Peng Ying, Schoenlaub Laura, Elliott Alexandra, Mitchell William J, Zhang Guoquan
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, Columbia, Missouri, USA.
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, Columbia, Missouri, USA
Infect Immun. 2014 Nov;82(11):4530-41. doi: 10.1128/IAI.01695-14. Epub 2014 Aug 11.
Our previous study demonstrated that treatment of Coxiella burnetii with the phase I lipopolysaccharide (PI-LPS)-targeted monoclonal antibody (MAb) 1E4 significantly inhibited C. burnetii infection in mice, suggesting that 1E4 is a protective MAb. To determine whether passive transfer of antibodies (Abs) can provide protection against C. burnetii natural infection, we examined if passive transfer of 1E4 would protect SCID mice against C. burnetii aerosol infection. The results indicated that 1E4 conferred significant protection against aerosolized C. burnetii, suggesting that 1E4 may be useful for preventing C. burnetii natural infection. To further understand the mechanisms of 1E4-mediated protection and to test the possibility of using humanized 1E4 to prevent C. burnetii infection, we examined whether the Fab fragment of 1E4 (Fab1E4), a recombinant murine single-chain variable fragment (muscFv1E4), and a humanized single-chain variable fragment (huscFv1E4) retained the ability of 1E4 to inhibit C. burnetii infection. The results indicated that Fab1E4, muscFv1E4, and huscFv1E4 were able to inhibit C. burnetii infection in mice but that their ability to inhibit C. burnetii infection was lower than that of 1E4. In addition, treatment of C. burnetii with Fab1E4, muscFv1E4, or huscFv1E4 can block C. burnetii infection of macrophages. Interestingly, treatment of C. burnetii with huscFv1E4 can significantly reduce C. burnetii infectivity in human macrophages. This report provides the first evidence to demonstrate that the humanized variable fragments of an LPS-specific MAb can neutralize C. burnetii infection and appears to be a promising step toward the potential use of a humanized MAb as emergency prophylaxis against C. burnetii exposure.
我们之前的研究表明,用靶向I相脂多糖(PI-LPS)的单克隆抗体(MAb)1E4处理伯氏考克斯体可显著抑制小鼠体内的伯氏考克斯体感染,这表明1E4是一种具有保护作用的单克隆抗体。为了确定抗体的被动转移是否能提供针对伯氏考克斯体自然感染的保护,我们检测了1E4的被动转移是否能保护SCID小鼠免受伯氏考克斯体气溶胶感染。结果表明,1E4对气溶胶化的伯氏考克斯体具有显著的保护作用,这表明1E4可能有助于预防伯氏考克斯体的自然感染。为了进一步了解1E4介导的保护机制,并测试使用人源化1E4预防伯氏考克斯体感染的可能性,我们检测了1E4的Fab片段(Fab1E4)、重组鼠单链可变片段(muscFv1E4)和人源化单链可变片段(huscFv1E4)是否保留了1E4抑制伯氏考克斯体感染的能力。结果表明,Fab1E4、muscFv1E4和huscFv1E4能够抑制小鼠体内的伯氏考克斯体感染,但它们抑制伯氏考克斯体感染的能力低于1E4。此外,用Fab1E4、muscFv1E4或huscFv1E4处理伯氏考克斯体可阻断巨噬细胞的伯氏考克斯体感染。有趣的是,用huscFv1E4处理伯氏考克斯体可显著降低其在人巨噬细胞中的感染性。本报告首次提供证据表明,LPS特异性单克隆抗体的人源化可变片段可中和伯氏考克斯体感染,这似乎是朝着潜在使用人源化单克隆抗体作为预防伯氏考克斯体暴露的紧急预防措施迈出的有希望的一步。