Urinary C-X-C Motif Chemokines 13:a Noninvasive Biomarker of Antibody-Mediated Renal Allograft Rejection

Objective: Since acute rejection remains one of the major complications which necessitate periodic surveillance, noninvasive diagnostic/prognostic methods are preferred by renal transplant recipients. Here, we explored whether urinary C-X-C motif chemokines 13(CXCL13) could be a potential candidate to reflect ongoing immune processes within the renal graft. Methods: We investigated urinary CXCL13 levels by a cross-sectional analysis of 146 renal allograft recipients and 40 healthy controls. Besides, a subset of patients(n=57) were followed-up for kinetic monitoring of immune status.Results: Urinary CXCL13/Cr was lower in normal transplants compared to those with acute tubular necrosis(ATN, P=0.001), chronic allograft nephropathy(CAN, P=0.01) and acute rejection(AR, P<0.0001), which yielded a good diagnosis performance of urinary CXCL13 for AR(AUC=0.818, P<0.0001). Interestingly, urinary CXCL13 further distinguished acute antibody mediated rejection(ABMR) from acute cellular rejection,with an AUC of 0.856. Besides, patients with steroid-resistant acute rejection had distinctly greater urinary CXCL13/Cr levels than patients with reversible acute rejection,P=0.001. Follow-up data revealed that urinary CXCL13/Cr varied in line with the occurrence of ABMR. Furthermore, elevated levels of urinary CXCL13/Cr within the first month was predictive of graft function at 3, 6 months, P=0.044 and 0.4. Conclusion: This study demonstrates that monitoring of urinary CXCL13/Cr might be a valuable noninvasive approach for the detection of AR, especially ABMR. Additionally, high urinary CXCL13/Cr levels related to the poor response to steroid treatment and predicted a compromised graft function after AR.

The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

Background: Recent studies indicate that C-X-C motif chemokine receptor 4(CXCR4) and its ligand, C-X-C motif chemokine ligand 12(CXCL12), stimulate expression of the cell cycle regulatory protein Cyclin D1 in neurofibromatosis 1-associated malignant peripheral nerve sheath tumor(MPNST) cells and promote their proliferation. In this study, we measured the expression of CXCR4, CXCL12, and Cyclin D1 proteins in sporadic MPNST tissues from Chinese patients and investigated their prognostic values.Methods: CXCR4, CXCL12, and Cyclin D1 protein expression in samples from 58 Chinese patients with sporadic MPNST was assessed with immunohistochemical staining.Their prognostic values were evaluated with Kaplan-Meier analysis and a log-rank test. Multivariate Cox regression analysis was used to identify independent prognostic factors.Results: High expression of CXCR4, CXCL12, and Cyclin D1 was observed in 19(32.8%), 32(55.2%), and 16(27.6%)samples, respectively. CXCR4 expression was positively correlated with CXCL12 expression(r = 0.334, P = 0.010) and Cyclin D1 expression(r = 0.309, P = 0.018). Patients with high CXCR4 expression showed longer overall survival than those with low CXCR4 expression(χ~2 = 4.642, P = 0.031).Conclusion: High CXCR4 expression may define a specific subtype of sporadic MPNST with favorable prognosis.

原发性肾病综合征患儿治疗前后外周血CXCL13和PD-L1~+B淋巴细胞水平变化及临床意义研究

目的 探讨原发性肾病综合征(primary nephrotic syndrome,PNS)患儿治疗前后外周血趋化因子C-X-C基序配体13(chemokine C-X-C motif ligand 13,CXCL13)及程序性死亡受体配体1(programmed death protein ligand 1,PDL1)+B淋巴细胞水平变化的临床意义。方法 选取2022年4~12月收治的激素敏感型初发PNS患儿52例,给予糖皮质激素治疗;以同期在医院体检的30例正常儿童作为健康对照。收集两组儿童临床实验室指标,流式细胞仪检测两组儿童外周血中总B细胞及其PD-L1~+B淋巴细胞比例;酶联免疫吸附法检测两组血清CXCL13,可溶性程序性死亡受体配体1(solubility programmed death protein ligand 1,s PD-L1)及细胞因子[转化生长因子-β1(transforming growth factor-β1,TGF-β1)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)和白细胞介素-1β(interleukin-1β,IL-1β)]水平;Pearson相关性分析CXCL13和PD-L1~+B淋巴细胞及其与实验室指标的相关性。结果与健康对照组比较,治疗前PNS组外周血总B细胞(12.54%±4.23%vs 4.95%±2.83%)和PD-L1~+B淋巴细胞比例(1.17%±0.38%vs 0.35%±0.12%),血清CXCL13(121.03±30.52 pg/ml vs 53.67±12.42 pg/ml)和s PD-L1(116.25±25.68pg/ml vs 47.27±8.14 pg/ml)水平及细胞因子TGF-β1(17.91±2.04 ng/ml vs 12.53±1.62 ng/ml),TNF-α(77.65±7.27ng/ml vs 52.43±4.68 ng/ml),IL-10(14.21±3.56 pg/ml vs 4.76±1.25 pg/ml),IL-1β(64.38±7.46 ng/ml vs 35.57±5.92ng/ml)水平均明显升高,差异具有统计学意义(t=-10.754,-11.468,-11.526,-14.271,-12.360,-17.048,-14.017,-18.103,均P<0.05)。与治疗前相比,PNS组患儿治疗后外周血总B细胞(6.20%±2.48%)和PD-L1~+B淋巴细胞比例(0.43%±0.25%),血清CXCL13(65.27±14.16 pg/ml),s PD-L1(55.63±11.44 pg/ml),TGF-β1(14.35±1.82ng/ml),TNF-α(56.48±4.16ng/ml),IL-10(5.15±1.09 pg/ml),IL-1β(39.38±4.05 ng/ml)均明显降低,差异具有统计学意义(t=9.324,11.731,11.951,15.549,9.930,18.226,17.548,21.237,均P<0.05)。PNS患儿PD-L1~+B淋巴细胞比例与血清ALB和Ig G水平呈负相关(r=-0.619,-0.587,均P<0.05),与Ig M水平呈正相关(r=0.563,P<0.05)。CXCL13表达水平与血清ALB和Ig G水平呈负相关(r=-0.574,-0.522,均P<0.05)。PD-L1~+B淋巴细胞比例与CXCL13表达水平呈正相关(r=0.632,P<0.05)。结论 外周血CXCL13和PD-L1~+B淋巴细胞比例升高与PNS患儿体液免疫紊乱相关。CXCL13可能通过促进外周血中B淋巴细胞趋化性,促进免疫细胞生产炎症细胞因子,加重PNS过度免疫炎症反应。

趋化因子配体13在常见感染性病原体感染中的表达及意义

趋化因子配体13(C-X-C motif chemokine ligand 13,CXCL13)又称B细胞吸引趋化因子1或B淋巴细胞趋化因子,通过与B细胞、辅助T细胞上的G蛋白偶联趋化因子受体结合,诱导其向生发中心运输,促进B细胞成熟和抗体应答,从而实现对病毒清除以及炎症的快速控制。然而,目前相关研究表明CXCL13水平过高可能导致免疫反应受损,促进疾病进展。CXCL13水平变化可作为反映疾病进展、抗病毒疗效有效指标,并且对疫苗研发有所帮助。因此,本文主要对CXCL13在常见感染性病原体感染如人类免疫缺陷病毒感染、乙型肝炎病毒感染、结核分枝杆菌感染以及SARS-CoV-2感染中的表达及其意义进行综述。

The miR-9-5p/CXCL11 pathway is a key target of hydrogen sulfide-mediated inhibition of neuroinflammation in hypoxic ischemic brain injury

We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.

CXCL-13通过激活PI3K-Akt信号通路对人骨髓间充质干细胞迁移的影响

目的验证趋化因子-13(CXCL-13)通过激活PI3K-Akt信号通路对人骨髓间充质干细胞(hBMSC)迁移的影响。方法无任何刺激的hBMSCs为对照组;80μmol/L CXCL-13刺激hBMSCs为CXCL-13组;hBMSCs先用25 nmol/L LY294002培养40 min,再添加80μmol/L CXCL-13为PI3K抑制剂组;hBMSCs先用50 nmol/L Triciribine培养30 min,再添加80μmol/L CXCL-13为Akt抑制剂组。细胞划痕实验和Transwell细胞迁移实验检测四组hBMSCs划痕面积闭合率和细胞迁移率;酶联免疫吸附试验检测四组人PI3K、Akt、P-Akt蛋白的表达。结果CXCL-13组hBMSCs划痕面积闭合率和迁移率均高于对照组,PI3K抑制剂组和Akt抑制剂组hBMSCs划痕面积闭合率和迁移率均低于CXCL-13组(P<0.05或P<0.01);CXCL-13组人PI3K、Akt、P-Akt蛋白含量均高于对照组,PI3K抑制剂组和Akt抑制剂组人PI3K、Akt、P-Akt蛋白含量低于CXCL-13组(均P<0.01)。结论CXCL-13激活PI3K-Akt信号通路促进hBMSCs迁移。

CCL5、CXCL10和CXCL13在重度脑外伤患者中的表达和意义

目的:观察趋化因子配体5(chemokine C-C motif ligand 5,CCL5)、趋化因子CXC配体[chemokine(C-X-C motif)ligand,CXCL]10、CXCL13在创伤性脑损伤(traumatic brain injury,TBI)患者外周血液中的表达变化,分析这些趋化因子对损伤程度和预后的影响。方法:选取39例重度TBI患者[格拉斯哥昏迷评分(Glasgow coma scale,GCS)3~8分]与13例健康对照者。酶联免疫吸附测定法检测TBI患者术后1、3、7 d和对照组外周血中CCL5、CXCL10、CXCL13蛋白含量;Spearman相关分析对不同时间点外周血中CCL5、CXCL10、CXCL13和入院GCS、术后30 d GCS和格拉斯哥结局量表评分(Glasgow outcome scale,GOS)进行相关性分析。结果:与对照组相比,重度TBI患者外周血液中CCL5、CXCL10、CXCL13的表达量均明显升高(均P<0.05)。重度TBI患者术后1 d血液中CCL5、CXCL13的蛋白浓度与入院GCS评分呈负相关(P<0.05),术后3 d CXCL10的浓度与术后30 d GCS评分呈负相关(P<0.05)。重度TBI患者术后1 d CCL5的蛋白含量、术后1 d和3 d CXCL10蛋白含量及术后7 d CXCL13的蛋白含量与术后30 d GOS评分呈负相关(P<0.05)。结论:趋化因子CCL5、CXCL10、CXCL13在TBI早期表达量迅速增加,且表达量越高,损伤程度越重,预后可能越差。

趋化因子配体13在三阴性乳腺癌中的表达及临床意义

目的探讨趋化因子配体13(CXCL13)表达水平对三阴性乳腺癌(TNBC)患者总体生存期(OS)的影响,分析CXCL13与肿瘤浸润性淋巴细胞(TIL)浸润程度及常见免疫检查点表达的关系。方法下载癌症基因组图谱数据库中TNBC患者的转录组测序结果及临床资料数据,分析CXCL13表达与TNBC患者OS的关系。应用基因集富集分析(GSEA)软件挖掘CXCL13在TNBC中的生物学功能;应用肿瘤免疫评估资源(TIMER)数据库分析CXCL13与TNBC中TIL浸润程度及常见免疫检查点表达量的相关性。结果CXCL13在TNBC中呈显著高表达(P<0.001),其表达量与患者OS具有显著相关性(P<0.05)。GSEA结果显示,CXCL13在TNBC中高表达可使关键免疫相关通路活化。TIMER数据库分析显示,CXCL13与TNBC中TIL的浸润程度及常见免疫检查点程序性死亡受体1、程序性死亡受体-配体1及细胞毒性T淋巴细胞相关蛋白4的表达量呈显著正相关(P<0.001)。结论CXCL13是TNBC患者预后的预测因子,或可在TNBC的免疫治疗中发挥重要作用。

Autocrine IL-8 Contributes to Propionibacterium Acnes-induced Proliferation and Differentiation of HaCaT Cells via AKT/FOXO1/Autophagy

Objective Proprionibacterium acnes(P.acnes)-induced inflammatory responses,proliferation and differentiation of keratinocytes contribute to the progression of acne vulgaris(AV).P.acnes was found to enhance the production of interleukin-8(IL-8)by keratinocytes.This study aimed to investigate the role of IL-8 in P.acnes-induced proliferation and differentiation of keratinocytes and the underlying mechanism.Methods The P.acnes-stimulated HaCaT cell(a human keratinocyte cell line)model was established.Western blotting and immunofluorescence were performed to detect the expression of the IL-8 receptors C-X-C motif chemokine receptor 1(CXCR1)and C-X-C motif chemokine receptor 2(CXCR2)on HaCaT cells.Cell counting kit-8(CCK-8)assay,5-ethynyl-20-deoxyuridine(EdU)assay and Western blotting were performed to examine the effects of IL-8/CXCR2 axis on the proliferation and differentiation of HaCaT cells treated with P.acnes,the IL-8 neutralizing antibody,the CXCR2 antagonist(SB225002),or the CXCR1/CXCR2 antagonist(G31P).Western blotting,nuclear and cytoplasmic separation,CCK-8 assay,and EdU assay were employed to determine the downstream pathway of CXCR2 after P.acnes-stimulated HaCaT cells were treated with the CXCR2 antagonist,the protein kinase B(AKT)antagonist(AZD5363),or the constitutively active forkhead box O1(FOXO1)mutant.Finally,autophagy markers were measured in HaCaT cells following the transfection of the FOXO1 mutant or treatment with the autophagy inhibitor 3-methyladenine(3-MA).Results The expression levels of CXCR1 and CXCR2 were significantly increased on the membrane of HaCaT cells following P.acnes stimulation.The IL-8/CXCR2 axis predominantly promoted the proliferation and differentiation of P.acnes-induced HaCaT cells by activating AKT/FOXO1/autophagy signaling.In brief,IL-8 bound to its receptor CXCR2 on the membrane of keratinocytes to activate the AKT/FOXO1 axis.Subsequently,phosphorylated FOXO1 facilitated autophagy to promote the proliferation and differentiation of P.acnes-induced keratinocytes.Conclusion This study demonstrated the novel autocrine effect of IL-8 on the proliferation and differentiation of P.acnes-induced keratinocytes,suggesting a potential therapeutic target for AV.

Foodborne toxin Aflatoxin B_(1)induced glomerular podocyte inflammation through proteolysis of RelA,downregulation of miR-9 and CXCR4/TXNIP/NLRP3 pathway

Aflatoxin B_(1)(AFB_(1))is a naturally-occurring mycotoxin and recognized as the most toxic foodborne toxin,particularly causing damages to kidney.Glomerular podocytes are terminally differentiated epithelial cells.AFB_(1)induces podocyte inflammation,proteinuria and renal dysfunction.Studying the mechanism of AFB_(1)-induced podocyte inflammation and murine kidney dysfunction,we detected that AFB_(1)increased ubiquitindependent degradation of the transcription factor RelA through enhanced interaction of RelA with E3 ubiquitin ligase tripartite motif containing 7(TRIM7)in mouse podocyte clone-5(MPC-5)and mouse glomeruli.Reduction of RelA resulted in decreasing microRNA-9(miR-9)and activating the chemokine receptor 4(CXCR4),thioredoxin interacting protein(TXNIP),and NOD-like receptor pyrin domain-containing 3(NLRP3)signaling axis(CXCR4/TXNIP/NLRP3 pathway),leading to podocyte inflammation.We also determined that downregulation of miR-9 led to CXCR4 expression and the downstream TXNIP/NLRP3 pathway activation.Overexpression of miR-9 or deletion of CXCR4 suppressed AFB_(1)-induced CXCR4/TXNIP/NLRP3 pathway,resulting in alleviating podocyte inflammation and kidney dysfunction.Our findings indicated that ubiquitin-dependent proteolysis of RelA,downregulation of miR-9,and activation of CXCR4/TXNIP/NLRP3 pathway played an essential role in AFB_(1)-induced glomerular podocyte inflammation.Our study revealed a novel mechanism,via RelA,for the control of AFB_(1)’s nephrotoxicity,leading to an effective protection of food safety and public health.

CXCL10 Induces Lytic Reactivation of EBV through EXTL1

Epstein-Barr virus (EBV) infects over 90% of the global population, establishing latent infections in most individuals. Under specific conditions like inflammation and immune suppression, EBV can be reactivated, leading to the initiation and progression of related diseases. While inflammation is known to induce EBV reactivation, the precise mechanisms underlying this phenomenon remain unclear. Chemokine (C-X-C motif) ligand (CXCL10), a key inflammatory factor, plays a significant role in various infectious diseases. In this study, we investigated how CXCL10 levels regulate the transition between the latent and lytic replication phases of the EBV lifecycle using cell culture, Western blot, fluorescent quantitative PCR, immunofluorescence, and flow cytometric apoptosis assays. Our findings indicate that CXCL10 induces EBV transition from latency to lytic replication through its receptor CXCR3 by regulating the downstream effector, exostosis-like glycosyltransferase 1. Additionally, CXCL10 activates the JAK2/STAT3 pathway. This study confirms the role of CXCL10 in promoting EBV lytic replication, providing crucial insights into the pathogenic mechanisms of inflammation-triggered EBV reactivation.

基于网络药理学探讨趋化因子-13对间充质干细胞增殖和迁移的影响

目的以网络药理学技术探讨趋化因子-13(CXCL-13)对人骨髓间充质干细胞(BMSCs)增殖和迁移的影响。方法在线数据库预测CXCL-13作用于BMSCs的靶点。Metascape数据库对靶点的基因本体论和京都基因与基因组百科全书(KEGG)信号通路进行富集分析。STRING 11.0数据库进行蛋白质相互作用分析,Cytoscape 3.8的cyto Hubba 0.1插件筛选核心基因编码的蛋白质。BMSCs分为对照组、CXCL-13组和PI3K抑制剂组。分别以MTT、流式细胞术和Transwell细胞小室迁移实验检测各组BMSCs的吸光度(A)值、细胞凋亡率和细胞迁移数目情况;ELISA检测各组BMSCs上清液表皮生长因子(EGF)和血管内皮生长因子(VEGF)蛋白含量。Western blotting检测各组BMSCs的Akt、磷酸化Akt(p-Akt)蛋白的表达。结果 CXCL-13作用于BMSCs 21个靶点。与细胞增殖相关的生物学过程包括干细胞增殖、调节内皮细胞增殖、正向调控平滑肌细胞增殖等32条;与细胞迁移相关的生物学过程包括调节细胞迁移、阿米巴状细胞迁移、调节内皮细胞迁移等22条。KEGG通路包括癌症途径、PI3K-Akt信号通路、MAPK信号通路等40条。核心蛋白包括肿瘤蛋白P53(TP53)、表皮生长因子受体(EGFR)、90kD热休克蛋白αB1(HSP90AB1)、蛋白激酶Cα(PRKCA)、雌激素受体2(ESR2)及前列腺素E受体4(PTGER4)。与其他组相比,CXCL-13组BMSCs的吸光度(A)值和细胞迁移数目均显著增高(P<0.01,n=15),细胞凋亡率明显降低(P<0.01,n=15);PI3K抑制剂组BMSCs的A值、细胞凋亡率和细胞迁移数目与CXCL-13组相比均呈相反变化(P<0.01,n=15)。相对于对照组,CXCL-13组BMSCs的EGF和VEGF蛋白含量显著提高(P<0.01,n=15),Akt和p-Akt相对表达均明显升高(P<0.01,n=9);而PI3K抑制剂组EGF和VEGF蛋白含量、Akt和p-Akt相对表达呈相反变化。结论 CXCL-13激活PI3K-Akt通路促进BMSCs旁分泌EGF和VEGF蛋白,提高BMSCs增殖和迁移,抑制BMSCs凋亡。

Functions and mechanisms of chemokine receptor 7 in tumors of the digestive system

Chemokine(C-X-C motif)receptor 7(CXCR7),recently termed ACKR3,belongs to the G protein-coupled cell surface receptor family,binds to stromal cellderived factor-1[SDF-1,or chemokine(C-X-C motif)ligand 12]or chemokine(CX-C motif)ligand 11,and is the most common chemokine receptor expressed in a variety of cancer cells.SDF-1 binds to its receptor chemokine(C-X-C motif)receptor 4(CXCR4)and regulates cell proliferation,survival,angiogenesis and migration.In recent years,another new receptor for SDF-1,CXCR7,has been discovered,and CXCR7 has also been found to be expressed in a variety of tumor cells and tumor-related vascular endothelial cells.Many studies have shown that CXCR7 can promote the growth and metastasis of a variety of malignant tumor cells.Unlike CXCR4,CXCR7 exhibits a slight modification in the DRYLAIV motif and does not induce intracellular Ca^2+release following ligand binding,which is essential for recruiting and activating G proteins.CXCR7 is generally thought to work in three ways:(1)Recruitingβ-arrestin 2;(2)Heterodimerizing with CXCR4;and(3)Acting as a“scavenger”of SDF-1,thus lowering the level of SDF-1 to weaken the activity of CXCR4.In the present review,the expression and role of CXCR7,as well as its prognosis in cancers of the digestive system,were investigated.

Overexpression of amplified in breast cancer 1 (AIB1) gene promotes lung adenocarcinoma aggressiveness in vitro and in vivo by upregulating C-X-C motif chemokine receptor 4

Background:We previously found that overexpression of the gene known as amplified in breast cancer 1(AIB1)was associated with lymph node metastasis and poor prognosis in patients with lung adenocarcinoma.However,the role of AIB1 in that malignancy remains unknown.The present study aimed to investigate the function of AIB1 in the process of lung adenocarcinoma cell metastasis.Methods:A series of in vivo and in vitro assays were performed to elucidate the function of AIB1,while real-time PCR and Western blotting were utilized to identify the potential downstream targets of AIB1 in the process of lung adenocarcinoma metastasis.Rescue experiments and in vitro assays were performed to investigate whether the invasive-ness of AIB1-induced lung adenocarcinoma was mediated by C-X-C motif chemokine receptor 4(CXCR4).Results:The ectopic overexpression of AIB1 in lung adenocarcinoma cells substantially enhanced cell migration and invasive abilities in vitro and tumor metastasis in vivo,whereas the depletion of AIB1 expression substantially inhibited lung adenocarcinoma cell migration and invasion.CXCR4 was identified as a potential downstream target of AIB1 in lung adenocarcinoma.The knockdown of AIB1 greatly reduced CXCR4 gene expression at both the transcription and protein levels,whereas the knockdown of CXCR4 in cells with AIB1 ectopic overexpression diminished AIB1-induced migration and invasion in vitro and tumor metastasis in vivo.Furthermore,we found a significant positive association between the expression of AIB1 and CXCR4 in lung adenocarcinoma patients(183 cases),and the co-overexpression of AIB1 and CXCR4 predicted the poorest prognosis.Conclusions:These findings suggest that AIB1 promotes the aggressiveness of lung adenocarcinoma in vitro and in vivo by upregulating CXCR4 and that it might be usable as a novel prognostic marker and/or therapeutic target for this disease.

重型再生障碍性贫血患者CXCR5^(+)CD8^(+)T细胞和CXCL13的异常表达及其与血液学指标的相关性分析

目的分析重型再生障碍性贫血(SAA)患者C-X-C趋化因子受体5(CXCR5)^(+)CD8^(+)T细胞比例和血浆C-X-C基序趋化因子13(CXCL13)的表达水平,及其与血液学指标的相关性。方法回顾性分析2018年1月至2021年9月天津医科大学总医院35例SAA患者的临床资料,根据患者是否接受过药物治疗,将患者分为2组:(1)初治SAA组:18例,患者未接受过药物治疗;其中男9例,女9例,年龄51(18~76)岁;(2)缓解期SAA组:17例,指经抗胸腺细胞球蛋白(ATG)联合环孢素A(CsA)免疫抑制治疗后,脱离成分血输注的患者;其中男7例,女10例,年龄46(16~70)岁。另外选取20名健康对照者,其中男8名,女12名,年龄45(15~72)岁。收集SAA患者外周血及骨髓标本,同时收集健康对照者外周血标本。采用流式细胞术检测外周血及骨髓标本中CXCR5^(+)CD8^(+)T细胞比例,采用酶联免疫吸附试验检测血浆中CXCL13表达水平。CXCR5^(+)CD8^(+)T细胞比例与CXCL13表达水平的相关性以及二者与血液学指标的相关性采用Spearman相关性分析。结果初治SAA组骨髓CXCR5^(+)CD8^(+)T细胞比例为(4.9±2.9)%,高于缓解期SAA组的(2.7±1.5)%(t=2.34,P=0.027)。初治SAA组、缓解期SAA组和健康对照组外周血CXCR5^(+)CD8^(+)T细胞比例分别为(8.4±4.2)%、(3.8±2.3)%、(2.6±2.0)%,初治SAA组外周血CXCR5^(+)CD8^(+)T细胞比例高于缓解期SAA组和健康对照组(均P<0.05)。初治SAA组血浆CXCL13表达水平为(97.2±46.8)ng/L,高于缓解期SAA组的(54.9±20.9)ng/L和健康对照组的(47.6±17.3)ng/L(均P<0.05)。SAA患者外周血CXCR5^(+)CD8^(+)T细胞比例与CXCL13表达水平呈正相关(r=0.545,P<0.001)。SAA患者外周血CXCR5^(+)CD8^(+)T细胞比例与白细胞、血小板、中性粒细胞百分比、中性粒细胞绝对值、网织红细胞百分比、网织红细胞绝对值、骨髓粒系百分比、骨髓红系百分比、骨髓巨核细胞数量均呈负相关(r=-0.556、-0.392、-0.617、-0.615、-0.395、-0.543、-0.432、-0.484、-0.523,均P<0.05),与外周血淋巴细胞百分比和骨髓淋系百分比均呈正相关(r=0.593、0.556,均P<0.05)。SAA患者外周血中CXCL13表达水平与白细胞、中性粒细胞绝对值、网织红细胞百分比、网织红细胞绝对值、骨髓红系百分比呈负相关(r=-0.447、-0.446、-0.498、-0.407、-0.456,均P<0.05),与骨髓淋系百分比呈正相关(r=0.384,P<0.05)。结论SAA患者CXCR5⁺CD8⁺T细胞比例及血浆CXCL13表达水平增高。外周血CXCR5^(+)CD8^(+)T细胞比例与CXCL13表达水平呈正相关,二者均与多项血液学指标具有相关性,可能在SAA免疫发病机制中具有重要作用。

The biological role of the CXCL12/CXCR4 axis in esophageal squamous cell carcinoma

Esophageal cancer is the eighth most common malignant tumor and the sixth leading cause of cancer-related death worldwide.Esophageal squamous cell carcinoma(ESCC)is the main histological type of esophageal cancer,and accounts for 90%of all cancer cases.Despite the progress made in surgery,chemotherapy,and radiotherapy,the mortality rate from esophageal cancer remains high,and the overall 5-year survival rate is less than 20%,even in developed countries.The C-X-C motif chemokine ligand 12(CXCL12)is a member of the CXC chemokine subgroup,which is widely expressed in a variety of tissues and cells.CXCL12 participates in the regulation of many physiological and pathological processes by binding to its specific receptor,C-X-C motif chemokine receptor type 4(CXCR4),where it causes embryonic development,immune response,and angiogenesis.In addition,increasing evidence indicates that the CXCL12/CXCR4 axis plays an important role in the biological processes of tumor cells.Studies have shown that CXCL12 and its receptor,CXCR4,are highly expressed in ESCC.This abnormal expression contributes to tumor proliferation,lymph node and distant metastases,and worsening prognosis.At present,antagonists and imaging agents against CXCL12 or CXCR4 have been developed to interfere with the malignant process and monitor metastasis of tumors.This article summarizes the structure,function,and regulatory mechanism of CXCL12/CXCR4 and its role in the malignancy of ESCC.Current results from preclinical research targeting CXCL12/CXCR4 are also summarized to provide a reference for the clinical diagnosis and treatment of ESCC.

Identification of differentially expressed genes in ulcerative colitis and verification in a colitis mouse model by bioinformatics analyses

BACKGROUND Ulcerative colitis(UC)is an inflammatory bowel disease that is difficult to diagnose and treat.To date,the degree of inflammation in patients with UC has mainly been determined by measuring the levels of nonspecific indicators,such as C-reactive protein and the erythrocyte sedimentation rate,but these indicators have an unsatisfactory specificity.In this study,we performed bioinformatics analysis using data from the National Center for Biotechnology Information-Gene Expression Omnibus(NCBI-GEO)databases and verified the selected core genes in a mouse model of dextran sulfate sodium(DSS)-induced colitis.AIM To identify UC-related differentially expressed genes(DEGs)using a bioinformatics analysis and verify them in vivo and to identify novel biomarkers and the underlying mechanisms of UC.METHODS Two microarray datasets from the NCBI-GEO database were used,and DEGs between patients with UC and healthy controls were analyzed using GEO2R and Venn diagrams.We annotated these genes based on their functions and signaling pathways,and then protein-protein interactions(PPIs)were identified using the Search Tool for the Retrieval of Interacting Genes.The data were further analyzed with Cytoscape software and the Molecular Complex Detection(MCODE)app.The core genes were selected and a Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis was performed.Finally,colitis model mice were established by administering DSS,and the top three core genes were verified in colitis mice using real-time polymerase chain reaction(PCR).RESULTS One hundred and seventy-seven DEGs,118 upregulated and 59 downregulated,were initially identified from the GEO2R analysis and predominantly participated in inflammation-related pathways.Seven clusters with close interactions in UC formed:Seventeen core genes were upregulated[C-X-C motif chemokine ligand 13(CXCL13),C-X-C motif chemokine receptor 2(CXCR2),CXCL9,CXCL5,C-C motif chemokine ligand 18,interleukin 1 beta,matrix metallopeptidase 9,CXCL3,formyl peptide receptor 1,complement component 3,CXCL8,CXCL1,CXCL10,CXCL2,CXCL6,CXCL11 and hydroxycarboxylic acid receptor 3]and one was downregulated[neuropeptide Y receptor Y1(NYP1R)]in the top cluster according to the PPI and MCODE analyses.These genes were substantially enriched in the cytokinecytokine receptor interaction and chemokine signaling pathways.The top three core genes(CXCL13,NYP1R,and CXCR2)were selected and verified in a mouse model of colitis using real-time PCR Increased expression was observed compared with the control mice,but only CXCR2 expression was significantly different.CONCLUSION Core DEGs identified in UC are related to inflammation and immunity inflammation,indicating that these reactions are core features of the pathogenesis of UC.CXCR2 may reflect the degree of inflammation in patients with UC.

Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs

In recent years,several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells.These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells.Interestingly,this notion has also led scientists to question the role of proteins in the secretome produced by cells,tissues or organisms under certain conditions or at a particular time of regenerative therapy.Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs.This could also facilitate the migration of immune,progenitor and stem cells within the body to the site of inflammation.Of these different paracrine factors present within the secretome,researchers have given proper consideration to stromal cell-derived factor-1(SDF1)that plays a vital role in tissue-specific migration of the cells needed for regeneration.Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration.Hence in this study,we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.

JNK in Spinal Cord Facilitates Bone Cancer Pain in Rats through Modulation of CXCL1

In patients with advanced cancer, cancer-induced bone pain（CIBP） is a severe and common problem that is difficult to manage and explain. As c-Jun N-terminal kinase（JNK） and chemokine（C-X-C motif） ligand 1（CXCL1） have been shown to participate in several chronic pain processes, we investigated the role of JNK and CXCL1 in CIBP and the relationship between them. A rat bone cancer pain model was established by intramedullary injection of Walker 256 rat gland mammary carcinoma cells into the left tibia of Sprague-Dawley rats. As a result, intramedullary injection of Walker 256 carcinoma cells induced significant bone destruction and persistent pain. Both phosphorylated JNK1（p JNK1） and p JNK2 showed time-dependent increases in the ipsilateral spinal cord from day 7 to day 18 after tumor injection. Inhibition of JNK activation by intrathecal administration of SP600125, a selective p JNK inhibitor, attenuated mechanical allodynia and heat hyperalgesia caused by tumor inoculation. Tumor cell inoculation also induced robust CXCL1 upregulation in the ipsilateral spinal cord on day 18 after tumor injection. Inhibition of CXCL1 by intrathecal administration of CXCL1 neutralizing antibody showed a stable analgesic effect. Intrathecal administration of SP600125 reduced CXCL1 increase in the spinal cord, whereas inhibition of CXCL1 in the spinal cord showed no influence on JNK activation. Taken together, these results suggested that JNK activation in spinal cord contributed to the maintenance of CIBP, which may act through modulation of CXCL1. Inhibition of the p JNK/CXCL1 pathway may provide a new choice for treatment of CIBP.

Follicular helper T lymphocytes in health and disease

A correct antibody response requires the participation of both B and T lymphocytes and antigen presenting cells. In this review we address the role of follicular helper T lymphocytes(T FH) in this reaction. We shall focus on the regulation of their development and function in health and disease. T FH can be characterized on the basis of their phenotype and the pattern of secretion of cytokines. This fact is useful to study their participation in the generation of antibody deficiency in primary immunodeficiency diseases such as common variable immunodeficiency, X-linked hyper Ig M syndrome orX-linked lymphoproliferative disease. Increased numbers of T FH have been demonstrated in several autoimmune diseases and are thought to play a role in the development of autoantibodies. In chronic viral infections caused by the human immunodeficiency virus, hepatitis B or C virus, increased circulating T FH have been observed, but their role in the protective immune response to these agents is under discussion. Likewise, an important role of T FH in the control of some experimental protozoan infections has been proposed, and it will be important to assess their relevance in order to design effective vaccination strategies.