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. 2024 Jan 29;36(1):33-43.
doi: 10.1093/intimm/dxad045.

Alcaligenes lipid A functions as a superior mucosal adjuvant to monophosphoryl lipid A via the recruitment and activation of CD11b+ dendritic cells in nasal tissue

Xiao Sun  1   2 , Koji Hosomi  1 , Atsushi Shimoyama  3   4 , Ken Yoshii  1   5 , Azusa Saika  1 , Haruki Yamaura  3 , Takahiro Nagatake  1   6 , Hiroshi Kiyono  7   8   9   10 , Koichi Fukase  3   4 , Jun Kunisawa  1   2   3   4   5   11   12   13   14
Affiliations

Alcaligenes lipid A functions as a superior mucosal adjuvant to monophosphoryl lipid A via the recruitment and activation of CD11b+ dendritic cells in nasal tissue

Xiao Sun et al. Int Immunol. .

Abstract

We previously demonstrated that Alcaligenes-derived lipid A (ALA), which is produced from an intestinal lymphoid tissue-resident commensal bacterium, is an effective adjuvant for inducing antigen-specific immune responses. To understand the immunologic characteristics of ALA as a vaccine adjuvant, we here compared the adjuvant activity of ALA with that of a licensed adjuvant (monophosphoryl lipid A, MPLA) in mice. Although the adjuvant activity of ALA was only slightly greater than that of MPLA for subcutaneous immunization, ALA induced significantly greater IgA antibody production than did MPLA during nasal immunization. Regarding the underlying mechanism, ALA increased and activated CD11b+ CD103- CD11c+ dendritic cells in the nasal tissue by stimulating chemokine responses. These findings revealed the superiority of ALA as a mucosal adjuvant due to the unique immunologic functions of ALA in nasal tissue.

Keywords: Alcaligenes; MPLA; chemokines; lipid A; nasal vaccine; stromal cells.

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Figures

Graphical Abstract
Graphical Abstract
Figure 1.
Figure 1.
For both intranasal and subcutaneous vaccination, Alcaligenes-derived lipid A (ALA) induced higher levels of antigen-specific antibodies than did monophosphoryl lipid A (MPLA). In particular, ALA was superior for intranasal vaccination. (A) OVA-specific IgG in the serum of mice vaccinated subcutaneously was evaluated by ELISA (n = 4). (B) OVA-specific IgA in the nasal wash of mice vaccinated intranasally was evaluated by ELISA (n = 4). (C) OVA-specific IgG in the serum of mice vaccinated intranasally was evaluated by ELISA (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by two-way analysis of variance (ANOVA; *P < .05, **P < .01, ***P < .001, ****P < .0001)..
Figure 2.
Figure 2.
Intranasal administration of ALA increased the generation of GCs and the numbers of GC B cells, IgA+ GL7hi B cells, and Tfh cells in NALT. (A) GCs (arrows) in NALT were analyzed through immunohistochemistry. PNA: GC marker; B220: B cell marker (n = 4) (scale bar = 200 μm). (B) The populations of GC GL7hi B cells (gated on: CD3ε− B220+ GL7hi) and IgA+ GL7hi B cells (gated on: CD3ε− B220+ GL7hi IgA+) in NALT were analyzed by flow cytometry (n = 4). (C) The population of Tfh cells (gated on: CD3ε+ CD8α− CD4+ PD-1hi) in NALT was analyzed by flow cytometry (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by one-way ANOVA (*P < .05, **P < .01, ****P < .0001).
Figure 3.
Figure 3.
Intranasal administration of ALA increased the generation of GCs and the numbers of GC B cells, IgA+ GL7hi B cells, and Tfh cells in CLNs. (A) GCs (arrows) in CLNs were analyzed by immunohistochemistry. PNA: GC marker; B220: B cell marker (n = 4) (scale bar = 50 μm). (B) The populations of GC GL7hi B cells (gated on: CD3ε− B220+ GL7hi) and IgA+ GL7hi B cells (gated on: CD3ε− B220+ GL7hi IgA+) in CLNs were analyzed by flow cytometry (n = 4). (C) The population of Tfh (gated on: CD3ε+ CD8α− CD4+ PD-1hi) in CLNs was analyzed by flow cytometry (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by one-way ANOVA (*P < .05, **P < .01, ***P < .001).
Figure 4.
Figure 4.
Intranasal administration of ALA induced greater infiltration of type 2 conventional DCs (cDC2s) in nasal tissue than did MPLA. (A) The numbers of CD103+ CD11b− cDC1s and CD11b+ CD103− cDC2s due to intranasal adjuvant administration were evaluated by flow cytometry (n = 3). (B) The density of CD103+ CD11b− cDC1s and CD11b+ CD103− cDC2s due to subcutaneous adjuvant administration were evaluated by flow cytometry (n = 3). (C) The populations of CD11b+ CD103− CLEC10A− cDC2As (*) and CD11b+ CD103− CLEC10A+ cDC2Bs (#) in nasal tissue were analyzed by flow cytometry (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by one-way ANOVA (*,#P < .05, **P < .01; ns, not significant).
Figure 5.
Figure 5.
Intranasally administered ALA induced greater expression of CD40 and CD80 than did MPLA. The expression of (A) CD40 and (B) CD80 on cDC2s after intranasal adjuvant administration was evaluated by flow cytometry (n = 3). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by one-way ANOVA (**P < .01, ***P < .001, ****P < .0001).
Figure 6.
Figure 6.
The gene expression of Csf2, Ccl2, Ccl3, Ccl4, and Tlr4 in sorted cells from nasal tissue. (A) The sorting strategy for CD45+ immune cells, EpCAM+ CD45− epithelial cells, and double-negative stromal cells (n = 4). (B) The gene expression of Tlr4 on sorted cells was evaluated by quantitative PCR (qPCR) analysis (n = 4). (C) The gene expression of Csf2, Ccl2, Ccl3, and Ccl4 from CD45+ cells due to stimulation by ALA or MPLA was evaluated by qPCR (n = 4). (D) The gene expression of Csf2, Ccl2, Ccl3, and Ccl4 from double-negative cells due to stimulation by ALA or MPLA was evaluated by qPCR (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by one-way ANOVA (**P < .01, ***P < .001, ****P < .0001).
Figure 7.
Figure 7.
Intranasal administration of ALA with anti-CCL2 or anti-CCL3 antibodies inhibited the infiltration of cDC2s in nasal tissue. The numbers of CD11b+ CD103− cDC2s in nasal tissue were analyzed by flow cytometry (n = 4). The data are shown as the mean ± 1 SD from two independent experiments; differences were analyzed by two-way analysis of variance (ANOVA; *P < .05, **P < .01; ns, not significant).
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References

    1. Ols S, Yang L, Thompson EA, et al. . Route of vaccine administration alters antigen trafficking but not innate or adaptive immunity. Cell Rep 2020;30:3964–3971.e7. 10.1016/j.celrep.2020.02.111 - DOI - PMC - PubMed
    1. Tang DCC, Nguyen HH.. The Yin-Yang arms of vaccines: disease-fighting power versus tissue-destructive inflammation. Expert Rev Vaccines 2014;13:417–27. 10.1586/14760584.2014.882775 - DOI - PubMed
    1. Zhang L, Wang W, Wang S.. Effect of vaccine administration modality on immunogenicity and efficacy. Expert Rev Vaccines 2015;14:1509–23. 10.1586/14760584.2015.1081067 - DOI - PMC - PubMed
    1. Calzas C, Chevalier C.. Innovative mucosal vaccine formulations against influenza a virus infections. Front Immunol 2019;10:1–21. - PMC - PubMed
    1. Afkhami S, D’Agostino MR, Zhang A, et al. . Respiratory mucosal delivery of next-generation COVID-19 vaccine provides robust protection against both ancestral and variant strains of SARS-CoV-2. Cell 2022;185:896–915.e19. 10.1016/j.cell.2022.02.005 - DOI - PMC - PubMed