Antimicrobial lytic factors of the coelomic fluid of Earthworms

Abstract

I. SUMMARY lnvertebratesare widely distrbuted animals.They can be found in almostany krnd ofhabitat. Their successfulsurvival shategiesare basedon short life span combinedwith numerousoffspring and, more importantly,all invertebratespecieshavedevelopeda wriety ofdefensemechanismselficientlyrecognizingand respondingto non-selfsubstances. The defensemechadsmsof earthwoms were studiedover the pastfou decades.It becameappdent thattheearthworms.aswell asotherinvertebrates,lackspecificimmunoglobulins,lymphoc)4esor otherfeatures ofthe adaptiveimmunesystemdescribedin verteblates,but possessirmatedefensecomponents In this thesis,we foousedon the detaildescriptionof somedefensemoleculesinvolvedin imate immunityof earthworms. l. Lysozyme is an erzyme with strong antibacterialactivity describedin many organisms. We characterizedthelysozymeof Erieniaandrei(formerly E.fetida andrei)earthwormbothstruchmlly and functionally.Moleoularcharactefiationof lyso4rne providesa new tool for monitoringof innateimmunity in earthworms. 2. A cltol''tic effect of the coelomicfluid of E. fetida was observedin experimentswith TNF-sensitive tumorL929 cell line. Subsequentisolationof l)tic proteinsled to theidentificationof 42-kDaprotein,which wasnamedcoelomiccytolyticfactor- CCF.CCFwasshom to bepresentalsoin coelomicfluid of mother...

2. SOUHRN Bezobratli jsou Siroce rozslienou skupinou iivodichri tijicich ve velice rodidnfch podminkdch. StrategiepfeZitije u bezobratlfchd6najak kdtkou dobouZivotaa veL[y'mmnoZstvlmpotomstva,tak piedeviim eistenci mnoZstvinejnlzrdjllch obrannjchmechanizrndridinndrozpoa{vajicich a eliminujicichcizorodCldtky. Objektemz6jmu imunologu se obrann6mechanizmylaoulkovcri staly vposlednich dtyficeti letech. U kroutkovcrl, stejnEjako u ostatnichbezobratlfch, nebyla zjiltdna pFltomnostimunoglobulinri,llmfocytri, ad jinfch sloZekadaptivniimunity. Ve svd disertadniprAci se v6nuji detailnlmupopisundktenfchobrannlch molekul piluozenCimunity kouZkovcri. 1. Lysorym je erzym sesilnouantibakteridlnlaktivitou,jehoi piitomnostbylapopsdnau iady organizmt. My jsme detekovalimolekululysozymuu ilrdy Eiseniaandrei (dftlveE.fetida anfuei) na rlrovnistruktury afunkce.Molekuldmicharakterizacetohotoproteinuumohluje sledov6nipfirozeni imunity u ffZal. 2. Cytolyticky efektcoelomovCtelr.nnyE.fetidabyl popsdnv experimentechs TNF-senzitivnlnedorovou bunddnoulinii L929. Ndsledn6izolacelytickich molekul vedlak identifikaci proteinuo velikosti 42-kDa, kter! byl nazv{ncoelomovjm cytolytickfm faktorem - CCF.Bylo zji5tdno,ZemolekulaCCFje piitomna tak6v coelomov6tekutinddalSihokouikovce - Lumbricastetestis. PfitomnostmolekulpodobnfchCCF byla vt6to...