Objective: Dendritic cells (DCs) are the most potent antigen presenting cells (APCs) of the immune system. We intent to block the expression of CD86 in DCs using antisense peptide nucleic acids (PNA), a novel synthetic structural DNA mimic, and interrupt the second signal transmission so that a suppression of corresponding T cell function can be achieved.
Methods: Human DCs grown up from peripheral blood monocytes in GM-CSF and IL-4 were collected. We investigated antisense PNA internalization with laser scan confocal microscope (LSCM). Fluorescence immunocytochemistry, flow cytometry and RT-PCR were used to determine the expression of CD86 protein and mRNA in DCs.
Results: LSCM proved that cultured immature DCs could internalized PNA efficiently, according to the specific internalization property of the immature DCs. Antisense PNA DC exhibited striking reductions in cell surface staining for CD86, but not MHC class II, and were poor stimulators of T cell proliferation. RT-PCR found that PNA depressed the amounts of CD86 mRNA in DCs.
Conclusion: Antisense PNA against CD86 could inhibit the expression of CD86 mRNA and protein in DCs. The blockade of B7/CD28 pathway may increase the potential of costimulatory molecule-deficient antisense PNA DCs of donor origin to induce long-lasting allograft survival.