Release of antigens and allergens during shake‐culture of Aspergillus fumigatus

Harvey, C; Longbottom, Joan L.

doi:10.1111/j.1398-9995.1987.tb02222.x

Cited by 7 publications

(9 citation statements)

References 10 publications

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“…In the present study, we observed fewer IgE bands in the CF extract as compared to EN spore-mycelial extract, although the number of IgG binding bands were larger in the CF extract. This was in contrast to many earlier studies that have reported greater number of allergenic proteins in CF extract of A. fumigatus and F. solani as compared to spore-mycelial extract [23,7]. Fewer allergenic bands have also been observed in CF extracts of four different strains of Alternaria tenuis as compared to their sporemycelial extracts [24].…”

Section: Discussioncontrasting

confidence: 54%

“…The use of peptone and other natural ingredients for preparing CF antigens have been refuted, and the use of synthetic media is suggested [11]. The CF components of A. fumigatus, Fusarium species, C. albicans, and Penicillium notatum are more potent and are easily obtained by cultivating them in the synthetic medium [23,13,7]. The growth of EN in synthetic medium was poor [1].…”

Section: Discussionmentioning

confidence: 99%

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Culture filtrate antigens and allergens of Epicoccum nigrum cultivated in modified semi-synthetic medium

Bisht

Singh

Kumar³

et al. 2002

Med Microbiol Immunol

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Epicoccum nigrum (EN) is an important fungal allergen for nasobronchial allergy. Fungal extracts should contain all the relevant allergen components from spores, mycelium and culture medium for the purpose of allergy diagnosis and therapy. EN extract from spore-mycelial mass has been standardized, but the culture filtrate (CF) allergens of EN have not been studied as EN grows poorly in synthetic medium. The objective of the present study was to obtain a standard CF extract of EN by cultivating the source material in a modified semi-synthetic medium and to compare this with the EN cellular extract. Sabouraud's medium containing yeast extract (50 mg/l) was filtered using 10-kDa cut-off membrane and the lower molecular mass media components were used to cultivate EN. The CF obtained after removing the spore-mycelia was dialyzed to remove media components. The CF extract was characterized by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot. It was compared with EN spore-mycelial extract by enzyme-linked immunosorbent assay (ELISA), ELISA inhibition and by intradermal testing on allergy patients. The CF extract of EN resolved into 30 protein bands on SDS-PAGE. About 27 IgG bands were detected using anti-EN rabbit antibodies and 12 IgE bands by EN-sensitive pooled patients' sera. Periodate modification of CF proteins showed that the carbohydrate moieties are not important for IgE binding. Protein components of 26, 34 and 43 kDa were recognized as the major CF allergens. Three different batches of CF extract required 7.5-9 ng of self protein for 50% inhibition of binding to anti-EN rabbit antibodies in ELISA. Intradermal testing with CF extract showed comparable allergenic potency to standardized EN spore-mycelial extract, although it contained some allergenic proteins in higher amounts as compared to the spore-mycelial extract. In summary, the semi-synthetic medium has been suitably modified for obtaining EN CF antigens. This medium can be an important substitute for producing potent CF allergens of fungi that grow poorly in synthetic medium. The EN CF extract elicited good allergenic reactivity and may be used for allergy diagnosis along with spore-mycelial extract.

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Section: Discussioncontrasting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Culture filtrate antigens and allergens of Epicoccum nigrum cultivated in modified semi-synthetic medium

Bisht

Singh

Kumar³

et al. 2002

Med Microbiol Immunol

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“…This paper demonstrates that the 88-kDa antigen, which has recently been purified and shown to be specific for antibody detection in aspergillosis (3), is indeed the so-called chymotryptic antigen of A. fumigatus used for the detection of specific anti-Aspergillus antibodies by immunodiffusion or counterimmunoelectrophoresis (15). The gene coding for the 88-kDa antigen was cloned and shown to contain homologies with dipeptidyl-peptidase genes.…”

mentioning

confidence: 86%

“…The chymotrypsin activity of the precipitin had been defined only on the basis of a colorimetric reaction resulting from the release of naphthol radicals from the hydrolysis of the substrate N-acetylphenylalanine naphthyl ester (NAPNE) 1 (13). Purification of the chymotryptic antigen has been attempted by affinity chromatography on ⑀-aminocaproyltryptophan methylester-agarose (14) or by immunoaffinity using rabbit antiserum directed against a precipitin band recovered from a two-dimensional immunoelectrophoresis gel (15). Preliminary gel filtration chromatography coupled to rocket immunoelectrophoresis experiments identified a fraction reactive to the specific antibody, allowing purification of this "chymotryptic" antigen to homogeneity for the first time.…”

mentioning

confidence: 99%

Biochemical and Antigenic Characterization of a New Dipeptidyl-Peptidase Isolated from Aspergillus fumigatus

Beauvais

Monod

Debeaupuis

et al. 1997

Journal of Biological Chemistry

117

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A novel dipeptidyl-peptidase (DPP V) was purified from the culture medium of Aspergillus fumigatus. This is the first report of a secreted dipeptidyl-peptidase. The enzyme had a molecular mass of 88 kDa and contained approximately 9 kDa of N-linked carbohydrate. The expression and secretion of dipeptidyl-peptidase varied with the growth conditions; maximal intra-and extracellular levels were detected when the culture medium contained only proteins or protein hydrolysates in the absence of sugars. The gene of DPP V was cloned and showed significant sequence homology to other eukaryotic dipeptidyl-peptidase genes. Unlike the other dipeptidyl-peptidases, which are all intracellular, DPP V contained a signal peptide. Like the genes of other dipeptidyl-peptidases, that of DPP V displayed the consensus sequences of the catalytic site of the nonclassical serine proteases. The biochemical properties of native and recombinant DPP V obtained in Pichia pastoris were unique and were characterized by a substrate specificity limited to the hydrolysis of X-Ala, His-Ser, and Ser-Tyr dipeptides at a neutral pH optimum. In addition, we showed that DPP V is identical to one of the two major antigens used for the diagnosis of aspergillosis.

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“…30, 31], culture conditions as medium. pH, temperature and type of culture [9,32,33], time of incuba tion [32,[34][35][36] and storage [33], Similar variations prob ably also determine the antigenic composition of other fun gal extracts. This also means that the molecular weights of A. fumigatus antigens that contain determinants in common with other moulds may vary in different preparations of A. fumigatus antigens.…”

Section: Penicilliummentioning

confidence: 99%

Cross-Reactivity between Aspergillus fumigatus and Penicillium

Brouwer

1996

Int Arch Allergy Immunol

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Rabbit anti-Aspergillus fumigatus, rabbit anti-Penicillium and sera from patients with precipitating antibodies against A. fumigatus or Penicillium were analyzed by means of inhibition experiments in ELISA (IgG) and (or) immunoblot analysis (IgG). Sera from healthy donors were also analyzed in ELISA inhibition and sera from patients with allergic bronchopulmonary aspergillosis (ABPA) in RAST (IgE) inhibition. Most sera from patients with precipitins against Penicillium gave anti-A. fumigatus ELISA titers in the same range as sera from patients with aspergillosis. Anti-Penicillium IgG reacted with several A.fumigatus antigens with molecular weights between 28 and 130 kD. This reaction could be inhibited by the carbohydrate fraction of A. fumigatus and by extracts of related fungi. The binding to Penicillium of IgE antibodies in sera from patients with ABPA could be inhibited by A. fumigatus almost as effectively as by Penicillium. The binding of these antibodies to A. fumigatus was only slightly affected by Penicillium antigens. The presence of IgG antibodies that cross-react with A. fumigatus may strongly affect the immunoblot patterns and ELISA results obtained with sera from patients with aspergillosis. Penicillium spp. probably belong to the major stimuli which induce the synthesis of so-called naturally occurring antibodies to A. fumigatus.

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Release of antigens and allergens during shake‐culture of Aspergillus fumigatus

Cited by 7 publications

References 10 publications

Culture filtrate antigens and allergens of Epicoccum nigrum cultivated in modified semi-synthetic medium

Culture filtrate antigens and allergens of Epicoccum nigrum cultivated in modified semi-synthetic medium

Biochemical and Antigenic Characterization of a New Dipeptidyl-Peptidase Isolated from Aspergillus fumigatus

Cross-Reactivity between <i>Aspergillus fumigatus</i> and <i>Penicillium</i>

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