辐照后基因修饰自然杀伤细胞的特征及对卵巢癌细胞杀伤活性的研究

Characterization of irradiated gene modified NK cells and cytotoxic activity against ovarian tumor cell

目的探讨800cGy辐照后人白细胞介素15基因修饰的自然杀伤细胞(NKG-IL15)表面分子特征及其对卵巢癌细胞的杀伤活性。方法收集NKG-IL15细胞,800cGy进行辐照,100cGy/分,流式法检测辐照前后NKG-IL15细胞表型;利用51cr释放法检测辐照前后NKG-IL15细胞对靶细胞的杀伤活性,同时3H掺入法检测照射后细胞的增殖能力。结果 800cGy辐照前后NKG-IL15细胞表型均为CD56强阳性,CD19和CD3为阴性;对卵巢癌HO-8910细胞杀伤结果显示,0、800cGy组杀伤率分别在70.0%～23.7%、69.4%～22.6%之间,杀伤无明显差别;800cGy照射后NKG-IL15细胞24h增殖能力为0。结论 800cGy辐照后的NKG-IL15细胞具有人自然杀伤细胞的特征,无增殖能力,对人卵巢癌细胞HO-8910有较高的杀伤活性,可成为卵巢癌过继性免疫治疗的方法 。

Objective To study the characterization of the irradiated interleukin-15 gene-modified human natural killer cells （NKG-IL15） at 800cGy,and cytotoxic activity against human ovarian tumor cells in vitro.Methods NKG-IL15 cells were harvested and irradiated at dose of 800cGy,100cGy/min;the surface phenotype of the irradiated NKG-IL15 cells were acquired by FACS;The cytotoxic activity of NKG-IL15 （800cGy） against target cells was measured by a standard 4-h 51Cr release assay;and the proliferation of irradiated NKG-IL15 cells was quantified using a standard 3H-thymidine incorporation assay.Results The phenotype of NKG-IL15 and irradiated cells at doses 800cGy were all expressed CD56＋ and CD3-CD19-;the cytotoxic of NKG-IL15 was approximately at 69.4%-22.6% for ovarian tumor cell line HO-8910 at 800cGy and it was 70.0%-23.7% at 0 cGy.By the 3H-thymidine incorporation analysis,it was found that there was no cell proliferation after the irradiation by 800cGy.Conclusion NKG-IL15 cells which were irradiated at 800cGy are highly cytotoxic to human ovarian tumor cells and have no proliferation in vitro.Thus,irradiated NKG-IL15 cells may be a potentially novel therapy for human ovarian carcinoma.