Influence Of Mannatide On The Activity Of Cytokine-induced Killer Cells

Objective Mannan peptide (mannatide) is a kind of biological response modifier,which is a kind of polysaccharide peptide substances, is obtained by submerged culture and fermented from Number33bacterial stain of alpha-hemolytic streptococcus (a-hemolytic Streptococcui) which is separated from the healthy people oral cavity, has both immunological competence and antitumor activity of dual biological characteristics. The CIK cells is a group of heterogeneous cell,which is induced by using human peripheral blood mononuclear cells in vitro with a variety of cytokines, included IL-2, IFN-y, and CD3monoclonal antibody in vitro. The chief effector cell of CIK cells is CD3+CD56+cell.ClK cells is a new style, highly efficient immune effector cells in tumor adoptive reveal the tremendous value of immunotherapy,which has a strong antitumor activity of T lymphocytes and non-MHC restricted of NK cells. Currently the research that mannatide enhance the amplication and cytotoxic activity of CIK cells is few. In this study, we observed the changes of cytotoxic activity of CIK cells for K562and Raji cell line after affected with mannatide and to explore the mechanism that Manatide affect people CIK cells cytotoxic activity function, to provide the theoretical and exprement basis for the apply that Mannatide be used in adoptive immunotherapy therapy.Method Mononuclear cells were isolated from peripheral blood of healthy honors,CIK cells were induced by culturing PBMC with regular method. CIK cells were collected on the fourteenth day and were induced with mannatide in different concentrations as experiment groups,the CIK cells which did not add mannatide were control group, the morphological changes of experiment groups and control group CIK cells were observated by using inverted microscope, the cells proliferation rate were assessed by MTT assay, the positive rate of CD3CD56"cells and the expression of the NKG2D content on the CIK cell surface were detected using flow cytometry, the expression of granzyme and perforin were detected by FCM assay,The cytotoxic activity of CIKs against K562, Raji cell lines were determined by Lactate dehydrogenase release assay in vitro.Results Different concentrations of mannan peptide could promote CIK cells proliferation, and were concentration dependent.The expression of the NKG2D receptors in CIK cells were raise.The ability of CIK cells secreting granzyme and perforin in experiment groups was significantly higher than the control group(P<0.05),and the cytotoxic activity of the CIK cells was siganificent enhanced by mannan peptide for K562and Raji cell lines (p<0.05).Conclusion Mannatide is capable become immunologic adjuvant which can promote the immunocompetence of CIK cells.