Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Li, Zhu-Nan; Cheng, Emily H.; Poirot, Eugenie; Weber, Kimberly M.; Carney, P.J.; Chang, Jae C.; Liu, Feng; Gross, F. Liaini; Holiday, Crystal; Fry, Alicia M.; Stevens, James; Tumpey, Terrence M.; Levine, Min Z.

doi:10.1111/irv.12695

Cited by 3 publications

(13 citation statements)

References 27 publications

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“…S1 and S2 in the supplemental material) (16). In our previous studies, we used a serum dilution of 1:40 to detect influenza virus antibody responses using the same assay platform (14,15,17). Here, the linearity analysis suggests that the 1:40 serum dilution falls well within the linear range for most seasonal and novel subtype influenza virus antigens and is suitable to capture the full scope of the antibody responses to most antigens while maintaining the assay throughput.…”

Section: Resultsmentioning

confidence: 97%

“…To determine seroconversion in the MIADA for paired serum samples, we used a $2-fold rise in MFIs after adjustment of low S1 MFIs to 1,000 (14,15). We found that one A(H1N1)pdm09-infected person and three A(H3N2)-infected persons achieved seroconversion ($2-fold rises in MFIs) against at least one novel subtype rHA (H5, H7, H9, or H13) (Tables S3 and S4), and nine persons showed high MFIs ($2,000) against a novel subtype rHA(s) in S2 and/or S1 sera, although no seroconversions in MFIs were achieved (Table S5 and data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…In influenza virus serologic studies, a $4-fold rise in HI/MN titers is often considered a positive antibody response (or seroconversion). Here, in the MIADA, we used a $2-fold rise in MFIs (S2/ S1) after adjustment of low MFIs in baseline S1 to 1,000 (to avoid an unreliable fold rise as described previously) as seroconversion (14,15). We also analyzed consistency in non-seroconversion cases measured by HI (,4-fold rise) and MIADA (,2-fold rise).…”

Section: Resultsmentioning

confidence: 99%

“…We previously developed a Magpix assay for serologic diagnosis of seasonal and novel influenza virus infection (14,15). In the current study, we expanded this platform and developed a high-throughput multiplex influenza antibody detection assay (MIADA) that can measure antibody responses to 42 HA antigens from 10 influenza virus subtypes simultaneously, plus a protein A (PA) control.…”

mentioning

confidence: 99%

“…In the current study, we expanded this platform and developed a high-throughput multiplex influenza antibody detection assay (MIADA) that can measure antibody responses to 42 HA antigens from 10 influenza virus subtypes simultaneously, plus a protein A (PA) control. Furthermore, we implemented an antibody adsorption technique using ectodomain (Ecto) HAs from A(H1N1) and A(H3N2) viruses to elucidate the effects of both homosubtypic (within-subtype) and heterosubtypic (cross-subtype) cross-reactivity (14,15).…”

mentioning

confidence: 99%

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Antibody Landscape Analysis following Influenza Vaccination and Natural Infection in Humans with a High-Throughput Multiplex Influenza Antibody Detection Assay

Liu

Gross

et al. 2021

mBio

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To better understand the antibody landscape changes following influenza virus natural infection and vaccination, we developed a high-throughput multiplex influenza antibody detection assay (MIADA) containing 42 recombinant hemagglutinins (rHAs) (ectodomain and/or globular head domain) from pre-2009 A(H1N1), A(H1N1)pdm09, A(H2N2), A(H3N2), A(H5N1), A(H7N7), A(H7N9), A(H7N2), A(H9N2), A(H13N9), and influenza B viruses. Panels of ferret antisera, 227 paired human sera from vaccinees (children and adults) in 5 influenza seasons (2010 to 2018), and 17 paired human sera collected from real-time reverse transcription-PCR (rRT-PCR)-confirmed influenza A(H1N1)pdm09, influenza A(H3N2), or influenza B virus-infected adults were analyzed by the MIADA. Ferret antisera demonstrated clear strain-specific antibody responses to exposed subtype HA. Adults (19 to 49 years old) had broader antibody landscapes than young children (<3 years old) and older children (9 to 17 years old) both at baseline and post-vaccination. Influenza vaccination and infection induced the strongest antibody responses specific to HA(s) of exposed strain/subtype viruses and closely related strains; they also induced cross-reactive antibodies to an unexposed influenza virus subtype(s), including novel viruses. Subsequent serum adsorption confirmed that the cross-reactive antibodies against novel subtype HAs were mainly induced by exposures to A(H1N1)/A(H3N2) influenza A viruses. In contrast, adults infected by influenza B viruses mounted antibody responses mostly specific to two influenza B virus lineage HAs. Median fluorescence intensities (MFIs) and seroconversion in MIADA had good correlations with the titers and seroconversion measured by hemagglutination inhibition and microneutralization assays. Our study demonstrated that antibody landscape analysis by the MIADA can be used for influenza vaccine evaluations and characterization of influenza virus infections. IMPORTANCE Repeated influenza vaccination and natural infections generate complex immune profiles in humans that require antibody landscape analysis to assess immunity and evaluate vaccines. However, antibody landscape analyses are difficult to perform using traditional assays. Here, we developed a high-throughput, serum-sparing, multiplex influenza antibody detection assay (MIADA) and analyzed the antibody landscapes following influenza vaccination and infection. We showed that adults had broader antibody landscapes than children. Influenza vaccination and infection not only induced the strongest antibody responses to the hemagglutinins of the viruses of exposure, but also induced cross-reactive antibodies to novel influenza viruses that can be removed by serum adsorption. There is a good correlation between the median fluorescence intensity (MFI) measured by MIADA and hemagglutination inhibition/microneutralization titers. Antibody landscape analysis by the MIADA can be used in influenza vaccine evaluations, including the development of universal influenza vaccines and the characterization of influenza virus infections.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Antibody Landscape Analysis following Influenza Vaccination and Natural Infection in Humans with a High-Throughput Multiplex Influenza Antibody Detection Assay

Liu

Gross

et al. 2021

mBio

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Multiplex Detection of Antibody Landscapes to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)/Influenza/Common Human Coronaviruses Following Vaccination or Infection With SARS-CoV-2 and Influenza

Liu

Jefferson

et al. 2022

Clinical Infectious Diseases

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Background SARS-CoV-2 and influenza viruses continue to co-circulate, representing two major public health threats from respiratory infections with similar clinical presentations. SARS-CoV-2 and influenza vaccines can also now be co-administered. However, data on antibody responses to SARS-CoV-2 and influenza co-infection, and vaccine co-administration remains limited. Methods We developed a 41-plex antibody immunity assay that can simultaneously characterize antibody landscapes to SARS-CoV-2/influenza/common human coronaviruses. We analyzed sera from 840 individuals (11-93 years), including sera from reverse transcription polymerase chain reaction (RT-PCR) confirmed SARS-CoV-2 positive (n = 218) and negative (n = 120) cases, paired sera from SARS-CoV-2 vaccination (n = 29) and infection (n = 11), and paired sera from influenza vaccination (n = 56) and RT-PCR confirmed influenza infection (n = 158) cases. Lastly, we analyzed sera collected from 377 individual that exhibited acute respiratory illness (ARI) in 2020. Results This 41-plex assay has high sensitivity and specificity in detecting SARS-CoV-2 infections. It differentiated SARS-CoV-2 vaccination (antibody responses only to spike protein) from infection (antibody responses to both spike and nucleoprotein). No cross-reactive antibodies were detected to SARS-CoV-2 from influenza vaccination and infection, and vice versa, suggesting no interaction between SARS-CoV-2 and influenza antibody responses. However, cross-reactive antibodies were detected between spike proteins of SARS-CoV-2 and common human coronaviruses that were removed by serum adsorption. Among 377 individual who exhibited ARI in 2020, 129 were influenza positive, none had serological evidence of SARS-CoV-2/influenza co-infections. Conclusions Multiplex detection of antibody landscapes can provide in-depth analysis of the antibody protective immunity to SARS-CoV-2 in the context of other respiratory viruses including influenza.

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A randomized controlled trial of antibody response to 2020 cell-based inactivated and egg-based live attenuated influenza vaccines in individuals ages 4-21 years

Martin

Williams

et al. 2022

Preprint

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Antibody responses to influenza vaccines may vary by many factors. Participants ages 4-21 were randomized to receive cultured inactivated influenza vaccine (ccIIV4; n =112) or live attenuated influenza vaccine (LAIV4; n=118). Hemagglutinin inhibition (HAI) reported in geometric mean titers (GMTs) and mean fold rise in titers (MFR) and IgG, IgA, and IgM antibodies reported in median fluorescence intensities (MFIs) were measured against cell-grown, egg-grown, and drifted A(H1N1) antigens, A(H1N1)pdm09, both vaccine B strains and the nucleoprotein (NP) from A(H3N2) on pre- and 28 days post vaccination. The HAI and immunoglobulin isotype response to ccIIV4 was greater than LAIV4. LAIV4 response was highest among the youngest participants. Prior LAIV4 vaccination was associated with a higher response to current season ccIIV4. Cross-reactive A/Delaware/55/2019(H1N1)pdm09 antibodies were present pre-vaccination and increased in response to ccIIV4, but not LAIV4. Immunoglobulin assays confirmed the findings of HAI titers to measure influenza vaccine immune response.Clinical Trials No.: NCT03982069

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Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

Cited by 3 publications

References 27 publications

Antibody Landscape Analysis following Influenza Vaccination and Natural Infection in Humans with a High-Throughput Multiplex Influenza Antibody Detection Assay

Antibody Landscape Analysis following Influenza Vaccination and Natural Infection in Humans with a High-Throughput Multiplex Influenza Antibody Detection Assay

Multiplex Detection of Antibody Landscapes to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)/Influenza/Common Human Coronaviruses Following Vaccination or Infection With SARS-CoV-2 and Influenza

A randomized controlled trial of antibody response to 2020 cell-based inactivated and egg-based live attenuated influenza vaccines in individuals ages 4-21 years

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