| Aims To prepare a cancer vaccine (H22-DC) selected expressing high levels of costimulation molecules based on fusion of the H22 cells with dendritic cells (DC) of mice and analyze the biological characteristics of H22-DC , and induce specific CTL activity. Methods By a method of density gradient centrifugation of freshly isolated procedure with metrizamide and by semi-adhesion to culture plates and characteristic of FcR ,and cytokine-induced expansion culture with Granulocyte-macrophage colony-stimulating factor (GM-C SF) and interleukine 4 (IL-4) , a large number of DC were developed, and then by polyethyleneglycol (PEG) fused with 1122 cells . The H22-DC were generated by selecting with MACS marked with CD 11 c(N4 18) MicroBeads .The techniques of cell culture , immunocytochemistry and light microscope were also used in the obtaining the characteristic of growth and morphology of H22-DC in vivtro . Directly with H22-DC as the immunogen were implanted subcutaneously into the right armpit of BALBIC mice ,and their characteristic of tumorigenicity in vivo were observed. Using a method called MTT examine the CTL activity of mice spleen in vitro. Results 瓺C which were isolated and generated are CD 11 c~ cells with irregular-shaped ,and expressing high levels of CD8O,CD86 and CD54 molecules ; 1122 are CD 11 & cells with spherical-shaped and bigger -3- volume ,and expressing no CD8O ,CD86 and CD54 molecules ; H22-DC are CD 11 c~ cells with bigger volume ,spherical ,fiat or irregular-shaped and expressing high levels of CD8O,CD86 and CD54 molecules too . ? H22-DC could divide and proliferate , but which significantly decreased as compared with H22 and their growth curve is more fiat than H22. ?Over 60 days after subcutaneous implantment of 1122-DC was observed no induced tumorigenesis , which were significantly different than compared with control groups (P<0 .01) . ~ The spleen CTL activity against H22 cells in mice implanted with fresh 1122-DC are significantly higher than control groups (P<0.0 1). Conclusions 0 Spleen-derrived DC of mice by density gradient centrifugation of freshly isolated procedure with metrizamide and cytokine-induced expansion culture with GM-CSF and IL-4 is matural DC with typical morphology and function . ?1422-DC obtained by PEG fusion , CD1 lc MicroBeads marking and Mini MACS selecting have some characteristics of both parental cells . The method is simple and reliable and has practical value . 0 The activity of dividing and proliferating of H22-DC are significantly decreased , and H22-DC induced no tumorigenisis in vivo and had losed the characteristic of malign growth of parental H22 cells. ?H22-DC could significantly stimulate the specific CTL activity of mice , which means the fusion cells have already obtained the antigen presenting function of parental DC and could present H22 specific antigen which has not been clear yet , and induce antitumor immune response ; H22 cells simple mixed with DC could stimulate the specific CTL activity , which could restrain the development of tumor in some degree , but can not prevent the generation of tumor. That is to say the DC vaccine is likely to became a new way of heptocarcinoma in~imunotherapy.
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