PS80 interferes with the antiallergic effect of Cry‐consensus peptide, a novel recombinant peptide for immunotherapy of Japanese cedar pollinosis, at very low concentration through modulation of Th1/Th2 balance

Abstract

SummaryPolysorbate 80 (PS80 or Tween-80) is often used as an additive to promote the rapid solubilization of pharmaceuticals in aqueous solutions. We investigated whether coinjection of a minimal amount of PS80 had a modulatory effect on the immunotherapeutic effects of Cry (Cryptomeria)-consensus peptide, a novel peptide developed for the therapeutic management of Japanese cedar pollinosis, using a Cry j 1-sensitized mouse model with experimental allergic rhinitis. Subcutaneous challenge with Cry-consensus peptide plus 50 lg/ml of PS80 did not affect the antigen-specific proliferation of splenocytes, but decreased the potency of Cry-consensus peptide to inhibit antigen-specific interleukin (IL)-5 production by the cells significantly in comparison with challenge with Cryconsensus peptide alone. However, there was no significant difference between the effect of Cry-consensus peptide administration on interferon (IFN)-c production in the presence and absence of PS80, indicating that PS80 interfered with the T helper 1 (Th1)-dominant T helper balance induced by Cry-consensus peptide challenge. Moreover, the increase in the level of antigen-specific immunoglobulin G2a (IgG2a) induced by Cry-consensus peptide challenge was inhibited slightly but unambiguously by PS80 coinjection. These in vitro experiments indicated that PS80 induces Th2-type differentiation of T helper cells through preferential inhibition of IFN-c expression relative to IL-5 expression in splenocytes in a concentration-dependent manner. In naïve mice, sensitization by Cry-consensus peptide with PS80 induced antigen-specific IL-5 production more potently than sensitization by Cry-consensus peptide alone, and when PS80 was added to bone marrow-derived dendritic cells, the endocytosis of fluorescence-labelled Cry-consensus peptide was dramatically inhibited in a concentration-dependent manner. Therefore, we conclude that PS80 has an immunomodulatory effect on the antigen-specific response resulting in a shift towards Th2 predominance with respect to the antigen recognition stage. Taken together, our findings suggest that PS80 might decrease the efficacy of Cry-consensus peptide through modulation of the efficiency of antigen endocytosis and/or of the direction of successive T helper cell differentiation.

Cry-Consensus Peptide, a Novel Peptide for Immunotherapy of Japanese Cedar Pollinosis, Induces Th1-Predominant Response in Cry j 1-Sensitized B10.S Mice

“…26) Statistical Evaluation For comparison between the two groups, the data were firstly analyzed for homogeneity of variance using the F test. If the variances were homogeneous, results were evaluated using Student's t test.…”

“…26) Cry j 1-specific serum IgG1 was also assayed according to the IgG2a measurement method using peroxidase-conjugated anti-mouse IgG1 monoclonal antibody (Roche, Indianapolis, IN, U.S.A.) instead of peroxidase-conjugated anti-mouse IgG2a monoclonal antibody.…”

“…21,22) Based on the above information, CCP has been newly designed as a linked peptide of T-cell epitopes. 26) CCP contains six T-cell epitopes chosen from Cry j 1 and Cry j 2 sequences based on the incidence of T cell response in patients suffering from JC pollinosis.…”

Allergen-Induced Basophil CD203c Expression as a Biomarker for Rush Immunotherapy in Patients with Japanese Cedar Pollinosis

“…Japanese cedar pollen (JCP) pollinosis represents a major clinical problem each year from late February to April in Japan, affecting as many as 10% of the Japanese population [3, 4]. Rush immunotherapy (RIT), in which a fast increment of allergen doses is allowed, has been reported to confer allergen-specific, rapid protection from the disease.…”

Anti-allergic potential of oligomannose-coated liposome-entrapped Cry j 1 as immunotherapy for Japanese cedar pollinosis in mice

“…Splenocytes from Cry j 1/OML-treated mice secreted much more IFN-γ than those from control mice in response to in vitro stimulation with the allergen, indicating that allergen-specific Th1 cells were induced in the treated mice. The IL4/IFN-γ and IL-5/IFN-γ ratios for cytokines secreted from spleen cells, which reflect the Th1/Th2 balance [20,21], were also lower in the Cry j 1/OML-treated mice than in PBS-and Cry j 1/alumtreated mice. Since the level of IgE is positively regulated by Th2 cytokines [19], repression of serum IgE elevation in Cry j 1/OMLtreated mice may be due to Th1-skewing of the immune response.…”