with 5 laboratory rabies virus strains of serotype 1 and DUV3 (Duvenhage, serotype 4). Western Blot analysis revealed fine specificity for the G glycoprotein (gp67) of rabies virus. HuMAb TW-1 neutralizes rabies virus in vitro (RFFIT) as well as in vivo and protects rabies infected mice. Compared to polyclonal human rabies immunoglobulins, huMAb TW-1 is advantageous, because of its defined specificity and the very low amounts of total protein needed for therapeutic effects.     

Monoclonal antibody to fungal glucosylceramide protects mice against lethal Cryptococcus neoformans infection

Glucosylceramides (GlcCer) are involved in the regulation of Cryptococcus neoformans virulence. In the present study, we demonstrate that passive immunization with a monoclonal antibody to GlcCer significantly reduces host inflammation and prolongs the survival of mice lethally infected with C. neoformans, revealing a potential therapeutic strategy to control cryptococcosis.     

Glycine protects cardiomyocytes against lethal reoxygenation injury by inhibiting mitochondrial permeability transition

Post-ischaemic reperfusion may precipitate cardiomyocyte death upon correction of intracellular acidosis due in part to mitochondrial permeability transition. We investigated whether glycine, an amino acid with poorly understood cytoprotective properties, may interfere with this mechanism. In cardiomyocyte cultures, addition of glycine during re-energization following 1 h of simulated ischaemia (NaCN/2-deoxyglucose, pH 6.4) completely prevented necrotic cell death associated with pH normalization. Glycine also protected against cell death associated with pH normalization in reoxygenated rat hearts. Glycine prevented cyclosporin-sensitive swelling and calcein release associated with re-energization in rat heart mitochondria submitted to simulated ischaemia or to Ca²⁺ stress under normoxia. NMR spectroscopy revealed a marked glycine depletion in re-energized cardiomyocytes that was reversed by exposure to 3 m m glycine. These results suggest that intracellular glycine exerts a previously unrecognized inhibition on mitochondrial permeability transition in cardiac myocytes, and that intracellular glycine depletion during myocardial hypoxia/reoxygenation makes the cell more vulnerable to necrotic death.     

Association of heat shock protein70-2 (HSP70-2) gene polymorphism with obesity

Abstract

Background: Obesity is a major risk factor of chronic-diseases, including cardiovascular-diseases (CVD). Increasing evidence is showing the association of heat-shock protein (HSP) with type-2 diabetes and CVD; however, there is little data on the relationship between the genetic-polymorphisms of HSP70-2 with obesity.

Aim: The present study has investigated the association between 1267HSP70-2 genetic polymorphism and obesity in an Iranian population with 317 subjects.

Subjects and methods: Anthropometric parameters and biochemical measurements were measured in all the samples, while genotypes were determined using PCR-RFLP. Univariate/multivariate analyses were conducted to explore the relationship between the genetic-polymorphisms and obesity.

Results: The data showed a significant association between 1267HSP70-2 polymorphism in obese subjects, compared to the non-obese group. Moreover, it was observed that this polymorphism was associated with obesity in the CAD?+?group, which had a high BMI compared to non-obese controls.

Conclusion: The 1267HSP70-2 polymorphism is associated with obesity in an Iranian population, supporting a possible potential genetic link between obesity and cardiovascular diseases.     

Adaptogen ADAPT modulates stress-induced HSP70 synthesis and improves organism's resistance to heat shock

The effect of the herbal adaptogen ADAPT on the basal and heat shock-induced synthesis of HSP70 and organism's resistance to heat shock is studied. It is shown that ADAPT decreases accumulation of these proteins in the myocardium and promotes their accumulation in the liver. ADAPT has no effect on arterial pressure but prevents its drop induced by heat shock and markedly increases the survival rate of experimental animals. These data agree with the hypothesis that the effect of ADAPT on the synthesis of HSP70 is an important component of its protective action. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 123, No. 6, pp. 629–631, June, 1997     

Heat shock protein 70 (HSP70) expression is associated with poor prognosis in intestinal type gastric cancer

Heat shock protein 70 (HSP70) is a molecular chaperone which plays an important role in cellular protection against various stressful stimuli and in the regulation of cellular growth and apoptosis. This study was conducted in gastric carcinoma (GC) to assess correlations of HSP70 expression with clinicopathological parameters and overall survival (OS). Tissue microarray blocks were constructed from 172 GCs and immunohistochemically stained for HSP70. Low HSP70 expression was found in 122 GCs (71 %), whereas 50 (29 %) had high expression. HSP70 expression was higher in tumours in the cardia (p?=?0.008), with non-signet ring cell histology (p?<?0.001), of intestinal type (p?=?0.045) and of higher pathological T stage (p?=?0.026). When considering the cohort as a whole, HSP70 expression did not correlate with OS (p?=?0.092). In intestinal type carcinomas, however, high HSP70 expression significantly correlated with worse OS (p?=?0.034). These results suggest that HSP70 expression might be an unfavourable prognostic factor in patients with GC, especially of intestinal type.     

Primary adult lactose intolerance protects against development of inflammatory bowel disease

We have compared the incidence of inflammatory bowel disease (IBD) to the prevalence of lactose malabsorption (LM) in several countries. Our observations indicate that IBD is rare where LM is highly prevalent. The correlation between incidence of Crohn's disease and LM is -0.93, p less than 0.01, the correlation between incidence of ulcerative colitis and LM is -0.89, p less than 0.01. We, therefore, propose that LM results in the formation of volatile fatty acids which may inhibit multiplication of potentially pathogenic organisms.