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PubMed2002

Purification and characterization of NAD(P)H quinone reductase from the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae).

NAD(P)H quinone reductase [NAD(P)H-QR] present in the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae) was purified to homogeniety from the B-serum fraction obtained by freeze-thawing of the bottom fraction of ultracentrifuged fresh latex. The purification protocol involved acetone fractionation, heat treatment, ion exchange chromatography and affinity chromatography. The M(r) determined by SDS-PAGE for the protein subunit was 21 kDa, and the molecular mass of the native enzyme estimated by gel filtration was 83 kDa, indicating that the native enzyme is a homotetramer. The enzyme showed pH stability over a range of 6 to at least 10 (with an optimum at pH 8) and thermal stability up to 80 degrees C. High NAD(P)H-QR activity (70%) was still retained after 10 h of preincubation at 80 degrees C. A comparable substrate specificity for this enzyme was observed among menadione, p-benzoquinone, juglone, and plumbagin, with only duroquinone generating a lower activity. Positive correlations between latex NAD(P)H-QR activity and rubber yield per tapping [fresh latex (r=0.89, P<0.01), dry rubber (r=0.81, P<0.01)] together with flow time (r=0.85, P<0.01) indicated that enzyme activity could possibly be used as a marker to predict the yield potential of selected clones.

Chromatography, Gel↗
PubMed2003

The patatin-like protein from the latex of Hevea brasiliensis (Hev b 7) is not a vacuolar protein.

Upon centrifugation, rubber latex is divided into a layer of rubber particles, the cytosol, and the lutoid-body fraction, which is of vacuolar origin. One of the proteins isolated from the lutoid-body fraction is a protein with a molecular mass of 43 kDa, which has esterase activity on p-nitrophenylpalmitate and which shows significant sequence similarity with patatin, a vacuolar protein with esterase activity from potato (Solanum tuberosum). This protein is a major allergen in rubber latex products (Hev b 7) and can also be isolated from the cytosol fraction of rubber latex. The mature protein isolated from lutoid-bodies has no structural features expected for a vacuolar protein: the N-terminal methionine in the cDNA-derived sequence is cleaved off, the second residue is N-acetylated, and the C-terminal sequence is identical to that in the cDNA-derived sequence. Thus the patatin-like protein in Hevea brasiliensis is not a vacuolar protein, but may be associated with not yet characterized particles in the cytoplasm, which either sediment with lutoid-bodies or remain in the cytosol fraction, depending on the centrifugation conditions.

Allergens↗
PubMed2003

Lipid transfer protein from Hevea brasiliensis (Hev b 12), a cross-reactive latex protein.

BACKGROUND: Latex-allergic individuals experience clinical cross-reactivity to a large number of fruits and vegetables. Much of the cross-reactivity can be attributed to Hev b 6, but evidence indicates that additional cross-reactive allergens may be present. A common pan-allergen, which has not previously been identified in latex, but may contribute to this cross-reactivity is lipid transfer protein (LTP). We sought to determine whether Hevea brasiliensis produces LTP and whether it would bind immunoglobulin E from latex-allergic patients. METHODS: LTP was identified in H. brasiliensis RNA by polymerase chain reaction using degenerate primers. The entire cDNA was obtained by polymerase chain reaction using rapid amplification of cDNA ends reactions. The complete coding sequence for LTP was determined and produced as a recombinant protein using the glutathione S-transferase and pET32 expression systems. Immunoblot analysis of sera from latex-allergic patients was used to determine whether patients recognize LTP as an allergen. RESULTS: We identified a 662-basepair cDNA with a 351-basepair open reading frame that encodes for a 116-amino acid protein. The protein has significant homology to the family of nonspecific LTPs. We expressed the protein as a mature LTP of 92 amino acids with a predicted isoelectric point of 10.8 and molecular weight of 9.3 kDa. Immunoblots demonstrated specific immunoglobulin E for LTP in the sera of 9 of 37 (24%) latex-allergic individuals. CONCLUSIONS: We describe the initial identification of rLTP in H. brasiliensis that may be important as a cross-reactive pan-allergen (Hev b 12).

Allergens↗
PubMedDate not supplied

Regulation of rubber biosynthetic rate and molecular weight in Hevea brasiliensis by metal cofactor.

Metal ion cofactors are necessary for prenyltransferase enzymes. Magnesium and manganese can be used as metal ion cofactor by rubber transferase (a cis-prenyltransferase) associated with purified rubber particles. The rubber initiation rate, biosynthetic rate, and molecular weight produced in vitro from Hevea brasiliensis rubber transferase is regulated by metal ion concentration. In addition, varies significantly with [Mg(2+)]. decreases from 8000 +/- 600 microM at [Mg(2+)] = 4 mM to 68 +/- 10 microM at [Mg(2+)] = 8 mM and increases back to 970 +/- 70 microM at [Mg(2+)] = 30 mM. The highest affinity of rubber transferase for IPP.Mg occurred when [Mg(2+)] = A(max) (metal concentration that gives highest IPP incorporation rate). A metal ion is required for rubber biosynthesis, but an excess of metal ions interacts with the rubber transferase inhibiting its activity. The results suggest that H. brasiliensis could use [Mg(2+)] as a regulatory mechanism for rubber biosynthesis and molecular weight in vivo.

Hemiterpenes↗
PubMed2002

Isolation of scopoletin from leaves of Hevea brasiliensis and the effect of scopoletin on pathogens of H. brasiliensis.

Scopoletin (7-hydroxy-6-methoxy coumarin) which inhibited the conidial germination of Corynespora cassiicola was isolated from the uninfected mature leaves of Hevea brasiliensis. Scopoletin was not detected in uninfected immature rubber leaves. The immature leaves produced scopoletin after being infected with C. cassiicola. The concentration of scopoletin in infected leaves was higher than in uninfected mature leaves. Scopoletin also inhibited the conidial germination of other fungal pathogens of H. brasiliensis. However, no correlation was observed between scopoletin accumulation and clonal resistance.

Antifungal Agents↗
PubMed2003

Association of decreased expression of a Myb transcription factor with the TPD (tapping panel dryness) syndrome in Hevea brasiliensis.

TPD (tapping panel dryness) is a complex physiological syndrome widely found in rubber tree (Hevea brasiliensis) plantations, which causes severe yield and crop losses in natural rubber-producing countries. The molecular mechanism underlying TPD is not known and there is presently no effective prevention or treatment for this serious disease. To investigate the molecular mechanism of TPD, we isolated and characterized genes for which the change of expression is associated with TPD. We report here the identification and characterization of a Myb transcription factor HbMyb1. HbMyb1 is expressed in leaves, barks, and latex of rubber trees, but its expression is significantly decreased in barks of TPD trees. Our results suggest that the expression of HbMyb1 is likely associated with TPD and that the function of HbMyb1 is associated with the integrity of bark tissue of rubber trees.

Amino Acid Sequence↗
PubMed2003

Transcriptome analysis reveals novel features of the molecular events occurring in the laticifers of Hevea brasiliensis (para rubber tree).

Latex of Hevea brasiliensis (Willd. ex A, Juss.) Mull. Arg. (Brazilian rubber tree) contains 30-50% (w/w) of natural rubber (cis-1,4-polyisoprene), which is an important raw material for many industrial uses. In order to gain insights into the molecular events occurring in latex, we analyzed more than 20,000 cDNA-AFLP-based TDFs (transcription-derived fragments) and 1176 ESTs. The results revealed several novel features of the latex transcriptome. First, the repertoire of the genes expressed in latex is unique. Only seven gene families accounted for more than 51% of the latex transcriptome. Among them, two of the most abundant ESTs were the genes encoding rubber particle proteins REF (rubber elongation factor) and SRPP (small rubber particle protein), comprising 29% of the total ESTs. Unexpectedly, several genes involved in the rubber biosynthesis were expressed at low levels in the latex. In fact, genes encoding cis -prenyltransferase (CPT), a potential candidate for rubber polymerase, were not present in the EST pool because of their low expression level. However, we were able to clone four full-length cDNAs by screening the same latex cDNA library used in the EST analysis and confirmed their enzyme activity in vitro. The second most abundant transcripts were defense- or stress-related genes, suggesting that defense is one of the functions of laticifers. Finally, the presence of the non-mevalonate DXP/MEP pathway for IPP synthesis in latex was noted by up-regulation of the 1-deoxy-D-xylulose 5-phosphate synthase gene.

Amino Acid Sequence↗
PubMed2003

Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102).

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.

Adolescent↗
PubMed2002

Antifungal effect of Hevea brasiliensis latex with various fungi. Its synergistic action with amphotericin B against Candida albicans.

Antifungal activity of latex from Hevea brasiliensis was observed with various fungi in macrobroth dilution assays. The strongest antifungal effect was obtained with Trichosporon cutaneum (MIC 80% = 40.615 microg protein ml(-1), Kaff = 0.075 microg(-1) protein ml) and Cryptococcus neoformans (MIC 80% = 56.078 microg protein ml(-1), Kaff = 0.059 microg(-1) protein ml). Amphotericin B was synergized with all H. brasiliensis latex concentrations tested. The rates of synergy were about 50, 44 and 55% with 15, 30 and 60 microg protein ml(-1) latex, respectively. The putative role of glycosidase activities measured in crude latex, especially alpha-d-mannosidase and N-acetyl-beta-d-glucosaminidase activities which are potentially able to split off intraparietal linkages between glycosidic residues, is discussed. The possible role of another antifungal factor associated with rubber particles of latex is suggested.

Amphotericin B↗
PubMed2003

In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex.

BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy. OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL). METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays. RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively. CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL.

Adult↗
PubMed2003

Observation of a short, strong hydrogen bond in the active site of hydroxynitrile lyase from Hevea brasiliensis explains a large pKa shift of the catalytic base induced by the reaction intermediate.

The hydroxynitrile lyase from Hevea brasiliensis (HbHNL) uses a catalytic triad consisting of Ser(80)-His(235)-Asp(207) to enhance the basicity of Ser(80)-O gamma for abstracting a proton from the OH group of the substrate cyanohydrin. Following the observation of a relatively short distance between a carboxyl oxygen of Asp(207) and the N delta(1)(His(235)) in a 1.1 A crystal structure of HbHNL, we here show by (1)H and (15)N-NMR spectroscopy that a short, strong hydrogen bond (SSHB) is formed between the two residues upon binding of the competitive inhibitor thiocyanate to HbHNL: the proton resonance of H-N delta 1(His(235)) moves from 15.41 ppm in the free enzyme to 19.35 ppm in the complex, the largest downfield shift observed so far upon inhibitor binding. Simultaneously, the D/H fractionation factor decreases from 0.98 to 0.35. In the observable pH range, i.e. between pH 4 and 10, no significant changes in chemical shifts (and therefore hydrogen bond strength) were observed for free HbHNL. For the complex with thiocyanate, the 19.35 ppm signal returned to 15.41 ppm at approximately pH 8, which indicates a pK(a) near this value for the H-N epsilon(2)(His(235)). These NMR results were analyzed on the basis of finite difference Poisson-Boltzmann calculations, which yielded the relative free energies of four protonation states of the His(235)-Asp(207) pair in solution as well as in the protein environment with and without bound inhibitor. The calculations explain all the NMR features, i.e. they suggest why a short, strong hydrogen bond is formed upon inhibitor binding and why this short, strong hydrogen bond reverts back to a normal one at approximately pH 8. Importantly, the computations also yield a shift of the free energy of the anionic state relative to the zwitterionic reference state by about 10.6 kcal/mol, equivalent to a shift in the apparent pK(a) of His(235) from 2.5 to 10. This huge inhibitor-induced increase in basicity is a prerequisite for His(235) to act as general base in the HbHNL-catalyzed cyanohydrin reaction.

Aldehyde-Lyases↗
PubMed2003

The micromorphology and protein characterization of rubber particles in Ficus carica, Ficus benghalensis and Hevea brasiliensis.

Rubber biosynthesis takes place on the surface of rubber particles. These particles are surrounded by a monolayer membrane in which the rubber transferase is anchored. In order to gain better insight into whether rubber particles from different plant species share common structural characteristics, the micromorphology of rubber particles from Ficus carica, Ficus benghalensis, and Hevea brasiliensis was examined by electron microscopy. Rubber particles of all three species were spherical in shape, and the size of rubber particles of H. brasiliensis was much smaller than those of F. carica and F. benghalensis. In addition, investigations were undertaken to compare the cross-reactivity of the antibody raised against either the H. brasiliensis small rubber particle protein (SRPP) which is suggested to be involved in rubber biosynthesis, or the cis-prenyltransferase (CPT) which has an activity similar to rubber transferase. Both western analysis and TEM-immunogold labelling studies showed that rubber particles of F. carica and F. benghalensis do not contain the SRPP. None of the rubber particles in F. carica, F. benghalensis and H. brasiliensis contained the CPT, suggesting that the CPT itself could not catalyse the formation of high molecular weight rubber. These results indicate that rubber particles in the three different plant species investigated share some degree of similarity in architecture, and that the SRPP and CPT themselves are not the core proteins necessary for rubber biosynthesis.

Blotting, Western↗
PubMed2005

The rubber tree (Hevea brasiliensis Muell. Arg.) homologue of the LEAFY/FLORICAULA gene is preferentially expressed in both male and female floral meristems.

The rubber tree (Hevea brasiliensis Muell. Arg.) is an important source of natural rubber in tropical regions and, as with many woody species, shows a long juvenile phase. To understand the genetic and molecular mechanisms underlying the reproductive process in rubber trees, H. brasiliensis RRIM600 flower and inflorescence development have been characterized, the rubber tree FLORICAULA/LEAFY (FLO/LFY) orthologue, HbLFY, cloned, and its expression patterns were analysed during vegetative and reproductive development. The rubber tree, similar to other Euphorbiaceae species, produces lateral inflorescences containing male, female, and bisexual flowers. HbLFY is expressed in lateral meristems that give rise to inflorescences and in all flower meristems, consistent with a role in reproductive development. Complementation studies using Arabidopsis lfy mutants indicated that the biological function of LFY might be conserved among Brassicaceae and Euphorbiaceae species.

Arabidopsis↗
PubMed2003

Laser diffraction: A new tool for identification and studies of physiological effectors involved in aggregation-coagulation of the rubber particles from Hevea latex.

Latex coagulation is the main limiting factor of rubber yield in Hevea brasiliensis (rubber tree). Using laser diffraction, we set up and optimized a new method for monitoring the kinetics of rubber particle (RP) aggregation, a prerequisite for latex coagulation. In contrast to any previous method used, laser diffraction allows continuous monitoring changes in size of RP aggregates, thereby allowing characterization and quantification of the processes involved in latex coagulation. Using this technique, we confirm that RP aggregating factors are proteins compartmentalized within latex cell vacuoles (lutoids), which, especially at relatively acidic physiological pH, can induce formation of RP aggregates large enough to induce plugging of severed latex vessel extremities. Conversely, latex cytosol was found to harbor anti-aggregating proteins. Further, we were able to titrate the RP-aggregating efficiency of the intralutoidic serum and the anti-aggregating efficiency of the cytosol. Preliminary assays showed that these two parameters were correlated with the yield potential of the tested rubber clones. This method will allow identification and characterization of proteins involved in latex coagulation, hence in rubber yield. We suggest that laser diffraction could be used to monitor the kinetics and characterize the physiological processes involved in aggregation of any particles, organelles or cells.

Chemical Precipitation↗
PubMed2003

Growth, photosynthetic performance and shade adaptation of rubber (Hevea brasiliensis) grown in natural shade.

We compared growth, photosynthetic performance and shade adaptation of rubber (Hevea brasiliensis Muell. Arg.) plants growing in natural shade (33, 55 and 77% reduction in incoming radiation) to control plants growing in full sunlight. Stem diameter and plant height, measured over a 15-month period, were greatest in plants grown in full sunlight, and both parameters decreased with increasing shade. At 7 and 14 months after planting (MAP), total plant dry mass was highest in control plants and lowest in plants in 77% shade. Expansion of the fourth leaf whorl, monitored at 5-6 MAP, was slowest in plants in 77% shade and fastest in unshaded plants, which had more leaves and higher leaf areas and inter-whorl shoot lengths. In response to increasing shade, specific leaf area increased, whereas leaf weight ratio and relative growth rate decreased. Chlorophyll a/b ratio decreased with increasing shade, indicating shade-induced partitioning of chlorophyll into light-harvesting complexes. Compared to the response in unshaded plants, CO2 assimilation saturated at lower photosynthetic photon flux densities in plants in 77% shade, with a lower upper-asymptote to the light response curve. Chlorophyll fluorescence revealed no evidence of sustained photoinhibitory damage in unshaded plants. Dynamic photoinhibition decreased with increasing shade, with the greatest depression in the ratio of variable to maximal fluorescence around midday. We conclude that shade adaptation and shade-induced reductions in dynamic photoinhibition account for the enhanced early growth of rubber in light shade.

Adaptation, Physiological↗
PubMed2005

Induction and differential expression of beta-1,3-glucanase mRNAs in tolerant and susceptible Hevea clones in response to infection by Phytophthora meadii.

Most cultivated rubber tree (Hevea brasiliensis Willd. ex A. Juss.) clones in India are susceptible to abnormal leaf fall disease (ALF), which is caused by various Phytophthora species and results in yield losses of up to 40%. Because the conventional breeding programs for this perennial tree crop are complex and time consuming, we attempted to find a molecular solution to increase the tolerance of rubber trees to ALF. The expression patterns of the gene coding for the pathogenesis-related beta-1,3-glucanase (beta-glu) enzyme in a tolerant (RRII 105) and a highly susceptible (RRIM 600) clone of rubber tree were examined, following infection with ALF-causing Phytophthora meadii McRae. Infected leaf samples were collected at different times after inoculation, and RNA was extracted and subjected to Northern blot hybridization and reverse transcriptase polymerase chain reaction (RT-PCR). On hybridization with a 1.25 kb beta-glu probe, Northern blots showed a marked increase in beta-glu transcript levels in both clones 48 h after inoculation. However, compared with the susceptible RRIM 600 clone, the tolerant RRII 105 clone had a higher rate of increase and a more prolonged induction, with beta-glu transcript levels remaining high for 4 days after inoculation. In RRIM 600, the mRNA levels decreased significantly 48 h after inoculation. On re-hybridization with an 18S rRNA probe, uniform signals were detected in all the lanes, indicating that an equal amount of total RNA was present in all samples. Similar results were obtained in relative quantitative RT-PCR experiments with the housekeeping actin gene as an internal control. Thus, although induction of the beta-glu gene occurred in both tolerant and susceptible clones, the predominant difference between clones was in the intensity and duration of the response. The tolerance of clone RRII 105 may be associated with the prolonged expression of the gene following infection. The antifungal activity of these hydrolase enzymes makes them rational candidates for overexpression by genetic transformation to produce disease resistant crops.

Enzyme Induction↗
PubMed2006

Effect of tapping activity on the dynamics of radial growth of Hevea brasiliensis trees.

Rubber tree (Hevea brasiliensis Müll. Arg.) radial growth dynamics were monitored with displacement sensors, together with latex production, to investigate three aspects of the dual production of latex and wood: (1) the usefulness of fine-scale dendrometric measurements as a physiological tool to detect water shortage through radial growth; (2) the dynamic aspects, both at the seasonal and at the multi-year scale, of the competition between latex and wood production; and (3) the spatial distribution of radial growth rates around the tapping cut. Radial growth of untapped control trees started with the onset of the rainy season and lasted until the onset of the dry season, ceasing completely during the driest period. Displacement sensors provided a sensitive means of detecting water shortage, with a clear correlation between diameter variations and changes in water availability (both daily evapotranspiration and monthly rainfall) over the whole annual cycle. However, the correlation was significantly disturbed in tapped trees. After resumption of tapping, the radial growth rate dropped sharply within two weeks and the effect persisted throughout the whole season, so that the cumulative growth of tapped trees was about half that of untapped trees, with the cumulative growth deficit reaching 80% for the period from mid-June to November. This long-known negative impact of tapping on growth was much stronger in the second year of tapping than in the first, whereas latex production increased significantly between the first and second year of tapping. The increased latex production, which could not be ascribed to climatic conditions, shows that the establishment of an artificial latex sink is a progressive, long-term process likely involving many aspects of metabolism. As expected, ethylene significantly increased latex production in both years; however, ethylene had no effect on the growth rates of tapped trees. Radial growth was differentially affected at different locations around the tapping cut, with growth rates significantly lower in the tapped panel than in the untapped panel, and higher above the cut than below the cut. Thus, caution is needed when deriving whole stem wood production from girth measurements at one location on the stem, especially from girth measurements made close to the tapping cut. This also provides new evidence for the location of the latex regeneration area in the tapped panel, below the cut.

Agriculture↗
PubMedDate not supplied

Allergen cross-reactivity between proteins of the latex from Hevea brasiliensis, seeds and pollen of Ricinus communis, and pollen of Mercurialis annua, members of the Euphorbiaceae family.

Allergen cross-reactions among three strongly sensitizing Euphorbiaceae species, i.e., the rubber tree (Hevea brasiliensis), castor bean (Ricinus communis), and the Mediterranean weed Mercurialis annua were studied in Finnish patients (n = 25) allergic to natural rubber latex (NRL), but with no known exposure to castor bean or M. annua, and French patients allergic to castor bean (n = 26) or to M. annua (n = 9), but not to NRL. In immunoglobulin E (IgE)-immunoblotting, 28% of NRL-allergic patient sera recognized castor bean seed and 48% reacted to castor bean pollen proteins. Likewise, 35% of the NRL-allergic patient sera bound to M. annua pollen allergens. Nineteen percent of castor bean-allergic patients showed IgE to NRL and 8% to M. annua proteins. Sera from patients allergic to M. annua reacted in 44% to NRL, in 56% to castor bean seed, and in 78% to castor bean pollen proteins. In immunoblotting, castor bean seed extract inhibited the binding of NRL-reactive IgE to 20 kDa, 30 kDa of NRL, and 55 kDa of proteins; NRL extract, in turn, inhibited the binding of castor bean-reactive IgE to 14, 21-22, 29, and 32-34 kDa of castor bean proteins. In ELISA inhibition, NRL extract inhibited 33% of the binding of M. annua--reactive IgE of pooled sera to M. annua pollen. In conclusion, allergen cross-reactivity in vitro was observed among three botanically related Euphorbiaceae members, H. brasiliensis, R. communis, and M. annua, but the molecular specificity of the observed cross-reactions as well as their clinical significance remains to be elucidated. Allergen cross-reactivity should be taken into account in diagnostic work.

Adolescent↗