A high-throughput multiplex assay to characterize flavivirus-specific immunoglobulins

Merbah, Mélanie; Wollen-Roberts, Suzanne E.; Shubin, Zhanna; Li, Yifan; Bai, Hongjun; Dussupt, Vincent; Mendez-Rivera, Letzibeth; Slike, Bonnie M.; Krebs, Shelly J.; Modjarrad, Kayvon; Michael, Nelson L.; Rolland, Morgane

doi:10.1016/j.jim.2020.112874

Cited by 8 publications

(7 citation statements)

References 20 publications

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“…We adapted assays from a previous study (15). Per 1 million beads, we coupled 10 µg of antigen for flavivirus proteins (15); 2.5 µg for coronavirus nucleocapsid (N) proteins; 5 µg for HIV-1 proteins; and 5 µg for coronavirus spike (S) proteins, including subunit 1 (S1), subunit 2 (S2), receptorbinding domain (RBD), and S-Trimer.…”

Section: Bead-based Multiplex Assaymentioning

confidence: 99%

Coronavirus Antibody Responses before COVID-19 Pandemic, Africa and Thailand

Li¹,

Merbah²,

Wollen-Roberts³

et al. 2022

Emerg. Infect. Dis.

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C OVID-19 clinical manifestations range from asymptomatic infection to death. Whether prior immune responses to human coronaviruses affect responses to SARS-CoV-2 remains unclear. At the population level, disparities in COVID-19 outcomes have been observed across geographic regions. For instance, countries in Africa have reported lower mortality rates than high-income countries, which can be attributed to the small percentage of persons in the oldest age groups and to underreporting (1,2). Previous responses to endemic coronaviruses also could influence how different populations responded to SARS-CoV-2.Findings conflict as to whether previous coronavirus antigen responses cross-react with SARS-CoV-2. Depending on the antigen and cohort tested, binding responses have been detected in prepandemic samples at varying frequencies, but neutralizing antibodies have been identified in fewer samples (3-8). Some studies of prepandemic samples indicated that neutralizing responses to endemic coronaviruses could protect against SARS-CoV-2 infection, but the effects of previous coronavirus responses on SARS-CoV-2 have not been clearly elucidated (6,7,9-13).To investigate coronavirus-specific antibody responses in different settings, we analyzed 2,565 samples collected during 2013 through early 2020 from persons living with HIV-1 (PLHIV) and persons without HIV in Kenya, Nigeria, Tanzania, Uganda, and Thailand. We profiled antibody binding responses to coronavirus antigens, including spike (S) and nucleocapsid (N) proteins of SARS-CoV-2, SARS-CoV-1, MERS-CoV, and 4 endemic coronaviruses. We further evaluated a subset of samples with strong binding responses for neutralizing, antibody-dependent cellular

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Section: Bead-based Multiplex Assaymentioning

confidence: 99%

Coronavirus Antibody Responses before COVID-19 Pandemic, Africa and Thailand

Li¹,

Merbah²,

Wollen-Roberts³

et al. 2022

Emerg. Infect. Dis.

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“…To evaluate how prior flavivirus vaccination shaped the immune response to the ZPIV vaccine, we characterized the antibody-mediated immune profile elicited by the vaccines and investigated the impact of potentially cross-reactive responses (due to prior vaccination) on ZPIV-induced immunity using a systems serology approach. 22 , 23 , 24 We found no evidence that priming with JEV or YFV vaccines improved the magnitude of the antibody response elicited by ZPIV, yet antibody-mediated responses showed qualitative differences across groups with more flaviviruses being recognized in the primed groups.…”

Section: Introductionmentioning

confidence: 57%

“…JEV, YFV and ZIKV E antigens matched the sequence used for the vaccine inserts; some proteins were produced in-house as previously described. 23 …”

Section: Methodsmentioning

confidence: 99%

“…Antibody responses were profiled using an assay adapted from previous studies. 23 , 24 Per million beads, 10 μg of antigen was coupled; 1200 conjugated beads of each antigen per well were used and samples were ran in triplicate at 2 dilutions, 1:200 and 1:800. Binding to seven antibody isotypes and subclasses (IgA, total IgG, IgG1-4 and IgM) and four Fc gamma receptors (FcγR) was measured.…”

Section: Methodsmentioning

confidence: 99%

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Priming with Japanese encephalitis virus or yellow fever virus vaccination led to the recognition of multiple flaviviruses without boosting antibody responses induced by an inactivated Zika virus vaccine

Li,

Merbah,

Wollen-Roberts

et al. 2023

eBioMedicine

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“…Bead-coupling and multiplex Luminex assays were performed as previously described, with modifications ( 79 ). Thirty µg of each S antigen and 15µg of NP antigen were covalently coupled to six million fluorescently coded carboxylated magnetic MagPlex beads (Luminex Corporation, Austin, TX).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Antibody-Dependent Fc-Mediated Viral Entry, as Compared With Neutralization, in SARS-CoV-2 Infection

et al. 2022

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Fc-mediated virus entry has been observed for many viruses, but the characterization of this activity in convalescent plasma against SARS-CoV-2 Variants of Concern (VOC) is undefined. In this study, we evaluated Fc-mediated viral entry (FVE) on FcγRIIa-expressing HEK293 cells in the presence of SARS-CoV-2 convalescent plasma and compared it with SARS-CoV-2 pseudovirus neutralization using ACE2-expressing HEK293 cells. The plasma were collected early in the pandemic from 39 individuals. We observed both neutralization and FVE against the infecting Washington SARS-CoV-2 strain for 31% of plasmas, neutralization, but not FVE for 61% of plasmas, and no neutralization or FVE for 8% of plasmas. Neutralization titer correlated significantly with the plasma dilution at which maximum FVE was observed, indicating Fc-mediated uptake peaked as neutralization potency waned. While total Spike-specific plasma IgG levels were similar between plasma that mediated FVE and those that did not, Spike-specific plasma IgM levels were significantly higher in plasma that did not mediate FVE. Plasma neutralization titers against the Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1) and Delta (B.1.617.2) VOC were significantly lower than titers against the Washington strain, while plasma FVE activity against the VOC was either higher or similar. This is the first report to demonstrate a functional shift in convalescent plasma antibodies from neutralizing and FVE-mediating against the earlier Washington strain, to an activity mediating only FVE and no neutralization activity against the emerging VOC, specifically the Beta (B.1.351) and Gamma (P.1) VOC. It will be important to determine the in vivo relevance of these findings.

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A high-throughput multiplex assay to characterize flavivirus-specific immunoglobulins

Cited by 8 publications

References 20 publications

Coronavirus Antibody Responses before COVID-19 Pandemic, Africa and Thailand

Coronavirus Antibody Responses before COVID-19 Pandemic, Africa and Thailand

Priming with Japanese encephalitis virus or yellow fever virus vaccination led to the recognition of multiple flaviviruses without boosting antibody responses induced by an inactivated Zika virus vaccine

Evaluation of Antibody-Dependent Fc-Mediated Viral Entry, as Compared With Neutralization, in SARS-CoV-2 Infection

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