中华放射医学与防护杂志

杨光,刘长安,贾廷珍.γ射线照射对脐血淋巴细胞增殖和NK细胞活性的影响[J].中华放射医学与防护杂志,2006,26(1):6-9

γ射线照射对脐血淋巴细胞增殖和NK细胞活性的影响

Effect of gamma-ray irradiation on cord blood lymphocyte proliferation and NK cell activity

投稿时间：2005-09-28

DOI：

英文关键词:Gamma-ray Cord blood Lymphocyte Natural killer cell Proliferation

基金项目:国家自然科学基金资助项目(39770238)

作者	单位	E-mail
杨光	100083 北京大学第三医院肿瘤中心肿瘤化疗放射病科	yanggg9002@yahoo.com.cn
刘长安	100083 北京大学第三医院肿瘤中心肿瘤化疗放射病科
贾廷珍	100083 北京大学第三医院肿瘤中心肿瘤化疗放射病科

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中文摘要:

目的观察γ射线照射对脐血淋巴细胞增殖和NK细胞活性的影响。方法取正常分娩的新生儿脐带血,体外肝素抗凝,应用常规密度梯度离心法分离脐血单个核细胞(MNC),计数并调整细胞浓度。GammaCell1000数控细胞照射仪体外照射脐血MNC细胞,吸收剂量率3.72Gy/min。①采用³H-TdR掺入法测量脐血淋巴细胞增殖:取已分离的单个核细胞,调至细胞浓度1×10⁶个/ml,接受0.248、0.496、0.992、1.984、3.9685、.952、7.936和15.872Gyγ射线照射。照射后继续培养56h后加³H-TdR(3.7×10⁷Bq/孔)再培养8h后收集细胞,用Beckman液体闪烁计数器计数放射性核素每分钟闪烁数(计数/min)。②采用³H-TdR释放法检测脐血NK细胞活性:取传代培养24~48h的K562细胞为靶细胞,调整浓度1×10⁶个/ml,加入³H-TdR(3.7×10⁸Bq/ml)孵育6h,洗涤2次去除游离的³H-TdR,再制备成5×10⁵个/ml备用。脐血单个核细胞调整细胞浓度为1×10⁷个/ml作为效应细胞(效靶比20∶1),给以上述不同剂量γ射线照射。将效应细胞与制备好的靶细胞继续培养18h后收集细胞,用Beckman液体闪烁计数器计数放射性核素每分钟闪烁数。结果在γ射线对脐血淋巴细胞增殖活性影响的研究中,0.248~15.872Gyγ射线照射显著降低淋巴细胞增殖能力,增殖活性抑制程度与辐射剂量呈正相关(r=0.839,P<0.05);3.968Gyγ射线照射组增殖活性仅为对照组的30.86%(P<0.01),SI=7.8,其中3.968~15.872Gyγ射线照射未发现其明显的增殖活性。而γ射线对脐血NK细胞活性的影响的观察中发现0.248~1.984Gyγ射线照射可引起脐血NK细胞活性显著增高(P<0.01);3.968Gyγ射线照射保持NK细胞活性,与对照组相比差异无统计学意义(P>0.05);5.952~15.872Gyγ射线可引起脐血淋巴细胞NK细胞活性显著降低(P<0.01)。结论 3.968Gyγ射线照射完全抑制脐血淋巴细胞增殖的同时仍呈现脐血NK细胞抗肿瘤活性。因此,在进行供者淋巴细胞输注或混合脐血移植时,应用3.968Gyγ射线照射淋巴细胞,再进行过继性细胞免疫治疗可能降低移植物抗宿主病效应,同时不影响移植物抗白血病效应;混合脐血移植时增强移植物抗白血病效应。

英文摘要:

Objective To investigate the effects of gamma-ray irradiation on cord blood lymphocyte proliferation and NK cell activity. Methods Freshly isolated mononuclear cells from human cord blood were irradiated with different doses（0.248～15.872 Gy） of gamma-rays.The lymphocyte proliferation and NK cell activity were measured using ³H-TdR incorporation assay and ³H-TdR release assay,respectively. Results In dose range of 0.248～15.872 Gy,lymphocyte proliferation was inhibited and the inhibition rate was positively correlated with the irradiation dose（r=0.839,P<0.05）.Lymphocyte proliferation was not found in dose range of 3.968～15.872 Gy.Irradiation doses from 0.248 to 1.984 Gy could enhance NK cell activity.The activity of NK cells was reserved after irradiated with 3.968 Gy.Within the dose range of 5.952～15.872 Gy,NK cell activity was significantly inhibited. Conclusion Lymphocyte proliferation is inhibited and the activity of NK cell is reserved when irradiated with the dose of 3.968 Gy gamma-rays.So if the lymphocytes are irradiated with such a dose before donor lymphocytes infusion or mix-cord blood transplantation,the effect of graft versus host disease（GVHD） could be decreased whereas the effect of graft versus leukemia（GVL） reaction could be reserved simultaneously during adoptive cellular immunotherapy.

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