T cell activation modulates retrovirus-mediated gene expression
| Autor: | E. R. Quinn, L. G. Lum, K. T. Trevor |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Interleukin 2
CD4-Positive T-Lymphocytes T cell T-Lymphocytes Gene delivery Biology CD8-Positive T-Lymphocytes Lymphocyte Activation Transfection Interleukin 21 Viral Proteins Proviruses Genetics medicine Humans Molecular Biology Cells Cultured Interleukin 3 Regulation of gene expression CD28 T lymphocyte Molecular biology medicine.anatomical_structure Retroviridae Gene Expression Regulation Molecular Medicine Cytokines medicine.drug Interleukin-1 |
| Zdroj: | Human gene therapy. 9(10) |
| ISSN: | 1043-0342 |
| Popis: | Important considerations for T lymphocyte-based gene therapy include efficient gene delivery and expression in primary, human T cells. In this study, retrovirus-mediated gene transfer and the fate of proviral gene expression were evaluated in human T cells activated using (1) immobilized anti-CD3 monoclonal antibody (MAb) plus interleukin 2, or (2) cis costimulation using beads carrying coimmobilized anti-CD3 and anti-CD28 MAbs. By cross-linking the CD3 and CD28 receptors, these MAbs mimic in vivo signaling events, leading to cytokine production and proliferation. A modified human interleukin 1beta (IL-1beta) cDNA inserted into the MFG retroviral vector served as an indicator gene. Retroviral transduction frequencies were similar for T lymphocytes activated by the respective methods. However, early after MAb stimulation and virus exposure, proviral gene expression was greater at the RNA and protein levels in optimized anti-CD3/anti-CD28 bead-activated T cells, corresponding with augmented endogenous cytokine responses and mitogenesis. Proviral gene expression was not regulated by extrinsic cell factors present in activated T cell supernatants. Regardless of the MAb stimulation method, proviral IL-1beta expression declined in later T cell cultures concomitant with a decrease in cellular cytokines. Restimulation by either method reinduced both T cell activity and vector expression. Our finding that proviral gene regulation is downmodulated in the absence of T cell signaling events has implications for clinical strategies using retrovirus-modified T cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|