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Title: Characterization of an antisense oligonucleotide targeted at Phospholipase C gamma 1 in the mouse T-cell line EL4.1L-2
Keywords: phospholipase C gamma 1, T cell activation, antisense oligonucleotides, calcium, IL-2, cell proliferation
Issue Date: 14-Jul-2006
Source: CHOO HUI HWA (2006-07-14). Characterization of an antisense oligonucleotide targeted at Phospholipase C gamma 1 in the mouse T-cell line EL4.1L-2. ScholarBank@NUS Repository.
Abstract: Phospholipase C gamma 1 (PLCgamma1) plays a prominent role in T cell activation. Therefore, in this study, we characterized an antisense oligonucleotide (ASO) against PLCgamma1 and study its effect on T cell activation. ASOs complementary to mouse PLCgamma1 mRNA were designed and screened in the mouse T cell line, EL4.IL-2. Among all, ASO3 showed the highest PLCgamma1 protein inhibition whereas scrambled control was without effect. In addition, ASO3 selectively reduced PLCgamma1 mRNA level in a concentration-dependent manner without altering PLCgamma2 expression. Time course study revealed maximum mRNA and protein downregulation at 6 h and 24 h posttransfection respectively. Subsequent measurement of IP3, intracellular calcium and IL-2 levels in ASO3-treated cells also showed significant inhibition. Further, our study demonstrated that ASO3-treated cells exhibited significant reduction in cell proliferation following TCR-CD28 costimulation. These findings implicate that antisense inhibition of PLCgamma1 may have therapeutic potential for the treatment of T cell-dependent disorders.
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