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Objective To investigate the killing effect of donor NK cells on recipient dentritic cells (DCs) by blocking KIR2DL1 and KIR2DL3 with monoclonal antibodies CD158a and CD158b,and the possible mechanism about donor NK cells reducing the incidence of graft versus host disease (GVHD).Methods SSP-PCR was used to examine the genotypic makeup of KIR in 15 pairs of patients and their HLA-identical sibling hematopoietic stem cell transplantation (HSCT) donors in our department.Peripheral blood mononuclear cells (PBMC) from 15 pairs of patients and their donors were extracted and preserved,and donor NK cells were isolated by DYNABEADS UNTOUCHED HUMAN NK sorting kit.Methyl thiazolyl tetrazolium (MTT) reduction assay was used to detect the killing activity of donor NK cells against recipients DCs.To observe the different NK killing activity before and after KIR receptors blockade,the anti-CD158a and CD158b monoclonal antibodies were used to block KIR inhibitory receptors.Results The killing effect of donor NK cells was significantly enhanced after inhibitory receptor KIRs blockades,the killing effect of NK cells in HVG KIR ligandmismatching pattern group was significantly higher than GVH or KIR receptor-ligand matched pattern group.The killing effect became stronger when the dose of antibody was increased.ConclusionIt is demonstrated in vitro that the killing effect of alloreactive NK cells could be enhanced by KIR functional modification,and the infusion of alloreactive NK cells could be used as a new strategy of immunotherapy for GVHD.
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Objective To investigate the relationship between CD158j expression and phosphorylated ERK (p-ERK) in CD4+ CD28null T cells in cerebral infarction (CI) patients- with carotid atherosclerosis and its effects on carotid atherosclerotic plaque stability. Methods Percentage of peripheral CD4+ CD28null and the expression of CD158j and perform on CD4+ CD28null cells was analyzed with flow cytometry in 106 CI patients with carotid atherosclerosis, 33 CI patients with normal carotid arteries and in 50 normal controls, respectively; p-ERK expression was assayed with flow cytometry in 36 CI patients with unstable plaque, and serum IFN-γ was detected with ELISA. The intima-media thickness (IMT) of bilateral carotid arteries in all subjects was confirmed by the colour Doppler ultrasonograph imagingResults Percentage of the CD4+ CD28null T cells, expression of CD158j and perform on CD4+ CD28null T cells and the serum IFN-γ levels was dramatically higher in CI patients than that in normal controls, respectively (all P <0.01), which was decreased in an order of CI patients with patients with unstable plaque, stable plaque, carotid artery IMT and with normal carotid artery. A strong positive correlation was observed between the CD158j expression and degree of p-ERK in CI patients with unstable plaque (P < 0. 01). Conclusion CD4+ CD28null T cells were significantly increased in CI patients with carotid atherosclerosis. CD158j might up-regulate p-ERK expression and induce the proliferation of the CD4+ CD28nullT cells; consequently, higher cytokine production such as IFN-γ produced by CD4+ CD28null T cells may cause the formation of unstable plaque.
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Objective To investigate the expression of CD158b in NK cells after allogeneic kidney transplantation.Methods In 62 patients with allogeneic kidney transplantation, blood samples were collected on the day before operation, first, 7th day after operation, the moment the graft (reco)-(vered) and the acute rejection occurred. The expression of CD158b was detected in peripheral blood NK cells. The ratio of CD3~-CD16/56~+CD158b~+ was measured.Results There were 38 patients without acute rejection during the whole transplantation period. The ratio of CD3~-CD16/56~+ cells and CD3~-CD16/56~+CD158b~+ cells were stable before and after the transplantation. Twenty-four patients experienced acute rejection post-transplantation. The ratio of CD3~-CD16/56~+ cells was increased significantly after acute rejection, the ratio of CD3~-CD16/56~+CD158b~+ cells decreased significantly, and the percentage of CD3~-CD16/56~+CD158b~+ cells of total NK cells decreased significantly.Conclusion There are not too much factors interfering with the expression of CD158b in NK cells, and the ratio of CD3~-CD16/56~+CD158b~+ cells had already decreased significantly before the clinical diagnosis of (acute) rejection. Monitoring of CD158b in NK cells is more accurate and sensitive for the evaluation of immune state.
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AIM: To investigate the sequence expression o f CD158 molecule after tacrolimus (FK506), mycophenolate mofetil (MMF) combined with methylprednisone (MP) treatment for refractory chronic graft-versus-host di seases (cGVHD). METHODS: The efficacy and the side effect were observed in 6 chi ld patients with extensive cGVHD after allogeneic hematopoietic stem cell transp lantation treated with the combination of FK506, MMF and MP, meanwhile the chang es of the CD158 expressions on T lymphocytes and NK cells in peripheral blood be fore and after treatment were observed. RESULTS: The expression of CD4+CD158a+ and CD4+CD158b+ were very low before and after transplantation and treatment, there was no stati stical significance. The expression of CD3+CD158b+ and CD3+CD8+CD158b + were 4.97%?2.36% and 4.58%?2.90% respectively in five patients with acut e GVHD, and there was statistical significance compared with that of before-tran splantation (P