| Dendritic cells (DCs) have the strong function of specific duty antigen presenting cells (APC) in now. It can be achieve immunofunction of antigen by some ways. Some study showed that DCs is a bridge linked antigen and immune response cells to activate the immuneresponse of effective antitumor. Tumor rejection antigen codded by MAGE family produces antigen peptided proceseed in cells. Tumor rejection antigen combined with HLA-I formed a compplex and it distinguished by self-cytotoxic T lymphocytes (CTL), induce it specially killing relevant tumor cells. MAGE more highly expresses in many tumor tissues but can't expresss in normal tissues (except testis and fetal tissue), it is a kind of ideal target to special immune mediated by CTL.1. The expansion and maturation of inducing dendritic cells derived from mouse bone marrow in vitroThe expansion and maturation of inducing dendritic cells derived from mouse bone marrow in vitro. The dendritic cells derived from mouse bone marrow were proliferated by using GM-CSF and IL-4 in vitro. A part of suspension cells were abserved on 2 days after mouse bone marrow cells were cocultured with IL-4 (500u/ml) and GM-CSF (1000u/ml), the suspension cells increased with short process with burr 3 days later, and most of cells were floated with long process with burr, at the same time large round and fusiform cells decreased on the bottom of bottles 5 days later. The results showed that these cells have the characteristic of typical morphology of dentritic cells. The expressions of surface marker CD83 and CD86 of CD maturation induced by TNF-α stimulating or freezing and thawing antigen increased significantly as compared with that in blank control group (P<0.05). There was significantly different between IL-4+GM-CSF+TNF-α group, IL-4+GM-CSF+carcinoma antigen group and IL-4+GM-CSF group, while there was not different between IL-4+GM-CSF+TNF-α and IL-4+GM-CSF+caicinoma antigen groups. The results showed that the expressions of CD83 and CD86 increased after dentritic cells were loaded with antigen and stimulated with TNF-α. Then the above mentioned mature DCs were used to perform mixture lymphocyte culture to stimulate T cells proliferation. The results showed that the proliferation of T cells was stimulated after DCs in IL-4+GM-CSF+TM-α group, IL-4+GM-CSF+freezing and thawing antigen group were cultured with T cells, CPM in which were significantly higher than that in blank control group and IL-4+GM-CSF group (P<0.001). The cpm of DCs cultured with T cells in IL-4+GM-CSF+freezing and thawing antigen group was significantly higher than that in IL-4+GM-CSF+TNF-α group (P<0.001). And the cpm in all the groups of mixture ratio of 1 to 5 was higher than that in ratio of 1:10 groups, but there was not significantly different. 2. DCs loaded with total RNA of tumorThe induction of DCsorigined from mononuclear leukocytes of periphery blood of lung cancer patient increased, DCs morphologically grew in a cluster of clony under invereted microscope, for 3 days in the culture of cells, the membrane in a part of them processed; for 6 days in culture, most of them grew in suspension, cell body was biger and irregular, cell membrane processed with branch tower, cytoplasm enriched, the cells present a typical DCs in morphology. There were a lot of microfolds and processes in the cell membrane, cell nucleus was biger and inclined to one side, mitochondria enriched in cells. After PBMC inducted with GM-CSF and IL-4 for 5 days in the culture, the positive rates of surface maker CD1a and CD83 detected with FCM were 11.10±1.08(%) and 7.24±2.25(%). Respectively, accompanied with the up-regulting expressions of HLA-ABC and CD86 (B7-2), down-regulating expression of CD14 and unclchange of HLA-DR. The results suggest that PBMC differentiated into DCs in the effecfs of GM-CSF and IL-4.The above-mentioned DCs loaded with total RNA of lung cancer cells to detect antigen presenting ability of DCs. We extracted the total RNA with two bands of 28 S and 18 S detected with dematured gel electr...
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