A synthetic 10-kD heat shock protein (hsp10) from Mycobacterium tuberculosis modulates adjuvant arthritis

The heat shock protein, hsp10, is an abundant protein in Mycobacterium tuberculosis (Mtb), its nucleotide sequence encoding a protein of 99 amino acids with a molecular mass of 10±7kD. This sequence is phylogenetically conserved, being represented by the GroES homologue of Escherichia coli. Hsp 10 and GroES are members of the chaperonin 10 family of molecular chaperones, and GroES is necessary for the optimal activity of GroEL, a member of the chaperonin 60 family and the E coli homologue of mycobacterial hsp65. Since hsp65 has been implicated in both experimental and human rheumatoid arthritis, we aimed to assess the immunomodulatory effects of its co-chaperonin, hsp10, in experimental arthritis. Our results show that an aqueous solution of a mycobacterial hsp10 delayed the onset and severity of adjuvant-induced arthritis in rodents when administered after disease induction but before joint involvement occurred. This biological activity was specific for the hsp10 of Mtb, since neither GroES nor the rat homologue was effective. Using synthetic hsp10 fragments, the activity was localized to the N-terminal region of the molecule. Assessment of circulating antibody levels to mycobacterial hsp10 and hsp65 indicated that all arthritic rats had increased litres to both hsp10 and hsp65: hsp10-treated rats showed further elevation of this humoral response not only to hsp10 but also to hsp65 when compared with the untreated arthritic control. This is the first report of the immunomodulatory activity of mycobacterial hsp10 in experimental arthritis, and exhibits a potential role for this co-chaperonin in pathophysiological situations.     

Egr-1基因剔除减少炎性相关因子在实验性胰腺炎中的表达

目的：探讨Egr-1基因剔除对实验性胰腺炎小鼠胰腺组织中炎性相关因子表达的影响。 〖HTH〗方法：利用Egr-1基因剔除小鼠，采用大剂量雨蛙素诱导的实验性胰腺炎模型，观察Egr-1基因剔除后，胰腺组织水肿、MPO水平、血清淀粉酶水平、肺组织MPO水平的改变。并利用定量PCR的方法，检测胰腺组织中炎症相关因子组织因子（TF）、纤维蛋白溶酶原激活因子抑制因子（PAI-1）、单核细胞趋化吸引蛋白1（MCP-1）、Gro-1、IL-6和细胞间黏附分子-1（ICAM-1） mRNA的表达。 〖HTH〗结果：Egr-1基因剔除小鼠胰腺组织水肿明显轻于野生型，但组织MPO水平与血清淀粉酶与野生型组间相比无明显差异；肺组织MPO水平明显低于野生型。定量PCR检测结果表明， Egr-1基因剔除组，胰腺组织中TF、PAI-1，以及MCP-1、ICAM-1和IL-6　mRNA的表达，明显少于野生型组。 〖HTH〗结论：Egr-1基因剔除可明显减轻急性胰腺炎的严重程度，其作用可能通过减少胰腺组织中TF、PAI-1，以及MCP-1、ICAM-1和IL-6的表达而实现。     

中药“神经再生素”促神经生长过程中基因差异性表达的初步研究

为了探索中药“神经再生素”促神经生长过程中基因水平的变化。本实验采用 dd-PCR方法 ,从体外培养背根神经节细胞加药组和不加药组中获得两者的差异表达片段 ,并经反杂交筛选、克隆测序、DNA序列检索分析、Northern验证。结果表明 ,差异显示共获得 8个差异条带 ,一个为下调基因 ,其余为上调基因 ,其中 2个 c DNA序列与 RRAJ5 16 1基因 (增殖相关基因 )、AF196 3 15基因 (锌指样蛋白 DDP2 ) 10 0 %同源 ,2个 c DNA序列与 AK0 0 175 7基因、STA5 SRR基因 (t RNA合成酶 )部分同源。结论是中药“神经再生素”在促神经生长过程中 ,对神经元基因的选择性表达起着重要的调控作用。     

Roles of tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), and IL-10 in the modulation of intercellular adhesion molecule-1 (ICAM-1) expression by macrophages during mycobacterial infection

Profiles of ICAM-1 expression on cultured murine peritoneal macrophages infected with Mycobacterium avium complex (MAC) were examined, with special reference to modulating roles of TNF-alpha, TGF-beta, and IL-10. When macrophages were infected with MAC, ICAM-1 expression, measured by microscopic counting of ICAM-1+ macrophages stained with anti-ICAM-1 antibody, ELISA, and flow cytometric analysis, was rapidly increased, peaking at day 3 (early-phase up-regulation) due to endogenous TNF-alpha, and thereafter gradually declined to the normal level within 1 week or more (late-phase down-regulation). The late-phase ICAM-1 down-regulation was also seen in macrophages phagocytosing heat-killed MAC and those stimulated with lipopolysaccharide but not in macrophages phagocytosing latex beads. ICAM-1 mRNA expression was augmented markedly at day 1 after MAC infection and thereafter decreased. While TNF-alpha and IL-10 production by MAC-infected macrophages was observed during the first 3 days, TGF-beta production was initiated from day 3 and continued until day 14. Exogenously added TGF-beta strongly inhibited the early-phase increase in ICAM-1 expression by infected macrophages, and the blockade of endogenous TGF-beta with anti-TGF-beta antibody markedly inhibited late-phase ICAM-1 down-regulation. Moderate blocking effect was also observed for anti-IL-10 antibody. On the other hand, late-phase ICAM-1 down-regulation was not prevented by the addition of exogenous TNF-alpha. Therefore, TGF-beta and IL-10, especially the former, appear to play active roles in the late-phase down-regulation of ICAM-1 in MAC-infected macrophages during long-term cultivation.     

Production of IL-6 by T cells from the femoral head of patients with rapidly destructive coxopathy (RDC)

RDC is a syndrome with unknown etiology that causes rapid destruction of a hip joint. We have investigated the production of osteoclast-activating cytokines (IL-6, IL-1α and tumour necrosis factor-alpha (TNF-α)), interferon-gamma (IFN-γ) and IL-8 by T cells in the affected joint. The level of IL-6 produced by the T cell lines (TCL) established from the femoral head was significantly higher than that from patients' or healthy donors' peripheral blood mononuclear cells (PBMC). IL-6 production by the TCL from synovial membrane or from patients' PBMC was also significantly higher than that from healthy donors' PBMC. IL-1α production by the TCL from the femoral head was significantly higher than any of the other groups when all the TCL were used for the analysis. TNF-α production was highest in the TCL from patients' PBMC. The levels of IFN-γ or IL-8 were not significantly different among these four groups. The plasma levels of all these cytokines except for IFN-γ, that was rather lower, in RDC patients were not significantly different from those in osteoarthrosis or trauma patients, or healthy donors. These results suggest that T cells at the affected femoral head, and also synovial membrane to some extent, are involved in bone resorption through the production of IL-6 and probably IL-1α in patients with RDC.     

Immunopathological features of palatine tonsil characteristic of IgA nephropathy: IgA1 localization in follicular dendritic cells

IgA nephropathy (IgAN) is generally thought to be mediated by the glomerular deposition of circulating immune complexes containing IgA as the major antibody component. Upper respiratory infections and tonsillitis often precede IgAN. and in some cases tonsillectomy is affective for the (treatment of IgAN. Thus, the tonsil seems to be a unique organ causing initial and/or progressive events to generate nephritogenic immune complexes in IgAN. in this study we focused on the analysis of immunopathological features of the palatine tonsil characteristic of IgAN patients by using an immunohistochemical technique. The IgAl subclass was demonstrated in follicular dendritic cells (FDC) of the tonsil of IgAN patients, but not in FDC of non-IgAN controls. On the other hand, IgA2, IgG, IgM and C3 did not show any differences in distribution between the two groups. Moreover, the expression of decay-accelerating factor (DAF), an inhibitor of homologous complement activation, and transforming growth factor-beta I (TGF-/β1). an inducer of antibody-producing ceils to IgA class switching, in FDC and interdigitating dendritic cells of the tonsil, respectively, which was also clarified in this study for the first time, was found to be identically distributed in the two groups. These findings may support the idea that IgA1. possibly in an immune complex form, is trapped by FDC and plays an important role in the persistent activation of particular B cell repertoires responsible for ihe onset and/or progression of IgAN.     

Cloned cellular reagents to define antigens encoded between HLA-DR and glyoxylase

Primed lymphocyte typing reagents have been used to define antigens encoded by genes of a locus (loci) mapping between HLA-DR and glyoxalase I. This locus, which we shall refer to as the third locus of the HLA-D region, has been variously referred to as D beta, PL beta, PL3, and SB. Generating discriminatory primed lymphocyte typing reagents which can be used to define these antigens, however, has been extremely difficult. Donors of responding and stimulating cells for the priming combinations have usually been matched not only for the DR, D, and MB/MT antigens but also for the HLA-A, -B, and -C antigens. Even under these very restricted conditions, not all bulk primed lymphocyte typing reagents that are generated are discriminatory enough to be useful for antigen definition. We have derived "clones" from bulk priming combinations in which stimulator and responder differed for known antigens of this third locus. Even though the bulk reagents that were prepared did not provide discriminatory results, approximately 7-12% of the clones derived from the bulk priming combination proved to be highly discriminatory. We have been able to obtain these results with regard to all three antigens of the third locus so far evaluated. The very ease of screening clones and deriving discriminatory reagents, as compared with screening responder-stimulator combinations, allows the ready derivation of cellular reagents that define the antigens of this third locus.     

糖尿病大鼠肾小管TGF-β1和MAPK1/3表达的动态观察

目的:动态观察糖尿病大鼠肾小管转化生长因子-β₁(TGF-β₁)、丝裂原活化蛋白激酶(MAPK_1/3)和纤维连接蛋白(FN)的变化,探讨其在肾小管间质病变发生发展中的作用。方法:将大鼠分为正常对照组;糖尿病1周、2周、4周和8周组。用链脲菌素复制糖尿病模型;免疫组化法检测肾小管间质TGF-β₁、MAPK_1/3和FN的表达;Western blot检测TGF-β₁蛋白质;HE和PAS染色,光镜观察动物肾组织形态;生化法测定血糖、血肌酐及尿蛋白。结果: 正常肾小管可见少量MAPK_1/3表达,而未见TGF-β₁表达。糖尿病1周可见肾小管上皮细胞表达TGF-β₁,并且随着病程发展而增加。MAPK_1/3和FN从糖尿病两周开始表达亦呈持续增加,并且与TGF-β₁及肾重/体重比呈正相关。糖尿病1周时MAPK_1/3与TGF-β₁无显著相关,但与FN呈正相关。 结论:大鼠糖尿病状态诱导肾小管表达TGF-β₁并激活MAPK_1/3,MAPK_1/3介导高糖和TGF-β₁的信号而促进FN的生成和沉积,在糖尿病肾脏肥大和纤维化中可能起重要作用。     

G-CSF对外周血树突状细胞亚群比例的影响

目的 :了解G CSF对小鼠外周血树突状细胞 (DC)亚群比例的影响。方法 :以不同剂量 (0、5 .0、10 .0或 15 .0 μg)的G CSF ,分别皮下注射于BALB/c小鼠 ,连续 6d ,每天 1次。第 6天分离外周血DC ,用流式细胞仪检测DC1(CD11c+ CD8a-)和DC2 (CD11c+ CD8a+ )的比例并计算其绝对值。结果 :经不同剂量的G CSF刺激后 ,外周血DC1的绝对值无显著变化 ,而DC2的绝对值增加 5倍 (9.6× 10 6/Lvs 5 5 .1× 10 6/L ,P <0 .0 1) ,DC1/DC2比值降低 (4.2vs 0 .7,P <0 .0 1)。结论 :G CSF在体内可提高外周血DC2的绝对数 ,对DC1的数无明显影响 ,从而使DC1/DC2的比例倒置