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

Changes of Peripheral Blood CXCL13 and PD-L1+B Lymphocyte Levels andTheir Clinical Significance in Children with Primary Nephrotic Syndromebefore and after Treatment

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。

Objective To investigate the clinical significance of changes in peripheral blood Chemokine C-X-C motif ligand 13(CXCL 13)and programmed death receptor ligand 1(PD-L1)+B lymphocyte level before and after treatment in children with primary nephrotic syndrome(PNS).Methods A total of 52 children with hormone-sensitive initial PNS treated from April 2022 to December 2022 were selected.Thirty normal children admitted for physical examination during the same period were taken as healthy control.Clinical laboratory indicators were collected and total B cells and PD-L1+B lymphocytes in peripheral blood were detected by flow cytometry.Serum CXCL13,solubility programmed death protein ligand 1(sPD-L1)and cytokines[transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interleukin-1β(IL-1β)]levels in the two groups were detected by enzyme-linked immunosorbent assay.CXCL13 and PD-L1+B lymphocytes and their correlation with laboratory indicators were analyzed by Pearson correlation.Results Before treatment,compared with healthy control group,the proportion of total B cells(12.54%±4.23%vs 4.95%±2.83%)and PD-L1+B lymphocytes(1.17%±0.38%vs 0.35%±0.12%)in peripheral blood,the levels of serum CXCL13(121.03±30.52 pg/ml vs 53.67±12.42 pg/ml)and sPD-L1(116.25±25.68 pg/ml vs 47.27±8.14 pg/ml),and the levels of serum cytokines TGF-β1(17.91±2.04 ng/ml vs 12.53±1.62 ng/ml),TNF-α(77.65±7.27 ng/ml vs 52.43±4.68 ng/ml),IL-10(14.21±3.56 pg/ml vs 4.76±1.25 pg/ml)and IL-1β(64.38±7.46 ng/ml vs 35.57±5.92 ng/ml)were significantly increased in PNS group,and the differences were statistical significance(t=-10.754,-11.468,-11.526,-14.271,-12.360,-17.048,-14.017,-18.103,all P<0.05).Compared with before treatment,the proportion of total B cells(6.20%±2.48%)and PD-L1+B lymphocytes(0.43%±0.25%)in peripheral blood and the levels of CXCL13(65.27±14.16 pg/ml),sPD-L1(55.63±11.44 pg/ml),TGF-β1(14.35±1.82ng/ml),TNF-α(56.48±4.16 ng/ml),IL-10(5.15±1.09 pg/ml)and IL-1β(39.38±4.05 ng/ml)in serum in the PNS group were significantly decreased after treatment,and the differences were statistical significance(t=9.324,11.731,11.951,15.549,9.930,18.226,17.548,21.237,all P<0.05).The proportion of PD-L1+B lymphocytes in PNS children was negatively correlated with serum ALB and IgG levels(r=-0.619,-0.5874,all P<0.05),and positively correlated with IgM level(r=0.563,P<0.05).CXCL13 expression level was negatively correlated with serum albumin and IgG levels(r=-0.574,-0.522,P<0.05).The proportion of PD-L1+B lymphocytes was positively correlated with CXCL13 expression level(r=0.632,P<0.05).Conclusion Increased proportions of CXCL13 and PD-L1+B lymphocytes in peripheral blood were associated with humoral immunity disorders in children with PNS.CXCL13 may promote the production of inflammatory cytokines by immune cells by promoting the chemotaxis of B lymphocytes in peripheral blood,and aggravate the excessive immune inflammatory response of PNS.