Heat shock protein 60 (HSP60) immunoreactivity in gastric epithelium associated with Helicobacter pylori infection: a pitfall in immunohistochemically interpreting HSP60-mediated autoimmune responses

Previous studies have suggested that heat shock proteins (HSP) of Helicobacter pylori (H. pylori) are involved in the induction of autoimmunity mediated gastritis. In the present report, the cross-reactivity between H. pylori-related HSP60 and gastric epithelial cells was investigated by the indirect immunoperoxidase method using two monoclonal antibodies (mAb) against H. pylori-derived HSP60, H9 and H20. H9 is reactive with an epitope common to bacterial HSP60, while H20 is specific to H. pylori HSP60. A total of 70 paraffin-embedded gastric biopsy specimens were analyzed after heat-induced epitope retrieval. Both mAb were cross-reactive with the gastric epithelial cells, with a higher frequency seen for the H9-reactive epitope. The frequency of positive epithelial decoration was not significantly different between H. pylori-positive and H. pylori-negative gastric mucosae. A variety of epithelial and non-epithelial cells were immunostained with mAb H9, while mAb H20 was cross-reactive only with small intestinal epithelia. Reactivity was mainly located in the Golgi area and rarely in the cytoplasm. These results suggest a noteworthy pitfall in immunohistochemical interpretations of HSP60-associated autoimmune reactions in the gastric mucosa.     

Induction of heat shock proteins in lymphocytes increases with mitogen stimulation

The effect of the growth state of a cell on the ability of hyperthermia to induce the synthesis of heat shock proteins (HSPs) was studied in resting and concanavalin A (ConA)-stimulated lymphocytes. Hyperthermia induced the synthesis of hsp 110, hsp 90, hsc 70, and hsp 70 in both resting and ConA-stimulated lymhocytes, and ConA-treatment induced the synthesis of the hsp 90 and hsc 70 at normal temperature. The induction of the synthesis of hsp 110 and hsp 70 by hyperthermia was 3- to 6-fold higher for lymphocytes cultured with ConA for 12 and 24 h than in non-stimulated lymphocytes. Thus, lymphocytes induced to undergo proliferation showed a greater response to hyperthermia than resting lymphocytes.     

LHRH messenger RNA in neurons in the intact and castrate male rat forebrain,studied by in situ hybridization

Summary The slow potential change (spc) accompanying spreading depression (SD) was studied in rats and in a seizure-sensitive strain of Mongolian gerbil under three different experimental paradigms, each involving the use of naloxone. Gerbils undergoing electroconvulsive shock treatment displayed SD during the post-ictal phase, which was blocked by the intraperitoneal (i.p.) administration of naloxone (20–50 mg kg^-1). Topical application of naloxone to the exposed cortex of the anaesthetized gerbil and rat blocked the spc of SD evoked by KCl. Microiontophoretic ejection of naloxone during extracellular recordings reversed cell refractoriness following the spc, demonstrated by the observation of a maintained sensitivity to iontophoretic pulses of glutamate. The results suggest a possible involvement of naloxone-sensitive processes in the mechanism responsible for cortical SD.     

热休克蛋白60对小鼠树突状细胞功能影响体外研究

目的：探讨动脉粥样硬化中重要炎性物质——热休克蛋白60（HSP60）体外对小鼠树突状细胞（mDC）功能影响.方法：小鼠骨髓提取DC,体外培养成熟后与两种浓度mHSP60孵育,动态观察DC突起改变;流式细胞仪检测孵育前后mDC表面标志改变;MLR测定孵育前后mDC刺激功能变化;ELISA法测定MLR上清液中细胞因子浓度.结果：孵育后,mDC突起增加明显;CD11c^＋、CD80及CD86表型显著增加;淋巴细胞刺激功能明显增强;分泌细胞因子IL-12、IFN-γ增加（P＜0.01）而IL-4增加不明显（P＞0.05）,IFN-γ/IL-4比值升高.结论：mHSP60体外可以促进mDC功能,作用呈剂量依赖性.     

ESWL实际焦点位置的理论和数值分析

从理论和数值模拟两个方面分析了ESWL中的非线性的动力学焦点和实际焦点的位置 ,给出三种典型的动力学焦点的数值结果 ,说明非线性动力学焦点和实际焦点一般不是几何焦点 ,指出正确确定实际焦点在ESWL技术中的重要性。     

Heat shock proteins HSP25, HSP60, HSP72, HSP73 in isoosmotic cortex and hyperosmotic medulla of rat kidney

The distribution of heat shock proteins (HSP) HSP60, HSP73, HSP72 and HSP25 in the isoosmotic cortex and the hyperosmotic medulla of the rat kidney was investigated using Western blot analysis and immunohistochemistry. HSP73 was homogeneously distributed throughout the whole kidney. The level of HSP60 was high in the renal cortex and low in the medulla. HSP25 and HSP72 were present in large amounts in the medulla. Only low levels of HSP25 and almost undetectable amounts of HSP72 were found in the cortex. HSP25 exists in one nonphosphorylated and several phosphorylated isoforms. Western blot analysis preceded by isoelectric focussing showed that HSP25 predominates in its nonphosphorylated form in the outer medulla but in its phosphorylated form in cortex and inner medulla. Although this intrarenal distribution pattern was not changed during prolonged anaesthesia (thiobutabarbital sodium), a shift from the nonphosphorylated to the phosphorylated isoforms of HSP25 occurred in the medulla. The characteristic intrarenal distribution of the constitutively expressed HSPs (HSP73, HSP60, HSP25) may reflect different states of metabolic activity in the isoosmotic (cortex) and hyperosmotic (medulla) zones of the kidney. The high content of inducible HSP72 in the medulla most likely is a consequence of the osmotic stress imposed upon the cells by the high urea and salt concentrations in the hyperosmotic medullary environment.     

Prevention of endotoxin shock by an antibody against leukocyte integrin {beta}2 through inhibiting production and action of TNF

Septic shock remains a serious disorder associated with highmortality. Accumulating evidence indicates that TNF is a majorand essential mediator of endotoxin shock. We report here thatadministration of an antibody against CD18 dramatically reducedendotoxin-induced shock inrabbits as revealed by preventionof severe hypotension, metabolic acidosis and a pathologicalchange suggestive of disseminated intravascular coagulationwith concomitant inhibition of elevation of plasma TNF activity.The anti-CD18 antibody also inhibited the hypotension inducedby administering recombinant TNF. Furthermore, an antibody againsta ligand for CD18 complexes, intercellular adhesion molecule-1,also prevented TNF-induced shock as well as endotoxin shockinrabbits. These observations suggest that adhesion of leukocytesto endothelium may be of primary importance in the action ofTNF as well as in the production of TNF in vivo and that theantibody against adhesion molecules could be of therapeuticbenefit in life-threatening septic shock in humans.     

空肠弯曲菌HSP43诱导自身免疫机理──分子模拟和抗原优势

我们曾发现空肠弯曲菌４３ｋＤ热休克蛋白（ＨＳＰ４３）能诱导小鼠产生自身免疫应答，本文则进一步分析了这种诱导作用的机理。二株针对ＨＳＰ６０保守区序列的单克隆抗体ＩＩＨ９和ＭＬ－３０不但能结合人及小鼠细胞中ＨＳＰ６０家族成员，而且也能结合ＨＳＰ４３，表明ＨＳＰ４３、人及小鼠ＨＳＰ６０三者均属同一家族成员，具有序列上的高度同源性。小鼠用空肠弯曲菌免疫后出现了针对１０种菌体蛋白的抗体，其中最早被诱导的抗体是针对ＨＳＰ４３的，并且该种抗体在随后８０ｄ观察期间内保持了较高活性，表明ＨＳＰ４３是优势抗原。本文结果提示，ＨＳＰ４３较易被免疫系统识别而产生应答，通过和宿主ＨＳＰ６０高度序列同源性而可能呈现分子模拟，从而诱导自身免疫损伤。     

Peripheral blood mononuclear cell proliferative responses to soluble and particulate heat shock protein 65 in health and inflammatory bowel disease

Objectives: The aims of this study were to determine, in peripheral blood mononuclear cells (PBMC), whether particulate antigen triggers (i) an amplified cell proliferative response compared to soluble antigen and (ii) a dysfunctional response in cells derived from patients with chronic inflammation and specifically in those with inflammatory bowel disease (IBD). Subjects: Healthy volunteers (n = 17), inflammatory controls (n = 8) and patients with IBD (n = 17) were recruited from St Thomas’ and Guys’ Hospital, London, UK. Methods: Following optimisation of experimental conditions (0.1–10.0 μg/ml antigen), PBMC were stimulated with (i) 10.0 μg/ml recombinant soluble heat shock protein 65 (hsp 65) and (ii) 1.0 and 10.0 μg/ml hsp 65 conjugated to microparticles (0.5 μm diameter). PBMC proliferative responses were measured by ³H-Thymidine incorporation at day 5 and results compared between groups using unpaired t-test. Results: Conjugation to microparticles of low dose hsp 65 significantly increased overall proliferative responses by 2–11 fold compared to soluble antigen alone (p < 0.05). However, no specific PBMC proliferative dysregulation was noted in cells from subjects with IBD. Conclusions: Low dose antigen, in microparticulate form, leads to amplified cell proliferation in primary human cells, as showed previously in cell lines and animal studies. However there is no abnormal proliferative response in cells from subjects with IBD. Received 8 February 2006; returned for revision 7 March 2006; accepted by G. Wallace 25 October 2006     

An Evaluation of Finger Pulse Volume as a Psychophysiological Measure of Anxiety

The present experiment explored the utility of finger pulse volume (FPV) as a measure of anxiety. Subjects were exposed to either a threatening or nonthreatening situation, and indices of physiological arousal (pulse rate (PR) and FPV) and self-report of anxiety (Affect Adjective Checklist (AACL)) were collected. Results indicated that FPV was responsive to changes in experimentally induced anxiety and significantly correlated with PR and AACL, although the strength of these relationships was not substantial. Relevance for psychophysiological theory and the clinical observation of anxiety is discussed.