Simmons Monika, Putnak Robert, Sun Peifang, Burgess Timothy, Marasco Wayne A
Viral and Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, Maryland.
Division of Viral Diseases, Walter Reed Army Institute of Research, Silver Spring, Maryland.
Am J Trop Med Hyg. 2016 Nov 2;95(5):1148-1156. doi: 10.4269/ajtmh.16-0319. Epub 2016 Sep 19.
Passive immunization with anti-dengue virus (DENV) immune serum globulin (ISG) or monoclonal antibodies (Mabs) may serve to supplement or replace vaccination for short-term dengue immune prophylaxis. In the present study, we sought to establish proof-of-concept by evaluating several DENV-neutralizing antibodies for their ability to protect rhesus macaques against viremia following live virus challenge, including human anti-dengue ISG, and a human Mab (Mab11/wt) and its genetically engineered variant (Mab11/mutFc) that is unable to bind to cells with Fc gamma receptors (FcγR) and potentiate antibody-dependent enhancement (ADE). In the first experiment, groups of animals received ISG or Mab11/wt at low doses (3-10 mg/kg) or a saline control followed by challenge with DENV-2 at day 10 or 30. After passive immunization, only low-titered circulating virus-neutralizing antibody titers were measured in both groups, which were undetectable by day 30. After challenge at day 10, a reduction in viremia duration compared with the control was seen only in the ISG group (75%). However, after a day 30 challenge, no reduction in viremia was observed in both immunized groups. In a second experiment to test the effect of higher antibody doses on short-term protection, groups received either ISG, Mab11/wt, Mab11/mutFc (each at 25 mg/kg) or saline followed by challenge with DENV-2 on day 10. Increased virus-neutralizing antibody titers were detected in all groups at day 5 postinjection, with geometric mean titers (GMTs) of 464 (ISG), 313 (Mab11/wt), and 309 (Mab11/mutFc). After challenge, there was complete protection against viremia in the group that received ISG, and a reduction in viremia duration of 89% and 83% in groups that received Mab11/wt and Mab11/mutFc, respectively. An in vitro ADE assay in Fcγ receptor-bearing K562 cells with sera collected immediately before challenge showed increased DENV-2 infection levels in the presence of both ISG and Mab11/wt, which peaked at a serum dilution of 1:90, but not in Mab11/mutFc containing sera. The results suggest that antibody prophylaxis for dengue might be beneficial in eliminating or reducing viral loads thereby minimizing disease progression. Our results also suggest that blocking FcγR interactions through Mab11 Fc engineering may further prevent ADE.
用抗登革病毒(DENV)免疫血清球蛋白(ISG)或单克隆抗体(Mab)进行被动免疫,可用于补充或替代疫苗接种,以进行短期登革热免疫预防。在本研究中,我们试图通过评估几种DENV中和抗体在活病毒攻击后保护恒河猴免受病毒血症侵害的能力来建立概念验证,包括人抗登革热ISG、一种人源单克隆抗体(Mab11/wt)及其基因工程变体(Mab11/mutFc),后者无法与具有Fcγ受体(FcγR)的细胞结合并增强抗体依赖性增强(ADE)。在第一个实验中,动物组接受低剂量(3 - 10mg/kg)的ISG或Mab11/wt或生理盐水对照,然后在第10天或第30天用DENV - 2进行攻击。被动免疫后,两组中仅检测到低滴度的循环病毒中和抗体滴度,到第30天时无法检测到。在第10天进行攻击后,仅在ISG组中观察到病毒血症持续时间与对照组相比有所缩短(75%)。然而,在第30天进行攻击后,两个免疫组均未观察到病毒血症的降低。在第二个实验中,为了测试更高抗体剂量对短期保护的效果,各组接受ISG、Mab11/wt、Mab11/mutFc(均为25mg/kg)或生理盐水,然后在第10天用DENV - 2进行攻击。注射后第5天,所有组均检测到病毒中和抗体滴度升高,几何平均滴度(GMT)分别为464(ISG)、313(Mab11/wt)和309(Mab11/mutFc)。攻击后,接受ISG的组对病毒血症有完全保护作用,接受Mab11/wt和Mab11/mutFc的组中病毒血症持续时间分别缩短了89%和83%。在攻击前立即采集的血清中,在携带Fcγ受体的K562细胞中进行的体外ADE测定显示,在ISG和Mab11/wt存在的情况下,DENV - 2感染水平升高,在血清稀释度为1:90时达到峰值,但在含有Mab11/mutFc的血清中未出现这种情况。结果表明,登革热的抗体预防可能有助于消除或降低病毒载量,从而将疾病进展降至最低。我们的结果还表明,通过Mab11 Fc工程阻断FcγR相互作用可能进一步预防ADE。