Platform development for human cytomegalovirus (HCMV) vaccine design to elicit cross-reactive neutralizing antibodies (nAbs)

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Thesis type
(Thesis) M.Sc.
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Author: Ang, Quiana
Human Cytomegalovirus cytomegalovirus (HCMV) can infectcauses intrauterine infections, which pregnant women, can lead toresulting in perinatal mortality and or deafness in the surviving infants. A protective vaccine is , therefore, highly desirable. Currently,Most HCMV vaccine strategies aim to elicit neutralizing antibodies (nAbs) targeting surface glycoproteins on the virus' outer surface, such as . Among them is the virus envelope glycoprotein B (gB), which is required for protein to induce cross-reactive neutralizing antibodies (nAbs) because it has a central role in virus entryinfection of all cell types. However,Previously assessed recombinant- forms of gB assessed to date haves not been fruitful in eliciting neutralizing antibodiesnAbs. To identify a potentially better gB-based immunogen, I constructed aA panel of 14 recombinant gB-based antigens were constructed and evaluated their antigenicity with 5 different a panel of nAbseutralizing and non-neutralizing antibodiesnAbs (2F12, HCMV37, 1G2, SM5-1, and ITC88), each against aspecific to different gB epitopee osn gB. I identified one promisingan antigen, dubbed sgB-SS-HIS, to whichwhereto which all neutralizing antibodiesnAbs were bound ly bound strongly but poorly bound by non-nAbs did notest, but that was bound poorly by non-neutralizing ones. I also report on my efforts to developI also developedattempted to develop an HCMV pseudovirus assay platform to rapidly screen and categorize anti-gB nAbs. From my results, one recombinant gB-antigen can present the nAb epitopes on gB and has a strong binding interaction: sgB-SS-HIS. Furthermore, a platform that is sensitive and amenable to high throughput might help to better categorize HCMV nAbs and, by extension, vaccine candidates. Here, weCo-transfection of plasmids encoding HCMV surface glycoproteins along with and plasmids to generate lentiviral particles yielded pseudotyped HCMV, which were able to infected representative epithelial and fibroblast cells. However, antibodies known to neutralize infection of replication-competent HCMV did not neutralize the the pseudotyped virus, suggesting that infection of target cells by pseudotyped HCMV may not mirror the infection with a replication-competent virus.
94 pages.
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Thesis advisor: Pantophlet, Ralph
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