Masafumi Yamamoto

A Nontoxic Chimeric Enterotoxin Adjuvant Induces Protective Immunity in Both Mucosal and Systemic Compartments with Reduced IgE Antibodies

A novel nontoxic form of chimeric mucosal adjuvant that combines the A subunit of mutant cholera toxin E112K with the pentameric B subunit of heat-labile enterotoxin from enterotoxigenic Escherichia coli was constructed by use of the Brevibacillus choshinensis expression system (mCTA/LTB). Nasal immunization of mice with tetanus toxoid (TT) plus mCTA/LTB elicited significant TT-specific immunoglobulin A responses in mucosal compartments and induced high serum immunoglobulin G and immunoglobulin A anti-TT antibody responses. Although TT plus native CT induced high total and TT-specific immunoglobulin E responses, use of the chimera molecule as mucosal adjuvant did not. Furthermore, all mice immunized with TT plus mCTA/LTB were protected from lethal systemic challenge with tetanus toxin. Importantly, the mice were completely protected from influenza virus infection after nasal immunization with inactivated influenza vaccine together with mCTA/LTB. These results show that B. choshinensis-derived mCTA/LTB is an effective and safe mucosal adjuvant for the induction of protective immunity against potent bacterial exotoxin and influenza virus infection.

Role of Gut-Associated Lymphoreticular Tissues in Antigen-Specific Intestinal IgA Immunity

This study assessed the roles of the postnatal lymphotoxin-β receptor (LTβR)-mediated signals in the gut-associated lymphoreticular tissues of mice for subsequent regulation of Ag-specific intestinal IgA responses. Blockade of LTβR-dependent events by postnatal administration of the fusion protein of LTβR and IgG Fc (LTβR-Ig) reduced both the size and numbers of Peyer’s patches (PP) without influencing the PP microarchitecture. Interestingly, inhibition of LTβR-dependent signaling revealed significant reductions in the formation of follicular dendritic cell clusters in mesenteric lymph nodes (MLN). Furthermore, these postnatal signaling events controlled the development of isolated lymphoid follicles (ILF) because treatment with LTβR-Ig eliminated the formation of ILF. LTβR-Ig-treated mice with altered microarchitecture of MLN and lacking ILF were still able to produce significant Ag-specific mucosal IgA responses after oral immunization; however, the levels were significantly lower than those seen in control mice. These results imply the importance of ILF for Ag-specific intestinal immunity. However, mice treated with both TNFR55-Ig and LTβR-Ig in utero, which lack PP and MLN, but retain intact ILF, failed to induce Ag-specific IgA responses after oral immunization. These findings demonstrate that ILF are not essential for induction of intestinal IgA Ab responses to orally administered Ag. Furthermore, the induction of intestinal IgA Ab responses requires the proper maintenance of the MLN microarchitecture, including a follicular dendritic cell network.

Chimeras of labile toxin one and cholera toxin retain mucosal adjuvanticity and direct Th cell subsets via their B subunit.

Native cholera toxin (nCT) and the heat-labile toxin 1 (nLT) of enterotoxigenic Escherichia coli are AB5-type enterotoxins. Both nCT and nLT are effective adjuvants that promote mucosal and systemic immunity to protein Ags given by either oral or nasal routes. Previous studies have shown that nCT as mucosal adjuvant requires IL-4 and induces CD4-positive (CD4+) Th2-type responses, while nLT up-regulates Th1 cell production of IFN-γ and IL-4-independent Th2-type responses. To address the relative importance of the A or B subunits in CD4+ Th cell subset responses, chimeras of CT-A/LT-B and LT-A/CT-B were constructed. Mice nasally immunized with CT-A/LT-B or LT-A/CT-B and the weak immunogen OVA developed OVA-specific, plasma IgG Abs titers similar to those induced by either nCT or nLT. Both CT-A/LT-B and LT-A/CT-B promoted secretory IgA anti-OVA Ab, which established their retention of mucosal adjuvant activity. The CT-A/LT-B chimera, like nLT, induced OVA-specific mucosal and peripheral CD4+ T cells secreting IFN-γ and IL-4-independent Th2-type responses, with plasma IgG2a anti-OVA Abs. Further, LT-A/CT-B, like nCT, promoted plasma IgG1 more than IgG2a and IgE Abs with OVA-specific CD4+ Th2 cells secreting high levels of IL-4, but not IFN-γ. The LT-A/CT-B chimera and nCT, but not the CT-A/LT-B chimera or nLT, suppressed IL-12R expression and IFN-γ production by activated T cells. Our results show that the B subunits of enterotoxin adjuvants regulate IL-12R expression and subsequent Th cell subset responses.

Specific antibodies induced by nasally administered 40-kDa outer membrane protein of Porphyromonas gingivalis inhibits coaggregation activity of P. gingivalis

In this study, we have assessed the efficacy of the 40-kDa outer membrane protein (40k-OMP) of Porphyromonas gingivalis as a nasal vaccine for the prevention of adult periodontitis. Mice nasally immunized with 40k-OMP and cholera toxin as mucosal adjuvant displayed significant levels of 40k-OMP-specific serum IgG1, IgG2b and IgA as well as mucosal IgA antibodies (Abs) in saliva and nasal secretions. Ab-forming cell (AFC) analysis confirmed the antibody titers by detecting high numbers of 40k-OMP-specific AFCs in spleen, salivary glands and nasal passages. Because 40k-OMP-specific IgG inhibited coaggregation of P. gingivalis vesicles and S. gordonii, it may be an important tool for the prevention of adult periodontitis.

Prenatal Blockage of Lymphotoxin β Receptor and TNF Receptor p55 Signaling Cascade Resulted in the Acceleration of Tissue Genesis for Isolated Lymphoid Follicles in the Large Intestine

Signaling by lymphotoxin (LT) and TNF is essential for the organogenesis of secondary lymphoid tissues in systemic and mucosal compartments. In this study, we demonstrated that the progeny of mice treated with fusion protein of LTβR and IgGFc (LTβR-Ig) or LTβR-Ig plus TNFR55-Ig (double Ig) showed significantly increased numbers of isolated lymphoid follicles (ILF) in the large intestine. Interestingly, double Ig treatment accelerated the maturation of large intestinal ILF. Three-week-old progeny of double Ig-treated mice showed increased numbers of ILF in the large intestine, but not in the small intestine. Furthermore, alteration of intestinal microflora by feeding of antibiotic water did not affect the increased numbers of ILF in the large intestine of double Ig-treated mice. Most interestingly, mice that developed numerous ILF also had increased levels of activation-induced cytidine deaminase expression and numbers of IgA-expressing cells in the lamina propria of the large intestine. Taken together, these results suggest that ILF formation in the large intestine is accelerated by blockage of LTβR and TNFR55 signals in utero, and ILF, like colonic patches, might play a role in the induction of IgA response in the large intestine.

Induction of cytotoxic T lymphocyte responses by cholera toxin-treated bone marrow-derived dendritic cells

Cholera toxin (CT), a powerful mucosal adjuvant, is a potent inducer of Th2-type responses via activation of co-stimulatory molecules for the induction of IgA antibody responses. Less appreciated is the ability of CT to induce and regulate cytotoxic T lymphocyte (CTL) responses. In order to help for clarifying mechanisms underlying the CTL-inducing ability of CT, we have examined the effects of CT on dendritic cells (DCs) that could lead to the induction of cytotoxic CD8(+) T cells. When bone marrow-derived DCs (BM-DCs) were cultured with CT in vitro, B7-1 but not B7-2 molecules were significantly enhanced and allogenic CTL responses were induced. Also, increased numbers of IFN-gamma-secreting CD8(+) T cells were elicited when CT-treated BM-DCs were co-cultured with allogenic CD8(+) CTLs. Antibody blockade of B7-1 on CT-treated BM-DCs suppressed allogenic CTL responses, further indicating the importance of CT-induced B7-1 molecules on DCs for the acquisition of cytolytic function by CTL precursors. CD40 signaling was proven not necessary for the CT-induced CTL response since CT-treated CD40(-/-) BM-DCs developed CTL responses equivalent to those detected in CT-treated BM-DCs derived from normal mice. Our results suggest that CT-treated DCs are effective inducers of CD8(+) CTL, and this induction is mediated through CTs ability to enhance B7-1 expression on DCs.

Service Annotation and Profiling by Review Analysis

With the increase in the number of user reviews on user review sites, useful tools for extracting good and bad points of services so that users can easily and intuitively understand the quality of the services are required. If the annotations are selected from the pre-defined list, there can always be missing keywords. Supervised annotation approaches would suffer from the same problem. In this paper, we present an unsupervised method for extracting unique aspects of services and user opinions on these aspects from plain user reviews and apply it to service annotation using Yelps Academic Dataset. Our method is simple and easy to extend to other languages and domains. By using only the term frequency (TF), general aspects such as whether the food or service is good can be extracted. Further, what is praised particularly to the specific service can be extracted by using the term frequency and inverse document frequency (TF-IDF).

A hyperbolic geometric flow for evolving films and foams

Simulating the behavior of soap films and foams is a challenging task. A direct numerical simulation of films and foams via the Navier-Stokes equations is still computationally too expensive. We propose an alternative formulation inspired by geometric flow. Our model exploits the fact, according to Plateaus laws, that the steady state of a film is a union of constant mean curvature surfaces and minimal surfaces. Such surfaces are also well known as the steady state solutions of certain curvature flows. We show a link between the Navier-Stokes equations and a recent variant of mean curvature flow, called hyperbolic mean curvature flow, under the assumption of constant air pressure per enclosed region. Instead of using hyperbolic mean curvature flow as is, we propose to replace curvature by the gradient of the surface area functional. This formulation enables us to robustly handle non-manifold configurations; such junctions connecting multiple films are intractable with the traditional formulation using curvature. We also add explicit volume preservation to hyperbolic mean curvature flow, which in fact corresponds to the pressure term of the Navier-Stokes equations. Our method is simple, fast, robust, and consistent with Plateaus laws, which are all due to our reformulation of film dynamics as a geometric flow.

Reproducing Spectral Reflectances From Tristimulus Colours: Reproducing Spectral Reflectances from Tristimulus Colours

Physically based rendering systems often support spectral rendering to simulate light transport in the real world. Material representations in such simulations need to be defined as spectral distributions. Since commonly available material data are in tristimulus colours, we ideally would like to obtain spectral distributions from tristimulus colours as an input to spectral rendering systems. Reproduction of spectral distributions given tristimulus colours, however, has been considered an ill‐posed problem since single tristimulus colour corresponds to a set of different spectra due to metamerism. We show how to resolve this problem using a data‐driven approach based on measured spectra and propose a practical algorithm that can faithfully reproduce a corresponding spectrum only from the given tristimulus colour. The key observation in colour science is that a natural measured spectrum is usually well approximated by a weighted sum of a few basis functions. We show how to reformulate conversion of tristimulus colours to spectra via principal component analysis. To improve accuracy of conversion, we propose a greedy clustering algorithm which minimizes reconstruction error. Using pre‐computation, the runtime computation is just a single matrix multiplication with an input tristimulus colour. Numerical experiments show that our method well reproduces the reference measured spectra using only the tristimulus colours as input.

Role of Gut-Associated Lymphoreticular Tissues in Intestinal IgA Immunity

Abstract Despite the fact that intestinal IgA responses are known to be regulated by gut-associated lymphoreticular tissues (GALT), the exact sites where this regulatory network is formed are only partially understood. Our study shows that antigen-specific intestinal IgA antibodies are induced after oral immunization in mice made deficient in Peyers patches (PP) by in utero treatment with lymphotoxin-β receptor and Ig (LTβR-Ig). These results suggest that the PP-independent pathways exist for antigen-specific intestinal IgA responses. On the other hand, postnatal LTβR-Ig-treated mice lacking isolated lymphoid follicles (ILF) were still able to produce significant mucosal IgA responses. Furthermore, mice treated with both TNF receptor p55 and Ig (TNFR55-Ig) and LTβR-Ig in utero , which lack PP and mesenteric lymph nodes (MLN) but retain intact ILF, failed to induce antigen-specific IgA responses after oral immunization. These findings demonstrated that ILF were not essential for induction of intestinal IgA responses. Interestingly, however, the IgA responses induced in the PP- or ILF- null mice were significantly lower than those seen in the control mice. Taken together, these findings suggest that aggregated lymphoid follicles residing in the intestinal lumen, e.g. PP and ILF, and draining MLN, may comprise an integrated regulatory network for the induction of maximum IgA antibody responses to orally administered antigens.