Construction of PSAspecific dendritic cell vaccine and its in vitro immune activity
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Supported by Foundation of Shanghai Science and Technology Committee(044119615).

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    Abstract:

    Objective:To construct of PSAspecific dendritic cell (DC) vaccine and to observe its in vitro antitumour activity, so as to pave a way for future study. Methods: Bone marrow precursors were isolated and bone marrow derived DCs were prepared. Mature DCs were pulsed by PSA, Lysate of cancer cells, OVA and PBS to yield PSADC, Lys DC, OvaDC, and NonDC, respectively. After primed by antigen, the changes of IL12 p70 and IL1β in the supernatant of dendritic cells were assessed by ELISA. The antigenspecific proliferation and cytotoxicity activity of T cellprimed by PSApulsed DCs were observed and the results were compared with those by Lys, Ova and PBSpulsed DCs. Results: Mature DCs were successfully derived from bone marrow cells with a purity higher than 95%. ELISA assay showed PSADC, LysDC and OvaDC group secreted high levels of IL12 p70 and IL1β than NonDC group (P<0.05). In addition, PSADCs and LysDCs had significantly stronger ability to stimulate the proliferation of CD4+T cells in 3day classic mixed lymphocyte reaction (MLR) compared with OvaDCs and NonDCs (P<0.01). Higher levels of IFNγ and IL2 were detected in PSADCs and LysDCs groups compared with the other two groups (P<0.01), whereas the levels of IL10 and IL4 were lower than the other two groups (P<0.05). Moreover, PSADCs and Lys DCs enhanced DTH responses of C57BL/6 mice after antigen immunization; the third antigen and control did not show the enhancement effect (P<0.05). To observe the in vitro antiPSA CTL reactions induced by PSADCs and LysDCs, the LNCaP cell line (PSA specific) was used as syngeneic target and the E.G7 cell line (H2b) was used as Ovaspecific target cells. Compared with OvaDCs and NonDCs, CTL cells induced by PSADCs, LysDCs had significantly enhanced antigenspecific CTL activity to LNCaP cells (P<0.05).Conclusion: DCbased PSAepitope vaccine can be prepared by pulsing DCs with PSA protein; the prepared vaccine has strong in vitro immune activity and can kill LNCaP cells.

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History
  • Received:December 14,2007
  • Revised:September 16,2008
  • Adopted:March 03,2009
  • Online: March 13,2009
  • Published:
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