Sagone A L, Husney R, Guter H, Clark L
J Immunol. 1984 Sep;133(3):1488-94.
Phorbol esters have been documented to stimulate the proliferation of human blood mononuclear cell cultures. In addition, these agents are also known to stimulate the production and release of reactive oxygen species by monocytes. We demonstrated previously that H2O2, one of these oxygen metabolites, impairs the proliferative capacity of human blood lymphocytes. Therefore, in these experiments, we determined whether or not the H2O2 released by monocytes after activation by PMA modifies the proliferation of lymphocytes to this agent. Human blood mononuclear cells (80% lymphocytes and 20% monocytes) were incubated with PMA, and lymphoblastic transformation (LBT) was quantitated at 3 and 5 days by pulsing the cultures with thymidine. Initial experiments established that the concentration of PMA required for optimal LBT was 50 ng/ml. We then demonstrated that this concentration of PMA also induces a burst in hexose monophosphate shunt activity and H2O2 production of mononuclear cells as indicated by the enhanced oxidation of 14C-glucose and 14C-formate, respectively. The amount of H2O2 released into the medium was substantial. Our measurements indicate that the concentration of H2O2 could reach values as high as 0.008 mM during the first 2 hr of the cultures. The addition of catalase to PMA-treated cultures in concentrations sufficient to scavenge the H2O2 released by the monocytes was associated with an enhanced thymidine uptake (mean 79%). These results indicate that the hydrogen peroxide released by the monocytes modifies the response of lymphocytes to the PMA. Paradoxically, mononuclear cell cultures depleted of monocytes also had a lower proliferation to PMA than mononuclear cell cultures. This observation indicates that monocytes also produce factors required for lymphocyte proliferation to PMA such as an interleukin. In contrast, to PMA cultures, catalase did not alter the proliferation of mononuclear cell cultures stimulated by PHA. We previously documented that PHA does not stimulate an immediate burst in the oxidative metabolism of mononuclear cultures. Therefore, the effect of catalase in these two culture systems appears to correlate with the capacity of the mitogen to stimulate the oxidative metabolism of mononuclear cells. These observations suggest that the release of reactive oxygen species by monocytes may modify the response of lymphocytes to antigens both in vitro and in vivo.
佛波酯已被证明可刺激人血单核细胞培养物的增殖。此外，这些物质还已知可刺激单核细胞产生活性氧并释放。我们之前证明，这些氧代谢产物之一的过氧化氢会损害人血淋巴细胞的增殖能力。因此，在这些实验中，我们确定了经佛波酯激活后的单核细胞释放的过氧化氢是否会改变淋巴细胞对该物质的增殖反应。将人血单核细胞（80%为淋巴细胞，20%为单核细胞）与佛波酯一起孵育，并在第3天和第5天通过用胸腺嘧啶核苷脉冲培养物来定量淋巴细胞转化（LBT）。初步实验确定，最佳LBT所需的佛波酯浓度为50 ng/ml。然后我们证明，该浓度的佛波酯还会诱导单核细胞的磷酸己糖旁路活性和过氧化氢产生爆发，分别表现为14C-葡萄糖和14C-甲酸氧化增强。释放到培养基中的过氧化氢量很大。我们的测量表明，在培养的最初2小时内，过氧化氢浓度可高达0.008 mM。向经佛波酯处理的培养物中加入足以清除单核细胞释放的过氧化氢的过氧化氢酶，与胸腺嘧啶核苷摄取增加有关（平均为79%）。这些结果表明，单核细胞释放的过氧化氢会改变淋巴细胞对佛波酯的反应。矛盾的是，去除单核细胞的单核细胞培养物对佛波酯的增殖也低于单核细胞培养物。这一观察结果表明，单核细胞还会产生淋巴细胞对佛波酯增殖所需的因子，如白细胞介素。相比之下，对于佛波酯培养物，过氧化氢酶不会改变由PHA刺激的单核细胞培养物的增殖。我们之前记录到，PHA不会刺激单核细胞培养物的氧化代谢立即爆发。因此，过氧化氢酶在这两种培养系统中的作用似乎与促有丝分裂原刺激单核细胞氧化代谢的能力相关。这些观察结果表明，单核细胞产生活性氧可能会在体外和体内改变淋巴细胞对抗原的反应。
