Nod2 is required for antigen-specific humoral responses against antigens orally delivered using a recombinant Lactobacillus vaccine platform.

Sara A Bumgardner,Chad B Frank,Lin Zhang,Gregg A Dean,Alora S Lavoy,Akinobu Kajikawa,Barbara Andre,Todd R Klaenhammer,Eliane N Miyaji

doi:10.1371/journal.pone.0196950

Abstract

Safe and efficacious orally-delivered mucosal vaccine platforms are desperately needed to combat the plethora of mucosally transmitted pathogens. Lactobacillus spp. have emerged as attractive candidates to meet this need and are known to activate the host innate immune response in a species- and strain-specific manner. For selected bacterial isolates and mutants, we investigated the role of key innate immune pathways required for induction of innate and subsequent adaptive immune responses. Co-culture of murine macrophages with L. gasseri (strain NCK1785), L. acidophilus (strain NCFM), or NCFM-derived mutants—NCK2025 and NCK2031—elicited an M2b-like phenotype associated with TH2 skewing and immune regulatory function. For NCFM, this M2b phenotype was dependent on expression of lipoteichoic acid and S layer proteins. Through the use of macrophage genetic knockouts, we identified Toll-like receptor 2 (TLR2), the cytosolic nucleotide-binding oligomerization domain containing 2 (NOD2) receptor, and the inflammasome-associated caspase-1 as contributors to macrophage activation, with NOD2 cooperating with caspase-1 to induce inflammasome derived interleukin (IL)-1β in a pyroptosis-independent fashion. Finally, utilizing an NCFM-based mucosal vaccine platform with surface expression of human immunodeficiency virus type 1 (HIV-1) Gag or membrane proximal external region (MPER), we demonstrated that NOD2 signaling is required for antigen-specific mucosal and systemic humoral responses. We show that lactobacilli differentially utilize innate immune pathways and highlight NOD2 as a key mediator of macrophage function and antigen-specific humoral responses to a Lactobacillus acidophilus mucosal vaccine platform.

Highlights

Induction of mucosal immunity by direct vaccination is an attractive strategy to combat pathogens that are transmitted at mucosal surfaces
nucleotide-binding oligomerization domain containing 2 (NOD2) and Toll-like receptor 2 (TLR2) have been implicated as key modulators of the innate immune response to lactobacilli, and TLR2 activation by L. gasseri and L. acidophilus has been demonstrated (Table 1) [15, 16, 35]
In this study we characterized the roles of TLR2, NOD2, and caspase-1 in macrophage phagocytosis, activation, and cytokine production in response to L. gasseri NCK1785, L. acidophilus NCFM, and the NCFM mutant strains NCK2025 and NCK2031

Summary

Introduction

Induction of mucosal immunity by direct vaccination is an attractive strategy to combat pathogens that are transmitted at mucosal surfaces. The majority of vaccines are parentally delivered due to the lack of robust mucosal vaccine platforms (reviewed in [1, 2]). Proof-of-principle studies have demonstrated that lactobacilli-based vaccine platforms are promising candidates for mucosal vaccination against a variety of pathogens including influenza, anthrax, severe acute respiratory syndrome (SARS), and human immunodeficiency virus (HIV) [3,4,5,6]. Lactobacilli are an attractive vaccine vehicle because they persist in the gastrointestinal tract, adhere to epithelial cells, and modulate the immune response through pattern recognition receptors (PRR) (reviewed in [7,8,9,10]). The challenge is to maintain the inherent safety of lactobacilli while maximizing immunogenicity against relevant pathogen epitopes

Methods

Results

Conclusion