Avaliação bioquímica, antigênica e alergênica de extractos brutos de Helminthosporium monoceras

Abstract

HeIiKineho-spors.usi aronacer-as, Dfffjst.erïjmyccït'ine subdivision -Fungus, is relatively -Frequent: in Brasil. And its dispersal is inade through atmospherical air. In previous íitudies per-Fonned in São Paulo city, it was veri-Fyed that the crude e-Kfcract -Fram that -Fungus has presented, in relation t:ü 42 -Fungus extracts, a tnajor cufcaneous reaction intensity in respiratory allergy patients. Based on this -Fact and in the non existence o-F data about this fungus in relavant literature, the purpose o-F the investigatioi) was estimating the crude extract cillergt.-tticila; anl igen icifc a and Biochemical composition, obtained by Tris-HCl in vegetative and conidial growth of slraiti used since Í973 in intradermic test -For dignostic o-F al l e?r sy . Ac.cording to the growth curve in Czapeck broth lïiodi-Fyed and wiacfo and inicromorphologic attendance -For ac qi-iiüií; ion o-F (.-rude extractü, the 4th day (logarithm and veyetïii ive phase); iôth day (estctcionary and vegetative' phase), iofch and 2£nd days (staic ionara and conidial phase) a.iid 28th day (declinina and conidial phase) of growth were chosen. 85 The maximuni values o-F total proteins and lipids were verifyed in the crude extract obtained in the 28th day o-F growth and maximum values of carbohidrats were observed in the extracts o-F the ioth, 22nd and 28th. The fractionated proteins by SDS-PAGE presented bands with molecular weights ü-F approximately between i4,4 to 67,0 KDa. The extracts obtained on the 4th and i0th days presented two bands of approxini.a.tely ó<S and 67 KDa, the ones obtained on the i6th and ££nd days, 5 bands of appruximatela i8,5, 30,0; 36,0; ó<&,0 and 67 KDa and the ones •From the 28th day, 7 bands, the previouü unes and two more with molecular weights of approximately 14,4 and 20,i KDa. The carbohidrats and dininoacids were characterized by Thin-Layer Cromatography, ctiid presented stains corresponding to sugars glucose, aã l ac t use and mannose, and the aminoacids asparagine, pfolirit, thyrosina, al an ine, leucine, isoleucine, f&'nilalanine, •v'aline, inethionine, trytophan, aicid slutamic, glycine, histidine e cystine. The antigenicity o-F the crude t-KÍrací.s was veti-Faecl by Itumunocl i-Pusion reaction in agar against i-abbifc hiperimmune sera. Precipitation lines were; observed in all studied extracts and common antigenic molecular populations. Finally, the allergenity o-F crude exfcracL obtained art the 28th day o-f growth was con-Firmed by lyE antibody pruductioit in isogenic mice immunized by the saine, and titled by Passive Cutaneous Anaphylaxis test in rat's. The maximum titles obtained were 80 and i60 in cAF-i «lice- and i.280 and 64^ in BALB/c mice.