Establishment of the tax gene expression cell lines and their biological activity.
- Author:
Yun XIA
1
;
Ying SHI
;
Yan GAO
;
Ji-Qiang GUO
;
Xiang-Feng SONG
;
Hui WANG
Author Information
1. Research Center for Immunology, Xinxiang Medical College, Xinxiang 453003, Henan Province, China.
- Publication Type:Journal Article
- MeSH:
Cell Line;
Gene Expression;
Gene Expression Regulation, Viral;
Genes, pX;
Human T-lymphotropic virus 1;
genetics;
Humans;
Jurkat Cells;
NF-kappa B;
metabolism;
Transfection
- From:
Journal of Experimental Hematology
2008;16(6):1425-1429
- CountryChina
- Language:Chinese
-
Abstract:
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.
