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Preparation of DCCIK using EBV peptidepulsed DC from peripheral blood mononuclear cells of posttransplant lymphoproliferative disorder patients after hematopoietic stem cell transplantation
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Abstract:
Objective:To construct a DCCIK (cytokineinduced killer cell) coculture system using peripheral blood mononuclear cells (PBMCs) derivedDC from hematopoietic stem cell transplantation (HSCT) patients with posttransplant lympholiferative disorder (PTLD) after pulsed with EBVspecial peptides, so as to lay a foundation for new adoptive immunotherapy of patients with PLTD after HSCT. Methods: PBMCs were obtained from patients with PTLD after HSCT; DC was induced from adherent cells; and CIK was induced from suspension cells. DC was further pulsed with EBVspecial peptides and cocultured with CIK to establish the DCCIK coculture system; the immunophenotype of cells in DCCIK system before and after coculture were determined by FACS, IFNγ secretion was assayed by ELISA, and TCRβ genealogy was examined by genetic analyzer. Results: The ratio of HLADR+CD86+DC increased from 125% to 91.17% after cytokine stimulation. After coculture with DC for 14 d, the numbers of CIK in two patients with PTLD increased to 5.3 and 6.8 times,respectively. The ratios of CD3+, CD8+, CD3+CD8+, and CD3+CD56+ cells were significantly increased after DCCIK coculture. IFNγ level in peptidepulsed DCCIK group was significantly higher than that in peptideunpulsed DCCIK group（\[1 332.6±92.38\] pg/ml vs \[693.42±62.41\] pg/ml，P<0.05\]）; TCRβ genealogy assay found the clone expansion peak of 5.2 TCRβ subfamily in DCCIK coculture system. Conclusion: EBV peptidepulsed DC can induce CD3+CD8+ and CD3+CD56+ cell expansion in DCCIK coculture system with high level of IFNγ. DCCIK can be used as a new adoptive immunotherapy to HSCT patients with EBV infection and PTLD.
Keywords:
dendritic cell (DC) ; cytokineinduced killer cell (CIK) ; EpsteinBarr virus(EBV) ; posttransplant lymphoproliferative disorder (PTLD) ; adoptive immunotherapy
Project Supported:
Project supported by Natural Science Foundation of Hebei Province (No.C2008001097), and the Research Foundation for the Doctoral Program of Higher Institution of China (No. 200800890011), and the Key Followup Program of Health Bureau of Hebei Province (N
