Study Of Gene Expression Profiles In Renal Transplantation Immunity By Gene Microarray

Objective: To explore and discover the relative genes with transplantal immunity.Method: By virtue of gene microarray to analyse the differential expression of the genes in the WBC of the patients having accepted renal transplantation and the high sensitive patients. The patient having accepted renal transplantation (4-6 weeks after operation) entered into the postoperation group (30 cases, pog), and the high sensitive patient with uremia (PRA>85%) was divided into the high sensitive group (30 cases, hsg), the normal blood donors were listed as control group (eg). The fresh blood was collected in all groups. By PVP natural sedimentation technique, WBC was collected and preserved in liquid nitrogen. Total RNA were extracted by one-step technique and purified?The total RNA of the two groups (pog and hsg) were labeled with Cy5-dUTP respectively, and total RNA of eg were labeled with Cy3-dUTP, then by reverse transcript way to label the cDNA probe. High throughout gene chip (16,920) was hybrided and scanned by ScanArray4000. Cy3/Cy5 image files of hsg and pog were copied. The area of point was determined with grids, the noise of background was filtered and fluorescent signal value of gene-expressing was obtained. ArrayPro Analyzer 4.0 computed and analysed thefluorecent signal intensity and ratio of Cy3 and Cy5.Results: In the postoperation group, there are differential expression 433 genes (2.55%), including up-regulated 26 (known 11 and unknown 15) and down-regulated 407(known 252 and unknown 155); hi the high sensitive group, there are differential expression 877 genes (5.18%), including up-regulated 88 (known 55 and unknown 33) and down-regulated 789(known 510 and unknown 279).Conclusions: cDNA microarray for analysis of gene expression proflies is a powerful method to identify novel associated genes with transplantal immunity.