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Antibody-Dependent Complement Activation and DENV3 Disease Severity

Nasser, Amro (2023) Antibody-Dependent Complement Activation and DENV3 Disease Severity. Master's Thesis, University of Pittsburgh. (Unpublished)

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Dengue Viruses (DENV), consists of a family with four serotypes. Dengue is a mosquito-borne agent responsible for causing Dengue Fever (DF), which in a small proportion of cases can further develop into a more severe disease, dengue hemorrhagic fever (DHF), characterized by blood plasma leakage, that can lead to cardiovascular shock and organ failure. Dengue disease overall is an inflammatory-driven pathology, with complement dysfunction having been implicated as playing a role in the progression to DHF. The extent to which antibody-dependent complement activation (ADCA) participates in this process, however, is not fully known. To investigate the ability of dengue antibodies specific to DENV3 NS1 to perform ADCA using a novel bead-based complement assay. In this assay, DENV3 NS1 is bound to fluorescent beads to incubate with patient samples, allowing for the formation of immune complexes. The beads are then incubated with a complement source, allowing for immune complexes to perform ADCA. The beads are then stained with an anti-complement factor 3 (C3) fluorophore-conjugated antibody, and flow cytometry is used to quantify the deposition of C3 fragments. This assay was utilized to quantify ADCA in a longitudinal cohort of primary and secondary DENV3 samples, from both DF and DHF cases. Additionally, antibody endpoint titers were measured by ELISA to determine correlation with ADCA. When comparing the ADCA capacity in the cohort, it is determined that secondary DENV3 infections have higher complement deposition than primary DENV3 infections and that secondary infections with DHF have the most potent antibodies to activate complement. Antibody titer was shown to have a moderate correlation with C3 deposition and ADCA, as expected. Data utilizing the complement assay supports the hypothesis that secondary DENV3 infections show greater complement deposition, particularly in secondary DENV3 DHF cases during the onset of severe symptoms. Further work will be done with this assay to further explore the potential relationship between ADCA and severe outcomes of dengue.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Nasser, AmroAMN84@pitt.eduAMN840009-0005-9974-5903
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorMarques, Ernestomarques@pitt.edumarques
Committee MemberHartman, Amyhartman2@pitt.eduhartman2
Committee MemberBain,
Date: 11 May 2023
Date Type: Publication
Defense Date: 21 April 2023
Approval Date: 11 May 2023
Submission Date: 27 April 2023
Access Restriction: 2 year -- Restrict access to University of Pittsburgh for a period of 2 years.
Number of Pages: 56
Institution: University of Pittsburgh
Schools and Programs: School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: Antibody Complement Dengue Severity
Date Deposited: 11 May 2023 16:10
Last Modified: 11 May 2023 16:10


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