Masayuki Kinbara

Lipopolysaccharide promotes and augments metal allergies in mice, dependent on innate immunity and histidine decarboxylase.

Background Few adequate murine models exist for metal allergies, it being especially difficult to induce Ni allergy in mice.

Nickel allergy-promoting effects of microbial or inflammatory substances at the sensitization step in mice

Microbial components stimulate innate immunity via Toll-like receptors (TLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), and/or IL-1. We recently reported that in mice, Escherichia coli lipopolysaccharide (LPS, TLR4-ligand) promotes allergic responses to nickel (Ni) at both the sensitization and elicitation steps. Here, we examined in mice the effects of administering other microbial or inflammatory materials at the Ni-sensitization step. A mixture of 1mM NiCl(2) and a test solution was injected into BALB/c mice intraperitoneally (0.1 ml/10 g body weight), and 10 days later 5mM NiCl(2) was challenged intradermally into the ear pinnas of the mice (20 μl/ear). The following preparations or substances exhibited adjuvant activities: Prevotella intermedia LPS, Saccharomyces cerevisiae mannan, a synthetic muramyl dipeptide (NOD2-stimulating cell-wall component of bacteria), Pam(3)Cys-SKKKK (TLR2-stimulating synthetic peptide), poly I:C (TLR3-stimulating double-stranded RNA), concanavalin A (a typical T-cell mitogen and T-cell-mediated hepatitis-inducer), heat-killed Propionibacterium acnes (Gram-positive bacterium that causes pimples and induces macrophage-mediated experimental hepatitis), and nitrogen-containing bisphosphonates (chemicals stimulating IL-1 production). Unexpectedly, P. intermedia LPS, which displayed the most potent adjuvant activity among the tested preparations, was effective in TLR4-dysfunctional mutant mice, but not in TLR2-deficient mice, whereas the reverse was true for S. cerevisiae mannan. These results suggest that (i) for the establishment of Ni-allergy in mice, stimulation of innate immunity (including TLRs, NLRs, IL-1 production, and/or other factors) may be important at the sensitization step, and (ii) P. intermedia may produce a substance(s) that potently promotes Ni-allergy via stimulation of TLR2.

Allergy‐inducing nickel concentration is lowered by lipopolysaccharide at both the sensitization and elicitation steps in a murine model

Background  Nickel (Ni) is the major cause of contact allergy. We previously found that lipopolysaccharide (LPS, a cell‐surface component of Gram‐negative bacteria) markedly promotes Ni allergy in a murine model. Establishing the minimum concentration or amount of Ni needed to induce allergic responses may help us to prevent or reduce such responses.

Biotin Status Affects Nickel Allergy via Regulation of Interleukin-1β Production in Mice

Biotin, a water-soluble B complex vitamin, is possibly involved in chronic inflammatory diseases, although the detailed mechanisms are unclear. In this study, we investigated the effects of biotin status on nickel (Ni) allergy in mice. Mice were fed a basal or biotin-deficient (BD) diet for 8 wk and sensitized with an intraperitoneal injection of NiCl(2) and lipopolysaccharide. Ten days after sensitization, NiCl(2) was intradermally injected into pinnas and ear swelling was measured. For in vitro analysis, we cultured a murine macrophage cell line, J774.1, under a biotin-sufficient (C, meaning control) or BD condition for 4 wk and analyzed interleukin (IL)-1 production. Significantly higher ear swelling was induced in BD mice than C mice. Adaptive transfer of splenocytes from both C and BD mice induced Ni allergy in unsensitized mice. Regardless of donor mice, ear swelling was significantly higher in BD recipient mice than C recipient mice. Ni allergy was not induced in either C or BD IL-1(-/-) mice. Splenocytes from BD mice produced a significantly higher amount of IL-1beta than those from C mice. Production and mRNA expression of IL-1beta were significantly higher in BD J774.1 cells than in C cells. Biotin supplementation inhibited the augmentation of IL-1beta production in vitro. In vivo supplementation of biotin in drinking water dose-dependently decreased ear swelling in C and BD mice. These results indicate that biotin status affects Ni allergy in the elicitation phase via the upregulation of IL-1beta production in mice, suggesting that biotin supplementation may have therapeutic effects on human metal allergy.

The elicitation step of nickel allergy is promoted in mice by microbe-related substances, including some from oral bacteria.

Microbial components activate the hosts innate immunity via interactions with molecules including TLRs and NODs. We previously reported that in mice (i) Escherichia coli lipopolysaccharide (LPS; TLR4 agonist) promotes Ni-allergy even in T-cell-deficient mice, (ii) E. coli LPS reduces the minimum allergy-inducing concentrations of Ni at both the sensitization and elicitation steps, and (iii) various microbe-related substances promote sensitization to Ni. Here, we examined the effects of microbe-related substances at the elicitation step. Mice (except for TLR4-mutated C3H/HeJ mice) were sensitized to Ni by intraperitoneal injection of NiCl(2) + E. coli LPS. Ten days later their ear-pinnas were challenged with 1 μM NiCl(2) with or without a test substance. Although NiCl(2) alone at this concentration does not induce Ni-allergy, its combination with the following substances induced Ni-allergy in BALB/c mice: LPS preparations from oral gram-negative bacteria (Prevotella intermedia and Porphyromonas gingivalis), a mannan preparation from a fungus (Saccharomyces cerevisiae), and synthetic NOD2 and TLR2 agonists. The effect of the mannan preparation was small in C3H/HeJ mice (sensitized with NiCl(2) + the P. intermedia preparation). The P. intermedia preparation promoted Ni-allergy in C3H/HeJ and nude mice, but not in mice deficient in either TLR2 or histidine decarboxylase. Intragingival injection of the P. intermedia preparation and later challenge with NiCl(2) alone to ear-pinnas also promoted Ni-allergy. These results indicate that (i) in Ni-allergy, a microbial milieu or innate immunity is important at the elicitation step, too, and (ii) some oral bacteria may promote Ni-allergy via TLR2-stimulant(s) production.

Resin Monomers Act as Adjuvants in Ni-induced Allergic Dermatitis in vivo

Resin monomers (RMs) are inflammatory agents and are thought to cause allergic contact dermatitis (ACD). However, mouse models are lacking, possibly because of the weak antigenicities of RMs. We previously reported that inflammatory substances can promote the allergic dermatitis (AD) induced by intradermally injected nickel (Ni-AD) in mice. Here, we examined the effects of RMs on Ni-AD. To sensitize mice to Ni, a mixture containing non-toxic concentrations of NiCl2 and an RM [either methyl methacrylate (MMA) or 2-hydroxyethyl methacrylate (HEMA)] was injected intraperitoneally or into ear-pinnae intradermally. Ten days later, a mixture containing various concentrations of NiCl2 and/or an RM was intradermally injected into ear-pinnae, and ear-swelling was measured. In adoptive transfer experiments, spleen cells from sensitized mice were transferred intravenously into non-sensitized recipients, and 24 h later NiCl2 was challenged to ear-pinnae. Whether injected intraperitoneally or intradermally, RM plus NiCl2 mixtures were effective in sensitizing mice to Ni. AD-inducing Ni concentrations were greatly reduced in the presence of MMA or HEMA (at the sensitization step from 10 mM to 5 or 50 µM, respectively, and at the elicitation step from 10 µM to 10 or 100 nM, respectively). These effects of RMs were weaker in IL-1-knockout mice and in macrophage-depleted mice. Cell-transfer experiments in IL-1-knockout mice indicated that both the sensitization and elicitation steps depended on IL-1. Challenge with an RM alone did not induce allergic ear-swelling in mice given the same RM + NiCl2 10 days before the challenge. These results suggest that RMs act as adjuvants, not as antigens, to promote Ni-AD by reducing the AD-inducing concentration of Ni, and that IL-1 and macrophages are critically important for the adjuvant effects. We speculate that what were previously thought of as “RM-ACD” might include ACD caused by antigens other than RMs that have undergone promotion by the adjuvant effects of RMs.

Promotion of arthritis and allergy in mice by aminoglycoglycerophospholipid, a membrane antigen specific to Mycoplasma fermentans

Mycoplasmas, which lack a cell wall and are the smallest self-replicating bacteria, have been linked to some chronic diseases, such as AIDS, rheumatoid arthritis (RA), and oncogenic transformation of cells. Their membrane components (lipoproteins and glycolipids) have been identified as possible causative factors in such diseases. Glycoglycerophospholipid (GGPL)-III, a unique phosphocholine-containing aminoglycoglycerophospholipid, is a major specific antigen of Mycoplasma fermentans, and has been detected in 38% of RA patients. Unlike those of lipoproteins, which induce inflammation via Toll-like receptor 2 (TLR2), the pathologic effects of GGPL-III are poorly understood. RA and metal allergies are chronic inflammatory diseases in which autoantigens have been implicated. Here, we examined the effects of chemically synthesized GGPL-III in murine arthritis and allergy models. GGPL-III alone exhibited little inflammatory effect, but promoted both collagen-induced arthritis and nickel (Ni) allergy, although less powerfully than Escherichia coli lipopolysaccharide. The augmenting effect of GGPL-III on Ni allergy was present in mice deficient in either T cells or active TLR4, but it was markedly weaker in mice deficient in macrophages, interleukin-1, or the histamine-forming enzyme histidine decarboxylase than in their control strains. These results suggest that GGPL-III may play roles in some types of chronic diseases via the innate immune system.

Cross-reactivity among some metals in a murine metal allergy model.

Background  Information concerning cross‐reactivity among metal allergens is scarce. We previously devised a murine metal allergy model using lipopolysaccharide (LPS) as an adjuvant. LPS reduces the minimum allergy‐inducing concentration (MAIC) of metals at both the sensitization and the elicitation steps.

CXCL4 is a novel nickel-binding protein and augments nickel allergy

Nickel (Ni) is the most frequent metal allergen and induces a TH1‐dependent type‐IV allergy. Although Ni2+ is considered to bind to endogenous proteins, it currently remains unclear whether these Ni‐binding proteins are involved in Ni allergy in vivo. We previously reported the adjuvant effects of lipopolysaccharide (LPS) in a Ni allergy mouse model. As LPS induces a number of inflammatory mediators, we hypothesized that Ni‐binding protein(s) are also induced by LPS.

Mast cell histamine-mediated transient inflammation following exposure to nickel promotes nickel allergy in mice.

We previously reported that allergic responses to nickel (Ni) were minimal in mice deficient in the histamine‐forming enzyme histidine decarboxylase (HDC‐KO), suggesting an involvement of histamine in allergic responses to Ni. However, it remains unclear how histamine is involved in the process of Ni allergy. Here, we examined the role of histamine in Ni allergy using a murine model previously established by us. Mice were sensitized to Ni by intraperitoneal injection of a NiCl2‐lipopolysaccharide (LPS) mixture. Ten days later, allergic inflammation was elicited by challenging ear‐pinnas intradermally with NiCl2. Then, ear‐swelling was measured. Pyrilamine (histamine H1‐receptor antagonist) or cromoglicate (mast cell stabilizer) was intravenously injected 1 h before the sensitization or the challenge. In cell‐transfer experiments, spleen cells from Ni‐sensitized donor mice were intravenously transferred into non‐sensitized recipient mice. In both sensitized and non‐sensitized mice, 1 mm or more NiCl2 (injected into ear‐pinnas) induced transient non‐allergic inflammation (Ni‐TI) with accompanying mast cell degranulation. LPS did not affect the magnitude of this Ni‐TI. Pyrilamine and cromoglicate reduced either the Ni‐TI or the ensuing allergic inflammation when administered before Ni‐TI (at either the sensitization or elicitation step), but not if administered when the Ni‐TI had subsided. Experiments on HDC‐KO and H1‐receptor‐KO mice, and also cell‐transfer experiments using these mice, demonstrated histamines involvement in both the sensitization and elicitation steps. These results suggest that mast cell histamine‐mediated Ni‐TI promotes subsequent allergic inflammatory responses to Ni, raising the possibility that control of Ni‐TI by drugs may be effective at preventing or reducing Ni allergy.