The Study Of Clinic Application Of DC Co-Cultured With CIK Derived From Peripheral Blood On Advanced Malignant Cancer

ObjectiveTurner is a complex process which occurrence and development involves multi-factors and multi-steps.The immune function and other factors play an important role in this process. Therefore,in the course of diagnosis and treatment of cancer,we stress correcting the systemic immunodeficiency,in addition to the traditional surgery,radio-therapy and chemotherapy.In this study context,the concept of cancer biotherapy has emerged,and becomes the fourth mode of cancer treatment. The antitumor immuno-therapy has become a research hotspot on the basic research of CIK and DC with their unique advantages at home and abroad in recent years.Dendritic cell (DC) is regarded as a powerful antigen-presenting cell,and can mediate specific immune response to specific antigen efficiently.The cytokine-induced killer cell (CIK) is a new and efficient immune effector cell with merits of non-MHC restricted killing activity,fasting proliferation,high killing activity. Combi-nation of DC which can recognize antigens and activate the immune system and effective killing cells of CIK to treat malignant tumors opens up new perspectives to tumor's biotherapy.In this study,we co-cultured the combination of antigen pulsed with DC and no-antigen pulsed with DC with CIK,and observated the variation of phenotype,proliferative activity and cytotoxicity of the culture. Fifty-five patients with advanced malignant cancer were treated with DC-CIK,and we evaluated the short-term clinical efficacy and variation of immune function,and provided the basis for clinical bio-immunotherapy.Methods1.DC and CIK cells were generated from culturing peripheral blood mononuclear cells(PBMC);A549 soluble antigen was prepared.The effector cells were co-cultured as:pure CIK,non A549 antigen pulsed DC with CIK and A549 antigen pulsed DC with CIK(Ag-DC-CIK).The proliferation and the killing activity of CIK with different E:T range were observed dynamicly. Phenotype of these effector cells were identified by flow cytometry analysis.2.Fifty-five patients with advanced malignant cancer were treated with two cycles'DC-CIK and the best nutritional support treatment.In the course of treatment, adverse reaction and general state of health of patients were closely observed.The level of T cell subsets,CD4+CD25+regulatory T cell,NK cells and relating tumor markers were detected before and after the treatment.The curative effect was evaluated by imaging examination.Results1.Antigen pulsed DC with CIK and non antigen pulsed DC with CIK cultured begined to proliferate quickly from daylO.Moreover the proliferating activity was higher,and had significant difference compared with CIK cells(P<0.05).2.Antigen pulsed DC with CIK had a higher percentage of CD3+CD8+ (89.68±2.42)%,CD3+CD56+(76.43±3.98)%,and had significant difference compared with CIK and DC-CIK(P<0.05). Antigen pulsed DC with CIK presented mature DC specific surface marker,including HLA-DR(87.71±6.30)%,CD86(84.23±3.84)%, CD83(92.10±3.52)%,CD40(78.41±4.21)%,and had significant difference compared with DC-CIK(P<0.05).3.The killing activity of antigen pulsed DC with CIK was the strongest in the E:T range of 20:1,and had significant difference compared with CIK,DC-CIK,P< 0.01and<0.05.4.In addition to three cases there were no obvious change,the remaining fifty-two cases had different levels ease,such as improving appetite and strength,alleviating cancer pain,gaining weight.5. The CD8+ T cell and CD4+CD25+ regulatory T cell decreased,meanwhile the CD3+,CD4+ T cell subsets,CD4+/CD8+ ratio and NK cell significantly increased one month after the DC-CIK therapy.6.The effective rate was 23.63%,the clinical benefit rate was 81.81% one month after therapy.CEA,CA125 and AFP decreased,and had significant difference(P< 0.05),while the variation of CA724 had no significant difference(P>0.05).7.In the course of treatment,six cases appeared fever,usually around 38℃,lasting 3～5 hours.Other serious adverse reactions such as allergy,shock,myelosuppression,liver and kidney failure didn't appeared.Conclusions1.The main effector cell of co-cultuerd DC and CIK are the CD3+CD8+ and CD3+CD56+ double positive cells;antigen-pulsed DC or non-antigen-pulsed DC both can enhance the proliferation activity of CIK;antigen-pulsed DC can strengthen the killing activity of CIK at low E:T.2.After two courses of treatment of the DC-CIK cell,patients with advanced cancer had significantly improved general state of health,the imbalance of T cell subsets has been improved to some extent;tumor has been effectively controlled;the quality of life has been improved obviously.