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Background: Chestnut and natural rubber latex (NRL) allergy are often associated in the latex-fruit syndrome., Aim of the Study: To establish whether the concurrent NRL and chestnut IgE antibody reactivity are the results of co-sensitization or cross-reactivity., Methods: Sera from 19 patients with chestnut- and NRL-specific IgE were selected and tested for reactivity with recombinant (r) latex allergens. Cross-reactivity was explored by IgE-inhibition experiments using chestnut or NRL allergens as solid phase on ImmunoCAP., Results: IgE-antibodies were detected to rHev b 6.01 (prohevein) in 58% of the sera, to rHev b 5 in 32%, to rHev b 12 in four of 13 sera, to rHev b 7.02 and rHev b 11 in four, and to rHev b 1 in two of 19 sera. rHev b 8-IgE antibodies were found in nine sera (47%), whereas six displayed mono-sensitization to rHev b 8 with regard to our test panel. Three of 16 sera showed IgE to cross-reactive carbohydrate determinants. In most sera recognizing rHev b 5 and/or rHev b 6.01 as major allergens the IgE-reactivity to NRL remained unaffected by chestnut extract and chestnut-IgE remained unaffected by NRL extract. Conversely, in sera with rHev b 8 as dominant allergen IgE-binding to NRL was nearly completely inhibited by chestnut and vice versa. IgE-binding to rHev b 8 was abolished by chestnut extract., Conclusions: Although patients have concomitant IgE antibody reactivity to chestnut and NRL, cross-reactivity could be demonstrated mainly in those patients with IgE to Hev b 8 (profilin) from NRL.

Background: Multiple immunoglobulin E (IgE)-binding proteins in natural rubber latex extracts have been identified. In the case of Hev b 6 a differentiation was made between the precursor protein prohevein (Hev b 6.01) and its two post-transcriptionally formed proteins, the N-terminal hevein (Hev b 6.02) and the C-terminal domain (Hev b 6.03). All three components act as independent allergens. The aim of this study was a detailed analysis of the T-cell responses and the IgE-binding capacity of Hev b 6.01, Hev b 6.02 and Hev b 6.03 by using these allergens as recombinant maltose-binding fusion (MBP) proteins and the usage of synthetic modified hevein peptides., Methods: Latex-allergic health care workers (HCWs) suffering from rhinitis, conjunctivitis, contact urticaria and/or asthma with increased specific IgE-antibodies to latex were tested for their IgE-binding capacity and T-cell reactivity (by proliferation response) to the recombinant MBP-rHev b 6.01, MBP-rHev b 6.02, MBP-rHev b 6.03, to native Hev b 6.02, to modified hevein peptides and wheat germ agglutinin (WGA). For testing of the human leucocyte antigen (HLA) class II restriction of MBP-rHev b 6.01 induced peripheral blood mononuclear cell (PBMC) responses, monoclonal antibodies against HLA-DR, HLA-DP or HLA-DQ were added., Results: Seventeen of 18 (94%) serum samples from latex-allergic HCWs had increased levels of specific IgE to MBP-rHev b 6.01, 16 (89%) to MBP-rHev b 6.02 and 13 (72%) to MBP-rHev b 6.03. A significant difference existed between the specific IgE-values of MBP-rHev b 6.02 and MBP-rHev b 6.03 (P < 0.01). Proliferation responses of PBMC of the same 18 latex-allergic patients were positive for MBP-rHev b 6.01 and MBP-rHev b 6.03 in 83 and 67% of the tested PBMC suspension, whereas the proliferation responses induced with MBP-rHev b 6.02 or native Hev b 6.02 were very low (5.6 and 22.2%). Sera from nine additional latex-allergic patients showed specific IgE binding to the native Hev b 6.02, but none of these sera showed specific IgE binding to the modified Hev b 6.02-peptides [whereby all eight cysteine residues were substituted by serine (C --> S) or by alanine (C --> A)]. Proliferation responses induced by the modified Hev b 6.02 peptides were not significantly different from that induced by Hev b 6.02. Potential HLA-DR4Dw4(DRB1*0401)-restricted T-cell epitopes of Hev b 6.01 predicted by two computer algorithms were only found in the Hev b 6.03-part of Hev b 6.01., Conclusion: In the Hev b 6.01 precursor the regions responsible for IgE binding and those for inducing the T-cell proliferation responses are settled in different parts of the protein. The Hev b 6.02 domain is responsible for IgE binding and carries discontinuous B-cell epitopes whereas Hev b 6.03 is a better inducer of a proliferation response and contains HLA-DR4-binding motifs.

Background: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana., Objectives: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity., Methods: A class I chitinase-specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex-allergic patients., Results: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex-allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE-reactivity of rHev b 11.0102., Conclusion: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens.

Background/objective: Latex allergy is a type 1 hypersensitivity reaction that mainly affects high-risk populations such as health care workers, spina bifida-affected or multiply-operated children. Ten molecules have so far been identified and registered as latex allergens (Hev b 1 to Hev b 10). The aim of the present investigation was to identify the major latex allergens by an individual analysis of the IgE response of latex-allergic patients to latex proteins separated by two-dimensional (2-D) gel electrophoresis., Materials and Methods: Latex proteins from a sap or a glove extract were separated by 2-D electrophoresis and transferred to a nitrocellulose membrane. Each membrane was incubated with the serum of one latex-allergic patient. The most frequently recognized latex allergens were characterized in sap and glove extracts using monoclonal antibodies or amino acid microsequencing., Results: The one-dimensional screening of 54 patient sera revealed 4 major bands recognized by IgE. The 2-D analysis of the sensitization to latex allergens allows the identification of allergen isoforms and the characterization of an individual response diversity. Hev b 6.01 was recognized by 88.9% of the patients. Protein spots around 14 kD were recognized by 48.1% of the patients and corresponded to Hev b 6.03 as well as other proteins. A not yet characterized doublet of acidic proteins with molecular masses of 43 and 94 kD was recognized by 20.4% of the sera. Only 5.5% of the sera did not recognize any of these 4 major allergens. Hev b 1 is the main protein from the glove extract but was not constantly found in sap extracts., Conclusions: One-dimensional electrophoretic analysis of the allergen is usually not sufficient to characterize the individual specificity of the IgE response to latex allergens. Latex-glove proteins which are allergens can be absent from the sap extracts and the sensitization to these allergens could be underestimated. Individual 2-D analysis of the sensitization to latex allergens is useful to define the best allergen mixture required for diagnosis and needed for individual therapy monitoring., (Copyright 2002 S. Karger AG, Basel)

Background: Type 1 hypersensitivity to natural rubber latex proteins is a well-recognized health problem. Recent data have shown that allergens can be extracted from natural latex mattresses. As Hev b 1 (rubber elongation factor) and Hev b 6.02 (hevein) were described as major allergens, the present work was carried out to evaluate their presence in latex mattresses as well as in latex gloves., Methods: Extracted proteins from latex mattresses and gloves were separated by SDS-PAGE or two-dimensional gel electrophoresis, transferred onto nitrocellulose and detected with monoclonal antibodies specific for Hev b 1 and Hev b 6.02., Results: The results showed that various forms of Hev b 1, as well as degradation products of Hev b 1 were detected in latex mattresses and gloves, whereas Hev b 6.02 was not detected either in mattresses or in gloves. In a standardized latex extract, Hev b 1 and Hev b 6.01 (prohevein) were identified by the monoclonal antibodies., Conclusion: The fact that only Hev b 1 was detected by immunoblot in latex articles indicates that Hev b 1 may be the last protein to be washed out of latex products and that the Hev b 1 content may be used as a criterion for the estimation of the allergenicity of the latex products., (Copyright 2000 S. Karger AG, Basel)

Background: Respiratory symptoms of natural rubber latex (NRL) allergy in health care workers (HCWs) have been reported in rooms with a detectable allergen load. Preventive measures have been proposed to reduce the risk of NRL sensitization., Objectives: Eliminating powdered NRL gloves from the workplace and giving NRL-free material to sensitized workers has been among proposed preventive measures. To appraise the success of such procedures among HCWs, a prospective study was carried out., Methods: Sensitization of HCWs to NRL was determined by skin prick tests and measurements of specific IgE antibodies. NRL allergen concentrations in room air were measured before and after substitution of powdered NRL gloves with powder-free or synthetic gloves in different departments of a hospital and determined by a competitive inhibition immunoassay., Results: The prevalence of HCWs with positive skin prick test responses and NRL-specific IgE-positive HCWs was 8% (n = 7) among the 90 examined staff members before the intervention started. All 7 reported glove-related allergic symptoms. Six of 7 sensitized subjects had a significant decrease of latex-specific IgE antibody concentrations during follow-up examinations in April and September 1997 (P <.003). Within 24 hours after substitution took place, NRL aeroallergen levels (up to 49.9 ng/m3) fell below the detection limit in areas with synthetic gloves or powder-free NRL gloves alike. Use of asthma medication and antiallergic drugs could be discontinued by 2 HCWs with NRL-related respiratory tract symptoms., Conclusions: Our results demonstrate that elimination of powdered NRL gloves is a useful device in reducing aerogen NRL allergen loads below the detection limit and permitting sensitized or allergic personnel to remain on the job.

Background: Recent studies demonstrated that allergy to natural rubber latex is frequently associated with hypersensitivity to avocado fruit. The responsible cross-sensitizing allergen has not been identified., Objective: The purpose of this study was to investigate the cross-reactivity of a latex major allergen, hevein, with avocado proteins., Methods: Serum samples from 118 health care workers (HCWs) allergic to latex (HCW group) and 78 patients with spina bifida (SB) allergic to latex (SB group) were included in this study. Anti-hevein and anti-avocado IgE antibodies were measured by enzyme-linked allergosorbent assay. Cross-reactivity of hevein to avocado proteins was assessed by inhibition of the IgE binding in individual patients' sera containing IgE antibodies to both hevein and avocado., Results: The prevalence of seropositive IgE antibodies to avocado was found to be strongly associated with the presence of hevein-specific IgE antibodies in subjects of both groups (P < .001). Sixty-seven of 91 (73%) subjects from the HCW group and all 19 subjects in the SB group with positive IgE antibodies to hevein also had elevated IgE values to avocado. Competitive RAST inhibition with 42 sera showed that IgE binding to avocado could be completely inhibited in 27 (64%) sera by preincubation with hevein. By contrast, the degrees of inhibition of IgE to hevein by avocado extract ranged from 0% to 36% (n = 16). These results indicate that sensitization to avocado in most patients allergic to latex is caused exclusively by IgE-binding epitopes present in hevein. Results of immunoblots and immunoblot inhibition with 11 serum samples confirmed that a 30-kd protein in avocado was the major IgE-binding component; the IgE-binding reactivity to this protein could be inhibited by hevein in all sera tested., Conclusion: Hevein is the major cross-reacting allergen with avocado in subjects with latex allergy.

A total of 109 subjects reporting symptoms indicating type I hypersensitivity reactions to natural rubber latex (NRL) gloves was included in this study, and 66 of them had latex-specific IgE antibodies. They underwent provocation tests by wearing two types of NRL gloves with high (n=103) and low (n=75) allergen contents. The first glove type caused positive skin reactions in 30% of IgE-positive and in 3% of IgE-negative subjects. After application of a commercially available skin protection (barrier) cream, the frequencies of positive skin responses in wearing tests increased to 41% and 7%, respectively. The gloves with low allergen content did not cause hypersensitivity without skin-protection cream but induced responses in 5% of IgE-positive subjects when this cream was applied. Corresponding findings were obtained in intraindividual comparisons of test results, which were possible in 69 cases. Of all wearing-test responders, 92% had latex-specific IgE antibodies. Our results demonstrate that high allergen contents in latex gloves frequently elicit skin responses in NRL-sensitized subjects, and that skin-protection creams may favor the uptake of allergens from gloves, thus increasing allergic reactions. The stipulation of a legally binding threshold limit value (TLV) for allergens in NRL products is urgently needed. This TLV should not be set above 2 microg allergen/g rubber.

Background: The rubber elongation factor in Hevea rubber (Hev b 1) is one of the important latex-allergenic proteins inducing the production of specific IgE antibodies in latex-exposed subjects. In addition, Hev b 1 induces lymphocyte proliferation responses. A study about detailed T-cell epitope analysis of major latex allergens has not been published yet., Objective: The aim of this study was to define the T-cell reactive regions on the latex allergen Hev b 1., Methods: Nine overlapping peptides with 19 or 17 amino acid lengths representing the complete sequence of Hev b 1 were used for T-cell epitope mapping. Peripheral blood mononuclear cells (PBMCs) of latex-sensitized patients and healthy subjects were isolated and stimulated with the synthetic peptides to determine the specific proliferation response. The examined patients were sensitized to latex by occupational exposure (n=14) and suffered from rhinitis, conjunctivitis, contact urticaria and/or asthma. Two control groups of non-sensitized subjects were studied, one occupationally exposed to latex (n=4), the other one not exposed to latex (n=4)., Results: Positive proliferation response induced by one or more peptides was detected in 10 of the latex-sensitized patients and in two of the latex-exposed non-sensitized subjects. Each patient exhibited an individual epitope reactivity pattern. The whole protein was found to be immunogenic, but the intensity of lymphocyte responses induced by the peptides was different. The majority of the patients' PBMCs (more than 66%) responded to the peptides 31-49 and 91-109. Using the peptide selection of Stille's algorithms, three epitopes were predicted covering region 38-74 and two others covering region 82-104; the two peptides 31-49 and 91-109 are part of these regions. A strong HLA-DR4Dw4 (DRB1*0401)-binding motif according to published DR4w4 binding motifs was predicted in the region of 102-110 including peptide 91-109 to which the majority of tested HLA-DR4-positive patients responded., Conclusions: Our data indicate that the whole 14 kDa protein Hev b 1 is immunogenic having regions that demonstrate individual and varying PBMC stimulation. Experimental data of T-cell reactive regions based on PBMC-stimulation complement the information on T-cell epitope prediction. In addition, the Hev b 1 molecule contains a HLA-DR4Dw4 (DRB1*0401)-binding motif.

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.

Background: Recent studies have caused much controversy about the prevalence of IgE antibodies to Hev b 1 among health care workers (HCWs) and patients with spina bifida (SB) who are allergic to latex. This investigation was carried out to verify the results reported., Method: Serum samples from 140 patients with SB as well as from 105 HCWs allergic to latex were tested by enzyme allergosorbest test (EAST) and EAST-inhibition assay to evaluate the rate and degree of sensitization to highly purified Hev b 1., Results: Eighty-one percent of patients with SB who were allergic to latex had IgE antibodies against Hev b 1. The prevalence of anti-Hev b 1 antibodies among HCWs allergic to latex was 52.3%. In 15 of 33 serum samples from patients with SB that were randomly tested, the IgE binding to commercial latex allergens could be completely inhibited by Hev b 1; in only six cases was the maximum inhibition of IgE binding to latex by Hev b 1 less than 50%. Testing two monoclonal anti-Hev b 1 antibodies with extracts of five brands of latex gloves revealed a predominant presence of Hev b 1 protein as a monomer or its aggregates. Molecular analysis of human leukocyte antigen-D region genes DRB and DQB1 suggested no statistically significant correlation between the human leukocyte antigen alleles tested and IgE responsiveness to Hev b 1., Conclusions: Our results indicate that Hev b 1 not only makes significant contributions to the IgE binding to latex, but it is also the unique sensitizer in about 45% of patients with SB who are allergic to latex.

Background and Objective: Type 1 sensitization to natural rubber latex occurs in up to 22% of health care workers. Most sensitizations are due to the use of powdered latex gloves. Work place-associated exposure tests were performed to ascertain how frequently persons who developed breathing difficulties, rhinitis/conjunctivitis or obstructive respiratory tract illness when using powdered latex gloves are allergic to latex., Patients and Methods: 60 persons (48 women and 12 men; mean age 29 +/- 7 years) with type 1 sensitization to natural rubber latex were studied. The past history was obtained through a questionnaire specially designed for those with latex allergy. Diagnostic measures included prick tests with different latex allergens, measuring the concentrations of total and latex-specific IgE and exposure trials, related to the work place, with powdered vinyl and latex gloves., Results: The prick tests with various latex allergens produced significant skin reactions in 59 subjects, 58 had latex-specific IgE antibodies (0.47 kU/l- > 100 kU/l). Exposure tests with powdered latex gloves produced rhinitis in 55 and conjunctivitis in 38 persons, urticaria in 3 and cough in 19. Whole-body plethysmography demonstrated a significant rise in respiratory tract resistance in 13, a reduction in 1-second expiratory volume of at least 20% in 15. Abnormal values in both tests were noted in 8 subjects., Conclusion: The breathing difficulty reported by all subjects was in most cases due not to bronchial obstruction but to obstructed nasal breathing. In cases with breathing difficulty on exposure to latex gloves a latex allergy should be considered as the cause. Powdered latex gloves present a danger to health care personnel and should no longer be used.

Background: Proteins of natural rubber latex cause IgE-mediated sensitization in 3% to 18% of health care workers and in up to 50% of patients with spina bifida., Objective: This study was aimed at the generation of a comprehensive latex protein database by two-dimensional electrophoresis (2-DE)., Methods: Proteins extracted from fresh Hevea brasiliensis latex were separated by 2-DE. IgE-reactive proteins were analyzed by immunoblotting with sera of health care workers with latex allergy. Protein microsequencing and monoclonal antibodies were used to identify the latex allergens., Results: The latex C-serum 2-DE map was very complex and exhibited about 200 distinct polypeptides. The proteins eluted from the latex particles consisted primarily of two groups of acidic proteins located in the 8 to 14 kd and 22 to 24 kd areas of the 2-DE map. Major IgE-reactivity was detected with C-serum proteins in the 56, 45, 30, 20, 14, and <6.5 kd areas of the immunoblots. The 8 to 14 kd particle proteins exhibited distinct IgE reactivity, whereas the 22 to 24 kd proteins were not stained. Seven of the soluble IgE-reactive protein spots showed high homology with enolase, superoxide dismutase, triosephosphate isomerase, proteasome subunit, and chitinase and represent previously undescribed latex allergens; whereas nine protein spots corresponded to known latex allergens, namely prohevein, hevein, prohevein C-domain, and hevamine. As identified by monoclonal antibodies, the IgE-reactive latex particle proteins mainly represent the allergenic rubber elongation factor., Conclusions: Two-dimensional electrophoresis, followed by immunoblotting and protein microsequencing, can rapidly identify a large number of IgE-binding latex proteins. The 2-DE latex maps generated will provide valuable information for the development of strategies to isolate the relevant latex allergens. Because the novel latex allergens are common plant enzymes, they may also act as cross-reacting proteins in various foods.

Background: Polypeptides in Hevea latex are known as the major cause of latex type I sensitivities. So far, only a few of them have been characterized., Methods: Proteins with a molecular weight lower than 10 kd in fresh Hevea latex were separated by ultrafiltration and further characterized by liquid chromatography on-line-coupled electrospray mass spectrometry. Hevein in this fraction was then purified by preparative reverse-phase high-performance liquid chromatography and characterized by matrix-assisted laser desorption ionization mass spectrometry and protein sequencing. Skin prick tests, enzyme-linked allergosorbent tests, and inhibition immunoblotting were performed to show the allergenicity of the purified hevein., Results: Hevein, a 4.7 kd polypeptide, is the predominant component in the fraction with latex proteins of smaller than 10 kd. Specific IgE antibodies to hevein were detected by enzyme-linked allergosorbent test in 48 of 64 (75%) sera from health care workers allergic to latex and in three of 11 (27%) sera from patients with spina bifida and hypersensitivity reactions to latex. Inhibition immunoblotting demonstrated that the preincubation of 14 sera and a serum pool from patients allergic to latex with purified hevein completely inhibited IgE binding to the 20 kd protein, which has been recently reported to be a major allergen in latex (prohevein). Skin prick testing showed a positive reaction to hevein in 17 of 21 (81%) patients with latex allergy., Conclusions: The results clearly demonstrate that hevein is an important latex allergen, and the IgE-binding capacity of prohevein in latex is mostly attributed to hevein, the N-terminal domain of prohevein.

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.

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.

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.

Background Hevea brasiliensis latex serum is commonly used as the in vivo and in vitro reference antigen for latex allergy diagnosis as it contains the full complement of latex allergens. Objective This study quantifies the concentrations of the significant allergens in latex serum and examines its suitability as an antigen source in latex allergy diagnosis and immunotherapy. Methods The serum phase was extracted from centrifuged latex that was repeatedly freeze-thawed or glycerinated. Quantitation of latex allergens was performed by two-site immunoenzymetric assays. The abundance of RNA transcripts of the latex allergens was estimated from the number of their clones in an Expressed Sequence Tags library. Results The latex allergens, Hev b 1, 2, 3, 4, 5, 6, 7 and 13, were detected in freeze-thawed and glycerinated latex serum at levels ranging from 75 (Hev b 6) to 0.06 nmol/mg total proteins (Hev b 4). Hev b 6 content in the latex was up to a thousand times higher than the other seven latex allergens, depending on source and/or preparation procedure. Allergen concentration was reflected in the abundance of mRNA transcripts. When used as the antigen, latex serum may bias the outcome of latex allergy diagnostic tests towards sensitization to Hev b 6. Tests that make use of latex serum may fail to detect latex-specific IgE reactivity in subjects who are sensitized only to allergens that are present at low concentrations. Conclusion Latex allergy diagnostics and immunotherapy that use whole latex serum as the antigen source may not be optimal because of the marked imbalance of its constituent allergens. [ABSTRACT FROM AUTHOR]

Several studies have shown that allergy to natural rubber latex is associated with cross reactivity to other plants. We have investigated a possible cross reactivity with a tree belonging to the Sterculiaceae family, Tryplochiton scleroxylon, in a patient allergic to obeche wood, that began to suffer symptoms of allergy in contact with latex. Determinations of specific IgE by CAP to obeche, natural rubber latex (NRL) and recombinant allergens of latex were done. Immunoblotting was performed to study allergens detected by serum of our patient. CAP-inhibition methods were used to study cross-reactive between NRL and obeche wood. In obeche extract, 4 bands were found of an apparent molecular weight of 18, 38, 75 and 199 kDa. In NRL extract the allergens have a molecular weight around 18, 21, 32, 38, 60, 70 and 199 kDa. IgE to latex was almost completely inhibited (96%) by the obeche extract in CAP-inhibition studies. Using obeche extract as inhibitor, IgE binding to native hevein (nHevb 6.02) was reduced by 25%. The results obtained in CAP-inhibition demonstrate the existence of cross-reactivity between proteins in NRL and obeche wood. In our patient, a band of apparent molecular weight of 38 kDa could be the most important allergen. [ABSTRACT FROM AUTHOR]

Recent studies have demonstrated that allergy to natural rubber latex (NRL) is associated with cross‐reactivity to certain foods. The aim of this study was to investigate the prevalence of NRL sensitization and allergy in children with atopic dermatitis (n=74). We also examined cross‐reactions between latex and foods, and compared the frequency of suspected latex cross‐reacting fruits in children with and without NRL‐specific immunoglobulin E (IgE). Twelve of the 74 atopic children studied (16.2%; 95% confidence interval (CI), 8.7–26.6%) had circulating IgE antibodies to latex. These NRL‐sensitized children were older and they showed significantly higher total IgE values (p<0.003) when compared with the group of children without NRL sensitization. Of the specific food IgE evaluations, 18.4% (93 out of 505) were positive, and 69.9% were observed in the group of children with latex‐specific IgE, most frequently to potato, tomato, sweet pepper, and avocado. An isolated latex‐specific IgE response without food‐specific IgE was never observed. Exclusively in the latex‐positive group, conformity with the report of allergic symptoms after ingestion of food and increased food‐specific IgE was found. Twenty children without proven latex sensitization showed increased food‐specific IgE, most frequently to potato, banana, and chestnut. Avocado‐specific IgE was never determined in this patient group. No significant differences were detected concerning the sensitization to potato, banana, and kiwi between NRL‐sensitized children and the group of 20 children without latex‐specific IgE. The competitive CAP inhibition using sera from children with specific IgE to both latex and food showed different cross‐reactivities between latex and the specific food. A close relationship existed between latex and avocado (median inhibition: 100%), whereas sensitization to latex... [ABSTRACT FROM AUTHOR]

Abstract: In the last few years many efforts have been made to reduce the incidence of latex allergy. The identification of the major latex allergens and their recombinant production are useful steps towards the understanding of latex allergy and its management. A clear advantage of the recombinant proteins in general is the possibility to produce large-scale quantities at highly reproducible quality. Recombinant allergens also facilitate the study of the molecular bases of the immune reactivity of the proteins. Nevertheless, they have to be validated against native proteins for equivalence in allergenicity before they can be more widely adapted for clinical use. Currently 13 latex allergens have been included in the latest nomenclature list of the International Nomenclature Committee of Allergens and assigned the official names Hev b 1–13. With the use of single native and/or recombinant allergens the determination of specific sensitization profiles for different patient groups dependent on their various routes of exposure is possible. The specific IgE-diagnostic was further improved significantly by addition of recombinant Hev b 5 to natural latex extract, which was underrepresented in the initial test extract. [Copyright &y& Elsevier]

The article examines the sera of healthcare workers (HCW) with symptomatic latex allergy and sera from Portuguese patients with multiple surgeries (MS) and spina bifida (SB) patients are being examined. The DNA fragment of 1167 base pair was the gene encoding for a Hevea brasiliensis (Hev b) 7.02 isoform. Sera of HCW are 33 percent of the SB sera showed Hev b 7.02-specific immunoglobulin E. MS sera are all negative.

The article presents information on latex allergy that affects well-recognized risk groups. The highest prevalence appears in children requiring multiple surgeries, especially those with spina bifida (SB). Natural rubber latex (NRL) contains more than 200 proteins and peptides, some of which are capable of eliciting immunoglobulin E (IgE) responses. It is known that manufacturing processing of latex can alter the structure of proteins; thus, ammoniated latex contains less allergenic determinants and has been proved to offer inferior results to those of nonammoniated extracts for the study of sensitization to latex