Screening Of Differentially Expressed Genes In Peripheral T Cells Of Monozygotic Twins Of Patients With Psoriasis

[Objective]To screen differentially expressed genes in peripheral T cells of monozygotic twins patientswith psoriasis vulgaris(PV), and further discuss the role of T cells in pathogenesis of psoriasis toconfirm the point “psoriasis is a immunological dermatosis mediated byT cells.”[Methods]1. T cells were isolated by density gradient centrifugation from peripheral blood in monozygotictwins patients with PV, lichen normal male control (N) and Atopic dermatitis patients (AD),cultured to get enough cells.2. Get the total RNA extraction, purifie and reverse the transcription of synthetic double-strandedcDNA.3. Prepare and sequence Taq.4. In accordance of the Reference Gene Database, build the Reference Tag Database by software.Select the tags with the single gene in correspondence in compurgation between clean tag andthe Reference Tag Database, and then have these tags signed.5. Referring to the methods of detecting digital gene differential expression profile, screen thedifferential gene expression among the three samples.6. Calculate the number of genes in each term, and then apply a hyper geometric test to find outcompared with the background of the entire genome; differentially expressed genes weresignificantly enriched Gene Ontology entries.[Results]1. T cells were cultured for amplification, amount of2-3×106cells were taking for puritymeasure by use of flow cytometry, which showed the purity of the cultured T cell was as highas96.47%.2.3.4-3.6million clean Taqs were screened by Taq sequencing, of which about47.65-51.71%clean Taqs were in correspondence with single gene, apprpximately18.52-24.21%of the Taqcompared to gene position.3. After analysis, a total of comments to9449genes, where P1and P2compared to no differencein expression of genes5438, P3and N2compared to the differences in the expression of genes 2432; differentially expressed genes in both disease twins accounted for57.55%of all expressedgenes, not the same disease in twins accounted for25.74%of all expressed genes, indicating thatthere were the differences in gene expression rate between P3N2and P1P2,and the former wassignificantly higher than the latter; psoriasis group and the control group compared to in theexpression of differences genes105; and with atopic dermatitis group compared differences ingene expression of40genes up regulated24.4. Monozygotic twins psoriasis group with control group and with atopic dermatitis groupcompare the differential expression of40genes, which the reduction in the minimum geneexpression25763,and the rest is not include in the form of the genes,is located only onchromosome,not yet have specific medical significance.5. Monozygotic twins psoriasis patients peripheral blood T-cell chemokine receptor7expressionincreased programmed cell death molecules down regulated, suggesting that thecyclin-dependent kinase inhibitor2B expression down with cyclinA2expression increased anegative correlation.[Conclusion]Many genes were differentially expressed in T cells of monozygotic twins psoriatic patients,which might play role in abnormal immune responses of T cells in psoriatic patients. Furtherstudy indicates that the model of T-cell differences in the expression are closely related withspecific immunal effection of T cells in the pathogenesis of psoriasis.