BACKGROUND & AIMS: :CCR9 + T helper (Th) cells can induce Sjögren-like symptoms in mice and both CCR9 + Th cells and their ligand CCL25 are increased in the salivary glands of primary Sjögren's syndrome (pSS) patients. Increased circulating CCR9 + Th cells are present in pSS patients. CCR9 + Th cells are hyperresponsive to IL-7, secrete high levels of IFN-γ, IL-21, IL-17 and IL-4 and potently stimulate B cells in both patients and healthy individuals. Our aim was to study co-expression of chemokine receptors on CCR9 + Th cells and whether in pSS this might differentially affect CCR9 + Th cell frequencies. Frequencies of circulating CCR9 + and CCR9- Th cells co-expressing CXCR3, CCR4, CCR6 and CCR10 were studied in pSS patients and healthy controls. CCL25, CXCL10, CCL17, CCL20 and CCL27 mRNA and protein expression of salivary gland tissue of pSS and non-Sjögren's sicca (non-SS) patients was assessed. Chemotaxis assays were performed to study migration induced by CXCL10 and CCL25. Higher expression of CXCR3, CCR4 and CCR6 but not CCR10 was observed on CCR9 + Th cells as compared to cells lacking CCR9. Decreased frequencies of circulating memory CCR9 + CXCR3+ Th cells were found in pSS patients, which was most pronounced in the effector memory subset. Increased salivary gland CCL25 and CXCL10 expression significantly correlated and both ligands functioned synergistically based on in vitro induced chemotaxis. Decreased memory CXCR3 + CCR9+ Th cells in blood of pSS patients may be due to a concerted action of overexpressed ligands at the site of inflammation in the salivary glands facilitating their preferential migration and positioning in the lymphocytic infiltrates.

背景与目标： : CCR9 T辅助 (Th) 细胞可以在小鼠中诱导sj ö gren样症状，并且原发性sj ö gren综合征 (pSS) 患者的唾液腺中CCR9 Th细胞及其配体CCL25均增加。pSS患者存在循环CCR9 + Th细胞增加。CCR9 + Th细胞对IL-7反应过度，分泌高水平的IFN-γ，IL-21，IL-17和IL-4，并有效刺激患者和健康个体的b细胞。我们的目的是研究趋化因子受体在CCR9 Th细胞上的共表达，以及在pSS中这是否可能差异影响CCR9 Th细胞频率。在pSS患者和健康对照中研究了共同表达CXCR3，CCR4，CCR6和CCR10的循环CCR9和CCR9- Th细胞的频率。评估了pSS和非sj ö gren sicca (非SS) 患者唾液腺组织的CCL25，CXCL10，CCL17，CCL20和CCL27 mRNA和蛋白表达。进行趋化性测定以研究CXCL10和ccl25诱导的迁移。与缺乏CCR9的细胞相比，在CCR9 Th细胞上观察到CXCR3，CCR4和CCR6的表达更高，但没有CCR10。在pSS患者中发现循环记忆CCR9 CXCR3 Th细胞的频率降低，这在效应记忆子集中最为明显。唾液腺CCL25和CXCL10表达的增加显着相关，并且两个配体基于体外诱导的趋化性协同作用。pSS患者血液中记忆CXCR3 CCR9 Th细胞减少可能是由于唾液腺炎症部位过度表达的配体协同作用，促进了它们在淋巴细胞浸润中的优先迁移和定位。

BACKGROUND & AIMS: :Chemokines and their receptors are essential for leukocyte trafficking, and are also involved in cancer metastasis to specific organs. Although the migration of tumor cells into the lymph nodes is an important aspect of cancer, the processes involved are poorly understood. Chemokine receptors CCR7 and CXCR3 have been shown to play an important role in tumor cell migration and lymph node metastasis. Therefore, the assessment of chemokine receptor expression on lung adenocarcinomas may improve the prediction of the spread of this carcinoma to the lymph nodes. In this study, we examined the expression and function of these two chemokine receptors (CCR7 and CXCR3) in lung adenocarcinoma. By using flow cytometry, they were detected in all of the lung adenocarcinoma cell lines examined. In the chemotaxis assays, A549 cells exhibited CCL21-induced migration, which was significantly suppressed by neutralizing anti-CCR7 antibody. The CXCL10-induced migration of A549 cells was also significantly suppressed by neutralizing anti-CXCR3 antibody. In clinical lung adenocarcinoma samples, we found the expression of CCR7 and CXCR3 in 65 and 90% cases, respectively, most of which had lymph node metastasis. Importantly, the expression of CCR7 was significantly associated with lymph node metastasis, although the expression of CXCR3 was not. These results suggest that the activation of CCR7 and CXCR3 with their ligands preferentially stimulates lung adenocarcinoma metastasis to the draining lymph nodes.

背景与目标： : 趋化因子及其受体对于白细胞运输至关重要，并且还参与癌症向特定器官的转移。尽管肿瘤细胞向淋巴结的迁移是癌症的重要方面，但对所涉及的过程知之甚少。趋化因子受体CCR7和CXCR3已被证明在肿瘤细胞迁移和淋巴结转移中起重要作用。因此，评估趋化因子受体在肺腺癌上的表达可能会改善该癌向淋巴结扩散的预测。在这项研究中，我们检查了这两种趋化因子受体 (CCR7和CXCR3) 在肺腺癌中的表达和功能。通过使用流式细胞仪，在所有检查的肺腺癌细胞系中检测到它们。在趋化性测定中，A549细胞表现出CCL21-induced的迁移，其被中和anti-CCR7抗体显著抑制。A549细胞的CXCL10-induced迁移也被中和anti-CXCR3抗体显著抑制。在临床肺腺癌样本中，我们发现CCR7和CXCR3分别在65和90% 例中表达，其中大多数有淋巴结转移。重要的是，CCR7的表达与淋巴结转移显着相关，尽管CXCR3的表达并非如此。这些结果表明，CCR7和CXCR3及其配体的激活优先刺激肺腺癌转移至引流淋巴结。

BACKGROUND & AIMS: AIMS:The CXC chemokine CXCL10 is up-regulated in the infarcted myocardium and limits cardiac fibrosis by inhibiting growth factor-mediated fibroblast migration. CXCL10 signals by binding to its receptor CXCR3; however, recently CXCR3-independent CXCL10 actions have been suggested. Our study explores the role of CXCR3 signalling in myocardial infarction and investigates its involvement in mediating the anti-fibrotic effects of CXCL10. METHODS AND RESULTS:Wild-type and CXCR3 null mice underwent reperfused infarction protocols. CXCL10 was markedly induced in the infarct; in contrast, expression of the other two CXCR3 ligands, CXCL9 and CXCL11 was extremely low. CXCR3 loss did not affect scar size, geometric ventricular remodelling, collagen deposition, and systolic dysfunction of the infarcted heart. CXCR3 null mice had increased peak neutrophil recruitment and delayed myofibroblast infiltration in the infarcted heart, but exhibited comparable myocardial expression of pro-inflammatory cytokines and chemokines. In vitro, CXCL10 did not modulate Transforming Growth Factor (TGF)-β signalling, but inhibited basic fibroblast growth factor (bFGF)-induced cardiac fibroblast migration in both wild-type and CXCR3 null cells. Treatment of fibroblasts with heparinase and chondroitinase to cleave glycosaminoglycan chains abrogated the inhibitory effects of CXCL10 on cell migration. CONCLUSION:CXCR3 signalling does not critically regulate cardiac remodelling and dysfunction following myocardial infarction. The anti-fibrotic effects of CXCL10 in the healing infarct and in isolated cardiac fibroblasts are CXCR3-independent and may be mediated through proteoglycan signalling. Thus, administration of CXCR3-defective forms of CXCL10 may be an effective anti-fibrotic strategy in the remodelling myocardium without activating a potentially injurious, CXCR3-driven T cell response.

背景与目标：

BACKGROUND & AIMS: :The chemokine receptor CXCR3 promotes the trafficking of activated T and NK cells in response to three ligands, CXCL9, CXCL10, and CXCL11. Although these chemokines are produced in the CNS in multiple sclerosis and experimental autoimmune encephalomyelitis (EAE), their role in the pathogenesis of CNS autoimmunity is unresolved. We examined the function of CXCR3 signaling in EAE using mice that were deficient for CXCR3 (CXCR3(-/-)). The time to onset and peak disease severity were similar for CXCR3(-/-) and wild-type (WT) animals; however, CXCR3(-/-) mice had more severe chronic disease with increased demyelination and axonal damage. The inflammatory lesions in WT mice consisted of well-demarcated perivascular mononuclear cell infiltrates, mainly in the spinal cord and cerebellum. In CXCR3(-/-) mice, these lesions were more widespread throughout the CNS and were diffused and poorly organized, with T cells and highly activated microglia/macrophages scattered throughout the white matter. Although the number of CD4(+) and CD8(+) T cells infiltrating the CNS were similar in CXCR3(-/-) and WT mice, Foxp3(+) regulatory T cells were significantly reduced in number and dispersed in CXCR3(-/-) mice. The expression of various chemokine and cytokine genes in the CNS was similar in CXCR3(-/-) and WT mice. The genes for the CXCR3 ligands were expressed predominantly in and/or immediately surrounding the mononuclear cell infiltrates. We conclude that in EAE, CXCR3 signaling constrains T cells to the perivascular space in the CNS and augments regulatory T cell recruitment and effector T cell interaction, thus limiting autoimmune-mediated tissue damage.

背景与目标： : 趋化因子受体CXCR3促进活化的T和NK细胞的运输，以响应三个配体CXCL9，CXCL10和cxcl11。尽管这些趋化因子是在多发性硬化症和实验性自身免疫性脑脊髓炎 (EAE) 的中枢神经系统中产生的，但它们在中枢神经系统自身免疫发病机理中的作用尚未解决。我们使用缺乏CXCR3(-/-)) 的小鼠检查了EAE中CXCR3信号传导的功能。CXCR3(-/-) 和野生型 (WT) 动物的发病时间和高峰疾病严重程度相似; 然而，CXCR3(-/-) 小鼠患有更严重的慢性疾病，脱髓鞘和轴突损伤增加。WT小鼠的炎性病变由边界明确的血管周围单核细胞浸润组成，主要在脊髓和小脑中。在CXCR3(-/-) 小鼠中，这些病变在整个中枢神经系统中更为广泛，并且弥散且组织不良，T细胞和高度活化的小胶质细胞/巨噬细胞散布在整个白质中。尽管在CXCR3(-/-) 和WT小鼠中浸润CNS的CD4 () 和CD8 () T细胞的数量相似，但在CXCR3(-/-) 小鼠中，Foxp3 () 调节性T细胞的数量显着减少并分散。在CXCR3(-/-) 和WT小鼠中，CNS中各种趋化因子和细胞因子基因的表达相似。CXCR3配体的基因主要在单核细胞浸润处和/或紧邻其周围表达。我们得出的结论是，在EAE中，CXCR3信号传导将T细胞约束到CNS中的血管周围空间，并增强调节性T细胞募集和效应T细胞相互作用，从而限制了自身免疫介导的组织损伤。

BACKGROUND & AIMS: :The CC chemokine known as 6Ckine (SLC, Exodus-2, or TCA4) has been identified as a ligand for CCR7. Mouse 6Ckine has also been shown to signal through mouse CXCR3 and share some of the activities of IFN-gamma inducible protein 10 and monokine induced by IFN-gamma. Nonetheless, human 6Ckine has not been shown to bind CXCR3 receptor or have angiostatic activity. In this study, we report that human 6Ckine does not induce a calcium flux in either human CXCR3 or mouse CXCR3 transfected cells, although it is an equally potent agonist as mouse 6Ckine and human macrophage inflammatory protein-3beta in human CCR7 transfected cells. Mouse 6Ckine (but not human 6Ckine) is capable of competing with radiolabeled IFN-gamma inducible protein 10 for human CXCR3. In addition, radiolabeled human 6Ckine does not bind to either human CXCR3 or mouse CXCR3. Together these data suggest that human CC chemokine 6Ckine is not a ligand for the human or mouse CXC chemokine receptor CXCR3.

背景与目标： : 被称为6ckine的CC趋化因子 (SLC，Exodus-2或TCA4) 已被确定为ccr7的配体。小鼠6Ckine也已显示通过小鼠CXCR3发出信号，并共享IFN-γ 诱导蛋白10和IFN-γ 诱导的单核因子的一些活性。尽管如此，尚未显示人6ckine结合CXCR3受体或具有血管抑制活性。在这项研究中，我们报告了人6ckine不会在人CXCR3或小鼠CXCR3转染的细胞中诱导钙通量，尽管它是与人CCR7转染的细胞中的小鼠6ckine和人巨噬细胞炎性protein-3beta同样有效的激动剂。小鼠6Ckine (但不是人6Ckine) 能够与放射性标记的IFN-γ 诱导蛋白10竞争人cxcr3。此外，放射性标记的人6ckine不与人CXCR3或小鼠CXCR3结合。这些数据一起表明，人CC趋化因子6ckine不是人或小鼠CXC趋化因子受体cxcr3的配体。

BACKGROUND & AIMS: BACKGROUND:Since human peripheral eosinophils have been shown to migrate to the CXC chemokine receptor 3 (CXCR3) ligands IFN-gamma-inducible protein 10 (IP10) and monokine induced by IFN-gamma (Mig), this confirms that CXCR3 is functionally expressed on these cells. IP10 expression has been shown to be increased in the airways of asthmatics. Eosinophil accumulations are found in bronchoalveolar lavage fluid (BALF) from patients with chronic eosinophilic pneumonia (CEP). To examine the contribution of IP10 and Mig in the pathogenesis of CEP, we measured the concentration of IP10 and Mig, and evaluated the expression of CXCR3 on eosinophils in BALF taken from patients with CEP. METHODS:The concentrations of IP10 and Mig in BALF were measured by ELISA. The proportion of CXCR3-expressing CD4+ T cells and CD16-negative eosinophils was determined by flow cytometry. RESULTS:The BALF concentrations of IP10 and Mig were higher in patients with CEP, as well as in patients with sarcoidosis, when compared to healthy controls. The absolute number of CXCR3+ CD4+ T cells was significantly higher in the BALF of patients with sarcoidosis, but not in the patients with CEP, when compared to healthy volunteers. There were higher percentages of CXCR3-expressing eosinophils in the BALF than in the peripheral blood of patients with CEP. CONCLUSIONS:Our findings suggest that IP10 and Mig contribute to the accumulation of CXCR3-expressing eosinophils in the lungs of patients with CEP, and modulate the eosinophilic inflammation of the lung.

背景与目标：

BACKGROUND & AIMS: :Chemokines are recognized as functionally important in many pathological disorders, which has led to increased interest in mechanisms related to the regulation of chemokine receptor (CKR) expression. Known mechanisms for regulating CKR activity are changes in gene expression or posttranslational modifications. However, little is known about CKR with respect to a third regulatory mechanism, which is observed among other seven-transmembrane receptor subfamilies, the concept of differential splicing or processing of heteronuclear RNA. We now report on the discovery of a variant human CKR, CXCR3, resulting from alternative splicing via exon skipping. The observed RNA processing entails a drastically altered C-terminal protein sequence with a predicted four- or five-transmembrane domain structure, differing from all known functional CKR. However, our data indicate that that this splice variant, which we termed CXCR3-alt, despite its severe structural changes still localizes to the cell surface and mediates functional activity of CXCL11.

背景与目标： : 趋化因子在许多病理疾病中被认为具有重要的功能，这导致人们对与趋化因子受体 (CKR) 表达调节有关的机制的兴趣增加。调节CKR活性的已知机制是基因表达的变化或翻译后修饰。然而，关于第三种调节机制的CKR知之甚少，在其他七个跨膜受体亚家族中，观察到这种调节机制，即异核RNA的差异剪接或加工的概念。我们现在报告发现了人类CKR变异体CXCR3，该变异体是通过外显子跳跃产生的。观察到的RNA加工过程需要急剧改变的C末端蛋白序列，具有预测的四或五跨膜结构域结构，与所有已知的功能CKR不同。然而，我们的数据表明，尽管这种严重的结构变化，但我们称之为CXCR3-alt的剪接变体仍位于细胞表面并介导cxcl11的功能活性。

BACKGROUND & AIMS: :We investigated possible cellular receptors for the human CXC chemokine platelet factor-4 variant/CXCL4L1, a potent inhibitor of angiogenesis. We found that CXCL4L1 has lower affinity for heparin and chondroitin sulfate-E than platelet factor-4 (CXCL4) and showed that CXCL10 and CXCL4L1 could displace each other on microvascular endothelial cells. Labeled CXCL4L1 also bound to CXCR3A- and CXCR3B-transfectants and was displaced by CXCL4L1, CXCL4, and CXCL10. The CXCL4L1 anti-angiogenic activity was blocked by anti-CXCR3 antibodies (Abs) in the Matrigel and cornea micropocket assays. CXCL4L1 application in CXCR3(-/-) or in wild-type mice treated with neutralizing anti-CXCR3 Abs, resulted in reduced inhibitory activity of CXCL4L1 on tumor growth and vascularization of Lewis lung carcinoma. Furthermore, CXCL4L1 and CXCL4 chemoattracted activated T cells, human natural killer cells, and human immature dendritic cells (DCs). Migration of DCs toward CXCL4 and CXCL4L1 was desensitized by preincubation with CXCL10 and CXCL11, inhibited by pertussis toxin, and neutralized by anti-CXCR3 Abs. Chemotaxis of T cells, natural killer cells, and DCs is likely to contribute to the antitumoral action. However, the in vivo data indicate that the angiostatic property of CXCL4L1 is equally important in retarding tumor growth. Thus, both CXCR3A and CXCR3B are implicated in the chemotactic and vascular effects of CXCL4L1.

背景与目标： : 我们研究了人类CXC趋化因子血小板因子4变体/CXCL4L1 (一种有效的血管生成抑制剂) 的可能细胞受体。我们发现CXCL4L1对肝素和硫酸软骨素E的亲和力低于血小板因子4 (CXCL4)，并表明CXCL10和CXCL4L1可以在微血管内皮细胞上相互取代。标记的CXCL4L1也与CXCR3A-和CXCR3B-transfectants结合，并被CXCL4L1、CXCL4和cxcl10取代。在Matrigel和角膜微袋试验中，CXCL4L1抗血管生成活性被anti-CXCR3抗体 (Abs) 阻断。CXCL4L1在CXCR3(-/-) 或用中和anti-CXCR3 Abs处理的野生型小鼠中的应用导致CXCL4L1对Lewis肺癌的肿瘤生长和血管形成的抑制活性降低。此外，CXCL4L1和CXCL4化学吸引活化的T细胞，人自然杀伤细胞和人未成熟树突状细胞 (dc)。通过与CXCL10和CXCL11预孵育，使dc向CXCL4和CXCL4L1的迁移脱敏，被百日咳毒素抑制，并被anti-CXCR3 Abs中和。T细胞，自然杀伤细胞和dc的趋化性可能有助于抗肿瘤作用。然而，体内数据表明，CXCL4L1的血管抑制特性在延缓肿瘤生长方面同样重要。因此，CXCR3A和CXCR3B都与CXCL4L1的趋化和血管作用有关。

BACKGROUND & AIMS: :Among the first cells to invade a site of infection, polymorphonuclear neutrophils (PMN) play an important role in the control of numerous infections. While PMN are considered critical for control of acute infections, their role in chronic infections remains less well understood. Here we report that PMN are essential for accurate early granuloma formation during chronic M. tuberculosis infection without influencing mycobacterial growth restriction. The PMN-mediated regulation of granuloma formation depended on chemokines signaling through CXCR3, in particular MIG, as indicated by immune histochemical analysis of lung sections from C57BL/6 wild-type and CXCR3(-/-) mutant mice and supported by microarray transcriptome analysis. Hence, PMN play a central role in regulating the focal granulomatous response in the lung, and this early granuloma formation can be segregated from long-term protection against pulmonary M. tuberculosis infection.

背景与目标： : 在第一批侵入感染部位的细胞中，多形核中性粒细胞 (PMN) 在控制多种感染中起着重要作用。虽然PMN被认为对于控制急性感染至关重要，但它们在慢性感染中的作用仍鲜为人知。在这里，我们报告了PMN对于慢性结核分枝杆菌感染期间准确的早期肉芽肿形成至关重要，而不会影响分枝杆菌的生长限制。PMN介导的肉芽肿形成的调节取决于通过CXCR3 (特别是MIG) 的趋化因子信号传导，如C57BL/6野生型和CXCR3(-/-) 突变小鼠的肺切片的免疫组织化学分析所表明的，并由微阵列转录组分析。因此，PMN在调节肺部局部肉芽肿反应中起着核心作用，这种早期肉芽肿的形成可以与针对肺结核分枝杆菌感染的长期保护措施分开。

BACKGROUND & AIMS: BACKGROUND:Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by repeated inhalations of finely dispersed organic particles or low molecular weight chemicals. The disease is characterized by an alveolitis sustained by CD8(+) cytotoxic T lymphocytes, granuloma formation, and, whenever antigenic exposition continues, fibrosis. Although it is known that T-cell migration into the lungs is crucial in HP reaction, mechanisms implicated in this process remain undefined. METHODS:Using flow cytometry, immunohistochemistry, confocal microscopy analysis and chemotaxis assays we evaluated whether CXCL10 and its receptor CXCR3 regulate the trafficking of CD8(+) T cells in HP lung. RESULTS:Our data demonstrated that lymphocytes infiltrating lung biopsies are CD8 T cells which strongly stain for CXCR3. However, T cells accumulating in the BAL of HP were CXCR3(+)/IFNgamma(+) Tc1 cells exhibiting a strong in vitro migratory capability in response to CXCL10. Alveolar macrophages expressed and secreted, in response to IFN-gamma, definite levels of CXCL10 capable of inducing chemotaxis of the CXCR3(+) T-cell line. Interestingly, striking levels of CXCR3 ligands could be demonstrated in the fluid component of the BAL in individuals with HP. CONCLUSION:These data indicate that IFN-gamma mediates the recruitment of lymphocytes into the lung via production of the chemokine CXCL10, resulting in Tc1-cell alveolitis and granuloma formation.

背景与目标：

BACKGROUND & AIMS: BACKGROUND:Adoptive therapy with tumour-infiltrating lymphocytes (TILs) induces durable complete responses (CR) in ∼20% of patients with metastatic melanoma. The recruitment of T cells through CXCR3/CCR5 chemokine ligands is critical for immune-mediated rejection. We postulated that polymorphisms and/or expression of CXCR3/CCR5 in TILs and the expression of their ligands in tumour influence the migration of TILs to tumours and tumour regression. METHODS:Tumour-infiltrating lymphocytes from 142 metastatic melanoma patients enrolled in adoptive therapy trials were genotyped for CXCR3 rs2280964 and CCR5-Δ32 deletion, which encodes a protein not expressed on the cell surface. Expression of CXCR3/CCR5 in TILs and CXCR3/CCR5 and ligand genes in 113 available parental tumours was also assessed. Tumour-infiltrating lymphocyte data were validated by flow cytometry (N=50). RESULTS:The full gene expression/polymorphism model, which includes CXCR3 and CCR5 expression data, CCR5-Δ32 polymorphism data and their interaction, was significantly associated with both CR and overall response (OR; P=0.0009, and P=0.007, respectively). More in detail, the predicted underexpression of both CXCR3 and CCR5 according to gene expression and polymorphism data (protein prediction model, PPM) was associated with response to therapy (odds ratio=6.16 and 2.32, for CR and OR, respectively). Flow cytometric analysis confirmed the PPM. Coordinate upregulation of CXCL9, CXCL10, CXCL11, and CCL5 in pretreatment tumour biopsies was associated with OR. CONCLUSION:Coordinate overexpression of CXCL9, CXCL10, CXCL11, and CCL5 in pretreatment tumours was associated with responsiveness to treatment. Conversely, CCR5-Δ32 polymorphism and CXCR3/CCR5 underexpression influence downregulation of the corresponding receptors in TILs and were associated with likelihood and degree of response.

背景与目标：

BACKGROUND & AIMS: BACKGROUND:We previously demonstrated the existence of two distinct subsets of T cell receptor (TCR)alphabeta+CD8alphabeta+ single positive (SP) cells in human postnatal thymus which express the chemokine receptor CCR7 or CXCR3 and migrate in vitro in response to their specific ligands. AIM:To investigate whether these two CD8+ thymocyte subsets had distinct peripheral colonisation. METHODS:TCRalphabeta+CD8+ SP cells were obtained from normal postnatal thymus, mesenteric lymph node (LNs), small bowel, and peripheral blood (PB) specimens. Cells were then evaluated for expression of surface molecules, cytolytic potential, telomere length, and profile of cytokine production. RESULTS:CD8+CCR7+CXCR3- thymocytes exhibited CD62L, in common with those which localise to LNs. In contrast, CD8+CCR7-CXCR3+ thymocytes lacked CD62L but exhibited CD103, similar to intraepithelial lymphocytes (IELs) present in the gut mucosa where the CXCR3 ligand, CXCL10, and the CD103 ligand, E-cadherin, are highly and consistently expressed. In addition, thymocytes and gut CD8+CXCR3+CD103+ cells showed comparable telomere length, which was higher than that of PB CXCR3+CD8+ T cells. However, both of these populations contained perforin and granzyme A, and displayed the ability to produce interferon gamma and interleukin 2. Of note, CXCR3 deficient, in comparison with wild-type C57Black/6, mice showed decreased proportions of CD3+CD8alphabeta+ and increased proportions of CD3+CD8alphaalpha+ lymphocytes at gut level. Moreover, adoptive transfer of CD3+CD8alphabeta+ thymocytes from wild-type into CXCR3 deficient mice resulted in a significant increase in CD3+CD8alphabeta+ T cells in the gut mucosa but not in other tissues. CONCLUSIONS:The results of this study demonstrate the existence of a previously unrecognised subset of TCRalphabeta+CD8alphabeta+ SP CXCR3+CD103+ thymocytes which share phenotypic and functional features with CD8+ IELs, thus suggesting the possibility of their direct colonisation of the gut mucosa.

背景与目标：

BACKGROUND & AIMS: PURPOSE:The cytokine milieu in pancreatic ductal adenocarcinoma (PDAC) promotes tumor progression and immune suppression, contributing to the dismal prognosis of patients with PDAC. The roles of many of these cytokines, however, have not been thoroughly investigated in PDAC. EXPERIMENTAL DESIGN:PDAC microarray and The Cancer Genome Atlas datasets were analyzed to identify cytokines and cognate receptors overexpressed in PDAC and associated with survival. Pathway and CIBERSORT analyses were used to elucidate potential mechanisms of altered patient survival. Comparative analysis of cytokine expression in KPC (K-rasG12D; TP53R172H; Pdx-1cre) and KC (K-rasG12D; Pdx-1cre) PDAC models and multicolor immunofluorescence (IF) staining of human PDAC-resected samples were used to validate these findings. RESULTS:CXCL9 and CXCL10 were among the most highly overexpressed cytokines by bioinformatics analyses, while their receptor, CXCR3, was significantly overexpressed by IHC analysis. Higher CXCR3 ligand expression was associated with shorter overall survival, while high CXCR3 expression was associated with better survival. The CXCR3 ligands, CXCL4, 9, and 10, were overexpressed in KPC compared with KC mice. Pathway analysis of CXCR3- and CXCR3 ligand-associated genes showed that CXCR3 is a marker of antitumor immunity, while its ligands may promote immunosuppression. CIBERSORT and IF studies of PDAC tissues demonstrated that high CXCR3 expression was associated with increased CD8+ T-cell and naïve B-cell signatures and loss of plasma cell signatures. CXCR3 ligand expression was associated with increased CD8+ T-cell signatures and loss of natural killer-cell signatures. CONCLUSIONS:CXCR3 ligands are overexpressed in PDAC and are associated with poor survival likely related to alterations in tumor immune infiltrate/activity.

背景与目标：

BACKGROUND & AIMS: :Chemokine-chemokine receptor interactions and the subsequent recruitment of T lymphocytes to the graft are believed to be among the initial events in the development of acute and chronic rejection of heart transplants. We sought to determine the role of chemokine receptor Cxcr3 on the development of acute and chronic rejection in a multiple minor Ag mismatched mouse heart transplant model. The frequencies and kinetics of immunodominant H60 (LTFNYRNL) miHA-specific CD8 T cells in wild-type or Cxcr3-/- C57BL/6 recipients were monitored using MHC class I tetramer after BALB/b donor hearts were transplanted. Acceptance of grafts, severity of rejection, and infiltration of T cells were not altered in Cxcr3-/- recipients. However, graft survival was moderately prolonged in Cxcr3-/- recipient mice undergoing acute rejection. Analyses of splenocytes, PBLs, and graft-infiltrating cells revealed increased alloreactive T cells (H60-specific CD8 T cells) in the peripheral blood and spleen but not in the graft. Adoptively transferred Cxcr3-/- CD8 T cells in the BALB/b heart-bearing B6 scid mice showed retention of alloreactive CD8 T cells in the blood but less infiltration into the graft. Cxcr3-/- recipients with long-term graft survival also showed a marked decrease of CD8+ T cell infiltration and reduced neo-intimal hyperplasia. These data indicate that Cxcr3 plays a critical role in the trafficking as well as activation of alloreactive T cells. This role is most eminent in a transplant model when a less complex inflammatory milieu is involved such as a well-matched graft and chronic rejection.

背景与目标： : 趋化因子-趋化因子受体的相互作用以及随后T淋巴细胞向移植物的募集被认为是心脏移植急性和慢性排斥反应发展的初始事件。我们试图确定趋化因子受体Cxcr3在多个次要Ag错配小鼠心脏移植模型中急性和慢性排斥反应的发展中的作用。BALB/b供体心脏移植后，使用MHC I类四聚体监测野生型或Cxcr3-/- C57BL/6受体中免疫显性H60 (LTFNYRNL) miHA特异性CD8 T细胞的频率和动力学。在Cxcr3-/-受体中，移植物的接受程度，排斥的严重程度和T细胞的浸润没有改变。然而，在接受急性排斥反应的Cxcr3-/-受体小鼠中，移植物存活率适度延长。对脾细胞，PBLs和移植物浸润细胞的分析显示，外周血和脾脏中的同种反应性T细胞 (H60-specific CD8 T细胞) 增加，但移植物中没有。在BALB/b心脏B6 scid小鼠中过继转移的Cxcr3-/- CD8 T细胞显示出血液中同种反应性CD8 T细胞的保留，但对移植物的浸润较少。具有长期移植物存活的Cxcr3-/-受体也显示出CD8 T细胞浸润的显着减少和新内膜增生的减少。这些数据表明Cxcr3在同种反应性T细胞的运输和激活中起关键作用。当涉及较不复杂的炎症环境 (例如匹配良好的移植物和慢性排斥反应) 时，这种作用在移植模型中最为突出。

BACKGROUND & AIMS: BACKGROUND AND PURPOSE:The chemokine receptor CXCR3 is a GPCR found predominantly on activated T cells. CXCR3 is activated by three endogenous peptides; CXCL9, CXCL10 and CXCL11. Recently, a small-molecule agonist, VUF10661, has been reported in the literature and synthesized in our laboratory. The aim of the present study was to provide a detailed pharmacological characterization of VUF10661 by comparing its effects with those of CXCL11. EXPERIMENTAL APPROACH:Agonistic properties of VUF10661 were assessed in a chemotaxis assay with murine L1.2 cells transiently transfected with cDNA encoding the human CXCR3 receptor and in binding studies, with [(125)I]-CXCL10 and [(125)I]-CXCL11, on membrane preparations from HEK293 cells stably expressing CXCR3. [(35)S]-GTPγS binding was used to determine its potency to induce CXCR3-mediated G protein activation and BRET-based assays to investigate its effects on intracellular cAMP levels and β-arrestin recruitment. KEY RESULTS:VUF10661 acted as a partial agonist in CXCR3-mediated chemotaxis, bound to CXCR3 in an allosteric fashion in ligand binding assays and activated G(i) proteins with the same efficacy as CXCL11 in the [(35)S]-GTPγS binding and cAMP assay, while it recruited more β-arrestin1 and β-arrestin2 to CXCR3 receptors than the chemokine. CONCLUSIONS AND IMPLICATIONS:VUF10661, like CXCL11, activates both G protein-dependent and -independent signalling via the CXCR3 receptor, but probably exerts its effects from an allosteric binding site that is different from that for CXCL11. It could stabilize different receptor and/or β-arrestin conformations leading to differences in functional output. Such ligand-biased signalling might offer interesting options for the therapeutic use of CXCR3 agonists.

背景与目标：