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Objective: mechanisms based on GVHD build vitro DC induced T cell regulation model, the ability to the observed donor immature dendritic cells (immature dendritic cell, imDC) stimulus White T cell proliferation, to explore the use of imDC Governance graft-versus-host disease (GVHD) feasibility of clinical application. Methods: from healthy donor peripheral blood mononuclear cells, using recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and interleukin (IL) -4 joint training five days induced differentiation into imDC; induced cultured for 7 days, and lipopolysaccharide (LPS) stimulation After 18 hours, the differentiation and maturation of dendritic cells (Mature dendritic cell, mDC). And cell morphology by inverted microscope and HE staining cells drops piece Immunohistochemical detection of the mDC surface of MHC-II molecules HLA-DR and CD86 expression. Utilize the flow cytometry imDC and the the mDC surface of the DC-specific molecular markers CDla and mature marker CD83 positive expression rate. A one-way MLR method, the donor and recipient of the mixed lymphocyte culture, build DC induced T-lymphocyte regulation model, the experiment is set to 3 groups: control group, imDC group and mDC group, a total of 48 hours of incubation after adding the CCK- 8, using a microplate reader to detect the OD values, the comparison for imDC and mDC ability to stimulate autologous T cell proliferation. Results: (1) training 5 days after the cells were semi-suspended growth surrounding burr coarse short, and shape, cell volume compared to monocytes increases the typical imDC morphological characteristics, flow cytometry CD1a, CD83, and The double anti expressed as (67.06 ± 0.93)%, (66.82 ± 5.06)% and (62.34 ± 1.94)%; significantly longer training, eight days after the cells were single suspension-like growth, cell body round, larger cells surrounding visible processes, morphological characteristics typical mDC, the cells drops piece immunohistochemistry results showed that HLA-DR, CD86-positive, flow cytometry CDla, CD83 and dual anti-expression (64.98 ± 2.99)%, respectively, (86.44 ± 4.10) % and (65.16 ± 0.55)%. CCK-8 detection the the OD value imDC group control group showed no statistical significance (P gt; 0.05): of mDC group and the control group, (2) one-way MLR mixed lymphocyte cultures were incubated for 48 hours, imDC significant group were statistically significant (P lt; 0.01), autologous T cell stimulation index greater than 2.00 (SI = 2.22), the difference was statistically significant (P lt; 0.05) compared with 2.00. Conclusion: The morphological observation, immunohistochemistry and flow cytometry cell phenotype test results prompted the joint success of rhGM-CSF IL-4-induced train imDC and of mDC; built in one-way MLR DC induced T lymphocyte CCK-8 assay results the prompt imDC can adjust the model induced white body low T-cell response, may become a method of prevention and treatment of GVHD.
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