| Objective:To study the antineoplasmic activity and mechanism of action of activated HLA haploidentical peripheral blood stem cells in vivo and in vitro,and further study the safety and feasibility in treatment of advanced intractable solid tumor patients.To analyze the correlative factors influencing the treatment effectiveness,and provid a new and safe immunotherapy method for the advanced intractable solid tumor patients.Methods:42 metastatic or chemoresistant solid tumor patients were enrolled with haploidentical relatives as donors,and the HLA match was analyzed by nested PCR-SSP typing.After mobilization with 10μg/kg·d of G-CSF for 3 days, haplo-PBSCs were collected and activated by high dose of rhIL-2,then transfused to patients.The common symptom improving and clinical responses were observed after therapy.The phenotypes of haplo-PBSCs before and after mobilization/activation were compared by flow cytometry.The GM-CFU colony formation and nonspecific lysis activity of haplo-PBSCs before and after rhIL-2 short-term activation were compared by semisolid agarose and LDH method.The multiple Th1/Th2 cytokines secreted in patients' serum before and after therapy were examined by ELISA.The patients were divided into several subgroups according to different influential factors, and then the remission ratio,clinical benefit ratio,overall survival,progression-free survival,life quality benefit ratio of different subgroups were analyzed.Results:In 42 patients,the purity of extracted genomic DNA was(1.843±0.136),with concentration of(213.7±116) ng/ul.(3.39±0.51)×1010 activated haplo-PBSCs cells each was transfused.After mobilization,the proportion of T cells,especially CD4+ T cells decreased and the proportion of NK cells increased which showed statistical significance(P＜0.05).After activation with high dose of rhIL-2,the proportion of activated cells remarkably elevated,which highly expressed CD69 and CD25,and the non-specific tumor lysis activity against diverse cell lines strengthened.After treatment,the levels of CD4+,CD4+/CD8+,CD3-CD16+56+ (NK) were increased significantly after treatment(P＜0.05),but CD4+CD25+ (Tregs) was decreased significantly(P＜0.05);Th1 cytokine(TNF-α,IFN-γ) obviously increased,in contrast the concentration of TGF-βdropped(P＜0.05).After treatment,10 patients showed improvent in life quality,22 patients stable,and the life quality benefit rate was 76.2%,the KPS score was medially elevated by 20.The clinical benefit rate(CR + PR + SD) was 73.8%,and 7 patients reached partial remission(PR),including 2 renal cancer patients,2 ovarian cancer,1 lung caner,1 breast cancer,1 colon cancer,and progression was observed in 11 cases.The clinical benefit rates of renal and ovarian cancer were 90.0%and 81.8%,and higher than other cancer types.The PFS of overall patients was 6.0 months,and the PFS of renal cancer,ovarian cancer,lung caner,breast cancer were 5.7,10.5,5.5,7.4 months respectively.The remission ratio,clinical benefit ratio,overall survival,progressionfree survival,life quality benefit ratio of patients with the mismatching of KIR/HLA-C in the GVH and HVG(or matching) direction were 35.3%vs 4.0%, 94.1%vs 60.0%,26.8±3.1 vs 17.4±3.0 months,13.4±1.3 vs 8.0±0.9 months, 89.5%vs 65.2%,P＜0.05.The one year follow-up treatment response ratio,overall survival,progression-free survival,life quality benefit ratio of maternal/children and paternal/children patients were 18/24 vs 7/18,25.1±2.7 vs 13.7±3.2 months,12.1±1.2 vs 7.5±1.3 months,81.5%vs 66.7%,P＜0.05.Conclusion:High dose of rhIL-2 short-time activated haplo-PBSCs possessed considerable antitumor effect,and the survivability was favorable with the high security.The immunotolerance could be reversed and overcome.The mismatching of KIR/HLA-C in the GVH direction and maternal/children relation patients generated more favorable alloreactive response than others,and also could gain access to better survival time and improving of life quality and immunization. |
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