ABSTRACT
Objective To investigate the mRNA expression level of neurogranin on peripheral blood mononuclear cells(PBMCs)from patients with systemic lupus erythematosus(SLE).Methods Top 20 tags of SLE PBMCs SAGE library were searched from normal lymphocytes SAGE library including navie-T,Th1,Th2, CD8+T, NK and B cells,and their abundance was compared.The mRNA expression level of neuro-granin,a differential over-expressed tag,was detected in 35 cases of SLE and 15 normal controls by reversetranscription-polymerase chain reaction (RT-PCR).Results Neurogranin tag could only be detected in SLE PBMCs SAGE library,but was hardly found in normal lymphocyte SAGE library.However,either SLE pa-tients or normal controls showed a detectable mRNA level of neurogranin on PBMCs by RT-PCR.The mRNA level of neurogranin in active SLE patients was significantly increased than those in controls(P＜0.001).but only slightly increased in inactive SLE patients (P＞0.05).Conclusion Neurogranin,as a novel proapototic factor,is overexpressed on PBMCs of SLE patients.It may be involved in the regulation of abnormal immune responses in lupus.
ABSTRACT
This study examined the gene expression patterns of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE) by using serial analysis of gene expression (SAGE) technology. Following the construction of serial analysis of gene expression (SAGE) library of PBMCs collected from 3 cases of familial SLE patients, a large scale of tag sequencing was performed. The data extracted from sequencing files was analyzed with SAGE 2000 V 4.5 software. The top 30 expressed genes of SLE patients were uploaded to http://david.niaid.nih.gov/david/ease.htm and the functional classification of genes was obtained. The differences among those expressed gene were analyzed by Chi-square tests. The results showed that a total of 1286 unique SAGE tags were identified from 1814 individual SAGE tags. Among the 1286 unique tags, 86.8% had single copy, and only 0.2% tags had more than 20 copies. And 68.4% of the tags matched known expressed sequences, 41.1% of which matched more than one known expressed sequence. About 31.6% of the tags had no match and could represent potentially novel genes. Approximately one third of the top 30 genes were ribosomal protein, and the rest were genes related to metabolism or with unknown functions. Eight tags were found to express differentially in SAGE library of SLE patients. This study draws a profile of gene expression patterns of PBMCs in patients with SLE. Comparison of SAGE database from PBMCs between normal individuals and SLE patients will help us to better understand the pathogenesis of SLE.
Subject(s)
Expressed Sequence Tags , Gene Expression Profiling/methods , Leukocytes, Mononuclear/metabolism , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/genetics , Sequence Tagged Sites , Transcription, GeneticABSTRACT
This study examined the gene expression patterns of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE) by using serial analysis of gene expression (SAGE) technology. Following the construction of serial analysis of gene expression (SAGE) library of PBMCs collected from 3 cases of familial SLE patients, a large scale of tag sequencing was performed. The data extracted from sequencing files was analyzed with SAGE 2000 V 4.5 software.The top 30 expressed genes of SLE patients were uploaded to http://david.niaid.nih. gov/david/ease.htm and the functional classification of genes was obtained. The differences among those expressed gene were analyzed by Chi-square tests. The results showed that a total of 1286 unique SAGE tags were identified from 1814 individual SAGE tags. Among the 1286 unique tags, 86.8% had single copy, and only 0.2% tags had more than 20 copies. And 68.4% of the tags matched known expressed sequences, 41.1% of which matched more than one known expressed sequence. About 31.6% of the tags had no match and could represent potentially novel genes. Approximately one third of the top 30 genes were ribosomal protein, and the rest were genes related to metabolism or with unknown functions. Eight tags were found to express differentially in SAGE library of SLE patients. This study draws a profile of gene expression patterns of PBMCs in patients with SLE. Comparison of SAGE database from PBMCs between normal individuals and SLE patients will help us to better understand the pathogenesis of SLE.