脓毒症患者应激性高血糖与免疫功能变化及预后的关系

目的： 探讨应激性高血糖在脓毒症患者病情评估和预后预测中的作用。方法:44例脓毒症患者根据入院24 h内血糖监测结果和既往病史分为3组：应激性高血糖组(SH, n=15)、糖尿病组(DM, n=10)和正常血糖组(NG, n=19)。确诊24 h内流式细胞仪检测CD4+/CD8+ 比值、Th1/Th2比值和。CD14+单核细胞人类白细胞抗原-DR (HLA-DR)表达率,记录感染相关器官衰竭评分系统（SOFA）评分和急性生理学与慢性健康状况评分系统Ⅱ（APACHEⅡ）,并随访28d临床结局。分析应激性高血糖与免疫功能变化及28 d死亡率的关系。结果: 应激性高血糖组死亡率高于糖尿病组(53.3% vs 10.0%)和正常血糖组(53.3% vs 21.1%);应激性高血糖组SOFA评分和APACHEⅡ评分均高于其余两组; HLA-DR活化水平明显低于糖尿病组（P<0.05）和正常血糖组（P<0.05）;CD4+/CD8+比值亦低于其余两组;死亡组SOFA评分和APACHEⅡ评分明显高于存活组,HLA-DR% (65.33±12.34 vs 91.54±9.89) 和Th1/Th2比值（5.54±2.81 vs 8.97±4.93）显著低于存活组。结论: 脓毒症患者应激性高血糖与免疫功能变化有一定相关性,可以作为病情评估和预后预测的指标之一。     

Graves' disease is associated with an altered CXCR3 and CCR5 expression in thyroid-derived compared to peripheral blood lymphocytes

The mechanisms by which T cells accumulate in the thyroid and support the autoimmune process in patients with Graves' disease (GD) are poorly understood. Chemokines and their receptors may be involved in this process. We have analysed the expression of CXCR3 and CCR5 as Th1-specific chemokine receptors, CCR3 as a marker for Th2 cells, CXCR4 (expressed on unprimed, naive T cells) and CCR2 (known to be involved in autoimmunity) on peripheral blood (PBL) and thyroid-derived lymphocytes (TL) using flow cytometry. Chemokine receptor expression on PBL of GD patients (n = 16) did not differ from that of normal controls (n = 10). In GD, CXCR3+ (67.3 +/- 4.0% versus 45.7 +/- 2.1%) and CCR5+ T cells (42.5 +/- 3.4% versus 18.8 +/- 2.1%) showed a significant enrichment in the TL compared to PBL. The positive cells were contributed mainly by the CD4+CD45R0+ subset. TL are mostly primed CD45R0+ T cells, but surprisingly, they had significantly higher levels of CXCR4+ cells among TL (96.2 +/- 1.0%) compared to PBL (66.8 +/- 4.2%). However, CXCR4 has been induced during in vitro isolation of TL. There was no correlation between chemokine receptors and the level of TSH-receptor and thyroid peroxidase autoantibodies. CCR3+ and CCR2+ cells remained unchanged in TL compared to PBL. We could confirm the results using RT PCR and immunohistology. In summary, TL showed a different chemokine receptor pattern compared to PBL from the same patient. This indicates a role for CXCR3 and CCR5 in the recruitment of T cells to the thyroid in GD.     

A CD40 Kozak sequence polymorphism and susceptibility to antibody-mediated autoimmune conditions: the role of CD40 tissue-specific expression

Previously, we and others have demonstrated the association of a C/T single nucleotide polymorphism (SNP), in the Kozak sequence of CD40, with Graves' disease (GD). Here, using an expanded data set of patients, we confirm the association of the CD40 SNP with GD (n=210, P=0.002, odds ratio (OR)=1.8). Subset analysis of patients with persistently elevated thyroid peroxidase (TPO) and/or thyroglobulin (Tg) antibodies (Abs), (TPO/Tg Abs), after treatment (n=126), revealed a significantly stronger association of the SNP with disease (P=5.2 x 10(-5), OR=2.5) than in GD patients who were thyroid antibody-negative. However, the CD40 SNP was not associated with TPO/Tg Abs in healthy individuals. Next, we tested the CD40 SNP for association with Myasthenia Gravis (MG), which, like GD is an antibody-mediated autoimmune condition. Analysis of 81 MG patients found no association of the SNP with disease. Functional studies revealed significant expression of CD40 mRNA and protein in the thyroid (target tissue in GD) but not in skeletal muscle (target tissue in MG). Combined, our genetic and tissue expression data suggest that the CD40 Kozak SNP is specific for thyroid antibody production involved in the etiology of GD. Increased thyroidal expression of CD40 driven by the SNP may contribute to this disease specificity.     

Adipose tissue of human omentum is a major source of dendritic cells, which lose MHC Class II and stimulatory function in Crohn's disease

Adipose tissue is reported to contain monocyte-like pre-adipocytes, which may mature into macrophages, contributing to local inflammation. Dendritic cells (DC) can be derived from monocytes and initiate and regulate primary immune responses. We hypothesized, therefore, that adipose tissue may provide DC involved in local immune activity. To test this, we studied cells from human omental adipose tissue samples from 17 patients with benign gynecological disease. The hypothesis that adipose tissue DC are involved in inflammatory disease was tested by comparing these cells with those from 18 patients with Crohn's disease, where hypertrophy of adipose tissue suggests involvement in disease. A high proportion of the 1.33 +/- 0.12 x 10(5) CD45-positive cells/mg, obtained from control omenta, expressed CD11c, CD1a, and CD83; costimulatory molecules CD40, CD80, and CD86; and major histocompatibility complex (MHC) Class II but little CD14, CD16, or CD33. Omental cells showing morphological characteristics of DC were also observed. Metrizamide gradient-enriched DC from these populations were potent stimulators of primary proliferation of allogeneic T cells in mixed leukocyte reactions. Increased numbers of CD45+ cells from omentum of Crohn's patients (4.50+/-1.08 x 10(5) CD45+ cells/mg) contained higher percentages of CD11c+ and CD40+ cells (80.8+/-3.8% vs. 63.4+/-6, P=0.032; 77.9+/-4% vs. 58.8+/-6.5, P=0.029, respectively), but MHC Class II and stimulatory capacity were almost completely lost (P= <0.001), suggesting innate activation but lost capacity to stimulate adaptive immune responses. Granulocytes were also present amongst the omental cells from Crohn's patients. Results indicated that omentum may provide DC, which could "police" local infections and contribute to and/or reflect local inflammatory activity.     

Long-Term Immunological Study in Graves’ Disease Treated with Thyroid Arterial Embolization

OBJECTIVE: The aim of this study was to investigate long-term immunological changes after the treatment of Graves' disease (GD) with thyroid arterial embolization and the effect of thyroid arterial embolization on the body's immunological functions. MATERIALS AND METHODS: Forty-one patients with clinically and laboratorily ascertained GD were treated with thyroid arterial embolization and followed up for 3-54 months following embolization. Prior to embolization and at 1, 3, 6, 12, and 36 months following embolization, thyroid autoimmune antibodies were tested respectively, including thyroid stimulating antibody (TSAb), thyrotropin antibody (TRAb), thyroglobulin antibody (TGAb), and thyroid microsomal antibody (TMAb), as well as subgroup lymphocytes of CD16+CD56+, CD19+, CD3+, CD3+CD4+ and CD3+CD8+. The autoimmune status of GD patients prior to embolization and the dynamic changes of the immunological function after embolization were analyzed. RESULTS: The therapy of thyroid arterial embolization could effectively decrease the activity/titer and positive rate of TRAb and the ratio of CD4+/ CD8+ to normal levels at 6 months following embolization, while the ratio of CD3+CD8+ increased gradually to normal level at 1 year following embolization. In patients with recurrence, TSAb and TRAb remained at a higher level, while the rate of CD3+CD8+ and the ratio of CD4+/CD8+ were not statistically significantly different from those before embolization. CONCLUSION: Immunological functional disorder exists in GD patients. The treatment method of thyroid arterial embolization can effectively resume the basic immunological function to normal range while patients with recurrence have no significant improvement, suggesting that thyroid arterial embolization has an effective role in adjusting the immunological function.     

Expression of mucosa-related integrin alphaEbeta7 on alveolar T cells in interstitial lung diseases.

The expression of alphaEbeta7 integrin has been related to the selective retention of lymphocytes in mucosal tissues of gut, urogenital tract and lung. To identify potential disease-associated alphaEbeta7 expression patterns on cells accounting for lymphocytic alveolitis in interstitial lung disease (ILD), alphaE expression on CD4+ and CD8+ T cell subsets was evaluated by dual-colour flow cytometry in peripheral blood and bronchoalveolar lavage fluid (BALF) of patients with idiopathic pulmonary fibrosis (IPF; n = 18), hypersensitivity pneumonitis (HP; n = 20) and sarcoidosis (n = 44) in comparison with healthy controls (n = 15). In both healthy individuals and all patient groups the proportion of alphaE-bearing T cells in peripheral blood was < 2%, whereas the vast majority of alveolar CD8+ T cells consistently co-expressed alphaE. Absolute alveolar CD8+alphaE+ cell numbers/ml were up to 30-fold increased in HP patients. Proportions of alphaE-bearing CD4+ cells in BALF were significantly elevated in IPF (74.0 +/- 2.7%) and HP (70.0 +/- 2.4%) compared with normals (30.0 +/- 1.8%) (mean +/- s.e.m.; P < 0.01). In sarcoidosis, the alphaE expression on BALF CD4+ cells displayed subgroup dependency: proportions significantly lower than normal were noted in chest radiographic stage I (14.3 +/- 1.5%), but increased proportions in stages II (50.0 +/- 3.8%) and III (64.0 +/- 4.8%). Correlations between common markers of T cell activation or BALF transforming growth factor-beta (TGF-beta ) bioactivity and alphaE expression were not noted. We conclude that the vast majority of alveolar CD8+ T cells consistently express alphaEbeta7 and that distinct patterns of alphaEbeta7 expression on alveolar CD4+ lymphocytes in sarcoidosis are related to the diverse manifestations of the sarcoid inflammatory process in the lung.     

Intrathyroidal accumulation of T cell phenotypes in autoimmune thyroid disease.

In this study we have correlated peripheral T cell subset phenotypes with intrathyroidal lymphocyte accumulation in patients with autoimmune thyroid disease (Graves' and Hashimoto's disease). Our study utilized euthyroid family members for one of our control groups (n = 48) thus significantly limiting familial, but not disease-specific, influences on these T cell phenotypes. Our principal new observations were found only in patients with Graves' disease. As previously reported, there was a decrease in CD8+ (suppressor/cytotoxic) T cells in the peripheral blood of patients with untreated hyperthyroid Graves' disease (n = 27) (mean +/- SEM, 19 +/- 1.1% in patients compared with 25 +/- 1.2% in controls, p = 0.03), a finding not observed in treated, euthyroid Graves' disease patients or their relatives. However, the relative number of CD8+ T cells, assessed by CD4:CD8 ratios, was increased in the intrathyroidal T cell populations (n = 10), when compared to normal and patient peripheral blood. There were no consistent changes in total CD4+ (helper) T cells in the peripheral blood of patients with treated and untreated Graves' disease but a reduction in CD4+2H4+ (suppressor-inducer) T cells was seen in patients undergoing surgery for Graves' disease (13 +/- 6.9% compared with 39 +/- 3.4%). Again, however, this T cell subset was increased within the target organ of the same patients (41 +/- 5.9%). These data point to either a selective accumulation, or a specific "homing", of certain T cell subsets within the thyroid gland of patients with Graves' disease where T cell differentiation may be strongly influenced by antithyroid drug treatment and the local immune environment.     

Costimulatory molecules and autoimmune thyroid diseases

At least two signals for proliferation and cytokine secretion by T-cells are required. The first signal is delivered through the interaction of the T-cell receptor with major histocompatibility complex (MHC) molecules expressed on the surface of antigen-presenting cells (APC). The second or costimulatory signal is delivered by cell surface molecules expressed by APC. The interaction of B7.1/B7.2 with CD28 provide the most potent costimulatory signal for T-cell activation. CD40 antigen and its ligand (CD40L) have been shown to play a major role in regulating both humoral and cellular immune responses. In autoimmune thyroid diseases autoantigen presentation could be provided by "professional" APC, such as dendritic cells, as well as "nonprofessional" APC, such as thyroid follicular cells (TFC). In fact, these cells aberrantly express MHC class II molecules in Graves' disease (GD) and Hashimoto's thyroiditis (HT), together with large amounts of MHC class I antigens: moreover, the expression of CD40 on TFC, has been demonstrated. On the other hand B7.1 has been demonstrated in HT, but not in GD TFC. This could provide in HT a local costimulatory signal for T-cell differentiation towards a type 1 cytokine secretion pattern and also result in rescue from apoptosis of infiltrating lymphocytes. The presence of ICAM-1 on the surface of HT TFC may further strengthen contact and facilitate cross-signaling between T-cells and TFC. In contrast, the absence of B7 and ICAM-1 antigens in most GD TFC may more easily be associated with anergy and apoptosis of infiltrating T-cells, preventing the perpetuation and expansion of a "destructive" autoimmune reaction.     

Costimulatory Molecules and Autoimmune Thyroid Diseases

At least two signals for proliferation and cytokine secretion by T-cells are required. The first signal is delivered through the interaction of the T-cell receptor with major histocompatibility complex (MHC) molecules expressed on the surface of antigen-presenting cells (APC). The second or costimulatory signal is delivered by cell surface molecules expressed by APC. The interaction of B7.1/B7.2 with CD28 provide the most potent costimulatory signal for T-cell activation. CD40 antigen and its ligand (CD40L) have been shown to play a major role in regulating both humoral and cellular immune responses. In autoimmune thyroid diseases autoantigen presentation could be provided by "professional" APC, such as dendritic cells, as well as "nonprofessional" APC, such as thyroid follicular cells (TFC). In fact, these cells aberrantly express MHC class II molecules in Graves' disease (GD) and Hashimoto's thyroiditis (HT), together with large amounts of MHC class I antigens: moreover, the expression of CD40 on TFC, has been demonstrated. On the other hand B7.1 has been demonstrated in HT, but not in GD TFC. This could provide in HT a local costimulatory signal for T-cell differentiation towards a type 1 cytokine secretion pattern and also result in rescue from apoptosis of infiltrating lymphocytes. The presence of ICAM-1 on the surface of HT TFC may further strengthen contact and facilitate cross-signaling between T-cells and TFC. In contrast, the absence of B7 and ICAM-1 antigens in most GD TFC may more easily be associated with anergy and apoptosis of infiltrating T-cells, preventing the perpetuation and expansion of a "destructive" autoimmune reaction.     

Luminal bacterial antigen-specific CD4+ T-cell responses in HLA-B27 transgenic rats with chronic colitis are mediated by both major histocompatibility class II and HLA-B27 molecules

Rats transgenic (TG) for the human major histocompatibility complex (MHC) class I HLA-B27 and beta2-microglobulin genes develop chronic colitis under specific pathogen-free (SPF) but not sterile (germ-free, GF) conditions. We investigated the role of antigen-presenting molecules involved in generating immune responses by CD4+ mesenteric lymph node (MLN) cells from colitic HLA-B27 TG rats to commensal enteric micro-organisms. All TG MLN cells expressed HLA-B27. A higher level of MHC class II was expressed on cells from TG rats, both SPF and GF, compared to non-TG littermates. In contrast, rat MHC class I expression was lower on TG than non-TG cells. Both TG and non-TG antigen presenting cells (APC) pulsed with caecal bacterial antigens induced a marked interferon-gamma (IFN-gamma) response in TG CD4+ T lymphocytes but failed to stimulate non-TG cells. Blocking MHC class II on both TG and non-TG APC dramatically inhibited their ability to induce TG CD4+ T cells to produce IFN-gamma. Blocking HLA-B27 on TG APC similarly inhibited IFN-gamma responses. When the antibodies against MHC class II and HLA-B27 were combined, no APC-dependent IFN-gamma response was detected. These data implicate both native rat MHC class II and TG HLA-B27 in CD4+ MLN T-cell IFN-gamma responses to commensal enteric microflora in this colitis model.     

The expression and distribution of S-100 protein and CD 83 in thyroid tissues of autoimmune thyroid diseases

To investigate the expression and distribution of S-100 protein and CD83 in the thyroid tissues of autoimmunethyroid diseases(ATDs),and to study the role of the dendritic cells in the pathogenesis of ATDs,immunohistochemical staining was used on pathological tissues of 20 patients with Hashimoto's thyroiditis(HT)and 20 patients with Graves' disease(GD) to check the expression and distribution of S-100 protein and CD83.Compared with control group(20 cases of thyroid follicular adenoma,TFA),the higher expressions of S-100 inHT(139.38±5.92 vs 59.47±11.69) and GD(119.42±14.48 vs 59.47±11.69) were observed respectively(p<0.001).The increased positive expressions of CD83 which is known as a marker of mature and activated DCs inHT(22.58±13.96 vs 5.19±8.08) and GD(29.92±14.43 vs 5.19±8.08) were also found respectively(p<0.001).Serum TPO antibody(TPO-Ab,67.3±11.6%) and Tg antibody(Tg-Ab,59.8±10.1%) in HT were higher thanthose in GD(28.4±5.7%,23.1±4.9%) and TFA(6.1±3.4%,7.2±4.6%)(p<0.01).Serum TR-Ab in GD(16.3±5.6 U/L) was higher than those in HT(4.8±2.3 U/L) and TFA(2.5±1.2 U/L)(p<0.01).Our findings suggestthat the high expression of DCs' markers may be related to the pathogenesis of HT and GD.The upregulationof both the number and the matured functions of DCs,may lead to present more antigens and to produce moreauto-antibodies(such as Tg-Ab and TPO-Ab in HT,TR-Ab in GD),which may be involved in pathogenesis ofthe autoimmune thyroid diseases.Cellular & Molecular Immunology.2004;1(5):378-382.     

Distribution of MHC class II antigens in feline tissues and peripheral blood

A new monoclonal antibody raised against gradient-purified feline immunodeficiency virus was found to recognize a bimolecular complex, comprising 27-29 kD and 32-35 kD subunits, on feline peripheral blood lymphocytes. Immunoperoxidase staining of feline tissues with this antibody, designated 43.2H2, demonstrated a reactivity pattern similar to that described for MHC II antigens of the dog, horse, and pig, but differed from human and mouse in having staining of T-cell zones in spleen and lymph nodes. Flow cytometric analysis revealed that 42.3H2 reacted with 88.97% +/- 16.00% of feline peripheral blood lymphocytes (n = 20). This high level of reactivity was found to be consistent by repeated sampling over a 4-month period. Two-color flow cytometric analysis was used to determined the reactivity pattern on lymphocyte subsets: 88.92% +/- 7.30% of CD4+ lymphocytes were 42.3H2-positive, while 85.99% +/- 11.46% of CD8+ cells were positive (n = 11 for both). B lymphocytes had the highest reactivity (99.47% +/- 0.45; n = 9) and also had the highest fluorescence intensity. By gating based on light scatter properties, 95.06% +/- 7.35% of monocytes were 42.3H2-reactive (n = 18), while granulocytes were negative.     

Decrease of peripheral large granular lymphocytes in Graves' disease.

Peripheral large granular lymphocytes (LGL) were enumerated in normal subjects and patients with autoimmune thyroid diseases. In normal subjects, the percentage of LGL was significantly lower in women (mean +/- s.d., 17.0 +/- 3.6%; n = 35; P less than 0.05) than in men (19.5 +/- 5.3%; n = 20). In untreated patients with thyrotoxic Graves' disease (GD), both the percentage (11.5 +/- 2.7%; n = 12; P less than 0.001) and the absolute count (244 +/- 102/mm3; P less than 0.01) of LGL were significantly lower than those in normal women (17.0 +/- 3.6% and 334 +/- 122/mm3; n = 35). No significant differences from normal controls were observed in the percentages or absolute counts of LGL in patients with euthyroid GD under treatment or in euthyroid or hypothyroid patients with Hashimoto's disease (HD). The percentage of LGL was inversely correlated with the serum levels of T4 and T3 and the free T4 index, but not with the titre of anti-thyroid antibodies, the size of goitre or the degree of proptosis in the group of untreated patients with GD and HD. The decreased levels of LGL in thyrotoxic patients with GD may be related to the self-perpetuation of thyrotoxicosis in patients with this disease.     

CD134 expression on CD4+ T cells is associated with nephritis and disease activity in patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is characterized by a deviation of the immune system that involves T cell-dependent autoantibody production. The aim of this study was to investigate the role of co-stimulatory markers on T cells in this disease. Twenty-eight patients with SLE as defined by the American College of Rheumatology (ACR) criteria and 11 healthy controls were included into the study. Eleven patients had biopsy-proven lupus nephritis while 17 patients had no clinical evidence of lupus nephritis. Clinical disease activity was assessed according to the systemic lupus erythematosus disease index (SLEDAI). CD4+ T cell populations in the peripheral blood were analysed for the expression of co-stimulatory markers CD45RO, CD70, CD80, CD86, CD137, CD137L, CD134, CD152, CD154 and ICOS. SLE patients showed an increased frequency of peripheral CD4+ T cells expressing high levels of CD80, CD86 and CD134 compared to healthy controls (7.1 +/- 1.5% versus 1.7 +/- 0.9%; P < 0.005; 2.3 +/- 0.4% versus 1.0 +/- 0.2%; P = 0.008, 20.2 +/- 2.0% versus 10.6 +/- 1.9%; P < 0.005, respectively). Significantly higher levels of CD80 on CD4+ T cells were detected in SLE patients with lupus nephritis compared to patients without nephritis (11.9 +/- 3.3% versus 4.0 +/- 0.7%; P < 0.005). There was an increased presence of CD134+ CD4+ cells in SLE patients with lupus nephritis (27.5 +/- 4.0% versus 15.5 +/- 1.3%; P < 0.005). CD80 and CD134 expression was significantly correlated with SLEDAI (r = 0.42, P = 0.03; r = 0.56, P < 0.005). Co-stimulatory molecules on CD4+ T cells are associated with renal disease and disease activity in patients with systemic lupus erythematosus.     

Th17细胞在Graves’病中的变化及意义

目的:探讨Th17细胞在中国人Graves’病(GD)中的变化及意义。方法:应用电化学发光法测定30例初发GD患者及30例健康体检者血清中促甲状腺激素(TSH)、游离三碘甲腺原氨酸(FT3)、游离甲状腺激素(FT4)、甲状腺球蛋白抗体(TgAb)和甲状腺过氧化酶抗体(TPOAb)的水平;采用流式细胞术检测外周血单个核细胞(PBMC)中CD4+IL-17+T(Th17)细胞的数量;用ELISA法测定初发GD患者和健康对照者PBMC产生IL-17的水平;应用实时定量PCR(qRT-PCR)检测PBMC中ROR-γt、IFN-γ、IL-4 mRNA的表达量;采用免疫组化染色检测甲状腺组织中IL-17+细胞的分布和数量。结果:初发GD组外周血中Th17细胞占CD4+T细胞的比例[(20.59±4.63)%]、PBMC分泌的IL-17水平[(83.22±34.28)pg/ml]以及PBMC中ROR-γt mRNA表达(1.67±0.98)均明显高于正常对照组Th17[(11.77±4.18)%](P<0.01)、IL-17[(19.74±5.99)pg/ml](P<0.01)和ROR-γt mRNA(0.92±0.18)(P<0.05)的水平。与正常对照组比较,GD患者PBMC中IFN-γmRNA的表达(0.31±0.07)、IL-4 mRNA的表达(2.53±0.70)均明显降低(P<0.01)。初发GD组患者PBMC中Th17细胞百分率与ROR-γt mRNA表达呈正相关(r=0.5047,P=0.01);与IFN-γmRNA表达量呈负相关(r=-0.5085,P<0.01);与IL-4 mRNA表达量呈负相关(r=-0.5372,P<0.01)。正常对照组甲状腺组织中未发现IL-17+细胞,GD组甲状腺组织滤泡间质内可见散在的IL-17+细胞。结论:初发GD患者PBMC中Th17细胞数量、PBMC分泌IL-17的水平和ROR-γt mRNA表达明显增高。GD患者甲状腺组织中有IL-17+细胞浸润,提示Th17细胞可能参与了中国人GD的发生。     

Leptine participation in the development of liver steatosis and biliar lithiasis

Obesity increases significantly the risk of developing several common gastrointestinal diseases such as gallstone disease (GD) and hepatic steatosis (HS). Elsewhere we have shown a relationship between HDL cholesterol, cholesterol saturation index, and leptin in obese patients loosing weight. Furthermore, leptin plays an important role facilitating HS and possibly in the associated inflammatory process. The aim of this study was to investigate the relationship between GD and HS. The sample was comprised by patients attending the unit for check-up. Subjects with visible stones or HS by ultrasound (cases) were compared with healthy controls. Demographic and body mass index (BMI) were recorded. Plasma leptin, insulin and serum lipids and lipoproteins levels were measured by standard methods. A total of 317 subjects were included in this study. They were divided in four groups as follows: GD (n=100), HS (n=84), GD + HS (n=33) and controls (n=100). The control group was significantly older (GD, 52.6+/-11.6; HS, 49.8+/-11.1; GD +HS, 51.6+/-10.5; 57.1+/-7.4), p< 0.05. BMI was higher in the HS groups (28.7 +/- 2.8) and GD +EH (29.0 +/- 3.8) than in the GD (27.4 +/- 4.3) and control (27.0 +/- 3.1) group, p<0.05. The GD group displayed the highest leptin levels (13.7 241 8.1), P < 0.05, whereas insulin levels were similar in all groups. Since GD and HS subjects have high plasma leptin levels compared with controls, our results suggest that leptin plays an important role in the pathophysiology of GD and HS.     

MHC class II compartment, endocytosis and phagocytic activity of macrophages and putative dendritic cells isolated from normal tissues rich in synovium

The endocytic and phagocytic activities of a population of MHC IIhi CD11c+ dendritic cell (DC)-like cells in synovium-rich tissues (SRTs) of normal rat paws were compared with CD163+ cells (putative macrophages) from the same tissues and pseudo-afferent lymph DCs, peritoneal macrophages and blood monocytes. Fifty percent of CD11c+ cells and 75% of CD163+ cells isolated from SRT internalized fluorescein-conjugated dextran (FITC-DX). Of these endocytic cells, half of those expressing CD11c, but only 30% of those expressing CD163, were surface MHC class II+ (sMHC II+). CD11c+ cells were more endocytic than monocytes or pseudo-afferent lymph DC, but some CD163+ cells (type A synoviocytes) were found to be highly endocytic. CD163+ cells from SRT were more phagocytic (25%) than the general MHC class II+ population (16%). Of phagocytic cells, 40% of CD163+ cells were sMHC II(variable) and they constituted 60% of all MHC class II+ phagocytic cells. Only 18% of phagocytic MHC II+ cells expressed CD11c and the most of these were MHC IIhi. In comparison, 60% of CD163+ peritoneal macrophages were phagocytic, while blood monocytes were poorly phagocytic. Intracellular MHC class II-rich compartments (MIIC) were prominent in sMHC IIhi cells in SRT but rare in CD163+ cells. Most MHC IIhi CD11c+ cells did not have a detectable MIIC.     

MHCII类分子反式激活因子突变体重组腺病毒治疗小鼠实验性自身免疫性甲状腺炎

评估IFN-γ调控型启动子(CIITA-pIV)驱动的MHC II类分子反式激活因子突变体(MHC class II transactivator mu-tant,CIITAm)重组腺病毒Ad-pIV-CIITAm对小鼠实验性自身免疫性甲状腺炎(experimental autoimmune thyroiditis,EAT)的治疗效果,并探讨其可能的作用机制。34只健康雌性CBA/J小鼠随机分成CIITAm治疗组(n=9)、GFP对照组(n=9)、EAT模型组(n=8)和正常对照组(n=8)共四组。正常对照组不做特殊处理,其余三组均以猪甲状腺球蛋白(porcinethyroglobulin,pTg)+弗氏佐剂(complete or incomplete Freund adjuvant,CFA/IFA)建立EAT小鼠模型,并分别静脉注射重组腺病毒Ad-pIV-CIITAm、Ad-GFP及等体积生理盐水。首次免疫后第29天处死小鼠,进行H-E染色观察甲状腺病理形态;免疫组织化学染色测定甲状腺MHC II类分子表达;分析pTg刺激下脾脏淋巴细胞的增殖及其上清液中IFN-γ的分泌水平;ELISA法检测血浆中抗-pTg自身抗体滴度;流式细胞术分析外周血和脾脏淋巴细胞中T细胞亚群。结果:H-E染色结果表明,CIITAm治疗组甲状腺淋巴细胞浸润指数(0.5±0.5)低于GFP对照组(1.5±0.2)和EAT模型组(1.4±0.4,P<0.01)。免疫组化结果显示,GFP对照组和EAT模型组甲状腺组织有弥漫性MHC II类分子表达,而CIITAm治疗组未见明显表达,正常对照组表达呈阴性。80μg/ml pTg刺激下,CIITAm治疗组小鼠淋巴细胞刺激指数(SI)明显低于GFP对照组或EAT模型组(P<0.01);培养上清各组IFN-γ分泌水平结果类似(P<0.01)。CIITAm治疗组血浆抗-pTg自身抗体滴度显著低于GFP对照组或EAT模型组(P<0.05);CIITAm治疗组外周血和脾脏CD4+T细胞百分率亦显著低于GFP对照组或EAT模型组(P<0.05)。重组腺病毒Ad-pIV-CIITAm能抑制EAT小鼠甲状腺组织MHC II类分子表达,抑制自身反应性T细胞增殖,减轻甲状腺炎性细胞浸润,降低自身抗体滴度,对EAT有一定的治疗作用。     

MHC class I pathway is not required for the development of crescentic glomerulonephritis in mice

MHC II and CD4+ T cells are required for anti-glomerular basement membrane (GBM) globulin-initiated crescentic glomerulonephritis (GN) in mice, but the role of MHC I and CD8+ T cells is unclear. The cytolytic function of CD8+ T cells requires recognition of peptide antigens presented on MHC I. CD8+ T cells can also perform helper functions via cytokine production. The contribution of MHC I to crescentic GN was investigated using TAP-1 gene knock out (TAP-1-/-) mice, which have deficient MHC I antigen presentation. Heterozygous TAP-1 mice have normal MHC I expression and developed GN with crescents in 42 +/- 4% of glomeruli (normal 0%), proteinuria (9.1 +/- 1.6 mg/20 h, normal 1.5 +/- 0.3 mg/20 h) and impaired renal function (creatinine clearance 110 +/- 8 microl/min, normal 193 +/- 10 microl/min) following administration of sheep anti-mouse GBM globulin. TAP-1-/- mice, which have extremely low MHC I expression and reduced CD8+ T cells, developed similar GN with 39 +/- 3% crescents, proteinuria (12.7 +/- 4.3 mg/20 h) and impaired renal function (creatinine clearance 123 +/- 20 microl/min). In vivo antibody-induced CD8 depletion did not attenuate crescent formation or protect renal function in C57Bl/6 mice developing GN, although significant reduction in proteinuria (5.3 +/- 1.2 mg/20 h, P = 0. 012) and glomerular recruitment of CD4+ T cells and macrophages were observed compared with control treated mice with GN. These data demonstrate that MHC I is not required for development of crescentic GN in mice. The MHC I-independent contribution of CD8+ T cells to proteinuria and inflammatory cell recruitment suggests that they may serve a 'helper' rather than cytolytic role in this disease.     

Increase in peripheral natural killer cell activity in patients with autoimmune thyroid disease.

Changes in the activity and number of natural killer (NK) cells in peripheral blood in patients with autoimmune thyroid disease were examined. NK activity was measured in a 4-hr 51Cr-release assay and the number of NK cells was analyzed with FITC-conjugated monoclonal antibodies by use of an automated flow cytometer. NK activity in patients with untreated Graves' disease (n = 25, 39.7 +/- 13.5%, P less than 0.05) and Hashimoto's thyroiditis (n = 18, 41.0 +/- 14.2%, P less than 0.05) was high compared to the activity in non-pregnant controls (n = 61, 32.6 +/- 15.0%). NK activity in patients with postpartum Graves' thyrotoxicosis (n = 11, 48.6 +/- 18.9%) was markedly increased compared to the activity in non-pregnant controls (P less than 0.01) and in postpartum controls (n = 29, 33.8 +/- 15.2%, P less than 0.05), although the mean ages of each group did not differ significantly. Moreover, NK activities in the thyrotoxic state were significantly higher than those in the euthyroid state in the same patients with postpartum Graves' thyrotoxicosis or with postpartum destructive thyrotoxicosis. The number of CD16 positive cells increased in patients with postpartum Graves' thyrotoxicosis. However the number of CD16 and CD57 positive cells were normal in all other groups of patients. These results indicate that an increase of NK activity is associated with exacerbation of autoimmune thyroid disease both in Hashimoto's thyroiditis and in Graves' disease and suggest that NK cells might have an important role for the control of disease activity in autoimmune thyroid disease.