AI Article Synopsis
- Researchers aimed to study how blocking the ICOS gene affects the activation and growth of rat T-lymphocytes using RNA interference.
- They designed oligonucleotides to target the ICOS gene, introduced them into rat lymphocytes, and measured changes in gene expression and proliferation through various methods.
- Results showed that silencing the ICOS gene led to decreased T-lymphocyte activation, reduced proliferation, and lower levels of immune markers IFN-gamma and IL-4, indicating potential implications for transplant immunity research.
Article Abstract
Objective: To observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.
Methods: Four interference sites targeting at rat ICOS gene were designed and four pairs of oligonucleotide fragments were cloned into the pSilencer 4.1-CMV neo plasmid vectors then transfected into rat lymphocytes with cationic liposome. The expression of mRNA and protein of ICOS was detected by RT-PCR and flow cytometry. The alteration of lymphocyte proliferation ability was evaluated by mix lymphocyte reaction, and the secretion levels of IFN-gamma and IL-4 were measured by ELISA procedure.
Results: After transfection, the expression of mRNA and protein of ICOS in test groups were lower than that in control groups (P < 0.05). The ability of T-lymphocytes in proliferation was poor and the levels of IFN-gamma and IL-4 were reduced with ICOS gene shut down.
Conclusions: RNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.
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