ABSTRACT
BACKGROUND: Recent investigations have shown that proteins, including Bet v 1a, are nitrated by exposure to polluted urban air. We have investigated immunogenic and allergenic properties of in vitro nitrated allergens in in vivo models. METHODS: Untreated and nitrated samples of ovalbumin or Bet v 1a were compared for their ability to stimulate proliferation and cytokine secretion in splenocytes from DO11.10 or from sensitized BALB/c mice, and for their ability to induce specific immunoglobulin (Ig)G1, IgG2a and IgE in sensitized mice. Additionally, sera from birch pollen-allergic individuals were analysed for IgE and IgG specific for nitrated Bet v 1a. RESULTS: Upon splenocyte stimulation with nitrated as compared with unmodified allergens, proliferation as well as interleukin 5 and interferon-gamma production were enhanced. Sera of mice sensitized with nitrated allergens showed elevated levels of specific IgE, IgG1 and IgG2a, compared with sera from mice sensitized with unmodified allergens. Moreover, cross-reactivity of antibodies against unrelated, nitrated allergens was observed in mice. We also found higher amounts of functional, specific IgE against nitrated than against untreated Bet v 1a in sera from birch pollen-allergic patients. CONCLUSIONS: Our findings suggest that nitration enhances allergic responses, which may contribute to an increased prevalence of allergic diseases in polluted urban environments.
Subject(s)
Allergens/immunology , Ovalbumin/immunology , Plant Proteins/immunology , Protein Processing, Post-Translational/immunology , Spleen/immunology , Tetranitromethane/pharmacology , Allergens/chemistry , Allergens/drug effects , Animals , Antigens, Plant , Cell Proliferation , Female , Food Hypersensitivity , Mice , Mice, Inbred BALB C , Ovalbumin/chemistry , Ovalbumin/drug effects , Plant Proteins/chemistry , Plant Proteins/drug effects , Spleen/cytology , Tetranitromethane/chemistry , Tyrosine/analogs & derivatives , Tyrosine/chemistry , Tyrosine/drug effects , Tyrosine/immunologyABSTRACT
Interaction of large unilamellar phosphatidylcholine vesicles (LUV, 75nm) and plasma high density lipoproteins (HDL) resulted in a non-leaky vesiculation of LUV. This vesiculation was detected by a HPLC-system consisting of a combination of three TSK-gel columns (6000PW, 5000PW, 3000SW). With increasing incubation time liposomal [14C]PC, entrapped [3H]inulin, and apoprotein of HDL origin decreased. The decrease was accompanied by a formation of new particles, consisting of liposomal PC and apoprotein. These particles also enclosed [3H]inulin, reflecting a hydrophilic inner space. The formation of the particles reached a maximum after one day of incubation. Retention time was 21 minutes for LUV, 28 minutes for the new particles, and 36 minutes for HDL. In vesicles with membranes consisting of phosphatidylcholine and 30% cholesterol no interactions were observed.
Subject(s)
Lipoproteins, HDL/metabolism , Liposomes/metabolism , Carbon Radioisotopes , Cholesterol/pharmacology , Chromatography, High Pressure Liquid , Humans , Inulin/metabolism , Kinetics , Phosphatidylcholines/metabolism , TritiumABSTRACT
The formation of the exoenzymes, neutral and alkaline proteinase as well as alpha-amylase of Bacillus subtilis, is characterized by the same time course. The exoenzyme formation starts in the exponential phase of growth by an excess of C and N sources. We assume that carbon metabolism of pyruvate is responsible for the exoenzyme formation during this growth phase. The proteinase formation at the transient and/or stationary phase of growth is related to amino acid limitation.
