The effect of deoxyguanosine on human lymphocyte function. I. Analysis of the interference with lymphocyte proliferation in vitro.
The effect of deoxyguanosine on human lymphocyte function. I. Analysis of the interference with lymphocyte proliferation in vitro.
复制标题
脱氧鸟苷对人淋巴细胞功能的影响。
DOI：
10.4049/jimmunol.132.5.2311
复制
发表时间：
1984
影响因子：
4.4
通讯作者：
B. Zegers
中科院分区：
文献类型：
--
作者：
L. Spaapen;G. Rijkers;G. Staal;G. Rijksen;S. K. Wadman;J. W. Stoop;B. Zegers
研究了脱氧鸟苷对有丝分裂原和抗原诱导的健康人外周血淋巴细胞增殖的影响。脱氧鸟苷可抑制细胞对有丝分裂原和抗原的增殖反应。引起50%增殖抑制的脱氧鸟苷浓度被证明依赖于培养基中使用的血清中分解代谢酶的活性，如嘌呤核苷磷酸化酶（PNP）。脱氧鸟苷和次黄嘌呤可以抑制脱氧鸟苷对植物血凝素（PHA）诱导的细胞增殖的抑制作用。通过测定细胞内（脱氧）核苷酸水平进一步分析了这些发现。在脱氧鸟苷存在的情况下，PHA刺激淋巴细胞导致dGTP在细胞内积聚。次黄嘌呤和脱氧鸟苷的存在消除了dGTP的抑制作用，但没有阻止dGTP的积累。另一方面，脱氧胞苷与脱氧鸟苷的联合添加不会导致细胞内dGTP可检测量的积累，而仅对毒性作用提供部分保护。此外，如果在培养基中添加预处理的胎牛血清，鸟苷对丝裂原诱导的细胞增殖的抑制程度与脱氧鸟苷相同。pnp缺陷或hgprt缺陷患者的外周血淋巴细胞在PHA或美洲商茅有丝分裂原刺激下对鸟苷的抑制作用有抵抗力，对脱氧鸟苷的敏感性低于正常供者的细胞。目前的结果清楚地表明，脱氧鸟苷介导的正常淋巴细胞增殖抑制涉及两条途径，即一方面PNP降解脱氧鸟苷，HGPRT回收鸟苷，并（可能）使GMP磷酸化最终产生GTP，另一方面脱氧胞苷激酶直接磷酸化脱氧鸟苷形成dGTP。
The effect of deoxyguanosine on mitogen- and antigen-induced proliferation of peripheral blood lymphocytes from healthy donors was studied. Deoxyguanosine was found to inhibit the proliferative response to mitogens and antigens. Concentrations of deoxyguanosine causing 50% inhibition of the proliferation proved to be dependent on the activity of catabolic enzymes, such as purine nucleoside phosphorylase (PNP), in sera used in the culture media. The inhibitory effect of deoxyguanosine on phytohemagglutinin (PHA)-induced cell proliferation was prevented by deoxycytidine as well as by hypoxanthine. These findings were analyzed further by determination of intracellular (deoxy)-nucleotide levels. Stimulation of lymphocytes by PHA in the presence of deoxyguanosine leads to intracellular accumulation of dGTP. The presence of hypoxanthine in addition to deoxyguanosine abolished the inhibitory effect but did not prevent dGTP accumulation. On the other hand, the addition of deoxycytidine in combination with deoxyguanosine did not lead to intracellular accumulation of detectable amounts of dGTP, but only gave partial protection against the toxic effect. Furthermore, guanosine inhibited mitogen-induced cell proliferation to the same extent as did deoxyguanosine provided that the culture media were supplemented with pretreated fetal calf serum. Peripheral blood lymphocytes of a PNP-deficient or a HGPRT-deficient patient in cultures stimulated with PHA or pokeweed mitogen were resistant to the inhibitory effects of guanosine and were less sensitive to deoxyguanosine than cells of normal donors. The present results clearly show the involvement of two pathways contributing to deoxyguanosine-mediated inhibition of the proliferation of normal lymphocytes, i.e., on the one hand degradation of deoxyguanosine by PNP, salvage of guanine by HGPRT, and (possibly) phosphorylation of GMP eventually leading to GTP, and on the other hand formation of dGTP by direct phosphorylation of deoxyguanosine by deoxycytidine kinase.