velikost textu

Antimicrobial lytic factors of the coelomic fluid of Earthworms

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Název:
Antimicrobial lytic factors of the coelomic fluid of Earthworms
Typ:
Disertační práce
Autor:
RNDr. Radka Josková, Ph.D.
Školitel:
RNDr. Martin Bilej, DrSc.
Oponenti:
prof. RNDr. Petr Horák, Ph.D.
prof. MUDr. Ludmila Prokešová, CSc.
Id práce:
82728
Fakulta:
Přírodovědecká fakulta (PřF)
Pracoviště:
Katedra buněčné biologie (31-151)
Program studia:
Imunologie (P1517)
Obor studia:
-
Přidělovaný titul:
Ph.D.
Datum obhajoby:
10. 12. 2009
Výsledek obhajoby:
Prospěl/a
Jazyk práce:
Čeština
Abstrakt:
2. SOUHRN Bezobratli jsou Siroce rozslienou skupinou iivodichri tijicich ve velice rodidnfch podminkdch. StrategiepfeZitije u bezobratlfchd6najak kdtkou dobouZivotaa veL[y'mmnoZstvlm potomstva,tak piedeviim eistenci mnoZstvinejnlzrdjllch obrannjch mechanizrnd ridinndrozpoa{vajicich a eliminujicich cizorodCldtky. Objektem z6jmu imunologu se obrann6mechanizmylaoulkovcri staly vposlednich dtyficeti letech. U kroutkovcrl, stejnEjako u ostatnich bezobratlfch, nebyla zjiltdna pFltomnostimunoglobulinri, llmfocytri, ad jinfch sloZekadaptivniimunity. Ve svd disertadniprAci se v6nuji detailnlmu popisu ndktenfch obrannlch molekul piluozenCimunity kouZkovcri. 1. Lysorym je erzym se silnou antibakteridlnl aktivitou, jehoi piitomnostbyla popsdna iady organizmt. u My jsme detekovalimolekulu lysozymu u ilrdy Eisenia andrei (dftlveE.fetida anfuei) na rlrovni struktury a funkce. Molekuldmi charakterizace tohoto proteinuumohluje sledov6ni pfirozeni imunity u ffZal. 2. Cytolyticky efekt coelomovC telr.nny E. fetidabyl popsdnv experimentech TNF-senzitivnlnedorovou s bunddnoulinii L929. Ndsledn6izolace lytickich molekul vedla k identifikaci proteinu o velikosti 42-kDa, kter! byl nazv{n coelomovjm cytolytickfm faktorem - CCF. Bylo zji5tdno,Ze molekulaCCF je piitomna tak6 v coelomov6tekutind dalSihokouikovce - Lumbricastetestis. Pfitomnostmolekul podobnfch CCF byla vt6to pnici popsdna tak6 u dalsich zistupcrl delediLumbricidae. Tyto molekuly byly charakterizovriny na uroni primdmi sekvence porovn6nyz hlediskabiologick6aktivity. a 3. CCF vykaaje firnkdnl analogii se savdimcytokinem tumor nekotizujicim faktorem (TNF), zaloZend na ro4roadvdni podobnych saoharidovjch strukhr. TNF i CCF indukuji zrdseni vodivosti a ndslednou depolarizaci membrdn nCkterlch savClch bun€k. V tdto prfci byla popsrfura interakce molekuly CCF s peritonedhimimakofdgy a nislednAaktivacetdchtobun€k" 4. Role molekuly kalretikulinu v obrannfch mechanizmech byla jiL dtive popsdnann6kterjch bezobratllch i obratlovctl. My jsme prokdzali plitomnost t6to molekuly v coelomove tekutine titaly E fetida, chanktenzovalijsmejejl primdmi sekvencia odli5nouexpresit€to molekuly v nizrfch orgrnech. 5. jsme rztah mezi dvdma hemolytickfmi molekulami- lyseninem afetidinem Charakterizovali jsme uroveiljejich expresev ooelomocyech a detekovali jednotlivori dnftu E.fetida. 6. kaskddapledstavujejeden z nejdileZitdj5ich obrannlch mechanizmtibezobratllch Profenoloxidriaov6 Zivodichti. Prokdaali jsme pfitomnostmolekulprofenoloxidizy a jejiho moln6ho inhibitoru v coelomovi teku(mEtialy E. fetlda, nicm€ndhladina profenoloxidazovd aktivity je v porovnAnl ostatnimibezobratlfmi s druhynizsla tak6jeji aktivace pomalejsi. je
Abstract v angličtině:
I. SUMMARY lnvertebrates widely distrbuted animals.They can be found in almost any krnd ofhabitat. Their are successfulsurvival shategies are based on short life span combined with numerous offspring and, more importantly, invertebrate all species havedeveloped wriety ofdefense a mechanisms elficientlyrecognizing and responding non-self to substances. The defensemechadsmsof earthwoms were studiedover the past fou decades. becameappdent It that the earthworms. well as other as invertebrates, lymphoc)4es other features lack specificimmunoglobulins, or ofthe adaptiveimmunesystemdescribed verteblates, possess in but irmatedefensecomponents In this thesis,we foousedon the detail description somedefense of moleculesinvolved in imate immunityof earthworms. l. Lysozyme is an erzyme with strong antibacterialactivity described in many organisms. We characterized lysozymeof Eri enia andrei (formerly E. fetida andrei) earthwormboth struchmlly and the functionally. Moleoular charactefiation of lyso4rne providesa new tool for monitoringof innate immunity in earthworms. 2. A cltol''tic effect of the coelomic fluid of E. fetida was observedin experiments with TNF-sensitive tumor L929 cell line. Subsequent isolation of l)tic proteinsled to the identification of 42-kDa protein, which wasnamed coelomiccytolytic factor - CCF.CCFwas shom to be present in coelomic also fluid of mother Lumbricustenestis. Thereforewe elucidatedtle presenoe CCF-like moleculesalso earthworm species, of inother earthworms from Lumbricidae family, characterized their primary sequence and compared properties thesemolecules. thebiological of 3. CCF was described sharefimctionalanalogies to recognitionspecificity basedon the similar saccharide with mammalian cltokine, tumor necrosis factor (TNF). Both TNF and CCF were shown to induce an increaseof membraneconductance some rnmmalim cells resulting in membranedepolarization. in We investigated interaction the macrophages subsequent ofCCF with peritoneal and activationofthesecells 4. The role of calreticulin in the defensemechanismswas previously describedin both vertebratesand invertebrates. proved the presence calreticulin in the coelomic fluid of E ftrda earthworms, We of characterized primary sequenoe determined expression different organs' the and fte in 5. between We elarifiedthe relationship - two hemolyticmolecules lysenin and fetidin md detemined the level oftheir expression coelomocytes individ.ual fetida earthworms' in of E. 6. The prophenoloxidase cascade represents of the most importmt defensemechmismin mmy one invertebrates. provedthe presence phenoloxidase We of and its putative inhibitor in the coelomicfluid to activity is lower as compared ofE. fetida earthworms, the level ofphenoloxidase but otherinvertebmte species.Moreover, activation the ofprophenoloxidasecascade slower' is
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