| Background and ObjectivesTraditional therapies of cancer are surgery, irradiation and chemotherapy. They can't clear cancer cells utterly. Otherwise, immunotherapy can stimulate the immune system to recognize and kill tumor cells by modulating reactions of tumor and host. The adoptive immunotherapy-infusion immunocompetent cells has anti-tumor effect to some extent. Compared with other anti-tumor drugs it can kill tumor cells directly, modulate and enhance the immune function without destroying the structure and function of immune system. Now it has become the important accessory therapy of tumor besides traditional methods. It supplies a new way of prophylaxising tumor relapse and improving the quality of life of patients. To obtain the best effect the immunocompetent cells should have stronger cytotoxicity and higher proliferation in the course of adoptive immunotherapy.Many experiments have proved that lymphokine-activated killer cells and tumor infiltrating lymphocytes have stronger cytotoxicity and lower proliferation in vivo and vitro. Recently a new type immunocompetent cell has been discovered. It iscytokine-induced killers. CIK cells have higher proliferation and cytotoxicity than LAK and TIL cells. Moreover it has an extended anti-tumor spectrum. CIK cells had anti-tumor and anti-tumor transference in Kunming mice bearing H-22 tumor and the mice bearing Si go sarcoma in vivo. It could restrict lymphoma occurring and get rid of lymphoma cells partly in severe combined immunodeficiency disease mice injected with human lymphoma cells. CIK cells generated from patients could kill K562 cells and lysis myeloid leukemia cells in vitro. After CIK cells transfected with a plasmid containing the interleukin-2 gene were used in patients with metastatic renal cell carcinoma, colorectal carcinoma and lymphoma, there were an increase in CD3 lymphocytes in the blood of patients and significant increase in serum level of interferon gamma, granulocyte-macrophage colony-stimulating factor and transforming growth factor beta during treatment. With the progression of studying CIK cells people improve culture conditions and disposal methods to enhance its proliferation and cytotoxicity. Phytohemagglutinin (PHA) is multicolony activator of T cells. PHA can stimulate silent T cells to lymphoblasts and be used as immune modulator. Using PHA stimulating peripheral blood mononuclear cells for twenty-four hours before cultured in CIK cells culture conditions to discuss whether PHA could enhance CIK cells' proliferation and cytotoxicity or not.MethodsPeripheral blood mononuclear cells were obtained from healthy persons. Every sample was divided into two groups. One group was cultured in traditional method. They were prepared and grown in complete medium consisting of RPMI1640 -, 10% autologus plasma 25mmol/L Hepes l00U/ml penicillin and l00U/ml streptomycin. l000U/ml rhIFN- Y was added on day 0. 25ng/ml MAb against CD3 and 300U/ml rhIL-2 were added after 24 hours of incubation. The other group was added PHA 20 u g/ml on day O.Then cultured them liketraditional culturing CIK cells after 24 hours. Cells were subcultured in fresh complete medium and 300U/ml rhIL-2 at 2 X 106 cells/ml every 3-4 days. During culturing cells morphological changes (cell volume, nucleus, cytoplasm) were observed. Cells' count, their proliferation fold and immunophenotype (CDS, CD19, CD4, CD8, CD56) were examined in two groups on 15th day. Their cytotoxicity against K562 cells, human esophagus carcinoma cells and primary acute leukemia cells were detected by modified MTT method.Results1. The cell volume and nucleus became larger and chromosome became thinner. Granule and vacuole appeared in some cytoplasm. The cytoplasm was filled with PAS positive granules in most of cells.2. The proliferation fold of CIK cells was 12.59 + 1.55.PHA-CIK cells' was 25.63 + 2.25.There was significant difference between them (P<0.05){Phe ratio of CD3+ CD8+and CD3"CD56+ cells changed markedly after culture (P<0.05), but no...
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