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06277cam a2200445 i 4500 |
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NSK01000230175 |
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HR-ZaNSK |
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20070917175508.0 |
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990223s1997 ci a m 000 0 hrv |
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|9 (HR-ZaNSK)230418
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|9 (HR-ZaNSK)990223002
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|a (HR-ZaNSK)000230175
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|a HR-ZaNSK
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|c HR-ZaNSK
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|a ci
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|a 578.2
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| 100 |
1 |
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|a Ambriović Ristov, Andreja
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| 245 |
1 |
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|a Utjecaj promotora na ekspresiju gena za glikoprotein D pseudorabiens virusa pri imunizaciji plazmidnim i adenovirusnim vektorima :
|b doktorska disertacija /
|c Andreja Ambriović.
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| 260 |
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|a Zagreb :
|b A. Ambriović,
|c 1997
|e ([s. l. :
|f s. n.])
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| 300 |
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|a 119 listova :
|b sheme, ilustr., table, graf. prikazi ;
|c 30 cm.
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| 500 |
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|a Doktor prirodnih znanosti - biologija
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| 500 |
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|a Mentor: Vladimir Delić; Komisija za ocjenu: Ivan Bašić, Tomo Naglić;
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| 502 |
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|a Sveučilište u Zagrebu, Prirodoslovno-matematički fakultet, Zagreb, 1997
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| 504 |
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|a Bibliografija: str. 99-119
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| 504 |
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|a Summary
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| 520 |
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|a Sažetak: Konstruirani su plazmidni vektori u kojima se nalazi gen za glikoprotein D (gD) pseudorabiens virusa (PRV) pod kontrolom tri promotora: promotora prvog ranog gena humanog citomegalovirusa (CMV), promotora dugog ponavljajućeg slijeda virusa Rousovog sarkoma (RSV) i mišićno specifičnog promotora humanog gena za desmin (DES). Plazmid pMLPgD, u kojem se gen za gD nalazi pod kontrolom glavnog kasnog promotora adenovirusa tip 2 (MLP), konstruiran je ranije. Pročišćenim plazmidnim vektorima pMLPgD, pCMVgD, pRSVgD i pDESgD vakcinirani su miševi, intramuskularnim putem, s tri različite doze, jednom ili dva puta. Enzim-imunološkim testom u serumima vakciniranih miševa, pokazana je prisutnost gD specifičnih protutijela u približno 80% mišev, bez velikih razlika u količini protutijela potaknutih različitim plazmidnim DNA.
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| 520 |
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|a Kod miševa koji su primili dvije doze gD specifična protutijela su prisutna duže nego kod miševa koji su primili samo jednu dozu. Serumi vakciniranih miševa nemaju neutralizacijsku sposobnost In vitro, a životinje nisu zaštićene od provokacije smrtnom dozom PRV. Konstruirani plazmidni vektori korišteni su za konstrukciju adenovirusa defektnih u replikaciji: AdCMVgD, AdRSVgD i AdDESgD. Metodom precipitacije monoklonskim protutijelima protiv gD, radioaktivno obilježenih proteina iz kulture stanica, pokazana je sposobnog konstruiranih adenovirusa (kao i ranije konstruiranog AdMLPgD) da u staničnoj liniji 293 koja podržava njihovu replikaciju, usmjeravaju sintezu velikih količina gD, bez znatnih razlika u količini gD u ovisnosti o umetnutom promotoru.
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| 520 |
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|a U staničnim linijama koje ne podržavaju replikaciju adenovirusa, primjećene su velike razlike u jačini promotora. Pokazano je da gotovo u svim istraženim kulturama stanica promotori CMV i DES potiču snažniju ekspresiju gena za gD, nego što to čine promotori MLP i RSV. Pretpostavljena je uključenost E1A pojačivača u gubitku mišićne specifičnosti ekspresije promotora DES. Vakcinacija miševa rekombinantnim AdMLPgD, AdCMVgD, AdRSVgD i AdDESgD učinjena je jednokratno, intramuskularnim putem. Dobivena protutijela u serumu imaju sposobnost neutralizacije In vitro, a vakcinirane životinje su zaštićene od provokacije smrtnom dozom PRV. Dobiveni rezultati In vitro u skladu su sa određenom zaštitom vakcinalnom dozom, koja je za AdCMVgD i AdDESgD znatno smanjena u odnosu na onu za AdMLPgD i AdRSVgD.
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| 520 |
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|a Summary: Plasmid vectors have been constructed, in which the glycoprotein D (gD) gene is under the control of three different promoters: cytomegalovirus immediate early gene promoter (CMV), Rous sarcoma virus-long terminal repeat promotor (RSV) and promotor of the human desmin gene (DES). The plasmid pMLPgD has been constructed previously. It contains the gD gene under the control of the major late promoter of adenovirus type 2 (MLP). Mice were vaccinated by intramuscular route, once or twice, with three different doses of purified plasmids pMLPgD, pRSVgD, pCMVgD and pDESgD. Sera from mice were analyzed by enzyme-immuno assay and the presence of gD specific antibodies was detected in 80% of the animals, without observed differences between different plasmid constructs.
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| 520 |
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|a Antibody responses against gD in mice that received two doses, persisted longer than in mice vaccinated only once. In all cases there was no neutralization titer In vitro detected in sera and there was no protection against lethal challenge. Plasmid vectors pCMVgD, pRSVgD i pDESgD have been used for the constructions of replication defective adenoviruses: AdCMVgD, AdRSVgD and AdDESgD. The ability of constructed adenovirus recombinants (as well as AdMLPgD constructed previously), to drive the expression of gD in 293 cells(that support virus replication) was tested by precipitation of radiolabelled proteins, using monoclonal antibodies against gD. All four adenoviruses were able to expresshigh levels of gD and the amount of gD was comparable. In cell that do not support virus replication vide variation in promoter stength In vitro were evident.
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| 520 |
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|a It was shown that in almost all cell types the CMV and DES promoters were better than MLP and RSV promoters, giving higher amount of gD. It is supposed that E1A enhancer is involved in long of cell type specificity of expression drived by DES promoter. Mice were vacinated once with AdMLPgD, AdCMVgD, AdRSVgD and AdDESgD, by the intramuscular route. The gD specific antibodies in sera of vaccinated mice have the neutralization abitiy In vitro, and mice were protected against lethal challenge. The results obtained In vitro could be correlated to the vaccinations protection dose obtained, which for AdCMVgD and AdDESgD decreased significantly compared with AdMLPgD and AdRSVgD.
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| 650 |
|
7 |
|a Herpesvirusne infekcije
|x Imunologija
|2 nskps
|
| 700 |
1 |
|
|a Delić, Vladimir,
|c biokemičar
|4 cns
|
| 700 |
1 |
|
|a Bašić, Ivan,
|c imunolog
|4 oth
|
| 700 |
1 |
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|a Naglić, Tomo
|4 oth
|
| 981 |
|
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|p CRO
|r HRB1997
|
| 998 |
|
|
|n DCD/97
|c sbno9903
|c dkrp9904
|
| 852 |
4 |
|
|j DCD-ZG-37/98
|
| 876 |
|
|
|e DCD
|a 37/1998
|
| 886 |
0 |
|
|2 unimarc
|b 06027iam0 2200373 450
|