SPECIFIC AMINO-ACID (L-ARGININE) REQUIREMENT FOR THE MICROBIOSTATIC ACTIVITY OF MURINE MACROPHAGES
SPECIFIC AMINO-ACID (L-ARGININE) REQUIREMENT FOR THE MICROBIOSTATIC ACTIVITY OF MURINE MACROPHAGES
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DOI：
10.1172/jci113427
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发表时间：
1988-04-01
影响因子：
15.9
通讯作者：
DURACK, DT
中科院分区：
文献类型：
--
作者：
GRANGER, DL;HIBBS, JB;DURACK, DT
用小鼠腹腔细胞毒巨噬细胞（CM）在含10%透析胎牛血清的Dulbecco培养基中对新生隐球菌（Cryptococcus neoformans）进行体外抑菌试验。在用去污剂溶解巨噬细胞后，使用电子颗粒计数测量真菌抑制。巨噬细胞抑真菌作用在仅缺乏L-精氨酸的培养基中失败。完全的真菌抑制作用被L-精氨酸恢复;恢复是浓度依赖性的，在200 μ M时最大。如果存在L-精氨酸，则所有其他必需氨基酸的缺失不会消除抑真菌作用。在包括D-精氨酸在内的20种胍基化合物中，只有3种（L-精氨酸、L-高精氨酸和L-精氨酸甲酯）支持抑真菌作用。已知的巨噬细胞细胞毒性的激活剂或介质（内毒素，干扰素γ，肿瘤坏死因子）不能取代L-精氨酸用于CM介导的真菌抑制。胍基类似物NG-单甲基-L-精氨酸是CM介导的真菌抑制的有效竞争性抑制剂，在抑制剂/L-精氨酸比例为1：27时抑制率为50%。虽然CM通过L-精氨酸依赖性机制完全阻断真菌繁殖，但大多数休眠真菌仍然存活。因此，该机制被视为与巨噬细胞的肿瘤抑制作用相似或相同的微生物抑制过程。这表明细胞毒性巨噬细胞消耗L-精氨酸后产生广谱生物抑制代谢物。
The microbiostatic action of macrophages was studied in vitro employing peritoneal cytotoxic macrophages (CM) from mice acting against Cryptococcus neoformans cultured in Dulbecco''s medium with 10% dialyzed fetal bovine serum. Fungistasis was measured using electronic particle counting after lysis of macrophages with detergent. Macrophage fungistasis failed in medium lacking only L-arginine. Complete fungistasis was restored by L-arginine; restoration was concentration dependent, maximal at 200 .mu.M. Deletion of all other essential amino acids did not abrogate fungistasis provided that L-arginine was present. Of twenty guanido compounds, including D-arginine, only three (L-arginine, L-homoarginine, and L-arginine methylester) supported fungistasis. Known activators or mediators of macrophage cytotoxicity (endotoxin, interferon gamma, tumor necosis factor) did not replace L-arginine for CM-mediated fungistasis. The guanido analogue NG-monomethyl-L-arginine was a potent competitive inhibitor of CM-mediated fungistasis giving 50% inhibition at an inhibitor/L-arginine ratio of 1:27. Although CM completely blocked fungal reproduction via an L-arginine-dependent mechanism, the majority of the dormant fungi remained viable. Thus, this mechanism is viewed as a microbiostatic process similar or identical to the tumoristatic effect of macrophages. This suggests the production of a broad spectrum biostatic metabolite(s) upon consumption of L-arginine by cytotoxic macrophages.