Dataset Information

The Effect of TAX-1 Gene of Human T-cell Leukemia Virus Type-1 on the Expression of CCR5 in K562 Cell Line.

ABSTRACT: Background:Tax-1 protein of Human T-cell Leukemia Virus type 1(HTLV-1) serves as a key transcriptional regulatory gene product and has a crucial role in transactivating genes of infected cells by employing their transcriptional factors. This modulation includes induction of genes which encode CC-chemokines and their receptors. In this study, a recombinant vector containing Tax-1 gene was made and tested for its ability to induce CCR5 (CC chemokine receptor 5) expression in K562 cell line. Methods:In order to perform this research, two blood samples of HTLV-1 positive were obtained from Urmia blood transfusion center. After DNA extraction, a complete sequence of Tax-1 gene was amplified by specific primers. Recombinant vectors carrying Tax-1 gene were synthesized and transformed into Escherichia coli (E. coli). After bacteria transformation, bacteria containing recombinant plasmid were selected and purified. Then, the recombinant shuttle vectors, pCDNA3.1-TAX, were transfected into the cell culture (K562 cell line). Expression of CCR5 was measured after 72 hr by Syber Green Real-Time PCR method compared to control cell culture. Normalization was done with GAPDH as a standard gene. Results:Cloning of Tax-1 gene in the vector, pCDNA3.1 was confirmed by colony PCR, restriction digestion, and sequencing methods. Expression of Tax-1 and CCR5 genes were confirmed by real time PCR and also, expression of CCR5 gene showed an 8-fold increase in comparison to mock-treated controls (p<0.05). Conclusion:Our data suggested that recombinant Tax-1 may have the enhancing effect on CCR5 expression rate at mRNA levels in K562 cell line. Further studies are necessary to evaluate the effect of pCDNA3.1-TAX on cell surface CCR5 expression.

SUBMITTER: Haghnazari Sadaghiani N

PROVIDER: S-EPMC6359701 | biostudies-literature | 2019 Jan-Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

The Effect of <i>TAX-1</i> Gene of Human T-cell Leukemia Virus Type-1 on the Expression of CCR5 in K562 Cell Line.

Haghnazari Sadaghiani Nasrin N Pirayeshfard Lila L Aghaie Afsaneh A Sharifi Zohreh Z

Avicenna journal of medical biotechnology 20190101 1

<h4>Background</h4>Tax-1 protein of Human T-cell Leukemia Virus type 1(HTLV-1) serves as a key transcriptional regulatory gene product and has a crucial role in transactivating genes of infected cells by employing their transcriptional factors. This modulation includes induction of genes which encode CC-chemokines and their receptors. In this study, a recombinant vector containing Tax-1 gene was made and tested for its ability to induce CCR5 (CC chemokine receptor 5) expression in K562 cell line ...[more]

PMID: 30800245

Similar Datasets

Project description:BackgroundHuman T-cell leukemia virus type I (HTLV-1) can cause an aggressive malignancy known as adult T-cell leukemia/lymphoma (ATL) as well as inflammatory diseases such as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). A transgenic mouse that expresses HTLV-1 Tax also develops T-cell leukemia/lymphoma and an inflammatory arthropathy that resembles rheumatoid arthritis. The aim of this study was to identify the primary T-cell subsets involved in the development of arthropathy in Tax transgenic mice.Principal findingsBy 24 months of age, Tax transgenic mice developed severe arthropathy with a cumulative incidence of 22.8%. The pathological findings of arthropathy in Tax transgenic mice were similar to those seen in human rheumatoid arthritis or mouse models of rheumatoid arthritis, with synovial proliferation and a positive rheumatoid factor. Before the onset of spontaneous arthropathy, young and old Tax transgenic mice were not sensitive to collagen and did not develop arthritis after immunization with type II collagen. The arthropathic Tax transgenic mice showed a significantly decreased proportion of splenic CD4(+) T cells, whereas the proportion of splenic CD8(+) T cells was increased. Regulatory T cells (CD4(+)CD25(+)Foxp3(+)) were significantly decreased and CD8(+) T cells that expressed the chemokine receptor CCR4 (CD8(+)CCR4(+)) were significantly increased in arthropathic Tax transgenic mice. The expression of tax mRNA was strong in the spleen and joints of arthropathic mice, with a 40-fold increase compared with healthy transgenic mice.ConclusionsOur findings reveal that Tax transgenic mice develop rheumatoid-like arthritis with proliferating synovial cells in the joints; however, the proportion of different splenic T-cell subsets in these mice was completely different from other commonly used animal models of rheumatoid arthritis. The crucial T-cell subsets in arthropathic Tax transgenic mice appear to resemble those in HAM/TSP patients rather than those in rheumatoid arthritis patients.

Dataset Information

The Effect of TAX-1 Gene of Human T-cell Leukemia Virus Type-1 on the Expression of CCR5 in K562 Cell Line.

Publications

The Effect of <i>TAX-1</i> Gene of Human T-cell Leukemia Virus Type-1 on the Expression of CCR5 in K562 Cell Line.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets