The Role of IgG Subclass in Antibody-Mediated Protection against Carbapenem-ResistantKlebsiella pneumoniae

Abstract

Monoclonal antibodies (Abs) have the potential to assist in the battle against multidrug-resistant bacteria such as Carbapenem-Resistant Klebsiella pneumoniae (CR-Kp). However, the characteristics by which these Abs function, such as the role of antibody subclass, must be determined before such modalities can be carried from the bench to the bedside. We performed a subclass switch on anti-capsular monoclonal murine IgG3 (mIgG3) hybridomas and identified and purified a murine IgG1 (mIgG1) hybridoma line through sib selection. We then compared the ability of the mIgG1 and mIgG3 antibodies to control CR-Kp ST258 infection both in vitro and in vivo. We found by ELISA and flow cytometry that mIgG3 has superior binding to CR-Kp CPS and superior agglutinating ability compared to mIgG1. The mIgG3 also predictably had better complement-mediated serum bactericidal activity than the mIgG1 and also promoted neutrophil-mediated killing at concentrations lower than the mIgG1. In contrast, the mIgG1 had marginally better activity in improving macrophage-mediated phagocytosis. Comparing their activities in a pulmonary infection model with wild type as well as neutropenic mice, both antibodies reduced organ burden in a non-lethal challenge, regardless of neutrophil status, with mIgG1 having the highest overall burden reduction in both scenarios. However, at a lethal inoculum, both antibodies showed reduced efficacy in neutropenic mice, with mIgG3 retaining the most activity. These findings suggest the viability of monoclonal Ab adjunctive therapy in neutropenic patients that cannot mount their own immune response, while also providing some insight into the relative contributions of immune mediators in CR-Kp protection.ImportanceCarbapenem-resistant Klebsiella pneumoniae is an urgent public health threat that causes life-threatening infections in immunocompromised hosts. Its resistance to nearly all antibiotics necessitates novel strategies to treat it, including the use of monoclonal antibodies. Monoclonal antibodies are emerging as important adjuncts to traditional pharmaceuticals, and studying how they protect against specific bacteria such as Klebsiella pneumoniae is crucial to their development as effective therapies. Antibody subclass is often overlooked but is a major factor in how an antibody interacts with other mediators of immunity. This paper is the first to examine how the subclass of anti-capsular monoclonal antibodies can affect efficacy against CR-Kp. Additionally, this work sheds light on the viability of monoclonal antibody therapy in neutropenic patients, who are most vulnerable to CR-Kp infection.

Innate Host Defense against <b><i>Klebsiella pneumoniae</i></b> and the Outlook for Development of Immunotherapies

“…Kobayashi et al [95] found that there is limited neutrophil phagocytosis of ST258 clinical isolates, a phenotype conferred largely by the CPS. Subsequent work by Diago-Navarro et al [96], Kobayashi et al [97], and Motley et al [98] demonstrated that phagocytosis and killing of ST258 are enhanced significantly by CPS-specific antibodies [96][97][98]. These results provide strong support to the idea that the CPS of ST258 can be used as vaccine antigen and/or target.…”

“…For example, designing antibodies that target only K. pneumoniae epitopes could potentially eliminate the pathogen and spare beneficial bacteria that constitute a healthy microbiome [154]. In vitro and in vivo studies indicate the use of specific antibodies dramatically increases uptake and killing of K. pneumoniae by immune cells, which consequently leads to improved recovery and increased survival in animal infection models [96][97][98]151]. Diago-Navarro et al [155] generated mAbs specific for the CPS of an ST23 hvKp strain (K1 capsule serotype) and then demonstrated the ability of these mAbs to protect against bacterial dissemination in a mouse infection model.…”