CAJ | 학술논문

目的冷冻手术联合过继细胞免疫可能提供一种新的口腔黏膜恶性黑色素瘤的综合治疗方法。本研究通过监测冷冻联合改良的细胞因子诱导杀伤细胞（ improved cytokine induced killer cells ，iCIK）治疗口腔恶性黑色素瘤患者的外周血T细胞免疫变化和临床疗效，探讨外周血来源的iCIK细胞辅助冷冻治疗的安全性、有效性和免疫学机制。方法应用流式细胞术对2009年10月以来中山大学附属口腔医院应用iCIK辅助冷冻手术治疗的5例Ⅲ期以上口腔黏膜恶性黑色素瘤患者进行定期复诊研究，分析iCIK辅助冷冻手术对患者外周血T细胞功能和体外T细胞激活的影响，评价其临床疗效及不良反应。结果5例患者治接受冷冻手术9次，iCIK细胞免疫治疗43次。本研究对冷冻手术治疗前后患者外周血T细胞激活的比较研究显示冷冻治疗显著增加了CD3＋T细胞，CD3＋CD4＋Th细胞及CD3＋CD8＋CTL的细胞比例，提高了IL2， IFN-γ，TNF-α等Th1细胞因子的分泌。 iCIK治疗前后的T细胞激活检测显示了类似的结果，表明这两种治疗可以促进患者T细胞免疫功能的激活（ P＜0.05）。这些患者外周血免疫分析显示iCIK辅助冷冻治疗显著地提高了血液中T，Th及CTL细胞比例和Th1细胞因子的分泌，降低Treg细胞比例，进一步提供了iCIK辅助冷冻治疗能有效地改善患者T细胞免疫功能的证据。临床随访资料显示冷冻手术结合iCIK治疗5名口腔黏膜恶性黑色素瘤中，除一例患者生存9个月以外，4例患者目前均存活，且生存期分别为41＋、39＋、39＋和31＋个月，无明显不良反应。结论冷冻结合iCIK治疗口腔恶性黑色素瘤可以显著改善患者的T细胞免疫状况，是一种安全有效的综合治疗。
목적냉동수술연합과계세포면역가능제공일충신적구강점막악성흑색소류적종합치료방법。본연구통과감측냉동연합개량적세포인자유도살상세포（ improved cytokine induced killer cells ，iCIK）치료구강악성흑색소류환자적외주혈T세포면역변화화림상료효，탐토외주혈래원적iCIK세포보조냉동치료적안전성、유효성화면역학궤제。방법응용류식세포술대2009년10월이래중산대학부속구강의원응용iCIK보조냉동수술치료적5례Ⅲ기이상구강점막악성흑색소류환자진행정기복진연구，분석iCIK보조냉동수술대환자외주혈T세포공능화체외T세포격활적영향，평개기림상료효급불량반응。결과5례환자치접수냉동수술9차，iCIK세포면역치료43차。본연구대냉동수술치료전후환자외주혈T세포격활적비교연구현시냉동치료현저증가료CD3＋T세포，CD3＋CD4＋Th세포급CD3＋CD8＋CTL적세포비례，제고료IL2， IFN-γ，TNF-α등Th1세포인자적분비。 iCIK치료전후적T세포격활검측현시료유사적결과，표명저량충치료가이촉진환자T세포면역공능적격활（ P＜0.05）。저사환자외주혈면역분석현시iCIK보조냉동치료현저지제고료혈액중T，Th급CTL세포비례화Th1세포인자적분비，강저Treg세포비례，진일보제공료iCIK보조냉동치료능유효지개선환자T세포면역공능적증거。림상수방자료현시냉동수술결합iCIK치료5명구강점막악성흑색소류중，제일례환자생존9개월이외，4례환자목전균존활，차생존기분별위41＋、39＋、39＋화31＋개월，무명현불량반응。결론냉동결합iCIK치료구강악성흑색소류가이현저개선환자적T세포면역상황，시일충안전유효적종합치료。
Objective Combined treatment of cryosurgery with adoptive T-cell transfer may offer novel tools for melanoma immunotherapy .This study was designed to evaluate the immune effect of peripheral blood-derived improved cytokine-induced killer cells ( iCIK ) combined with cryosurgery in treating advanced oral malignant melanoma .Methods Peripheral blood mononuclear cells ( PBMC) of five patients with oral malignant melanoma were collected pre-or post-cryosurgery and expanded for iCIK cells with the timed addition of RetroNectin ,OKT3 mAb, IFNγand IL-2.Over 1 ×1010 iCIK cells were injected intravenously into patients after cryosurgery , once every 3 months or longer .Follow-up of these patients was carried out using clinical records .Results The five patients received a total of 9 cryosurgeries and 43 rounds of iCIK treatments .There were a significant increase in the population of CD3 +CD4 + Th,CD3 +CD8 + CTL and the production of Th1-type cytokine (IL-2,TNF-α) in iCIK cultures from peripheral blood mononuclear cells of these patients not only in post -cryosurgery but also in post-therapy with iCIK transfer.In addition,the combination treatment of cryosurgery and iCIK rescued these patients from an abnormal elevation of the population of circulating CD 4 +CD25 +Treg cells.All of the five patients achieved an objective clinical response according to the Response Evaluation Criteria in Solid Tumors , including three ongoing complete remissions ( 39 +, 39 + and 31 + months ) and two partial responses ( progression-free survival:41 +, 9 months).Four responders are currently alive .Conclusions The combination treatment of cryosurgery and iCIK infusion seems to be a promising treatment for oral malignant melanoma based on the improvement of anti -tumor immune response in patients .