In vitro and in vivo biological activities of old and fresh Cupressus arizonica pollen.

BACKGROUND: Respiratory allergy to the pollen of Cupressaceae is becoming more and more common every year in the Mediterranean area. OBJECTIVE: The purpose of this study was to see whether the allergenic potency of Cupressus arizonica pollen diminished after a 6-year period (1994-2000). MATERIALS AND METHODS: Among the Cupressaceae, we selected the pollen of C arizonica. The mode of sampling in 1994 and in 2000 was the same and the pollen was collected on the same tree and stored at room temperature. To compare its biological and allergenic activities data was collected with the following methods: cytohistology of Alexander, 2,3,5-triphenyltetrazolium chloride enzyme staining, skin testing, nasal provocation test, radioallergosorbent test (RAST), RAST inhibition, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and immunoblotting to detect protein content. Thirty-eight patients with respiratory allergy to Cupressaceae were selected. RESULTS: We found no decrease in the allergenic potency of the pollen, but did find that viability and germinating power had disappeared completely after 30 to 40 days. Moreover, the amount of protein in the old pollen was half the amount found in the fresh one. Skin prick testing showed identical results with the old and the fresh pollens. CONCLUSIONS: The allergenic in vivo and in vitro activity of cypress pollen is retained for years after its collection. This activity seems to be independent of the viability of pollen grains and of the total protein content. This may explain the presence of clinical symptoms in patients out of the pollen season.

Occupational respiratory allergy to cyclamen pollen: a case report.

A case of cyclamen (Cyclamen persicum) pollen allergy is described on a flower cultivator. The diagnosis was assessed by skin test, methacoline test, nasal provocation test and RAST. SDS-PAGE and immunoblotting have shown an intense response against a 18 kDa component and a weaker one at 100 Kda. A RAST inhibition against Cupressus sempervirens and Acacia floribundia showed no identity with cyclamen. This case confirm the cyclamen allergenity, in professional exposure. The nasal challenge prove to be very useful to confirm allergic sensitizations an we emphasize the necessity of preventive measures of protection for those florists who cultivate cyclamen plants.

Efficacy of sublingual coseasonal immunotherapy with a monomeric allergoid in Cupressaceae pollen allergy--preliminary data.

BACKGROUND: Respiratory allergy to conifers pollens is a growing phenomenon all over the world (Mediterranean area, USA, Japan). In France and Italy Cupressus sempervirens is the commonest offender. OBJECTIVE: To check the efficacy of specific immunotherapy by the sublingual route using a monomeric allergoid of Cupressus Arizonica pollen. MATERIAL AND METHODS: 20 patients and 10 controls were randomly selected. The diagnosis was assessed on a clinical hivernal symptomatology of allergic rhinitis, and/or asthma, positive skin test and RAST, and nasal provocation test to Cupressus arizonica. The extract was a monomeric allergoid titrated in allergenic units (AU). The treatment was coseasonal from the beginning of december to the end of march. It was not conducted double blind. RESULTS: The evaluation of the allergenic potency of native and modified Cupressus arizonica extracts have shown that the modified extract has a decreased skin test IgE binding capacity. According to symptom, drug consumption scores and nasal provocation test, there was a statistical improvement in the active group (compared to the control group) without any side effect. DISCUSSION: The use of a modified Cupressus arizonica extract, which is easier to prepare than Cupressus sempervirens and which is a monomeric allergoid, has shown that by the sublingual route, this therapeutic approach is safe and that the first results were satisfactory without any side effect. This good tolerance was anticipated by the fact that, checked on 16 volunteers by skin test, the allergoid gave a less important local reaction on skin testing than the native extract. CONCLUSION: This study has shown that: The KCNO modified Cupressus arizonica extract has a reduced IgE binding capacity; This monomeric allergoid is safe and effective administrated by the oromucosal route in clinical respiratory allergy to Cupressaceae and devoid of any side effect.

Evaluation of cross-reactivity between Holoptelea integrifolia and Parietaria judaica.

BACKGROUND: Holoptelea integrifolia and Parietaria judaica belong to the family Urticaceae, but are geographically distantly located. H. integrifolia is an important pollen allergen of India and sensitizes almost 10% of the atopic population in Delhi. P. judaica, on the other hand, is a very dominant pollen allergen of the Mediterranean region, sensitizing almost 80% of the allergic population. Since both these important pollen allergens belong to the family Urticaceae, the objective of the present study was to assess cross-reactivity between these two pollen allergens from different geographical regions. METHODS: Cross-reactivity between these two pollen allergens was assessed on the basis of skin prick tests and ELISA, ELISA inhibition and immunoblot inhibition studies. RESULTS: Out of 44 atopic Indian patients skin prick tested with H. integrifolia extract, 34% were found to be sensitized. All the patients sensitized to H. integrifolia also showed varying degrees of skin positivity to P. judaica pollen extract. ELISA and ELISA inhibition studies suggested strong cross-reactivity between H. integrifolia and P. judaica pollen. Immunoblot inhibition studies revealed that 14-, 16-, 28-, 38-, 42- and 46-kDa proteins are the cross-reactive proteins in H. integrifolia and P. judaica. However, Par j 1, the major allergen of P. judaica, is absent in H. integrifolia pollen. CONCLUSION: H. integrifolia and P. judaica pollens share cross-reactive as well as unique epitopes. The major allergen of P. judaica, Par j 1, seems to be absent in H. integrifolia pollen allergen.

Cross-reactivities between date palm (Phoenix dactylifera L.) polypeptides and foods implicated in the oral allergy syndrome.

BACKGROUND: Date fruit and pollen antigens share a number of cross-reactive epitopes. Date pollen has been shown to cross-react with antigens from Artemisia, cultivated rye (Secale cereale), Timothy grass (Phleum pratense), Sydney golden wattle (Acacia longifolia) and Bermuda grass (Cynodon dactylon) pollen. The present study was carried out to examine any cross-reactivities between date palm polypeptides and antigens of some common foods and vegetables that have been implicated in the oral allergy syndrome (OAS). Because most of such cross-reactivities in other allergens are attributable to the presence of carbohydrate chains and profilin, their role was also investigated. METHODS: Fresh extracts of 20 common fruits and vegetables were prepared. Putative date profilins were isolated by affinity chromatography using a poly L-proline column. Date fruit extracts were digested by various endoglycosidases and the immunoglobulin (Ig)E binding of the postdigest products was assessed in immunoblots. Rabbit antisera to whole date fruit extracts, Timothy grass profilin and putative date profilins, as well as human sera from date sensitive individuals were used in immunoblotting, ELISA and in inhibition experiments. RESULTS: IgG, ELISA and immunoblot results with the different rabbit antisera and date-sensitive atopic sera showed several antigenic cross-reactivities and similar cross-reactivities were seen with birch, date and timothy grass profilins. IgE, ELISA and immunoblot experiments with pooled date sensitive human sera showed a range of cross-reactivities with some food extracts. A number of the IgE cross-reactivities could be inhibited after preabsorption of pooled sera with date extracts. Sixty-six percent of individual date hypersensitive human sera bound IgE in putative date fruit profilin and their pooled sera bound IgE in birch pollen profilin. IgE-binding of the endoglycosidase digested date fruit extracts to atopic serum pool was restricted to only a very low molecular weight band of 6.5-8 kDa. CONCLUSION: These results indicate that date palm polypeptides share cross-reactive IgG and IgE epitopes with a number of foods implicated in the oral allergy syndrome, bind to birch and Timothy grass profilins and bind IgE through glycosyl residues. The clinical relevance of these cross-reactivities needs to be further elucidated.

Efficacy of sublingual specific immunotherapy in Cupressaceae allergy using an extract of Cupressus arizonica. A double blind study.

BACKGROUND: non-injective routes of immunotherapy in respiratory allergy have being proposed as an alternative to conventional immunotherapy. We carried out a study to evaluate the clinical efficacy and effects sublingual immunotherapy (SLIT) in patient with Cupressaceae pollen respiratory allergy. METHODS: twenty patients with Cupressaceae pollen (C. sempervirens) rhinoconjunctivitis, mild asthma or both were randomly chosen for sublingual immunotherapy (10 patients) or placebo treatment (10 patients) using a double blind placebo controlled technique. We have used an extract of Cupressus arizonica because of its better stability in solution and its crossreactivity with Cupressus sempervirens. The patients underwent treatment for 12 months (from april 1999 to april 2000). Symptoms and drug scores as well as nasal provocation tests were recorded. The pollen counts were carried out, during all the same period. RESULTS: we found significantly lower symptom scores (p < 0.05) and drug consumption scores (p < 0.05) in the immunotherapy group than in the placebo group, during the pollen season, after the first year of therapy. Besides threshold of allergen reactivity in nasal provocation tests was increased in the active group (p < 0.01). No untoward reactions have been observed. CONCLUSIONS: SLIT with Cupressus arizonica (pollen extracts) produced, after one year treatment, a significant improvement of allergic symptoms and a decrease of drug consumption scores and allergen-specific nasal reactivity. SLIT with Cupressus arizonica appeared to be effective and safe in the treatment of Cupressaceae pollen respiratory allergy.

New clinical data and therapeutic prospects in Cupressaceae pollen allergy.

The authors emphasize, thanks to their clinical data over the last 20 years: The steady increase of allergy to C. sempervirens pollen compared to other pollinosis and to atopy. The frequency of cross-reactivity between C. sempervirens and other pollens belonging to Cupressaceae and Taxodiaceae families, in order, in the absence of a very good standardization and of a purification of this pollen, to try to undertake a sound specific immunotherapy using the most cross-reactive pollen mixed with C. sempervirens extracts. As a matter of fact the partially purified extracts of C. japonica gave the higher incidence (91%) of cross-reactivity. Consequently, a mixture of C. sempervirens and C. japonica extracts was used since 1993 with good results in a double blind study.

Allergy to date fruits: characterization of antigens and allergens of fruits of the date palm (Phoenix dactylifera L.).

BACKGROUND: Date-palm (Phoenix dactylifera L.) fruits are eaten daily by most inhabitants of the Middle East and the neighboring countries. Recent reports have indicated that dates are allergenic. This study aimed to investigate the antigenic and allergenic potential of date fruits. METHODS: Date-fruit extracts from eight cultivars were evaluated in skin prick tests (SPT) in an atopic population, used to produce antisera, analyzed by SDS-PAGE, and fractionated by gel-filtration chromatography. Sera from SPT-positive individuals were evaluated by ELISA and RAST, and in anti-igE immunoblot experiments. RESULTS: About 13% of patients were SPT-positive for at least two extracts. SDS-PAGE of whole extracts revealed 15-18 protein bands of 6.5->100 kDa, and Sephacryl S-200 fractions gave distinct peptide bands. RAST and anti-IgE ELISA gave a range of positive results, which could be abrogated when sera were preabsorbed with fruit extracts. IgE immunoblots of different extracts with pooled positive sera revealed different anti-IgE-binding immunoprints. All the positive sera from fruit-allergic and pollen-allergic individuals bound strongly to two anti-IgE reactive bands of 6.5 to 12-14 kDa and 28-33 kDa, respectively, and about 50% of sera bound to a 54-58-kDa band. CONCLUSIONS: These results strongly indicate that 1) date-palm fruit is a potent allergen 2) sera from fruit-allergic as well as pollen-allergic patients recognize common fruit-specific epitopes 3) there is heterogeneity in patient responses to the different extracts.

Efficacy and safety of oral immunotherapy in respiratory allergy to Parietaria judaica pollen. A double-blind study.

Two groups of 15 patients with respiratory allergy due to the pollen of Parietaria judaica were studied; one received a treatment of oral specific hyposensitization, the other one a placebo. All of them were monosensitized and clinically matched according to age, gender, score symptoms, drug consumption, skin test and RAST, and nasal or bronchial provocation test. The pollen counts were the same during this period. The study was double-blind and lasted 2 years. The clinical tests including score symptoms, drug consumption, and nasal or bronchial provocation test, revealed a statistically significant improvement in the treated group as compared to the placebo group. No side effects were observed.

Late onset asthma clinical and immunological data: importance of allergy.

In people over 65 years of age (mean age 72) late onset asthma (LOA) is often mistaken for chronic obstructive pulmonary disease (COPD). Furthermore, it is generally believed that true allergic asthma starts very seldom in old age and that, in this case, it is actually intrinsic asthma i.e., without any sensitization to aeroallergens. The aim of this study was to show that LOA is not an uncommon entity and is often allergic. Among a population of 1,888 subjects (over 65 years of age) in a general medicine ward we found 21 patients (1.1%) corresponding to this entity. The diagnosis of asthma was assessed according to clinical data, pulmonary function and nasal provocation tests, and blood eosinophilia. The diagnosis of atopy was assessed on total IgE, prick tests and RAST for the commonest allergens of our area. This group of 21 patients was compared to three groups of age-matched subjects including: 68 patients afflicted with chronic bronchitis, a group of 14 patients who had asthma before the age of 65 (presenile asthma), and 20 normal controls. A group of 15 asthmatics under age 65 was also included (young asthma mean age 37). We found a rather high incidence of allergy (72%), mostly to Dermatophagoides pteronyssinus, but also to the pollens Parietaria judaica, grasses and Olea europea. Although the average age (except in the young asthma group) and the severity of symptoms were the same in the four groups, unexpectedly mean RAST and skin test values were higher in LOA than in the two groups of young asthmatics and presenile asthmatics. It was concluded that LOA is not uncommon among elderly patients over 65, which is in contrast with most previous studies; 72% of our cases were allergic, the main allergens being mite, and Parietaria judaica pollen.

Ric c 1 and Ric c 3, the allergenic 2S albumin storage proteins of Ricinus communis: complete primary structures and phylogenetic relationships.

The 2S albumin storage protein of Ricinus communis consists of the two heterodimeric proteins Ric c 1 and Ric c 3 each of which is composed of a small and a large subunit linked together by disulphide bridges. The complete primary structures of both heterodimeric proteins were determined by enzymatic degradation and automated Edman degradation. The sequences of all four chains correspond to the known cDNA sequence of the gene of a presumed precursor molecule and to the previously determined partial sequences for Ric c 1 and Ric c 3. In addition, few differences in amino acid positions were found which seem to be related to different varieties of R. communis. Sequence comparisons with 2S albumin from other plant genera revealed high degrees of homology and support the view of a common genetic origin of this protein family. Ric c 1 and Ric c 3 which have 11,212 and 12,032 daltons, respectively, share a similar molecular size, biological function and allergenicity with the 2S albumins from Brassica juncea (Braj 1E) and Sinapis alba L (Sin a 1). Ric c 1 and Ric c 3 may be classified as isoallergens if, additionally, the high degree of similarity in the position of polar residues is taken into account.

Specific IgE to castor bean (Ricinus communis) pollen in the sera of clinically sensitive patients to seeds.

Human sensitization to castor bean seeds in occupational workers and people living close to oil processing factories has been acknowledged for a long time. In view of the cross-reactivity among different plant parts of the same species, we studies crossreactivity between seeds of castor bean and its pollen at the molecular level. Sera from 26 seed-positive atopics, when analyzed for ELISA against seed and pollen extracts of castor bean, showed binding with both seed and pollen extracts, but binding was stronger with seed extracts as compared to pollen. ELISA inhibition revealed partial similarity as pollen extract could not achieve 90% inhibition even at 100 micrograms/ml, and remained the same after protein concentrations of 40 micrograms/ml. Antigenic extracts of seeds and pollen separated into 12 and 20 fractions on SDS-PAGE, respectively. The 26 sera studied for specific IgE binding to different fractions of seed and pollen extracts showed IgE binding in 17 and 16 cases respectively, but with weak binding to pollen fractions. The crossreactivity was confirmed with pooled sera by blot inhibition. Seed antigen completely inhibited the sera for specific IgE at 10 mg/ml protein while pollen antigen showed only partial inhibition. Crossreactivity and presence of common epitopes between seed and pollen extracts are confirmed.

Double-blind placebo controlled specific immunotherapy with mixed Cupressaceae taxodiaceae pollens in respiratory allergy to Cupressus sempervirens.

Twenty adult patients presenting a respiratory allergy to the pollen of Cupressus sempervirens were randomly divided into two groups; one received a placebo the other one an equal mixture of Cupresssus sempervirens and Cryptomeria japonica pollen during a period of 3 years (1993-1996). According to the following criteria: symptoms score, drug consumption, skin test and RAST, nasal provocation test, the results for the treated group were statistically better than those for the placebo group. No side effects were observed.

Monitoring of specific IgG4 antibodies in respiratory allergy due to the pollen of Parietaria judaica. Evidence for a protective role.

Two groups of 20 patients having an allergic rhinitis due to the pollen of Parietaria judaica were studied, one received a treatment of specific hyposensitization, the other one a placebo. All of them were monosensitized. The study was carried out using double blind methodology and was conducted for one year. Each case was assessed by clinical tests: case history, skin tests, score symptoms, drug consumption, nasal provocation test and by in vitro tests (FAST technique): total and specific IgGE, specific IgG4. The clinical tests revealed a statistically significant improvement in the treated group as compared to those receiving the placebo. We found a statistically significant regular and important increase of specific IgG4 values in the treated group. In this case, the specific IgG4 antibodies most likely play a protective role and act as blocking antibodies.

Local intranasal immunotherapy with allergen in powder in atopic patients sensitive to Parietaria officinalis pollen.

Local Intranasal Immunotherapy (LII) is a new approach to the treatment of allergic rhinitis due to the pollen of Parietaria officinalis. The aim of this study was to verify the usefulness of LII and to determine the proper doses. Twenty adult patients all presenting a sensitization to the pollen of Parietaria officinalis were randomly divided into two groups: 15 received the treatment and five were the control group. Treatment started before the beginning of the pollinic season of Parietaria officinalis and continued during the season. The extract used was an active extract of macronized powder Parietaria officinalis pollen in increasing doses. Doses were determined periodically with specific nasal provocation tests. Results in the treated group compared to the control group were statistically significant if one considers the increase in threshold sensitivity in the nasal provocation test, the diminution of clinical symptomatology and the quantity of drug required to control symptoms in the treated group. The side effects were few; it was only in one case that the treatment had to be interrupted. LII for allergic rhinitis due to the pollen of Parietaria officinalis seems an effective and practicable method even during the pollinic season.

Screening for atopy in a coffee processing factory.

Screening for blood IgE mediated allergy (atopy) by the RAST technique among 76 people working in a coffee processing factory showed rather unexpected findings: although we found a prevalence of positivity (17.1%) to the common airborne antigens (pollens, mites, cat, Alternaria tenuis) which was close to the prevalence of atopy among normal adults in our area (19.3%), only one case of allergy to green coffee and two cases to castor bean came to the fore. Specific IgG4 antibodies were measured only for castor bean and green coffee, and rather elevated figures were found: green coffee 17.1%, castor bean 13.1%. The occurence of positive RASTs to castor bean is more likely to be due to contamination of the bags containing green coffee. The low prevalence of RAST positivities to green coffee, and the elevated specific IgG4 antibodies to both castor bean and green coffee antigens, raise several possibilities which are discussed; however, all the subjects but one having elevated specific IgG4 levels to green coffee worked in more exposed areas. Probably IgG4 antibodies in this particular case are acting as blocking antibodies.

Pollinosis in a Mediterranean area (Riviera Ligure, Italy): ten years of pollen counts, correlation with clinical sensitization and meteorological data.

We present in this study the results of 10 years of pollen counts in the atmosphere of San Remo, Italy. These results were compared with clinical sensitizations in 5481 pollen-allergic patients of both sexes (children and adults) living in the same area. Pollen counts showed that the pollens found in greatest quantities were the Cupressaceae (22.4%), followed by the Urticaceae (Parietaria officinalis, 17.4%), the Oleaceae (Olea europaea, 10.8%) and the Pinaceae (10.1%), the Poaceae being present in smaller quantities (5.2%). In contrast, the clinical importance of these pollens, as assessed by anamnesis, skin tests and, in some instances, nasal or bronchial provocation tests, showed a high incidence of clinical sensitization to P. officinalis (41.8%) and Poaceae (29.5%); the other pollens, i.e., O. europaea (15.4%) and Cupressaceae (5.7%), were of lesser importance. These findings differ from data obtained in similar studies in other European countries, including other parts of Italy. A statistically significant positive correlation between pollen counts and some meteorological factors, especially temperature, was observed; for instance, the higher the average temperature in July of the previous year, the greater the airborne presence of Palmaceae, P. officinalis and Cupressaceae.

Prevalence of patent and latent atopy among a general normal adult population in the south east of France by RAST investigation and correlation with circulating total IgE levels.

A population of 512 adults (men: 266, 52%, women: 246, 48%) with mean age 41.4 years (S.D. 10.7) was investigated. This population was subdivided into five groups: Marseilles city, Manosque a small town in a prealpine area, the area surrounding the Etang de Berre, a rural area in Northern Bouches du Rhône and Chateaurenard, a small town in an intensive agricultural area. RASTs to a panel of eight allergens (Lolium perenne, Parietaria officinalis, Cupressus sempervirens, Olea europea, Cat dander, Alternaria tenuis, Dermatophagoides pteronyssinus, and Castor bean seed) were done. 99 subjects were found to have a RAST positive to one or several allergens. The main positives found by RAST were a grass pollen (Lolium perenne) and the mite Dermatophagoides pteronyssinus with respectively 10.1% and 11.3% of the results followed by the pollens of Olea Europea 4.4%, Parietaria officinalis 3.1% and Cupressus sempervirens 1.9%, Castor bean seed 1.7% Cat dander 0.9% and Alternaria tenuis 0.9%. Allergy to cstor bean (Ricinus communis) seed was found mostly in the Chateaurenard area where castor castor bean pomace is used in the fields as a fertilizer. There is an almost equal number of RAST positivities to mites (13.3%) than to pollens (11.1%). The prevalence of RAST positive was higher in men (24.1%) than in women (14.2%), p < 0.01). This prevalence is lower when age is increasing. Thirty four subjects (6.6%) had clinical symptoms: This group has a higher prevalence of positive RAST in cases of allergic rhinitis (p < 0.01) and of asthma (p > 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)