Alcaligenes faecalis is the predominant Gramnegative bacterium inhabiting gut-associated lymphoid tissues, Peyersp atches.W ep reviously reported that an A. faecalis lipopolysaccharide (LPS) acted as aweak agonist for Toll-like receptor 4( TLR4)/myeloid differentiation factor-2 (MD-2) receptor as well as ap otent inducer of IgA without excessive inflammation, thus suggesting that A. faecalis LPS might be used as as afe adjuvant. In this study,w ec haracterized the structure of both the lipooligosaccharide (LOS) and LPS from A. faecalis.W esynthesized three lipid Amolecules with different degrees of acylation by an efficient route involving the simultaneous introduction of 1-and 4'-phosphates.H exaacylated A. faecalis lipid As howed moderate agonistic activity towards TLR4-mediated signaling and the ability to elicit adiscrete interleukin-6 release in human cell lines and mice.It was thus found to be the active principle of the LOS/LPS and apromising vaccine adjuvant candidate.

Alcaligenes spp. are identified as commensal bacteria and have been found to inhabit Peyer’s patches in the gut. We previously reported that Alcaligenes-derived lipopolysaccharides (LPS) exerted adjuvant activity in systemic vaccination, without excessive inflammation. Lipid A is one of the components responsible for the biological effect of LPS and has previously been applied as an adjuvant. Here, we examined the adjuvant activity and safety of chemically synthesized Alcaligenes lipid A. We found that levels of OVA-specific serum IgG antibodies increased in mice that were subcutaneously immunized with ovalbumin (OVA) plus Alcaligenes lipid A relative to those that were immunized with OVA alone. In addition, Alcaligenes lipid A promoted antigen-specific T helper 17 (Th17) responses in the spleen; upregulated the expression of MHC class II, CD40, CD80, and CD86 on bone marrow-derived dendritic cells (BMDCs); enhanced the production of Th17-inducing cytokines IL-6 and IL-23 from BMDCs. Stimulation with Alcaligenes lipid A also induced the production of IL-6 and IL-1β in human peripheral blood mononuclear cells. Moreover, Alcaligenes lipid A caused minor side effects, such as lymphopenia and thrombocytopenia. These findings suggest that Alcaligenes lipid A is a safe and effective Th17-type adjuvant by directly stimulating dendritic cells in systemic vaccination.

Effective and safe vaccine adjuvants are needed to appropriately augment mucosal vaccine effects. Our previous study demonstrated that lipopolysaccharide (LPS) from Peyer’s patch resident Alcaligenes stimulated dendritic cells to promote the production of mucosal immunity-enhancing cytokines (e.g., IL-6 and BAFF), thus enhancing antigen-specific immune responses (including IgA production and Th17 responses) without excessive inflammation. Here, we chemically synthesized Alcaligenes lipid A, the biologically active part of LPS, and examined its efficacy as a nasal vaccine adjuvant for the induction of protectively immunity against Streptococcus pneumoniae infection. Mice were nasally immunized with pneumococcal surface protein A (PspA) as a vaccine antigen for S. pneumoniae, together with Alcaligenes lipid A. Alcaligenes lipid A supported the generation of high levels of PspA-specific IgA and IgG responses through the augmentation of germinal center formation in the nasopharynx-associated lymphoid tissue and cervical lymph nodes (CLNs). Moreover, Alcaligenes lipid A promoted PspA-specific CD4+ Th17 responses in the CLNs and spleen. Furthermore, neutrophils were recruited to infection sites upon nasal infection and synchronized with the antigen-specific T and B cell responses, resulting in the protection against S. pneumoniae infection. Taken together, Alcaligenes lipid A could be applied to the prospective adjuvant to enhance nasal vaccine efficacy by means of augmenting both the innate and acquired arms of mucosal immunity against respiratory bacterial infection.

Alcaligenes faecalis is the predominant Gramnegative bacterium inhabiting gut-associated lymphoid tissues, Peyersp atches.W ep reviously reported that an A. faecalis lipopolysaccharide (LPS) acted as aweak agonist for Toll-like receptor 4( TLR4)/myeloid differentiation factor-2 (MD-2) receptor as well as ap otent inducer of IgA without excessive inflammation, thus suggesting that A. faecalis LPS might be used as as afe adjuvant. In this study,w ec haracterized the structure of both the lipooligosaccharide (LOS) and LPS from A. faecalis.W esynthesized three lipid Amolecules with different degrees of acylation by an efficient route involving the simultaneous introduction of 1-and 4'-phosphates.H exaacylated A. faecalis lipid As howed moderate agonistic activity towards TLR4-mediated signaling and the ability to elicit adiscrete interleukin-6 release in human cell lines and mice.It was thus found to be the active principle of the LOS/LPS and apromising vaccine adjuvant candidate.

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