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Due to the limitations of traditional surgery, chemotherapy and radiotherapy, cancer treatment is an urgent need for new means to open up new avenues. In recent years, with the rapid development of molecular biology, cell biology, and biotechnology, and promote the development of biological therapeutics in the medical field, the tumor biological treatment of more and more people's attention to one of the cells in adoptive immunotherapy The research focus in recent years. Adoptive immunotherapy is the most critical to select ideal effector cells from the eighties of the last century, since LAK cells, there have been TIL A-LAK, CTL, TAK, CIK cells were found and used in clinical trials, but there different defects and deficiencies. We explore new tumor the adoptive immune effector cell-PHA-CD3AK cells, this new effector cells are superior to other effector cell proliferation times, killing activity in vitro survival time. In this study, a preliminary study of its biological characteristics and clinical application. The main content: of PHA-by cord blood CD3AK were cells Preparation and biological characteristics of the initial research purposes: research of PHA-CD3AK cells induced in vitro methods and their biological characteristics, compared with LAK cells. : Isolated from human peripheral blood mononuclear cells (PBMC), phytohemagglutinin hormone (PHA), a monoclonal antibody (anti-CD3McAb) and recombinant human interleukin 2 (of rhIL-2), and co-induced preparation of PHA-CD3AK cells; application flow analysis of PHA-CD3AK cell immune phenotype, lactate dehydrogenase (LDH) detection the PHA-CD3AK cell killing activity observed morphology; Giemsa staining karyotype. Results: trace amounts of anti-CD3McAb (0.05ug/m1) supplemented with a small amount of rhIL-2 (300u/m1) and PHA (100ug/mL), co-cultured the induction and amplified of PHA-CD3AK cells, the amplification multiples significantly higher than LAK cells maintain high amplification is also much higher than LAK cells lasting; effector to target cell ratio of 80:1, PHA-by cord blood CD3AK were cell killing of tumor cells in vitro (K562) was 56.5%; immune the CD3 phenotype detection of PHA-CD3AK cells, of CD4 ~~, CD8 ~~ cell ratio, respectively (86.5 ± 5.89)%, (38.20 ± 5.27)%, (42.63 ± 3.50)%; karyotype was normal diploid chromosome number 46. Conclusion: PHA-CD3AK cells in CD3 to CD4 ~~, CD8 ~~ cell-based heterogeneous cell populations, and a lymphoblastoid cell-like characteristics The cells of PHA-by cord blood CD3AK were normal diploid cells. The of PHA-CD3AK cells are easy to induced in vitro amplification ability, live outside the body for a long time, a broad application prospects tumor adoptive immune effector cells tumoricidal activity. 2, PHA-CD3AK cell adoptive immunotherapy in advanced malignancies preliminary clinical study Objective: To observe the anti-CD3 monoclonal antibody activated killer cells (anti-CD3 monoclonal antibody activated killer cell, the PHA-cord blood CD3AK) Adoptive immunotherapy with advanced malignancies effect. Methods: 51 patients with malignant tumors of autologous peripheral blood mononuclear cells were isolated, adding of PHA, anti-CD3 mAb, recombinant human IL-2 (rhIL-2) were cultured in vitro induction, prepared from body the PHA-CD3AK cells. After the cells cultured for 12 to 14 days sampling for quality control testing, collection of quality control testing qualified the PHA-CD3AK cells adjusted to a final concentration (4.0 to 6.0) × 10 9 / L, intravenous infusion to the patient each patient treatment three courses, each course of reinfusion 5 times (2 d reinfusion 1) number of cells, each reinfusion of not less than 1 × 10 9, each course reinfusion cells the total number is greater than 5 × 10 ~ 9. Application of human T cell subsets detection kit and flow cytometry in peripheral blood of patients before and after treatment CD3 ~~, CD4, CD8 ~~ T cells and CD16 ~ 56 ~ cells (NK cells) ratio; seized observation of patients before and after treatment indicators of quality of life, and adverse reactions; evaluate the efficacy and calculation efficiency and clinical benefit rate. The results: separation and preparation of PHA-CD3AK cells meet the the active cell quality control requirements, and can be used to further the experimental treatment. The of PHA-CD3AK cells after adoptive immunotherapy, patients CD3 to of CD4 ~~, CD8 ~~ T cells and CD16 ~ 56 ~ cells (NK cells) rates were (48.88 ± 9.42)%, (35.09 ± 4.11)%, ( 28.17 ± 4.97)%, (20.31 ± 6.98)%, compared with before treatment were significantly increased (all P lt; 0.05). 51 patients, 45 patients after treatment, systemic symptoms improved significantly; related laboratory parameters were abnormal change after treatment, all patients, nor other systemic toxicity; including six cases of complete remission, partial remission in 14 cases, minor the 10 cases, 12 cases of stable progress in nine cases, the total effective rate of 58.8%, clinical benefit rate of 82.4%. Conclusion: PHA-CD3AK cells in adoptive immunotherapy of malignant tumors efficacy, effectively improve and enhance the patient's immune function, but also safe, with no side effects. In this study, Preparation and biological characteristics of the induced by of PHA-CD3AK cells in vitro and in vivo clinical application of a preliminary study of the system, and provides important experimental basis for further clinical application and useful exploration. Preliminary indication of PHA-CD3AK cells is a safe, effective, and has broad application prospects adoptive anti-tumor immune effector cells.
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