Gripas vīrusa hemaglutinīna fragmenta strukturāla izpēte un eksponēšana uz vīrusiem līdzīgo daļiņu virsmas
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Latvijas Universitāte
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Abstract
Iekļaujot vakcīnā konservatīvus vīrusa proteīnu domēnus, būtu iespējams nodrošināt plaša spektra aizsardzību pret dažādiem gripas vīrusu apakštipiem. Viens no šādiem konservatīviem rajoniem ir hemaglutinīna (HA) stalka peptīds HA2.3. Bakalaura darba mērķis bija noteikt HA2.3 struktūru, tā eksponēšanas iespējas uz dažādu vīrusiem līdzīgo daļiņu (VLP) virsmas, un pētīt šo daļiņu imunoloģiskās īpašības. Pētījumā tika veikta HA2.3 ekspresija, attīrīšana, kristalizācija. Izmantojot rentgenstaru difrakciju, tika noteikts, ka HA2.3 struktūra vides pH 5,5 atbilst natīvajai HA stalka trimēra post fusion konformācijai. HA2.3 tika ķīmiski konjugēts pie dažādu VLP virsmas, kuras pēc tam izmantoja peļu imunizācijai. Himēro VLP izraisītā antivielu atbilde pret HA2.3 bija zemāka nekā HA2.3 kontrolei, kas, visticamāk, izskaidrojams ar HA2.3 konformācijas izmaiņām.
Including conserved domains of virus proteins in influenza vaccine could elicit broad-spectrum protection against multiple subtypes of influenza virus. One of such evolutionary conserved regions is HA2.3 peptide within hemagglutinin (HA) stalk. The main objective of this work was to determine the structure of HA2.3, to test the potential of exposing it on the surface of virus like particles (VLPs) and to determine the immunological properties of these particles. Expression, purification and crystallization of the HA2.3 were accomplished in the study. It was determined by the X-ray crystallography that the structure of HA2.3 at pH 5,5 match the post-fusion form of native HA stalk. HA2.3 was chemically coupled to the surface of various VLPs, which were further used to immunize mice. Antibody response against HA2.3 elicited by chimeric VLPs was lower than by HA2.3 control that most likely can be explained by conformational changes of HA2.3.
Including conserved domains of virus proteins in influenza vaccine could elicit broad-spectrum protection against multiple subtypes of influenza virus. One of such evolutionary conserved regions is HA2.3 peptide within hemagglutinin (HA) stalk. The main objective of this work was to determine the structure of HA2.3, to test the potential of exposing it on the surface of virus like particles (VLPs) and to determine the immunological properties of these particles. Expression, purification and crystallization of the HA2.3 were accomplished in the study. It was determined by the X-ray crystallography that the structure of HA2.3 at pH 5,5 match the post-fusion form of native HA stalk. HA2.3 was chemically coupled to the surface of various VLPs, which were further used to immunize mice. Antibody response against HA2.3 elicited by chimeric VLPs was lower than by HA2.3 control that most likely can be explained by conformational changes of HA2.3.