Evaluation of Burkholderia mallei Δ tonB Δ hcp1 (CLH001) as a live attenuated vaccine in murine models of glanders and melioidosis

Abstract

Background

Glanders caused by Burkholderia mallei is a re-emerging zoonotic disease affecting solipeds and humans. Furthermore, B. mallei is genetically related to B. pseudomallei, which is the causative agent of melioidosis. Both facultative intracellular bacteria are classified as tier 1 select biothreat agents. Our previous study with a B. mallei Δ tonB Δ hcp1 (CLH001) live-attenuated vaccine demonstrated that it is attenuated, safe and protective against B. mallei wild-type strain s in the susceptible BALB/c mouse model.

Methodology/Principal finding

In our current work, we evaluated the protective efficacy of CLH001 against glanders and melioidosis in the more disease-resistant C57BL/6 mouse strain. The humoral as well as cellular immune responses were also examined. We found that CLH001-immunized mice showed 100% survival against intranasal and aerosol challenge with B. mallei ATCC 23344. Moreover, this vaccine also afforded significant cross-protection against B. pseudomallei K96243, with low level bacterial burden detected in organs. Immunization with a prime and boost regimen of CLH001 induced significantly greater levels of total and subclasses of IgG, and generated antigen-specific splenocyte production of IFN-γ and IL-17A. Interestingly, protection induced by CLH001 is primarily dependent on humoral immunity, while CD4 ⁺ and CD8 ⁺ T cells played a less critical protective role.

Conclusions/Significance

Our data indicate that CLH001 serves as an effective live attenuated vaccine to prevent glanders and melioidosis. The quantity and quality of antibody responses as well as improving cell-mediated immune responses following vaccination need to be further investigated prior to advancement to preclinical studies.

Author summary

Glanders (caused by Burkholderia. mallei) and melioidosis (caused by B. pseudomallei) are severe infectious diseases of concern worldwide because of the rising number of cases and mortality rate. The low infectious doses of these two pathogens along with their amenability for aerosolization are factors that could be exploited as potential biothreat agents. Once the diseases have developed in humans and animals, intrinsic resistance to broad classes of antibiotics becomes a challenge for treatment and increases the risk for relapse. The progress in vaccine development demonstrates that live attenuated vaccine strains are the most effective in protection and providing long-lasting immune responses against both diseases. Our data indicate that the B. mallei double mutant (Δ tonB Δ hcp1) strain CLH001, is a feasible vaccine candidate to prevent glanders and melioidosis, especially for biodefense and public health purposes.