GENE-TRANSFER INTO HUMANS - IMMUNOTHERAPY OF PATIENTS WITH ADVANCED MELANOMA, USING TUMOR-INFILTRATING LYMPHOCYTES MODIFIED BY RETROVIRAL GENE TRANSDUCTION
GENE-TRANSFER INTO HUMANS - IMMUNOTHERAPY OF PATIENTS WITH ADVANCED MELANOMA, USING TUMOR-INFILTRATING LYMPHOCYTES MODIFIED BY RETROVIRAL GENE TRANSDUCTION
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DOI：
10.1056/nejm199008303230904
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发表时间：
1990-08-30
影响因子：
158.5
通讯作者：
ANDERSON, WF
中科院分区：
文献类型：
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作者：
ROSENBERG, SA;AEBERSOLD, P;ANDERSON, WF
背景和方法：肿瘤浸润淋巴细胞（TIL）加白细胞介素-2治疗可以介导约一半患者的转移性黑色素瘤消退。为了优化这种治疗方法并确定TIL的体内分布和存活率，我们使用逆转录病毒介导的基因转导将编码新霉素抗性的基因引入人TIL中，然后将其输注到患者体内，从而使用新基因作为输注细胞的标记。结果：5例患者接受了基因修饰的TIL。所有患者均耐受良好，未发现因基因转导引起的副作用。Southern杂交和新霉素磷酸转移酶检测均证实了新霉素耐药基因的存在和表达。来自五名患者中的四名患者的细胞在高浓度的G418中成功生长，G418是一种新霉素类似物，否则对真核细胞有毒。通过聚合酶链反应分析，在所有五名患者的循环中连续三周发现基因修饰细胞，在两名患者中长达两个月。在细胞施用后多达64天从肿瘤沉积物中回收细胞。根据所有标准，包括TIL和患者中不存在感染性病毒，该程序是安全的。结论：这些研究证明了逆转录病毒基因转导用于人类基因治疗的可行性和安全性，并对设计具有改善抗肿瘤效力的TIL以及可能使用淋巴细胞进行其他疾病的基因治疗具有意义。
Background and Methods: Treatment with tumor-infiltrating lymphocytes (TIL) plus interleukin-2 can mediate the regression of metastatic melanoma in approximately half of patients. To optimize this treatment approach and define the in vivo distribution and survival of TIL, we used retroviral-mediated gene transduction to introduce the gene coding for resistance to neomycin into human TIL before their infusion into patients-thus using the new gene as a marker for the infused cells. Results: Five patients received the gene-modified TIL. All the patients tolerated the treatment well, and no side effects due to the gene transduction were noted. The presence and expression of the neomycin-resistance gene were demonstrated in TIL from all the patients with Southern blot analysis and enzymatic assay for the neomycin phosphotransferase coded by the bacterial gene. Cells from four of the five patients grew successfully in high concentrations of G418, a neomycin analogue otherwise toxic to eukaryotic cells. With polymerase-chain reaction analysis, gene-modified cells were consistently found in the circulation of all five patients for three weeks and for as long as two months in two patients. Cells were recovered from tumor deposits as much as 64 days after cell administration. The procedure was safe according to all criteria, including the absence of infectious virus in TIL and in the patients. Conclusions: These studies demonstrate the feasibility and safety of using retroviral gene transduction for human gene therapy and have implications for the design of TIL with improved antitumor potency, as well as for the possible use of lymphocytes for the gene therapy of other diseases.