Barrera L F, Kramnik I, Skamene E, Radzioch D
Department of Medicine, Montreal General Hospital Research Institute, Quebec, Canada.
Immunology. 1994 Jul;82(3):457-64.
Reactive nitrogen intermediates (RNI) have been implicated in the interferon-gamma (IFN-gamma)-induced anti-microbial action of macrophages against a wide variety of pathogens. We have been studying the production of NO2- by macrophage lines derived from the bone marrow of either B10.A (Bcgs) strain mice (B10S cell lines), or their congenic BCG-resistant partners of the B10A.Bcgr (Bcgr) strain (B10R cell lines). We have discovered that there is a significant difference in the production of NO2- of B10S compared with B10R macrophages in response to IFN-gamma. By 48 hr following treatment with 10 U/ml IFN-gamma, B10R macrophages had produced an approximately threefold higher level of NO2- than B10S macrophages. Similar results were obtained when experiments were performed with total splenic cells harvested from the spleens of B10.A.Bcgr and B10.A strain mice. The bacteriostatic activity, as assessed by the [3H]uracil incorporation by Mycobacterium bovis BCG, was higher in B10R macrophages compared to B10S macrophages. The bacteriostatic activity of B10R and B10S macrophages correlated with the amount of nitric oxide produced by the macrophages. The anti-mycobacterial activity was inhibited by NgMMLA, a specific inhibitor of nitrite and nitrate synthesis from L-arginine. Addition of L-arginine to IFN-gamma-stimulated macrophages in the presence of NgMMLA restored nitrite production and bacteriostatic activity of macrophages. Northern blot analysis of macrophage nitric oxide synthase (iNOS) revealed that the difference in NO2- production by IFN-gamma-treated B10S and B10R lines was reflective of the difference in iNOS mRNA expression.
反应性氮中间体（RNI）与干扰素-γ（IFN-γ）诱导的巨噬细胞对多种病原体的抗菌作用有关。我们一直在研究源自B10.A（Bcgs）品系小鼠骨髓的巨噬细胞系（B10S细胞系）或其B10A.Bcgr（Bcgr）品系的同基因卡介苗抗性伙伴（B10R细胞系）产生NO2-的情况。我们发现，与B10R巨噬细胞相比，B10S巨噬细胞在响应IFN-γ时产生NO2-的量存在显著差异。在用10 U/ml IFN-γ处理48小时后，B10R巨噬细胞产生的NO2-水平比B10S巨噬细胞高出约三倍。当用从B10.A.Bcgr和B10.A品系小鼠脾脏中收获的全脾细胞进行实验时，也得到了类似的结果。通过牛分枝杆菌卡介苗的[3H]尿嘧啶掺入评估的抑菌活性，B10R巨噬细胞高于B10S巨噬细胞。B10R和B10S巨噬细胞的抑菌活性与巨噬细胞产生的一氧化氮量相关。抗分枝杆菌活性被NgMMLA抑制，NgMMLA是一种从L-精氨酸合成亚硝酸盐和硝酸盐的特异性抑制剂。在存在NgMMLA的情况下，向IFN-γ刺激的巨噬细胞中添加L-精氨酸可恢复巨噬细胞的亚硝酸盐产生和抑菌活性。巨噬细胞一氧化氮合酶（iNOS）的Northern印迹分析表明，IFN-γ处理的B10S和B10R细胞系产生NO2-的差异反映了iNOS mRNA表达的差异。
