Objective To detect the level of soluble programmeddeath 1 (sPD-1) and soluble programmed death ligand 1(sPD-L1) in serum and urine of children with primarynephrotic syndrome (PNS),and explore its clinical significanc...Objective To detect the level of soluble programmeddeath 1 (sPD-1) and soluble programmed death ligand 1(sPD-L1) in serum and urine of children with primarynephrotic syndrome (PNS),and explore its clinical significance.Methods From July 2017 to November 2017,children with PNS admitted to the Children's Hospital Affiliatedto Soochow University were divided into onsetgroup (36 cases) and remission group (33 cases). Thirtyhealthy children who underwent medical examinationfor enrollment,undersize or overweight in the outpatientdepartment of pediatric health care and inpatient departmentof Endocrinology were selected as healthy controlgroup. Serum and urine samples were collected, inwhich the levels of sPD-1 and sPD-L1 were detected byenzyme-linked immunosorbent assay (ELISA). The correlationbetween serum and urine sPD-1,sPD-L1 levelsand lymphocyte subsets,urinary protein were analyzedby Pearson and Spearman correlation analysis.展开更多
Increasing evidence points to a role for killer immunoglobulin-like receptors(KIRs)in the development of autoimmune diseases.In particular,a positive association of KIR3DS1(activating receptor)and a negative associati...Increasing evidence points to a role for killer immunoglobulin-like receptors(KIRs)in the development of autoimmune diseases.In particular,a positive association of KIR3DS1(activating receptor)and a negative association of KIR3DL1(inhibitory receptor)alleles with ankylosing spondylitis(AS)have been reported by several groups.However,none of the studies analyzed these associations in the context of functionality of polymorphic KIR3DL1.To better understand how the KIR3DL1/3DS1 genes determine susceptibility to AS,we analyzed the frequencies of alleles and genotypes encoding functional(KIR3DL1*F)and non-functional(KIR3DL1*004)receptors.We genotyped 83 AS patients and 107 human leukocyte antigen(HLA)-B27-positive healthy controls from the Russian Caucasian population using a two-stage sequence-specific primer PCR,which distinguishes KIR3DS1,KIR3DL1*F and KIR3DL1*004 alleles.For the patients carrying two functional KIR3DL1 alleles,those alleles were additionally genotyped to identify KIR3DL1*005 and KIR3DL1*007 alleles,which are functional but are expressed at low levels.KIR3DL1 was negatively associated with AS at the expense of KIR3DL1*F but not of KIR3DL1*004.This finding indicates that the inhibitory KIR3DL1 receptor protects against the development of AS and is not simply a passive counterpart of the segregating KIR3DS1 allele encoding the activating receptor.However,analysis of genotype frequencies indicates that the presence of KIR3DS1 is a more important factor for AS susceptibility than the absence of KIR3DL1*F.The activation of either natural killer(NK)or T cells via the KIR3DS1 receptor can be one of the critical events in AS development,while the presence of the functional KIR3DL1 receptor has a protective effect.Nevertheless,even individuals with a genotype that carried two inhibitory KIR3DL1 alleles expressed at high levels could develop AS.展开更多
目的构建可定量检测杀伤细胞免疫球蛋白样受体(KIR)转录水平的方法。方法筛选KIR3DS1和KIR3DL1保守区特异性引物,利用RT-PCR和熔解曲线法对引物特异性进行检验,建立可定量的标准内参,应用SYBR Green RT-PCR方法检测标本中KIR3DS1和KIR3...目的构建可定量检测杀伤细胞免疫球蛋白样受体(KIR)转录水平的方法。方法筛选KIR3DS1和KIR3DL1保守区特异性引物,利用RT-PCR和熔解曲线法对引物特异性进行检验,建立可定量的标准内参,应用SYBR Green RT-PCR方法检测标本中KIR3DS1和KIR3DL1的mRNA含量。结果通过电泳和实时定量PCR的熔解曲线明确了KIR3DS1和KIR3DL1引物,其特异性好,无杂带。应用TA克隆建立的KIR定量标准品线性关系好,可检测待测物的范围大。应用临床标本可检测出两组标本有显著差异,符合临床预期。结论本文建立的KIR mRNA定量检测方法,可以定量检测活化和抑制性KIR,可以进一步研究不同疾病进展的机制以及预测疾病的临床转归。展开更多
文摘Objective To detect the level of soluble programmeddeath 1 (sPD-1) and soluble programmed death ligand 1(sPD-L1) in serum and urine of children with primarynephrotic syndrome (PNS),and explore its clinical significance.Methods From July 2017 to November 2017,children with PNS admitted to the Children's Hospital Affiliatedto Soochow University were divided into onsetgroup (36 cases) and remission group (33 cases). Thirtyhealthy children who underwent medical examinationfor enrollment,undersize or overweight in the outpatientdepartment of pediatric health care and inpatient departmentof Endocrinology were selected as healthy controlgroup. Serum and urine samples were collected, inwhich the levels of sPD-1 and sPD-L1 were detected byenzyme-linked immunosorbent assay (ELISA). The correlationbetween serum and urine sPD-1,sPD-L1 levelsand lymphocyte subsets,urinary protein were analyzedby Pearson and Spearman correlation analysis.
基金by the State Support of the Leading Scientific Schools NS-2395.2008.4,Molecular and Cell Biology Program RAS,Rosnauka 02.512.12.2053,Rosobrazovanie P 256 and Basic Research for Medicine RAS.
文摘Increasing evidence points to a role for killer immunoglobulin-like receptors(KIRs)in the development of autoimmune diseases.In particular,a positive association of KIR3DS1(activating receptor)and a negative association of KIR3DL1(inhibitory receptor)alleles with ankylosing spondylitis(AS)have been reported by several groups.However,none of the studies analyzed these associations in the context of functionality of polymorphic KIR3DL1.To better understand how the KIR3DL1/3DS1 genes determine susceptibility to AS,we analyzed the frequencies of alleles and genotypes encoding functional(KIR3DL1*F)and non-functional(KIR3DL1*004)receptors.We genotyped 83 AS patients and 107 human leukocyte antigen(HLA)-B27-positive healthy controls from the Russian Caucasian population using a two-stage sequence-specific primer PCR,which distinguishes KIR3DS1,KIR3DL1*F and KIR3DL1*004 alleles.For the patients carrying two functional KIR3DL1 alleles,those alleles were additionally genotyped to identify KIR3DL1*005 and KIR3DL1*007 alleles,which are functional but are expressed at low levels.KIR3DL1 was negatively associated with AS at the expense of KIR3DL1*F but not of KIR3DL1*004.This finding indicates that the inhibitory KIR3DL1 receptor protects against the development of AS and is not simply a passive counterpart of the segregating KIR3DS1 allele encoding the activating receptor.However,analysis of genotype frequencies indicates that the presence of KIR3DS1 is a more important factor for AS susceptibility than the absence of KIR3DL1*F.The activation of either natural killer(NK)or T cells via the KIR3DS1 receptor can be one of the critical events in AS development,while the presence of the functional KIR3DL1 receptor has a protective effect.Nevertheless,even individuals with a genotype that carried two inhibitory KIR3DL1 alleles expressed at high levels could develop AS.
文摘目的构建可定量检测杀伤细胞免疫球蛋白样受体(KIR)转录水平的方法。方法筛选KIR3DS1和KIR3DL1保守区特异性引物,利用RT-PCR和熔解曲线法对引物特异性进行检验,建立可定量的标准内参,应用SYBR Green RT-PCR方法检测标本中KIR3DS1和KIR3DL1的mRNA含量。结果通过电泳和实时定量PCR的熔解曲线明确了KIR3DS1和KIR3DL1引物,其特异性好,无杂带。应用TA克隆建立的KIR定量标准品线性关系好,可检测待测物的范围大。应用临床标本可检测出两组标本有显著差异,符合临床预期。结论本文建立的KIR mRNA定量检测方法,可以定量检测活化和抑制性KIR,可以进一步研究不同疾病进展的机制以及预测疾病的临床转归。
