Relationship between HMGB1 content and MHC-Ⅱ expression in circulating monocytes and spleen of mice challenged with zymosan

Objective: To observe the regularity of change in high mobility group protein box 1 (HMGB1) content in serum and spleen of mice with multiple organ dysfunction syndrome (MODS), to analyze the correlation between HMGB1 content and major histocompatibility complex (MHC)-Ⅱ-I-Ab expression on monocytes in blood and spleen, and to explore the effect of HMGB1 on immune function of circulating monocytes and splenocytes. Methods: One hundred 8-week-old male 57BL/6 mice were randomly divided into normal group and experimental group subdivided into 8 subgroups: 3, 8, 12 hours, 1, 2, 3, 5-7 days and 10-12 days post zymosan injection (PZI). MODS model was replicated by injecting zymosan into the peritoneal cavity. At each time point, blood and spleen were collected to detect HMGB1 content and the rate of I-Ab positive monocytes. Results: In normal and PZI 3-hour, 8-hour mice, serum HMGB1 was not detected, but it significantly increased at PZI 12 hours. In spleen of normal mice, there was low level of HMGB1 expression. In zymosan-treated mice, HMGB1 started to rise in spleen at PZI 3 hours. Subsequently, HMGB1 content in both serum and spleen significantly increased, and it reached the peak level in 1-2 days, decreased in 5 days, and then increased in 10-12 days. The number of I-Ab positive monocytes in circulating blood and spleen decreased at 1-2 days (t=9.589, 4.432, P<0.01) and 10-12 days following the challenge, forming a two trough like decrease, just corresponding with two-peak increase of HMGB1. However, at 3 hours after zyrnosan challenge, I-Ab expression on circulating monocytes was downregulated (t=5.977, P<0.01), while that in spleen upregulated (t=4.814, P<0.01). Conclusion: In mice with MODS, up-regulated HMGB1 expression can regulate I-Ab expression on monocytes to depress their ability of presenting antigen, which results in immune disturbance contributing development of MODS.     

Depletion of macrophages in mice results in higher dengue virus titers and highlights the role of macrophages for virus control

Monocytes and macrophages are target cells for dengue infection. Besides their potential role for virus replication, activated monocytes/macrophages produce cytokines that may be critical for dengue pathology. To study the in vivo role of monocytes and macrophages for virus replication, we depleted monocytes and macrophages in IFN‐αβγR knockout mice with clodronate liposomes before dengue infection. Although less virus was first recovered in the draining LN in the absence of macrophages, monocyte/macrophage depletion eventually resulted in a ten‐fold higher systemic viral titer. A massive infiltration of CD11b⁺CD11c^lowLy6C^low monocytes into infected organs was observed in parallel with increasing virus titers before viremia was controlled. Depletion of monocytes in the blood before or after local infection had no impact on virus titers, suggesting that monocytes are not required as “virus‐shuttles”. Our data provide evidence that systemic viremia is established independently of tissue macrophages present at the site of infection and blood monocytes. Instead, we demonstrate the importance of monocytes/macrophages for the control of dengue virus.     

Aberrancies in Antigen-presenting Cells and T Cells in Autoimmune Thyroid Disease. A Role in Faulty Tolerance Induction

Various thyrocyte, monocyte, macrophage, DC and T cell abnormalities exist in the animal models of spontaneously developing autoimmune thyroiditis and in patients with autoimmune thyroid disease. An aberrant interaction between such abnormal thyrocytes, abnormal professional antigen-presenting cells (APC) and abnormal T cells forms the basis for the atypical autoimmune reaction targeting thyroid antigens. In the atypical interaction more than one gene and various environmental factors are involved. The genetic and environmental factors must act together to induce full-blown disease.

Although there is a general blueprint for the development of destructive autoimmune thyroiditis, thyrocyte and immune cell abnormalities differ between the various animal models and the various forms of autoimmune thyroid disease (either associated with type 1 diabetes, associated with bipolar disorder or not associated). This tells us that there are different etio-pathogenic forms of destructive autoimmune thyroiditis. Whether such heterogeneity is also the case for the etio-pathogenesis of Graves' disease remains unknown. Animal models of spontaneously developing Graves' disease would be helpful in unraveling this question.

If indeed there are various etio-pathogenic routes in different patients that lead to destructive autoimmune thyroiditis, then tailor-made therapeutic approaches need to be carried out in attempts to correct the underlying immune abnormalities in individual patients or to prevent the development of destructive autoimmune thyroiditis in individuals at risk. While in some forms of destructive autoimmune thyroiditis (f.i. those associated with bipolar disorder) immune suppression should be the first choice of intervention, other forms (f.i. those associated with type 1 diabetes) may benefit from immune stimulation in certain pre-stages of the disease (to restore f.i. the faulty APC function characteristic of this condition). Obviously a more precise determination of the spectrum of cell-mediated immune abnormalities is required in individual cases of destructive autoimmune thyroiditis, before therapies that aim at correcting the immune abnormalities can be tested successfully.     

Correlation between Profile of Circulating Mononuclear Cells and Clinical Manifestations in Patients with Pemphigus Vulgaris

Phenotypes of 38 samples of mononuclear (PBMC) cells from 11 different patients with pemphigus vulgaris (PV) at different stages of the disease were explored looking for a possible relationship between cell immunity, mucocutaneous or mucosal lesion intensity and capacity of serum autoantibodies to elicit the disease in mice. PBMC from 5 patients with mucocutaneous lesions and sera with IgG capable of inducing the disease in neonatal mice had a high proportion of mature monocytes with CD14^lowDR ^gh and co-expresing CD16 and CDllb. In addition, a high proportion of CD19⁺CD5⁺ activated B cells and a very low proportion of naive CD4⁺CD45RA⁺ and CD8⁺CDllb⁺ T lymphocytes was observed. Monocytes from these patients expressed inducible nitric oxide synthase (iNOS). In contrast, PBMC from 6 patients, with lesions restricted to mucosal membranes and IgG lacking the capacity to induce the disease in mice, contained a high proportion of CD14^hlg DR^low co-expressing CD16 circulating macrophages, CD8⁺CDllb⁺ T cells, and a low proportion of activated B lymphocytes. The results suggest a possible association between proportion of different antigen presenting cells (monocytes with high HLA-DR and low CD 14 expression and activated B lymphocytes, or differentiated monocytes/macrophages), type of PV and capacity of serum autoantibodies to elicit the disease in mice     

白细胞介素6诱导的组织因子表达在慢性排斥反应发病机制中的作用

目的研究白细胞介素6（IL-6）对人外周血单核细胞（PBMCs）中组织因子（TF）的表达及活性的诱导作用。方法分别用人重组白细胞介素6（rhIL-6）100ng／L和rhIL-6100ng／L＋rhIL-6单克隆抗体（MoAb）10μg／L培养刺激PBMCs24h，将未经刺激的PBMCs作为对照组，观察各组细胞中TF抗原含量、mRNA的表达及TF活性的变化。结果经rhIL-6培养刺激24h后，PBMCs的TF抗原含量、mRNA的表达及TF活性均较对照组和rhIL-6＋rhIL-6 MoAb组显著增加（P〈0．01）。结论rhIL-6可诱导PBMCs的TF抗原表达和活性，并使mRNA的表达增加，rhIL-6 MoAb能有效抑制rhIL-6所致的PBMCs的TF表达及活性，提示炎症因子与凝血机制的启动存在内在联系，并在慢性排斥反应中起重要作用。     

ATG16L1 and NOD2 polymorphisms enhance phagocytosis in monocytes of Crohn’s disease patients

AIM:To investigate if the presence of relevant genetic polymorphisms has effect on the effectual clearance of bacteria by monocytes and granulocytes in patients with Crohn’s disease(CD).METHODS:In this study,we assessed the differential responses in phagocytosis by measuring the phagocytic activity and the percentage of active phagocytic monocytes and granulocytes in inflammatory bowel disease patients as well as healthy controls.As both autophagy related like 1(ATG16L1)and immunityrelated guanosine triphosphatase gene are autophagy genes associated with CD and more recently nucleo-tide-binding ligomerization domain-containing protein2(NOD2)has been identified as a potent inducer of autophagy we genotyped the patients for these variants and correlated this to the phagocytic reaction.The genotyping was done with restriction fragment length polymorphisms analysis and the phagocytosis was determined with the pHrodo?Escherichia coli Bioparticles Phagocytosis kit for flowcytometry.RESULTS:In this study,we demonstrate that analysis of the monocyte and granulocyte populations of patients with CD and ulcerative colitis showed a comparable phagocytic activity(ratio of mean fluorescence intensity)between the patient groups and the healthy controls.CD patients show a significantly higher phagocytic capacity(ratio mean percentage of phagocytic cells)compared to healthy controls(51.91%±2.85%vs 37.67%±7.06%,P=0.05).The extend of disease was not of influence.However,variants of ATG16L1(WT:2.03±0.19 vs homozygoot variant:4.38±0.37,P<0.009)as well as NOD2(C-ins)(heterozygous variant:42.08±2.94 vs homozygous variant:75.58±4.34(P=0.05)are associated with the phagocytic activity in patients with CD.CONCLUSION:Monocytes of CD patients show enhanced phagocytosis associated with the presence of ATG16L1 and NOD2 variants.This could be part of the pathophysiological mechanism resulting in the disease.     

人单核吞噬细胞应答鼠伤寒沙门氏菌的体外研究

目的　为了探讨鼠伤寒沙门氏菌对人类的致病机理以及机体对鼠伤寒沙门氏菌的防御机制。方法　采用体外培养的人单核吞噬细胞 ,测定鼠伤寒沙门氏菌在PMA分化的THP - 1细胞中的存活率以及诱导该细胞产生TNF -α和IL -12的量。结果　鼠伤寒沙门氏菌在人类单核吞噬细胞中的存活数量远远低于伤寒沙门氏菌的存活数量。鼠伤寒沙门氏菌和伤寒沙门氏菌同样可以诱导人类单核吞噬细胞产生TNF -α和IL - 12 ,然而 ,随着时间延长 ,鼠伤寒沙门氏菌诱导产生TNF-α和IL - 12的量明显低于伤寒沙门氏菌的诱导产生量。结论　鼠伤寒沙门氏菌的致病性可能部分与它诱导人类单核吞噬细胞释放前炎症细胞因子有关     

类风湿关节炎外周血与滑液细胞CD147分子表达的研究

目的　为了探讨关节内CD14 7表达细胞的来源与类风湿关节炎 (RA)的关系 ,研究CD14 7分子在RA患者及正常健康人外周血与滑液细胞中的表达水平。方法　取 10例活动期RA患者用药治疗前后外周血、治疗前滑液及 10名正常健康对照外周血 ,外周血和滑液分离后细胞悬液采用双色免疫荧光标记 ,流式细胞术测量淋巴细胞 (CD3+ T)、CD14 + 单核细胞和中性粒细胞表达CD14 7阳性率及表达荧光强度水平。结果　①与正常对照相比 ,RA患者外周血CD3+ T淋巴细胞与CD14 + 单核细胞的表面CD14 7表达强度较高 (P <0 0 1) ,而其表达率比较差异无显著性。中性粒细胞表面CD14 7的表达率与表达强度与正常人相比差异均无显著性 ;②正常对照外周血细胞膜表面CD14 7的分子的表达在IFN γ刺激后有显著增多 ,而RA患者在刺激前后差异无显著性 ;③关节滑液中淋巴细胞、单核细胞与中性粒细胞上的CD14 7表达强度均比外周血细胞表达高 ,且淋巴细胞的表达率较高。结论　CD14 7在RA中各种炎细胞上均有较高表达 ,提示其在RA的病理损伤中起到重要作用。     

急性冠状动脉综合征患者单核细胞过氧化体增殖物激活型受体δ的表达与转化生长因子β1和白细胞介素1β的相关性

目的探讨急性冠状动脉综合征患者单核细胞中过氧化体增殖物激活型受体δ的表达水平与血清转化生长因子β1和白细胞介素1β水平的相关性。方法纳入55例急性冠状动脉综合征患者和47例健康对照者,分离其周围血单核细胞,应用逆转录聚合酶链反应法检测过氧化体增殖物激活型受体δmRNA的表达;应用酶联免疫吸附试验检测血清转化生长因子β1和白细胞介素1β的水平。结果急性冠状动脉综合征患者血单核细胞中过氧化体增殖物激活型受体δmRNA的表达水平显著低于健康对照组(0.13±0.09比0.34±0.08,P0.01);血清白细胞介素1β水平高于健康对照组(142.7±39.3 ng/L比81.5±23.9 ng/L,P0.01),而转化生长因子β1低于健康对照组(11.4±4.6μg/L比26.3±10.5μg/L,P0.01);过氧化体增殖物激活型受体δmRNA表达水平与白细胞介素1β呈负相关(r=-0.437,P0.05),与转化生长因子β1呈正相关(r=0.598,P0.05)。结论过氧化体增殖物激活型受体δ可能在急性冠状动脉综合征的发病中具有重要作用。     

阿托伐他汀降低急性冠脉综合征患者外周血CD_(14)~+单核细胞表面TLR4的表达

目的 Toll 样受体4(TLR4)的激活与动脉粥样斑块的进展及斑块的不稳定导致临床并发症的发生有关。他汀类调脂药物临床获益可能与其调脂以外的作用尤其是抗炎作用有关,但抗炎作用的确切机制目前还不清楚。推测其抗炎作用可能是通过抑制 TLR4炎症信号通路起作用的。方法入选者为健康志愿者(n=22)及2006-07-2007-09在门诊和住院患者共121例[稳定性心绞痛(SAP)患者17例,急性冠脉综合征(ACS)患者82例],入院即刻抽取外周静脉血;将其中的41例 ACS 患者随机分为两组:在常规抗心绞痛治疗的基础上服用阿托伐他汀10 mg 组(20例)及阿托伐他汀40 mg 组(21例),治疗1月后抽取外周静脉血。观察指标为血脂、高敏 C 反应蛋白(hsCRP)、外周血 CD_(14)~+单核细胞表面 TLR4的表达,单核细胞表面 TLR4的表达采用流式细胞仪方法测定。结果 ACS 患者血 hsCRP 及外周血 CD_(14)~+单核细胞表面 TLR4的表达明显高于 SAP 患者(P<0.05)及正常对照组(P<0.05),hsCRP 与 TLR4的相关性分析结果是两者轻度相关(r=0.261,P=0.002)...