ANTIBODY-COMPLEMENT INTERACTIONS WITH PURIFIED LYMPHOCYTIC CHORIOMENINGITIS VIRUS
ANTIBODY-COMPLEMENT INTERACTIONS WITH PURIFIED LYMPHOCYTIC CHORIOMENINGITIS VIRUS
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DOI：
10.1016/0042-6822(76)90060-x
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发表时间：
1976-01-01
期刊：
影响因子：
3.7
通讯作者：
OLDSTONE, MBA
中科院分区：
文献类型：
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作者：
WELSH, RM;LAMPERT, PW;OLDSTONE, MBA
对淋巴细胞性脉络膜脑膜炎病毒(LCMV)阿姆斯特朗株的免疫复合体形成和免疫病毒解进行了研究。从急性感染的BHK细胞中无血清培养获得LCMV，用甲醇沉淀法和蔗糖密度梯度离心法进行纯化。在放线菌素D处理细胞的条件下标记了番茄花叶病毒的31-和23-S核糖体，阻止了病毒粒子中细胞核糖体的标记。豚鼠抗LCMV抗体的加入降低了病毒的感染力，提高了病毒的沉降率，但不能提高[U-~3H]LCMV和125I表面蛋白标记的LCMV的密度。这表明，抗体将病毒粒子聚集成更快的沉积复合体。补体(C)的加入进一步降低了LCMV-Ab的感染性，提高了[125I]LCMV的沉降率和密度。当[U-~3H]LCMV与抗体和C反应并进行速率区带沉淀时，RNA标记保持在梯度的顶部，而不是与125I-表面蛋白膜标记沉淀。这一观察结果表明，抗体和C裂解了病毒，经EM证实，首先在病毒膜上包覆了电子致密物质，然后膜肿胀、破裂，病毒核心物质释放。使用缺乏特定C组分的血清表明，C介导的失活是通过经典的C途径进行的。缺乏后C组分的血清(C3以外)可增强LCMV-Ab复合体的失活，但当使用完整的C源时，这种失活要广泛得多。有证据表明，免疫和持续感染的小鼠产生了针对LCMV表面的C固定抗体。[这些研究使用的是小鼠、兔、豚鼠和人的C组分。]。
Immune complex formation and immune virolysis of the Armstrong strain of lymphocytic choriomeningitis virus (LCMV) were examined. LCMV, harvested from acutely infected [baby hamster kidney] BHK cells propagated in the absence of serum, was purified by methanol precipitation and sucrose density gradient centrifugation. The 31- and 23-S RNA species of LCMV were labeled under conditions of actinomycin D treatment of cells that prevented the labeling of cell ribosomes in the virion. The addition of guinea pig antibody (Ab) to LCMV lowered viral infectivity and enhanced the sedimentation rate but not the density of [U-3H]LCMV and 125I surface protein-labeled LCMV. This suggested that Ab aggregated the virions into faster sedimenting complexes. The addition of complement (C) to the LCMV-Ab complex further reduced the infectivity and increased the sedimentation rate and density of [125I]LCMV. When [U-3H]LCMV was reacted with Ab and C and subjected to rate zonal sedimentation, the RNA label remained at the top of the gradient rather than sedimenting with the 125I-surface protein-membrane label. This observation, which indicated that Ab and C lysed the virus, was confirmed by EM showing first coating of the viral membrane with electron dense material, then swelling and rupture of the membrane and release of viral core material. The use of sera deficient in specified C components indicated that the C-mediated inactivation proceeded via the classical C pathway. Sera deficient in latter C components (beyond C3) augmented the inactivation of LCMV-Ab complexes, but this inactivation was far more extensive when a complete C source was used. Evidence suggesting that immune and persistently infected mice produce C-fixing Ab to the surface of the LCMV was documented. [Mouse, rabbit, guinea pig and human C components were used in these studies.].