Type 1 and type 2 tumor infiltrating effector cell subpopulations in progressive breast cancer

Effector T cells fall into two subpopulations based on cytokine-secretion. Type 1 cells secrete IFN-gamma, whereas type 2 cells secrete IL-4, IL-10, and GM-CSF. NKT cells represent a third subpopulation that secretes similar cytokines and have been associated with immunoregulation. Using the TS/A adenocarcinoma, we assessed the phenotype and kinetics of tumor-infiltrating lymphocytes (TIL) in mice challenged subcutaneously in the mammary region. Flow cytometric analysis shows that T cells do not infiltrate the primary tumor site until days 7-14 following tumor challenge. Both CD4 and CD8 TILs were predominantly CD44(High) and expressed CD25, CD69, and CD95 cell surface activation markers. Activated CD4/CD44(High) TIL numbers reached peak levels at day 21 that precipitously decreased by day 28 whereas corresponding CD8 cell numbers progressively increased, however, at lower levels and with later kinetics. Intracellular cytokine staining showed that greater numbers of IL-4-producing Th2 cells were elicited and with earlier kinetics than that of IFN-gamma-producing Th1 cells. T cells co-expressing DX5 (CD3(+)/DX5(+)) emerged (>21 days), suggesting a recruitment of NK-like T cells at later stages of tumor progression. Moreover, tumors selectively up-regulated TGF-beta, MIF, and IP-10 gene expression at times as early as day 4, with peak levels at day 7 in vivo. Such gene expression remained elevated and correlated with a continued progression in tumor growth suggesting that preferential effector cell recruitment and production of select factors during different stages of tumor maturation may aid in regulating effective endogenous antitumor responses in progressive breast cancer.     

Cytokines and fibrinolytic enzymes in tuberculous and parapneumonic effusions

Tuberculous (TB) pleurisy and parapneumonic effusion (PPE) are common causes of pleural fibrosis. The mechanisms underlying fibrin deposition may be different since involved inflammatory cells are distinct. In this study, we measured various cytokines and fibrinolytic enzymes and compared the differences between the two effusions. PPE was further divided into noncomplicated PPE and complicated PPE/empyema subgroups. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, IL-8, macrophage inflammatory protein (MIP)-1beta, monocyte chemoattractant protein (MCP)-1, plasminogen activator inhibitor type 1 (PAI-1) and tissue type plasminogen activator (tPA) were measured using enzyme-linked immunosorbent assays. Significantly higher values of PAI-1, PAI-1/tPA ratio, IL-1beta, IL-8 and MIP-1beta and significantly lower values of TNF-alpha, IL-6 and MCP-1 were observed in PPE/empyema than in TB effusions. Compared to noncomplicated PPE, complicated PPE/empyema had significantly higher levels of TNF-alpha, IL-1beta, IL-8 and MIP-1beta. TB pleurisy patients who had higher effusion levels of TNF-alpha, IL-1beta and IL-8 were predisposing to residual pleural thickening. The underlying mechanisms of fibrin formation and deposition between the two effusions studied (PPE/empyema and TB pleurisy) could not be fully explained by the results of the present study. More studies are needed to explore this further.     

疏风宣肺方和解表清里方体外干预甲型流感病毒H1N1诱导炎性细胞因子分泌作用的研究

目的：研究疏风宣肺方和解表清里方对甲型流感病毒H1 N1感染的人肺腺癌上皮细胞（A549）中炎性细胞因子的影响。方法培养 A549,甲型流感病毒 H1N1感染 A549后,分为细胞对照组、H1V1感染组、奥司他韦对照组、疏风宣肺组及解表清里组。采用实时定量聚合酶链式反应（RT －PCR）和Western blotting 法检测各组细胞中炎症相关的白细胞介素（IL）1、肿瘤坏死因子α（TNF －α）、IL －6、IL －10、单核细胞趋化蛋白1（MCP －1）、调节活化正常 T 细胞表达和分泌的趋化因子（RANTES）的 mRNA 及蛋白表达的变化。结果基因芯片结果提示,与细胞对照组比较,H1N1感染组差异表达基因 Il1β、Thf、Ccl5、Il10、Il6、Ccl2明显上调。与 H1N1感染组比较,奥司他韦组、疏风宣肺组和解表清里组差异基因 Thf、Il1β、Ccl5、Il10、Il6、Ccl2表达显著下调。 RT －PCR 结果显示,与细胞对照组比较,H1N1感染组 IL －1、TNF －α、IL －6、IL－10、MCP －1、RANTES 的 mRNA 表达均显著升高（P ＜0．01）。与 H1N1感染组比较,疏风宣肺组 IL －1、TNF－α、IL －6、IL －10、MCP －1的 mRNA 表达均明显降低（P ＜0．01,P ＜0．05）,解表清里组 IL －1、TNF －α、IL －6、MCP －1的 mRNA 表达均明显降低（P ＜0．01,P ＜0．05）。 Western blotting 结果显示,H1N1感染组 IL －1、TNF －α、IL －6、IL －10、MCP －1、RANTES 蛋白表达较细胞对照组显著升高（P ＜0．05）;与 H1N1感染组比较,疏风宣肺组及解表清里组 IL －1、TNF －α、IL －6、IL －10、MCP －1、RANTES 蛋白表达均显著降低（P ＜0．05,P＜0．01）。结论疏风宣肺方和解表清里方均可抑制流感病毒感染后炎性细胞因子 IL －1、TNF －α、IL －6、MCP －1的 mRNA 过表达及蛋白分泌,减轻炎症反应,并恢复机体免疫功能的稳定和平衡。     

单核细胞趋化蛋白3、白介素8及干扰素γ对寻常型银屑病患者外周血中性粒细胞趋化功能的影响

目的研究单核细胞趋化蛋白3(MCP-3)及其他银屑病相关细胞因子对银屑病患者中性粒细胞(PMN)趋化功能的影响。方法体外分离银屑病患者外周血PMN,用Transwell趋化小室测定MCP-3、IL-8、IFN-γ对其趋化功能的影响。结果MCP-3组与对照组比较差异无统计学意义,IL-8组对PMN有较强趋化作用,MCP-3 IL-8组趋化性明显高于IL-8组。各组细胞用IFN-γ处理前后比较,除MCP-3 IL-8组外,其他组差异均无统计学意义。结论MCP-3本身对PMN无趋化作用,但它能协同IL-8趋化PMN,IFN-γ可进一步放大这种协同趋化作用。*     

Herpesvirus-encoded GPCRs rewire cellular signaling

Viral G-protein-coupled receptors (vGPCRs) are chemokine receptor homologues encoded by the Herpes- and Capripoxviridae. They are thought to have been hijacked from the host genome during the course of evolution. These vGPCRs play different roles in the viral lifecycle and associated pathologies. Three members of the Herpesviridae, Kaposi sarcoma-associated herpesvirus (KSHV), Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) are capable of setting up persistent latent infections in humans. Two of the herpesviruses, KSHV and EBV, are associated with cancer, while HCMV may have an oncomodulary effect.     

Leishmania donovani: Dynamics of L. donovani evasion of innate immune cell attack due to malnutrition in visceral leishmaniasis

ObjectiveMalnutrition may be significant in the modulation of immune responses in visceral leishmaniasis (VL). Data on the relationship between malnutrition and innate immune response in VL are limited. The aim of this study was to examine the effect of malnutrition on the profile of innate immune functions of polymorphonuclear neutrophil granulocytes (PMNs) and monocytes through comparison of well-nourished and malnourished Indian patients with VL.MethodsForty individuals were enrolled comprising 20 active and untreated cases of VL and 20 non-VL individuals from the endemic region of Bihar, India. The patients with VL were segregated into two groups of 10 well-nourished and 10 malnourished participants. Patients' blood samples were directed against a crude Leishmania donovani extract (soluble leishmanial antigen) and phorbol 12-myristate-13-acetate plus ionomycin. The transendothelial cell adherence migration abilities of the PMNs and monocytes directed against these antigens were determined in whole-blood assays by flow cytometry. The chemokine (interleukin [IL]-8, macrophage inflammatory protein [MIP]-1 α) and cytokine support (tumor necrosis factor -α, interferon [IFN]-γ, IL-10), which could be significant in transendothelial cell migration, and efficacies of antileishmanial phagocytic function and reactive oxygen species (ROS) generation were also determined.ResultsSevere hindrance in the adherence of innate immune cells to the endothelial wall due to Leishmania parasites, as revealed by decreased shedding of l-selectin (CD62 L) and down-regulation of CD11 b expression on the surface of the PMNs and monocytes, occurred in malnourished VL patients. The production of MIP-1 α and IL-8 in response to L. donovani antigen was reduced in malnourished patients. In contrast, malnutrition in VL patients significantly reduced the IFN-γ and TNF-α produced by these immune cells, whereas the levels of IL-10 were significantly elevated. Malnourished VL patients were observed with severely dysfunctional PMNs and monocytes in terms of ROS activity that could not be recovered by stimulation with L. donovani antigen.ConclusionsMalnutrition linked to VL can be a decisive factor in the dynamics of L. donovani evasion of innate immune cell function in VL patients.     

Chemokine receptor expression on infiltrating lymphocytes from abdominal aortic aneurysms: role of CXCR4-CXCL12 in lymphoid recruitment

This is a study to define the profile of chemokine receptors expressed on isolated infiltrating lymphocytes in human abdominal aortic aneurysms (AAAs), and to examine their role in lymphoid recruitment. AAA T-lymphocytes were CXCR4-positive, CCR7-negative and partially CXCR3 and CCR5-positive. Functionally, AAA T-cells were proinflammatory effector cells, as they produced IFN-γ and granzyme A. AAA B-lymphocytes were CXCR4-positive and exhibited low CXCR5 expression. A relevant feature of infiltrating T- and B-lymphocytes was the high intensity of CXCR4 expression and the capability to migrate to CXCL12. CXCL12-producing cells were found in the adventitia of AAA. These cells were CD45-negative and partially VCAM-1 and DR-positive. In summary, the present results suggest that CXCR4, expressed on infiltrating lymphocytes, and CXCL12, expressed on stromal cells, is involved in the recruitment of lymphocytes within the arterial wall in AAA.     

CXCR3阻断剂Genistein对大鼠胰腺移植后排斥反应的影响

目的 探讨Genistein对大鼠移植胰腺急性排斥反应的影响以及作用机制.方法 以Wistar和SD大鼠为供体,SD大鼠为受体,建立胰腺移植模型.分成同种系移植组、异种系移植组和Genistein治疗组(在异种系移植后每天腹腔注射Genistein 10 mg/kg体重,共7 d),每组lO只.术后7 d切除移植物行病理学观察及CXCR3免疫组化染色,抽血检测血清IFN-γ、IL-2浓度.结果 同种系移植组移植物无急性排斥反应,无CXCR3表达.血清IFN-γ、IL-2浓度分别为(2.76±0.66)pg/ml和(11.83±1.86)pg/ml;异种系移植组的移植胰腺发生强烈的镜下排斥反应,CXCR3强阳性表达,血清IFN-γ、IL-2浓度分别为(32.64±2.52)PS/ml和(29.59±1.71)PS/ml,较同种系移植组显著升高(P<0.05);Genistein治疗组的移植胰腺排斥反应减轻,CXCR3弱阳性表达,血清IFN-γ、IL-2浓度分别为(15.94±1.95)Pg/ml和(22.62±1.76)ps/ml,较异种系移植组显著降低(P<0.05),但高于同种系移植组(P<0.05).结论 Genistein通过阻断CXCR3的表达,可有效减轻胰腺移植后急性排斥反应.     

Evolution and function of chemokine receptors in the immune system of lower vertebrates

Chemokine receptors and their counterpart ligands are one of the evolutionary innovations of vertebrates. They play a guiding role in the coordination of cell trafficking in many biological processes. Comparative syntenic and phylogenetic analyses provide insight into the evolution of chemokine receptors and suggest that the repertoire of chemokine receptors varies in each species, regardless of the evolutionary position of the species. Despite the rapid evolution of chemokine receptors, the expression and function of orthologous chemokine receptors in lower and higher vertebrates are very similar. This is also true for the chemokine ligands that have been examined so far, such as CXCL8, CXCL12, and CCL25. As examples, this review will discuss how the evolution of the chemokine receptor CXCR4 is coincident with the emergence of lymphocytes in jawless vertebrates (lamprey); and that, in jawed vertebrates, CXCR4 and CCR9 are involved in thymus colonization. In myeloid cells, the function of CXCR1 in neutrophils and the expression of CXCR3 in macrophages and DCs are evolutionarily conserved between fish and mammals. In this context, medaka and zebrafish are outstanding models for studying the function of chemokines and their receptors.