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Biomedical subjects

M Raulf-Heimsoth

Publications and source records attributed to M Raulf-Heimsoth.

65 records · Page 4Linked to original sources

Flow-cytometric analysis of T-cell receptor expression in peripheral blood lymphocytes.

BACKGROUND: Lymphocytes of subjects sensitized to the insect-derived allergen Chi t 1-9 are, in response to the allergen, characterized by an elevated proliferation and increased expression of activation markers such as HLA-DR and CD25 in vitro. A restriction for HLA-DR B1 in monosensitized patients was found. OBJECTIVE: The aim of this study was to investigate whether the response to the allergen Chi t 1-9 involves the preferential cell surface expression of a specific alpha/beta T-cell receptor type. METHODS: The T-cell receptor repertoire was measured with 7 monoclonal antibodies to epitopes on the variable region of the alpha- and beta-chain by flow cytometry. Cell lines of 9 patients were established with Chi t 1-9, 6 with tetanus toxoid and 6 with phytohaemagglutinin in the presence of interleukin 2. In addition, non-stimulated lymphocytes as well as lymphocytes of 5 non-sensitized controls exposed to Chi t 1-9 were examined. RESULTS: Each of the 9 sensitized and 5 control subjects studied showed an individual pattern of lymphocyte expression for each T-cell receptor specificity. However, after stimulation specific to Chi t 1-9 for 2 weeks, a significant increase in V beta 8-expressing cells was measurable only in patients sensitized to Chi t 1-9. The cells of the 5 control subjects showed no significant changes due to the allergen stimulation. CONCLUSION: These data suggest that the expression of certain T-cell receptor types plays an important role in the development of Chi t 1-9 allergy.

Allergens↗

Differentiation between cosensitization and cross-reactivity in wheat flour and grass pollen-sensitized subjects.

BACKGROUND: To diagnose baker's asthma, occupational sensitization to wheat flour should be distinguished clearly from influences of cosensitization such as it may exist in pollinosis patients. To define the route of sensitization, the cross-reactivity between wheat flour and grass pollen allergens was investigated. METHODS: Two groups of atopic individuals with or without professional contact to wheat flour were screened by skin prick test for their sensitization to wheat flour and grass pollen and, in the case of hints for cosensitization, by Enzyme Allergo Sorbent Test (EAST). Cross-reactivity between wheat flour and grass pollen allergens was investigated by IgE binding inhibition assay using sera of 20 cosensitized individuals and by immunoblots. RESULTS: The immunological cross-inhibition between wheat flour and grass pollen proves some proteins to share common allergenic determinants. Significant differences between bakers and individuals not occupationally exposed to flour could be seen in the inhibition of IgE binding to wheat flour. While the IgE binding to wheat flour allergens was only slightly inhibited by grass pollen proteins in baker's sera its inhibition was nearly complete by this extract in sera of nonexposed atopics. Immunoblots indicate that wheat proteins with a molecular weight of about 8-12 and 22 kD preferentially react with sensitized bakers' IgE and not with IgE of other individuals. IgE binding to grass pollen allergens in immunoblots and inhibition experiments showed no differences between bakers and other subjects. CONCLUSION: In the group of atopics without professional contact to flour, the positive test results from wheat flour obviously ensue from contact to cross-reacting grass pollen proteins whereas bakers are exposed and sensitized to allergens of both sources. Such cross-inhibition experiments help to identify the source of sensitization which may be difficult to obtain in case of cross-reacting allergens.

Asthma↗

[Latex allergens: review of current knowledge].

Both health care workers and spina bifida patients are at risk to develop type I latex allergy since allergenic proteins of natural rubber latex are present in a variety of latex products. Natural latex contains more than 250 polypeptides and approximately 60 of them show IgE-binding characteristics. A multitude of these latex allergens has been identified, and their sensitization potency for both risk groups has been determined. While hevein (Hev b 6.02) is the major allergen in latex-allergic health care workers, 80% of latex sensitized-spina bifida children have IgE antibodies to the "rubber elongation factor" (Hev b 1). Almost all relevant latex allergens have been identified in natural latex products: however, no data on the allergen profiles of individual latex products exist. The knowledge of the major latex allergens enables improved diagnosis and to monitor the success of prevention strategies for decreasing the high prevalence of latex allergies.

Allergens↗

Humoral and cellular immune responses in asthmatic isocyanate workers: report of two cases.

Two workers (23 and 28 years old) developed rhinitis and bronchial asthma after occupational contact with the isocyanate MDI. Positive skin prick test results for MDI-HSA and IgE antibodies to all isocyanate-HSA conjugates were obtained in both cases, and the inhalation challenge test with MDI produced immediate and late asthmatic reactions. In the patch test and the stimulation assay of peripheral mononuclear blood cells, a specific sensitization to the diamine MDA (in both cases) and to further amines (in one case), as well as to hydrolysates of the respective diisocyanates, was seen, which appears to be independent of the IgE response to isocyanate-HSA conjugates. Our results offer evidence of IgE-mediated, as well as lymphocyte, responses induced by exposures to isocyanate products over several months or years.

Adult↗

Lymphocyte proliferation response to extracts from different latex materials and to the purified latex allergen Hev b 1 (rubber elongation factor).

BACKGROUND: Type I allergy to latex is a growing problem, especially among health care workers. A detailed study of the peripheral blood cell responses to latex allergens has not been reported. METHODS: Peripheral blood mononuclear cells of patients and healthy subjects were isolated and stimulated with protein extracts from latex sap and latex gloves and the purified latex allergen Hev b 1 (rubber elongation factor) at different concentrations to determine the antigen-specific proliferation response. The examined patients were sensitized to latex by occupational exposure (n = 23) and had rhinitis, conjunctivitis, contact urticaria, and/or asthma. Two control groups of nonsensitized subjects were studied: one occupationally exposed to latex (n = 8), and the second, not exposed to latex (n = 8). RESULTS: In general, only latex-exposed subjects responded to the different latex antigen preparations. Lymphocyte proliferation responses to latex sap extract were found in 65% of latex-sensitized subjects and in 37.5% of the latex-exposed healthy subjects. Latex glove extract induced a significant proliferative responses in 47.8% of latex-sensitized patients and in 25% of the latex-exposed individuals. Hev b 1 induced lymphocyte proliferation responses in 52% of the latex-sensitized patients and in 25% of the latex-exposed subjects indicating that Hev b 1 is relevant antigen in these latex-sensitized and latex-exposed groups. Peripheral blood mononuclear cells of 39.1% of the latex-sensitized subjects responded to all three allergen preparations (latex sap and latex glove extract, as well as Hev b 1). We could find no correlation between latex-specific IgE level and latex-induced lymphocyte proliferation response. CONCLUSION: Our data indicate that the 14 kd protein Hev b 1 is a relevant allergen in health care workers. It can be detected by specific IgE antibodies to Hev b 1, as well as in lymphocyte proliferation assay. In addition, our study suggests that antigen-specific proliferation response to latex is associated with exposure to latex, but not with the level of specific latex IgE. This may be useful for the evaluation and prediction of latex hypersensitivity development.

Adult↗

EAST and CAP specificity for the evaluation of IgE and IgG antibodies to diisocyanate-HSA conjugates.

Sera of 54 symptomatic workers showing sensitization to isocyanate-human serum albumin (HSA) conjugates were subjected to parallel enzyme allergosorbent test (EAST) and CAP measurements to determine IgE antibodies to diphenyl-methane diisocyanate-HSA, toluene diisocyanate-HSA and hexamethylene diisocyanate-HSA. Results of both methods correlated rather well with each other. In comparison to the EAST results, the CAP values were twice as high, and 4, 17 and 13% more frequently positive findings were obtained with the three different antigens. Autoinhibition performed with both methods proved the specificity of IgE binding in 92% of sera in EAST and in 89% of sera in CAP if values of > or = 0.35 kU/l were considered. The total IgE level in sera influenced the antibody results. Four of 20 sera studied by autoinhibition had a total IgE of > 700 kU/l, and two of them did not show significant autoinhibition with all conjugates by CAP and one serum by EAST. In addition, three of these sera showed an elevated binding to control HSA only (0.31-0.5 kU/l), and two revealed only a slightly increased IgE binding when compared with the HSA control (ratio of isocyanate-HSA to HSA, < 2). Only 2 of the 16 sera with a total IgE level of < 700 kU/l yielded a noninhibitory positive CAP result, whereas all positive EAST values of these sera could be significantly blocked by autoinhibition. Therefore, we suggest regarding EAST and CAP IgE results to isocyanate-HSA as positive if they exceed HSA control by 100% and are above 0.35 kU/l. Weak positive CAP results (< or = kU/l), especially of sera with total IgE > 700 kU/l, should by confirmed by inhibition experiments. Twelve of 40 symptomatic isocyanate workers exhibited borderline or weakly increased IgG values for diisocyanate-HSA conjugates in the CAP system and IgG-EAST. HSA tested in EAST as a reference showed nearly the same results as the isocyanate-HSA conjugates. In the 23 inhibition experiments, IgG-binding specificity was not confirmed. These findings imply IgG measurement to be of no diagnostic value in isocyanate-induced airway disorders.

Binding, Competitive↗

Allergenic and antigenic determinants of latex allergen Hev b 1: peptide mapping of epitopes recognized by human, murine and rabbit antibodies.

BACKGROUND: The rubber elongation factor in Hevea rubber (Hev b 1) is one of the most important latex allergen and is leading cause of latex type 1 hypersensitivity in children with spina bifida. OBJECTIVE: The aim of this study was to define the allergenic and antigenic epitopes of Hev b 1. METHODS: The immunoglobulin- (Ig)E and IgG antibody binding sites on Hev b 1 allergen were delineated by enzyme linked immunosorbent assay (ELISA) using synthetic overlapping peptides covering the whole Hev b 1 sequence. In order to improve the binding capacity and specificity all peptides were biotinylated at the N-terminal end via a 6-aminohexanoic acid as spacer and then adsorbed to streptavidin pre-coated microtitre plates. Fine mapping to define the essential amino acid residues for the antibody binding was achieved by using overlapping peptides with one amino acid offset. RESULTS: It was demonstrated that the IgE epitopes were located in different regions of Hev b 1 including the C-terminal segment (121-137) and the segments with amino acid residues of 30-49 and 46-64. Two monoclonal antibodies (MoAbs) II2F3 and II4G9 raised against purified Hev b 1 recognized the C-terminal segment only. The results of epitope mapping with three rabbit antisera revealed that five positive peptides, including the epitope peptides 31-49, 46-64 and 121-137, were involved in the antibody-binding sites. Fine mapping on the segments 46-64 and 121-137 showed that the two MoAbs reacted with the peptide 125-134 in the C-terminal region, whereas the peptide with amino acids 124-134 was essential for recognition by human IgE antibodies. Epitopes to rabbit polyclonal IgG and human IgE were also found to be involved in the amino acid residues of 47-59. CONCLUSION: Our results indicate that the most allergenic/antigenic portions of Hev b 1 allergen are the C-terminal region and the region with amino acid residues of 31-64. In both regions, the minimal IgE-binding epitope is almost identical with the IgG-binding epitope.

Allergens↗

Cross-reacting IgE antibodies recognizing latex allergens, including Hev b 1, as well as papain.

The cross-reactivity of IgE antibodies recognizing epitopes of latex allergens and papain was studied in sera of 36 latex-exposed subjects and 22 papain workers. Eight out of 24 latex-sensitized persons also showed a positive reaction to papain in the CAP assay (mostly of low or moderate degree). On the other hand, six out of the 12 sensitized papain workers also revealed IgE binding to latex allergen(s). Reciprocal inhibition experiments confirmed that groups monosensitized to one of the two allergens can be separated from a group showing partial or nearly complete immunologic cross-reactivity. Papain inhibited IgE binding by 20-33% in these subjects, whereas IgE binding to papain was strongly blocked in most cases. Comparison between the primary sequences of Hev b 1, a major latex allergen, and papain suggests that the cross-reactivity may be due to several identical trimers and tetramers.

Allergens↗

[Components of hair bleaches as triggers of bronchial hypersensitivity--studies using a model for occupationally-induced lung damage].

Ammonium persulfate (APS) is used as an oxidant in many industrial branches and is contained in bleaching substances used in hair cosmetic industry. In addition to skin diseases, respiratory disorders have been frequently reported. We examined whether APS can induce bronchial hyperresponsiveness (BHR) which has to be regarded as a first step in the development of obstructive respiratory disorders. In a rabbit model we tested the influence of aerosolized APS-solutions in concentrations of 0.1, 0.01 and 0.001 M (50, 5, 0,5 ppm in air) for about four hours on the contractile bronchial reaction. Exposure to APS-aerosols does not alter basal airway Resistance (RI), dynamic Elastance (Edyn), slope of inspiratory pressure generation (delta Poes/tI), arterial blood pressure and blood gases (PaO2, PaCO2). Before, after two and after four hours the influence of APS application on the bronchial reactivity to 0.2%- and 2% acetylcholine-aerosols (ACH) was tested. Exposure to an aerosol from 0.1 M APS solution, which is equivalent to a concentration of 50 ppm APS in air for two hours induces a significant increase in bronchoconstrictory responses to ACH. A nebulized 0.01 M APS solution causes an increased ACH response after four hours of exposure. However, inhalation of 0.001 M APS (0.5 ppm in air) does not alter sensitivity to ACH within four hours of exposure. In bronchoalveolar lavage an increased number of inflammatory cells was found. Our results demonstrate, aerolized APS-solutions to cause bronchial hyper-responsiveness after few hours of exposure presumably under the participation of inflammatory cells.

Aerosols↗

Respiratory and other hazards of isocyanates.

Isocyanates are increasingly being used for manufacturing polyurethane foam, elastomers, adhesives, paints, coatings, insecticides, and many other products. At present, they are regarded as one of the main causes of occupational asthma. The large number of workers who are exposed to these chemicals have a concentration-dependent risk of developing chronic airway disorders, especially bronchial asthma. Different pathophysiologic mechanisms are involved. Immunoglobulin E (IgE)-mediated sensitization and irritative effects have been clearly demonstrated in both exposed subjects and animals. Presumably, neural inflammation due to neuropeptide release of capsaicin-sensitive afferent nerves is crucial. We collected data on 1780 isocyanate workers who had been examined by our groups. Of them 1095 (including subjects from outpatient departments) had work-related symptoms, predominantly of the respiratory tract. Specific IgE antibodies were found in 14% of the 1095 subjects. The methacholine challenge test was shown to be an inadequate predictor of the results of inhalative isocyanate provocation tests in workers and in asthmatic controls. Isocyanate (toluene diisocyanate TDI) air concentrations of 10 ppb (0.07 mg/m3) and 20 ppb (0.14 mg/m3), respectively, did not cause significant bronchial obstruction in the majority of previously unexposed asthmatics with bronchial hyperreactivity. IgG-mediated allergic alveolitis, a rare disease among isocyanate workers, was found in approximately 1% of the symptomatic subjects. Experimental studies exhibit dose-dependent toxic effects and give evidence for tachykinin-mediated bronchial hyperreactivity after exposure to isocyanates. The clinical role of genotoxic effects of isocyanates and their by-products demonstrated here in vitro and in vivo has yet to be clarified.

Air Pollutants, Occupational↗