Immunodetection of Pectic Epitopes, Arabinogalactan Proteins, and Extensins in Mucilage Cells from the Ovules of Pilosella officinarum Vaill. and Taraxacum officinale Agg. (Asteraceae).

The main aim of this study was to compare the cytological difference between ovular mucilage cells in two Asteraceae species-Pilosella officinarum and Taraxacum officinale-in order to determine whether pectic epitopes, arabinogalactan proteins, or extensins are present. The immunocytochemical technique was used. Both the Taracacum and Pilosella genera have been used recently as models for understanding the mechanisms of apomixis. Knowledge of the presence of signal molecules (pectic epitopes, arabinogalactan proteins, and extensins) can help better understand the developmental processes in these plants during seed growth. The results showed that in Pilosella officinarum, there was an accumulation of pectins in the mucilage, including both weakly and highly esterified pectins, which was in contrast to the mucilage of Taraxacum officinale, which had low amounts of these pectins. However, Taraxacum protoplasts of mucilage cells were rich in weakly methyl-esterified pectins. While the mucilage contained arabinogalactan proteins in both of the studied species, the types of arabinogalactan proteins were different. In both of the studied species, extensins were recorded in the transmitting tissues. Arabinogalactan proteins as well as weakly and highly esterified pectins and extensins occurred in close proximity to calcium oxalate crystals in both Taraxacum and Pilosella cells.

Allergens of weed pollen: an overview on recombinant and natural molecules.

Weeds represent a botanically unrelated group of plants that usually lack commercial or aesthetical value. Pollen of allergenic weeds are able to trigger type I reactions in allergic patients and can be found in the plant families of Asteraceae, Amaranthaceae, Plantaginaceae, Urticaceae, and Euphorbiaceae. To date, 34 weed pollen allergens are listed in the IUIS allergen nomenclature database, which were physicochemically and immunologically characterized to varying degrees. Relevant allergens of weeds belong to the pectate lyase family, defensin-like family, Ole e 1-like family, non-specific lipid transfer protein 1 family and the pan-allergens profilin and polcalcins. This review provides an overview on weed pollen allergens primarily focusing on the molecular level. In particular, the characteristics and properties of purified recombinant allergens and hypoallergenic derivatives are described and their potential use in diagnosis and therapy of weed pollen allergy is discussed.

Immunotoxic sesquiterpene lactone from Carpesium rosulatum Miq.

The whole plants of Carpesium rosulatum were chloroform extracted and the isolated sesquiterpene lactones and immunotoxicity effects were studied. The structures and stereochemistry of these compounds were established on the basis of analysis of spectra including mp, [α](D)(25), IR, UV, EI-MS, MS, (1)H-NMR, (13)C-NMR and some chemical transformations as follows: 1 (4ß,10α-dihydroxy-guaia-8α,12-olide), 2 (4ß,10α-dihydroxy-1(2),11 (13)-guaiadien -8α,12-olide), 3 (3ß,8ß-dihydroxy-1α,5α-guaian-10(14)-ene-6α,12-olide). 4 (2ß,5-epoxy-5,10-dihydroxy-6α,9ß-diangeloyloxy-germacran-8α,12-olide) The chloroform extracted had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L with an LC(50) value of 13.11 ppm and an LC(90) value of 20.33 ppm. The results could be useful in search for newer, safer, and more effective natural immunotoxicity agents against A. aegypti.

Study of pro- and anti-inflammatory cytokine profile in the patients with parthenium dermatitis.

BACKGROUND: Parthenium dermatitis is a common airborne allergic contact dermatitis induced by exposures to the weed Parthenium hysterophorus. The disease manifests as itchy erythematous papules, papulovesicular and plaque lesions on exposed areas of the body. OBJECTIVES: The aim of this study was to show the alterations in pro/anti-inflammatory cytokines in parthenium dermatitis. METHODS: The study included 50 patients with parthenium dermatitis confirmed by patch testing using aqueous extracts of P. hysterophorus and 50 age-matched healthy controls. The levels of pro-inflammatory [tumour necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-17] and anti-inflammatory (IL-4 and IL-10) cytokines were estimated by commercially available high sensitivity enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: All the dermatitis patients showed significantly (P < 0.001) elevated levels of TNF-α, IL-6, IL-8, and IL-17 levels as compared to healthy controls. In contrast, the anti-inflammatory cytokine IL-4 showed an insignificant decrease (P < 0.217) and a decrease in level of IL-10 was statistically significant (0.001) compared with controls. CONCLUSIONS: The present study suggests the involvement of pro-inflammatory cytokines in the pathogenesis of parthenium dermatitis. A decrease in levels of anti-inflammatory cytokines was demonstrated, which could not downregulate pro-inflammatory cytokines in parthenium dermatitis.

Anti-allergic effects of fermented Ixeris sonchifolia and its constituent in mice.

To evaluate the antiallergic effect of fermented Ixeris sonchifolia (IS, family Compositae), we prepared IS Kimchi, isolated Lactic acid bacteria (LAB) from it, fermented IS with these LAB, and investigated their antiallergic effects. IS Kimchi more potently inhibited the passive cutaneous anaphylaxis (PCA) reaction induced by an IgE-antigen complex as well as the scratching behavior induced by compound 48/80 or histamine than IS. When IS was fermented with LAB isolated from IS Kimchi, its antiallergic effects was also increased. Of LAB used for fermentation, Lactobacillus brevis more potently increased the antiallergic effects. Its main constituents, chlorogenic acid and luteolin potently inhibited PCA reaction induced by IgE-antigen complex as well as pruritus induced by compound 48/80 or histamine. These constituents inhibited the expression of proinflammatory and allergic cytokines, TNF-alpha and IL-4, and transcription factor, NF-kappaB, activation induced by IgE-antigen complex in RBL-2H3 cells, as well as the degranulation of RBL-2H3 cells induced by an IgE-antigen complex. Luteolin more potently inhibited these allergic reactions than chlorogenic acid. These findings suggest that antiallergic effect of IS can be increased by LAB fermentation and fermented IS might improve allergic reactions, such as pruritus, anaphylaxis, and inflammation.

Asteraceae species as potential environmental factors of allergy.

The statistics from Europe and the USA have proven a high risk for skin diseases associated with plant contact. Therefore, plant-induced dermatitis is of increasing attention in dermatology. The focus of this paper was to present the current knowledge on aspects of contact allergy related to Asteraceae (Compositae) species. The Asteraceae family is one of the largest in the world with members across all continents. The PubMed/Medline databases have been searched. The Asteraceae representatives consist of diverse secondary metabolites, which exhibit various advantageous effects in humans. In particular, sesquiterpene lactones (SLs) may cause sensitization resulting in skin irritation and inflammation. In this study, we tried to reveal the allergenic potential of several Asteraceae species. The Asteraceae-related allergy symptoms involve eczema, hay fever, asthma, or even anaphylaxis. Furthermore, the evidence of severe cross-reactivity with food and pollen allergens (PFS) in patients sensitive to Asteraceae allergens have been announced. Further identification and characterization of secondary metabolites and possible allergens in Asteraceae are necessary for the better understanding of Asteraceae-related immune response. The Asteraceae allergy screening panel (the SL mix and the Compositae mix of five plant species) is a promising tool to improve allergy diagnostics and therapy.

Allergenicity and cross-reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach.

BACKGROUND: The genus Senecio is the largest genus of the family Asteraceae (Compositae). The allergenicity of Senecio has not been assessed previously. OBJECTIVE: The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance. METHODS: Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one-dimensional electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross-reactivity with other pollen. RESULTS: Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human-allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two-dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross-reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies. CONCLUSION: S. jacobea constitute a newly discovered allergenic source. It shows cross-reactivity with other members of the Asteraceae plant family as well as with P. judaica.

Immunogold electron microscopic localization of the 2 EF-hand calcium-binding pollen allergen Phl p 7 and its homologues in pollens of grasses, weeds and trees.

BACKGROUND: The 2 EF-hand calcium-binding allergen from timothy grass pollen, Phl p 7, contains the majority of relevant IgE epitopes among calcium-binding allergens occurring in pollen species of different plants. OBJECTIVE: To describe the ultrastructural localization of Phl p 7 allergen in timothy grass pollen and its homologues in a broad spectrum of allergologically relevant pollens from grasses (timothy grass, rye grass), trees (birch, alder, olive) and weeds (mugwort, ribwort, ragweed) commonly growing in Europe. MATERIALS AND METHODS: Mature pollens from 8 different plant species were collected and anhydrously prepared for transmission electron microscopy. In ultrathin sections, allergens were localized using an antibody prepared against a Phl p 7-derived peptide comprising the C-terminal half of the Phl p 7 wild-type molecule in combination with a secondary antibody coupled to 10-nm colloidal gold particles. RESULTS: Phl p 7 and Phl p 7 homologues were detected in pollen from each of the 8 pollen species investigated. The allergens were found in the cytoplasm of the pollen grains (cytoplasmic matrix, mitochondria, nuclei) and in the pollen wall (preferably the exine). Reserve materials were unlabeled. CONCLUSIONS: The 2 EF-hand calcium-binding allergen Phl p 7 from timothy grass and its homologues can be localized in all pollen species under investigation. This finding confirms that Phl p 7 is a marker allergen for sensitization of patients to a novel family of 2 EF-hand calcium-binding pollen allergens occurring in a number of important allergenic plants in Europe.

Lactic acid bacterial fermentation increases the antiallergic effects of Ixeris dentata.

Ixeris dentata (ID, family Asteraceae), called Seumbakuy in Korea, was fermented with lactic acid bacteria (LAB) and their antiallergic activities were investigated. Fermentation of ID with Bifidobacterium breve or Lactobacillus acidophilus increased its inhibition of degranulation in RBL-2H3 cells induced by the IgE-antigen complex. Oral administration of these extracts to mice inhibited the passive cutaneous anaphylaxis (PCA) reaction induced by the IgE-antigen complex and scratching behaviors induced by compound 48/80. The fermented ID more potently inhibited the PCA reaction and scratching behaviors than the non-fermented one. These extracts also inhibited mRNA expression of TNF-alpha and IL-4, as well as NF-kappaB activation in RBL-2H3 cells induced by the IgE-antigen complex. These findings suggest that LAB fermentation improves ID-mediated inhibition of IgE-induced allergic diseases such as rhinitis and asthma, and that ID works by inhibiting degranulation and NF-kB activation in mast cells and basophils.

Associations among pollen sensitizations from different botanical species in patients living in the northern area of Madrid.

OBJECTIVE: To determinate the existence of associations among sensitizations to antigens produced by pollen grains of different botanical species as assessed by skin prick tests in patients with respiratory disorders. METHODS: Six hundred twenty nine consecutive patients living in the northern area of Madrid who underwent clinical evaluation because of rhinoconjunctivitis, and/or asthma were studied. All patients were tested with a skin prick test using a battery of inhalants including pollens, dust mites, molds and danders. The exploratory multivariate technique of Multiple Correspondence Analysis was used to compare the homogeneity of sensitizations between groups. Of the 629 patients, 459 (73.0%) had positive skin prick tests to pollen and were selected as the study group. RESULTS: The most prevalent pollen sensitization was to Gramineae pollen (83.7%) followed by Oleaceae sensitisation (75.8%). Multiple Correspondence Analysis revealed the existence of an association among pollen sensitizations, showing that they clustered two groups: sensitizations to Gramineae, Oleaceae, Cupressaceae, Chenopodiaceae, Plantaginaceae (group I), and sensitizations to Betulaceae, Platanaceae, Compositae (group II). Sensitization to Parietaria was not included in any of the sensitization groups and showed an independent behaviour. CONCLUSION: Pollen sensitizations in our area cluster into two association groups which have not previously been reported.

Susceptibility of gerbera and chrysanthemum varieties (Gerbera jamesoni and Chrysanthemum morifolium) to Fusarium oxysporum f.sp. chrysanthemi.

In 2002, gerbera plants (cv Kaliki) were observed exhibiting symptoms of a wilt in a soilless cultivation at Albenga area (Northern Italy). A similar wilt was also observed in the Sanremo area (Northern Italy) on cv Red Bull, Anedin and Gud finger grown in soil. The same observations were carried out in 2004 in SW Spain where gerbera plants showing wilt symptoms were observed in soilless crops. In all cases, the planting material originated from the Netherlands. Recently on the base of experimental trials F. oxysporum f. sp. chrysanthemi was recognized as the causal agent of wilts of gerbera both in Italy and in Spain. The aim of this experimental work was the evaluation of the resistance/susceptibility of available cultivars of chrysanthemum and gerbera to the Fusarium wilt. The pathogenicity of two isolates of Fusarium chrysanthemi obtained from infected gerbera (Gerbera jamesonii) and chrysanthemum (Chrysanthemum morifolium) plants was tested on several varieties both of gerbera and chrysanthemum in 2004-2006. In 2004 and 2005 respectively 54 and 30 cultivars of chrysanthemum and 57 and 55 of gerbera were tested, while in 2006 only 53 cultivars of gerbera were tested. The results showed that respectively in 2004 and 2005 67 and 33 % of chrysanthemum cultivars were highly resistant to F. chrysanthemi obtained from chrysanthemum while 57 and 53 % were highly resistant to strain isolated from gerbera. In 2004, 2005 and 2006 47, 65 and 75 % of gerbera cultivars were highly resistant to F. chrysanthemi obtained from chrysanthemum and 48, 56 and 72 % were highly resistant to the strain isolated from gerbera.

Anti-inflammatory and antioxidant potential of Guaianolide isolated from Cyathocline purpurea: Role of COX-2 inhibition.

BACKGROUND: Inflammation activated by oxidative stress can cause various diseases, such as asthma, rheumatoid arthritis, cancer, diabetes, etc. Plant constituents with sesquiterpene lactones possess antioxidant and anti-inflammatory properties. AIM: To determine the antioxidant and anti-inflammatory potential of isolated phytoconstituent from Cyathocline purpurea Buch-Ham ex D (CP). Don in laboratory animals. Furthermore, to understand the interactions involved in the binding of this compound to cyclooxygenase-2 (COX-2) via computational docking. METHODS: Phytoconstituent was isolated, purified and well characterized (using IR, NMR, and MS) from ethyl acetate fraction of CP methanolic extract. It was then evaluated for its in-vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) and hydroxyl (OH) radical assays as well as in-vivo anti-inflammatory potential against carrageenan-induced paw edema model in rats. The molecular docking study was performed against the crystal structure of COX-2 to evaluate the binding potential of phytoconstituent towards this enzyme. RESULTS: The isolated compound 6α-hydroxy-4 [14], 10 [15]-guainadien-8α, 12-olide (HGN) showed significant (p<0.001) antioxidant activity with IC50 values of 76µg/mL. Administration of HGN (10 and 20mg/kg) significantly (p<0.001) reduced the increased paw volume after subplantar administration of carrageenan. It also exhibits good binding affinity towards with COX-2 with a docking score of -8.98 and Glide binding energy of -36.488kcal/mol shedding light on the potential mechanism of anti-inflammatory action. CONCLUSIONS: The presence of hydroxyl group in HGN provides a credential to its in-vivo anti-inflammatory and in-vitro antioxidant activities. Furthermore, the good binding affinity of HGN for the active site of COX-2 may open novel vistas in therapeutic option with natural antioxidants like Cyathocline purpurea to treat various inflammatory disorders.

Characterization of polyclonal allergen-specific IgE responses by affinity distributions.

Polyclonal IgE responses have been previously characterized by allergen-specific antibody levels and by identification of amino acid sequences related to immunodominant epitopes. However, the binding affinities related to these antibody families are not well known. Using sera from donors with known sensitivities to ragweed or house dust mite allergens, we studied the binding reactions between the purified allergens Amb a 1 and Der p 1 and allergen-specific IgE's by determining affinity distribution functions. The distributions of binding affinities only exhibited a few dominant reactions indicated by peaks in an affinity distribution display. In all the donors tested, there were two dominant peaks and in 2/3 of the cases there was a third peak for both Amb a 1 and Der p 1. We further characterized the polyclonal interactions between IgE and Der p 1 by inhibiting the specific binding of IgE using peptide fragments known to be constituents of Der p 1 epitopes. Each peptide inhibited only a single peak in the affinity distributions. It would appear that the peaks in the affinity distribution represent antibodies directed to single epitopes. These results suggest that in our atopic population the response is surprisingly uniform. The bulk of the IgE response (70-80%) is of high affinity (10(8)-10(11) M(-1)) and directed towards a few epitopes. The relative affinities towards epitopes seem to be determined by the structure of the epitope and not variations of individuals' immune responses.

Expression of IgH chain genes in antibody response to ragweed and purified Amb a 1: evidence for differential regulation of isotype production.

A survey of the work with Ig response to allergens carried out previously reveals an allergen-specific response both by IgE and all of IgG subclasses. Response of non-sensitive people is characterized by the appearance of a variety of the IgG subclasses. We have reexamined ragweed and Amb a 1 specific Ig response in 54 nonsensitive and 147 atopic or atopic-allergic people using a new inverse sandwich immunoassay allowing discrimination based on antibody affinity. We show that non-sensitive people present no, 0 out of 54, Ig response with affinities higher than Ka 10(7) M(-1). The subpopulation of 66 atopics who never have experienced desensitization responds vigorously and solely (56 out of 66) with genes of the sequence gamma2-alpha2. Only ten showed an additional weak response from gamma1-alpha1. This suggests a possible association between the atopic state and selective activation of part of the gene sequence.

Performance of a commercially available plant allergen series in the assessment of suspected occupational contact dermatitis to plants in north Indian patients.

BACKGROUND: Parthenium hysterophorus is the leading cause of phytogenic allergic contact dermatitis in India. The Indian Standard Series currently supplied by Systopic Laboratories Ltd and manufactured by Chemotechnique Diagnostics ® contains parthenolide as the only allergen representing plant allergens. AIM: The study was conducted to assess the performance of the Chemotechnique plant series (PL-1000), consisting of 14 allergens, in patients with clinically suspected occupational contact dermatitis to plant allergens. METHODS: Ninety patients were patch tested with the Chemotechnique plant series from 2011 to 2013. Demographic details, clinical diagnosis and patch test results were recorded in the contact dermatitis clinic proforma. RESULTS: Of 90 patients, 24 (26.7%) showed positive reactions to one or more allergens in the plant series. Positive patch tests were elicited most commonly by sesquiterpene lactone mix in 19 (78.6%) patients, followed by parthenolide in 14 (57.1%), Achillea millefolium in 10 (42.9%) and others in decreasing order. CONCLUSION: The plant allergen series prepared by Chemotechnique Diagnostics is possibly not optimal for diagnosing suspected allergic contact dermatitis to plants in north Indians. Sesquiterpene lactone mix should replace parthenolide as the plant allergen in the Indian Standard Series until relevant native plant extracts are commercially available for patch testing.

Genetic endowment of proinflammatory cytokine tumor necrosis factor-α (-) 308 G > A polymorphism to Parthenium hysterophorus-induced airborne contact dermatitis in an Indian cohort.

BACKGROUND: Parthenium dermatitis is a common airborne allergic health problem that induces a cell-mediated hypersensitivity immune response involving activated T lymphocytes, which culminates in injury to the skin. The disease is manifested as itchy erythematous papules and plaques and primarily affects the exposed areas and flexures. This study aimed to identify the role of tumor necrosis factor (TNF)-α (-) 308 G>A polymorphism in the pathogenesis of parthenium dermatitis. MATERIALS AND METHODS: A total of 120 subjects, including 60 patients exclusively diagnosed for parthenium dermatitis and 60 healthy individuals, were included in the study. The genotyping of the TNF-α (-) 308 G>A region was carried out by the amplification refractory mutational system. RESULTS: In the present study, we demonstrated that polymorphism of the TNF-α (-) 308 position (A and/or G) was not statistically significant, and there was no difference in the distribution of any alleles of this locus in cases and controls. CONCLUSION: The present study suggests that there is a lack of association of potent proinflammatory cytokine TNF-α (-) 308 G>A polymorphism in parthenium dermatitis in the Indian cohort. It interprets genetically endowed transcriptional capacity due to this particular single nucleotide polymorphism but does not support the prevalence of high serum levels of TNF-α in parthenium-induced skin allergic inflammation.

The effect of 4α,5α-epoxy-10α,14-dihydro-inuviscolide, a novel immunosuppressant isolated from Carpesium abrotanoides, on the cytokine profile in vitro and in vivo.

The plant Carpesium abrotanoides (CA) is used in Asian herbal medicines as an insecticide and to treat bruises. However, the effect of single compounds from CA blooms and the mechanism of its immunosuppressive effect remain poorly understood. The aim of this study was to investigate the mechanism of the immunosuppressive effect in the three kinds of immune cells, and the immunosuppressive effect of CA bloom extract (CAE) in acute inflammation models (LPS and ConA-induced inflammation). Interleukin-6, IL-4, IL-13, IFNγ, and IL-10-but not TNFα-were significantly reduced in a dose-dependent manner by 4α,5α-epoxy-10α,14-dihydro-inuviscolide (INV). Furthermore, INV inhibited NF-κB transcriptional activation and IL-10 promoter activity in the same manner as for Bay11. Meanwhile, treatment with dexamethasone reduced the levels of IFNγ, but not IL-10, and resulted in no change in NF-κB transcriptional activation or the IL-10 promoter. INV did not affect PMA-induced IκB kinase complex phosphorylation, IκB degradation, or MAPK and the nuclear translocation of p65, as with DEX. The in vivo, CAE has an immunosuppressive effect on the LPS-induced inflammation response model by inhibiting the plasma level of IFNγ and IL-6 levels. CAE treatment also tends to attenuate the plasma level of IFNγ, IL-4, and IL-6 in ConA-induced inflammation. These findings indicate that INV causes the reduction of the cytokine profile by blocking the NF-κB transcription factor activation and the molecular mechanism by which INV operates could provide new insights into the unique mechanisms responsible for NF-κB inhibition, in contrast to established immunosuppressants, as a therapeutic agent for immunopathological treatment.