ABSTRACT
Dendritic cells (DC) are important antigen-presenting cells in the development of anti-leukemic T-cells responses and it' s reported that DC can differentiate from monocytes or granulocytes in vitro. In the present study, the human leukemic DC were generated from acute promyelocytic leukemic( APL)cells after treatment with cytokines. M3 APL cells were isolated from peripheral blood of patients and incubated with rhGM-CSF( 100ng/ml)or rhGM-CSF( lOOng/ml) plus rhIL-4(500U/ml)for 14 days and the cells were co-cultured wirh TNF-?(100ng/ml)during last 3 days. The results demonstrated that both proliferation and differentiation of APL cells were induced by GM-CSF as the number of cultured cells increased and the cells expressed high level of CD45. Some cells displayed the characteristics of monocytes which expressed CD14 and some were leukemic DC as they expressed GDI a. The co-culturation of APL cells with GM-CSF and TNF-? augmented the differentiation of cells and about 35 percent of leukemic DC generated (was obtained). The maturation of APL cells could also be induced by GM-CSF plus IL-4 but there were few monocytes generated and about 10 percent of the cells were leukemic DC. If cultured for 3 weeks, the number of leukemic DC increased to 60 percent. The addition of TNF-? could augmented GM-CSF and IL-4 induced maturation of cells significantly and 90 percent of leukemic DC generated. Analysis of the cells with electric microscopy indicated that these leukemic DC displayed the similarly morphologic characteristics as monocyte-derived DC and there were still a few granules in their cytoplasm. The leukemic DC expressed high levels of HLA-DR, B7-1, B7-2 and CD54 molecules and could stimulate allo-T cells to proliferate in vitro. Such kind of leukemic DC might be useful tools for the generation of specific anti-tumor CTL and play important roles in immunotherapy of APL.
ABSTRACT
Objective:To observe the expression of ATPase F1? in colorectal cancer(CRC) tissues and in LoVo cells,and to discuss its clinical significance.Methods:Expression of ATPase F1? protein in 44 CRC specimens and their adjacent normal tissues(August 2007 to December 2007,Changhai Hospital) and ATPase F1? mRNA in 8 colorectal cancer tissues and their adjacent normal tissues were examined by immunohistochemistry EnVision assay and RT-PCR,respectively.Expression of ATPase F1? on the cell surface of LoVo cells was observed by immunofluorescence.The inhibitory effect of anti-ATPase F1? antibody on the proliferation of LoVo cells was evaluated by CCK-8 assay.Results:Expression of ATPase F1? in the 44 CRCs were significantly higher than those in the adjacent normal tissues as detected by immunohistochemistry(P0.05).Expression of ATPase F1? was observed on the cell surface of LoVo cells,and anti-ATPase F1? antibody significantly inhibited the proliferation of LoVo cells(P
ABSTRACT
TNF gene was transfected into murine LAK cells by retrovirus. Our results showed that TNF gene-transfected LAK cells secreted TNF more than normal LAK cells and control gehe-transfected LAK cells. The in vitro growth ability and cytotoxicity of TNF gene-transfected LAK cells were augmented significantly.The cytotoxicity of ,TNF gene-transfected LAK cells was markedly inhibited by anti - TNF monoclonal antibody, indicating that the, above augmentation was mediated by TNF secreted by transfected LAK cells. Significant therapeutic effect on the ascitic liver carcinoma.-bearing mice was achieved by i.p. injection of low dosage TNF gene transfected LAK cells and IL - 2.
ABSTRACT
Peripheral blood mononuclear cells (PBMNC) isolated from patients with acute leukemia (AL) and from normal controls were cultured in medium containing 1000 units/ml of recombinant interleukin 2 (IL-2). Marked LAK activity was induced on the third culture day in the normal controls,, with the highest cytotoxicity appearing between day 3 and 5 whereas induction of LAK activity in the AL patients began on the 5th day of culture, with the peak level appearing at day 15, showing that the peak of LAK activity was significantly delayed in AL. LAK cells surface phenotyping tests showed that CD8+ and CD16+ positive cells began to increase significantly from day 5 and reached the highest level at week 3, whereas CD4+ subclass began to decrease on day 5 and dropped to the nadir at week 3. The proportion of CD8+ and CD16+ cells were positively cor related with LAK activity, but, that of the CD4+ cell was inversely related with the LAK activity.