Establishment of the tax gene expression cell lines and their biological activity / 中国实验血液学杂志

This study was aimed to establish the cell lines of human T-cell leukemia virus type-1 (HTLV-1) tax gene expression and their biological activity. The eukaryotic vector pCMV-tax and pCMV-neo-Bam were transfected into Jurkat E6-1 by using liposome, following screening with G418, the tax gene expression cell line-TaxP and the negative cell lines-TaxN were selected and enriched. Then, the mRNA expressions of LAT, SLP70, ZAP70 and NF-kappaB (p65) were measured by using RT-PCR; the NF-kappaB bioactivity was tested after transfecting the pNF-kappaB-Luc plasmids into TaxP and TaxN cells; meanwhile CD25, CD69 expressions on surface of the two cell lines were tested by flow cytometry. The results showed that TaxP and TaxN cell lines were established and the tax gene expression was detected; as compared with TaxN, the mRNA expressions of LAT, SLP70 and ZAP70 in TaxP were enhanced (p<0.05), while the NF-kappaB bioactivity in TaxP cells was stronger than that in TaxN cells (p<0.01). The CD25, CD69 on TaxP cells surface was highly expressed. It is concluded that TaxP and TaxN cell lines are established, TAX protein could promote the activation of T cells and activate the NF-kappaB pathway.

Effection of Prolactin on Immunoresponsiveness of Activated T Lymphocytes Induced by Concanavalin A / 实用儿科临床杂志

Objective To study the effect of prolactin(PRL) on the activation of T lymphocytes stmiulated by concanavalin A(ConA),and to explore the action of PRL in the activation of T lymphocytes. Methods After CD4 +T cell line JurkatE6-1 cells were respectively stmiulated by 5 mg/L ConA,25 ?g/L PRL and 500 ?g/L bromocriptine(Brc).The blank control group,the ConA group,the PRL and ConA group(PRL group),the Brc and ConA group(Brc group),the PRL and Brc group(PRL-Brc group) were set in the experiment.The total RNA was extracted by Trizol after 48 hours and was reversed transcription immediately.The expression of tumor necrosis factor receptor associated factor 6(TRAF6) mRNA of T lymphocytes was checked by PCR.The expressions of tumor necrosis factor(ligand) super family 4(TNFSF4) and Killer specific secretory protein of 37 000(KSP37) mRNA of T lymphocytes were detected by real-time polymerase chain reaction. Results The PRL group and the Brc group could inhibit the expressions of TRAF6,TNFSF4,and KSP37 mRNA of the activated T lymphocyte compared with the blank control group and the ConA group(P a0.05).The PRL-Brc group could inhibit significantly the expressions of TRAF6,TNFSF4,and KSP37 mRNA of the activated T lymphocyte compared with the ConA group(P a