Molecular studies of louping ill virus

<p>The genomic RNA encoding the structural proteins of louping ill, a tickborne flavivirus, was cloned and sequenced. Sequence comparisons of louping ill envelope protein showed greater homology with tick-borne than mosquito-borne flaviviruses and greater homology with the western than the fa...

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Main Authors: Shiu, S, Shiu, Stephen Yuen Wing
Other Authors: Gould, E
Format: Thesis
Language:English
Published: 1991
Subjects:
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author Shiu, S
Shiu, Stephen Yuen Wing
author2 Gould, E
author_facet Gould, E
Shiu, S
Shiu, Stephen Yuen Wing
author_sort Shiu, S
collection OXFORD
description <p>The genomic RNA encoding the structural proteins of louping ill, a tickborne flavivirus, was cloned and sequenced. Sequence comparisons of louping ill envelope protein showed greater homology with tick-borne than mosquito-borne flaviviruses and greater homology with the western than the far eastern subtype of tick-borne encephalitis virus. Louping ill and tick-borne encephalitis viruses are probably varieties of a common tick-borne ancestral virus. The average amino acid sequence diversity between members of the tick-borne serogroup was significantly lower than that of mosquito-borne serogroups, suggesting that tick-borne flaviviruses have been subjected to different evolutionary immune selection pressure from the mosquito-borne viruses. Using the published model of tick-borne encephalitis envelope protein and the derived sequence data on louping ill virus, three discontinuous peptides (amino acids 81-88, 207-212 and 230-234) which may represent critical molecular determinants within the receptor binding site of tick-borne flaviviruses, were identified. These peptides may provide a specific genetic marker for these viruses.</p> <p>Recombinant baculoviruses and vaccinia viruses containing cloned DNA, encoding either the envelope protein or the structural proteins of louping ill virus, were constructed. Glycosylated envelope protein, presented both inside and on the surface of insect and mammalian cells, was expressed by all four recombinant viruses. Differences in antigenic presentation of envelope protein were observed between envelope protein and structural protein constructs as well as between insect cell and mammalian cell expression systems. Despite the expression of epitopes known to elicit neutralizing and protective antibodies when present in authentic antigen, the recombinant envelope protein expressed by either baculovirus or vaccinia virus failed to induce, under the experimental conditions employed, either neutralizing or protective antibodies in both mice and rabbits against louping ill virus. Hence, louping ill envelope protein expressed by baculoviruses and vaccinia viruses was antigenically reactive but immunogenically inert.</p>
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spelling oxford-uuid:949ac059-3386-4435-90e1-15166075fd892022-03-26T23:40:36ZMolecular studies of louping ill virusThesishttp://purl.org/coar/resource_type/c_db06uuid:949ac059-3386-4435-90e1-15166075fd89Louping illMolecular aspectsTick-borne encephalitis virusesEnglishPolonsky Theses Digitisation Project1991Shiu, SShiu, Stephen Yuen WingGould, EGould, E<p>The genomic RNA encoding the structural proteins of louping ill, a tickborne flavivirus, was cloned and sequenced. Sequence comparisons of louping ill envelope protein showed greater homology with tick-borne than mosquito-borne flaviviruses and greater homology with the western than the far eastern subtype of tick-borne encephalitis virus. Louping ill and tick-borne encephalitis viruses are probably varieties of a common tick-borne ancestral virus. The average amino acid sequence diversity between members of the tick-borne serogroup was significantly lower than that of mosquito-borne serogroups, suggesting that tick-borne flaviviruses have been subjected to different evolutionary immune selection pressure from the mosquito-borne viruses. Using the published model of tick-borne encephalitis envelope protein and the derived sequence data on louping ill virus, three discontinuous peptides (amino acids 81-88, 207-212 and 230-234) which may represent critical molecular determinants within the receptor binding site of tick-borne flaviviruses, were identified. These peptides may provide a specific genetic marker for these viruses.</p> <p>Recombinant baculoviruses and vaccinia viruses containing cloned DNA, encoding either the envelope protein or the structural proteins of louping ill virus, were constructed. Glycosylated envelope protein, presented both inside and on the surface of insect and mammalian cells, was expressed by all four recombinant viruses. Differences in antigenic presentation of envelope protein were observed between envelope protein and structural protein constructs as well as between insect cell and mammalian cell expression systems. Despite the expression of epitopes known to elicit neutralizing and protective antibodies when present in authentic antigen, the recombinant envelope protein expressed by either baculovirus or vaccinia virus failed to induce, under the experimental conditions employed, either neutralizing or protective antibodies in both mice and rabbits against louping ill virus. Hence, louping ill envelope protein expressed by baculoviruses and vaccinia viruses was antigenically reactive but immunogenically inert.</p>
spellingShingle Louping ill
Molecular aspects
Tick-borne encephalitis viruses
Shiu, S
Shiu, Stephen Yuen Wing
Molecular studies of louping ill virus
title Molecular studies of louping ill virus
title_full Molecular studies of louping ill virus
title_fullStr Molecular studies of louping ill virus
title_full_unstemmed Molecular studies of louping ill virus
title_short Molecular studies of louping ill virus
title_sort molecular studies of louping ill virus
topic Louping ill
Molecular aspects
Tick-borne encephalitis viruses
work_keys_str_mv AT shius molecularstudiesofloupingillvirus
AT shiustephenyuenwing molecularstudiesofloupingillvirus