三种脑炎甲病毒,即,委内瑞拉,东方,和西方马脑炎病毒(VEEV,EEEV,和WEEV),被疾病控制和预防中心(CDC)归类为生物试剂。目前,没有针对这些病毒的许可医疗对策(MCM)可用于人类.中和抗体(NAb)是快速起效且高效的MCM,用于在暴露前和暴露后设置中针对生物抗菌药。虽然已经做了大量工作来识别反VEEVNAb,在识别针对EEEV和WEEV的NAb方面做得很少。为了开发抗EEEV或-WEEVNAb,使用各种不同EEEV或WEEV免疫原的互补策略对小鼠进行免疫,以最大限度地产生针对这些病毒的NAb。在产生的杂交瘤中,鉴定出3种抗EEEV和7种抗WEEV单克隆抗体具有体外中和活性。进一步评估了最有效的中和剂(两种抗EEEVNAb和三种抗WEEVNAb)对其他EEEV菌株的中和活性,一种Madariaga病毒(以前的南美EEEV),或WEEV。其中,G1-2-H4和G1-4-C3中和了所有三种EEEV毒株和Madariaga病毒毒株,而G8-2-H9和12WA中和了8株WEEV中的6株。为了确定这些NAs的保护功效,在各自的小鼠气溶胶攻击模型中评估了五种最有效的中和剂。当通过气溶胶途径在各自的病毒暴露之前24小时以2.5mg/kg或10mg/kg的剂量施用时,所有五种NAb均表现出不同水平的保护。其中,抗EEEVNAbG1-4-C3和抗WEEVNAb8C2在两种剂量下提供了100%的保护,并且所有存活的小鼠在整个研究中没有临床症状.此外,病毒暴露后14天,大脑中未检测到病毒。一起来看,开发了有效的NAb,证明了开发针对EEEV和WEEV感染的基于跨菌株抗体的MCM的潜力。
The three encephalitic alphaviruses, namely, the Venezuelan, eastern, and western equine encephalitis viruses (VEEV, EEEV, and WEEV), are classified by the Centers for Disease Control and Prevention (CDC) as biothreat agents. Currently, no licensed medical countermeasures (MCMs) against these viruses are available for humans. Neutralizing antibodies (NAbs) are fast-acting and highly effective MCMs for use in both pre- and post-exposure settings against biothreat agents. While significant work has been done to identify anti-VEEV NAbs, less has been done to identify NAbs against EEEV and WEEV. In order to develop anti-EEEV or -WEEV NAbs, mice were immunized using complementary strategies with a variety of different EEEV or WEEV immunogens to maximize the generation of NAbs to each of these viruses. Of the hybridomas generated, three anti-EEEV and seven anti-WEEV monoclonal antibodies were identified with in vitro neutralization activity. The most potent neutralizers (two anti-EEEV NAbs and three anti-WEEV NAbs) were further evaluated for neutralization activity against additional strains of EEEV, a single strain of Madariaga virus (formerly South American EEEV), or WEEV. Of these, G1-2-H4 and G1-4-C3 neutralized all three EEEV strains and the Madariaga virus strain, whereas G8-2-H9 and 12 WA neutralized six out of eight WEEV strains. To determine the protective efficacy of these NAbs, the five most potent neutralizers were evaluated in respective mouse aerosol challenge models. All five NAbs demonstrated various levels of protection when administered at doses of 2.5 mg/kg or 10 mg/kg 24 h before the respective virus exposure via the aerosol route. Of these, anti-EEEV NAb G1-4-C3 and anti-WEEV NAb 8C2 provided 100% protection at both doses and all surviving mice were free of clinical signs throughout the study. Additionally, no virus was detected in the brain 14 days post virus exposure. Taken together, efficacious NAbs were developed that demonstrate the potential for the development of cross-strain antibody-based MCMs against EEEV and WEEV infections.