Multiplexed fluorospot for the analysis of dengue virus- And zika virus-specific and cross-reactive memory B cells

Authors

Awadalkareem Adam, University of Rhode Island
Marcia Woda, University of Rhode Island
Sonia Kounlavouth, University of Rhode Island
Alan L. Rothman, University of Rhode Island
Richard G. Jarman, Walter Reed Army Institute of Research
Josephine P. Cox, National Institute of Allergy and Infectious Diseases (NIAID)
Julie E. Ledgerwood, National Institute of Allergy and Infectious Diseases (NIAID)
Gregory D. Gromowski, Walter Reed Army Institute of Research
Jeffrey R. Currier, Walter Reed Army Institute of Research
Heather Friberg, Walter Reed Army Institute of Research
Anuja Mathew, University of Rhode Island

Document Type

Article

Date of Original Version

12-15-2018

Abstract

Dengue virus (DENV) and Zika virus (ZIKV) are mosquito-borne pathogens that have a significant impact on human health. Immune sera, mAbs, and memory B cells (MBCs) isolated from patients infected with one DENV type can be cross-reactive with the other three DENV serotypes and even more distantly related flaviviruses such as ZIKV. Conventional ELISPOTs effectively measure Ab-secreting B cells but because they are limited to the assessment of a single Ag at a time, it is challenging to distinguish serotype-specific and cross-reactive MBCs in the same well. We developed a novel multifunction FluoroSpot assay using fluorescently labeled DENV and ZIKV (FLVs) that measures the cross-reactivity of Abs secreted by single B cells. Conjugation efficiency and recognition of FLVs by virus-specific Abs were confirmed by flow cytometry. Using a panel of DENV immune, ZIKV immune, and naive PBMC, FLVs were able to simultaneously detect DENV serotype-specific, ZIKV-specific, DENV serotype cross-reactive, and DENV/ZIKV cross-reactive Abs secreted by individual MBCs. Our findings indicate that the FLVs are sensitive and specific tools to detect specific and cross-reactive MBCs. These reagents will allow the assessment of the breadth as well as the durability of DENV/ZIKV B cell responses following vaccination or natural infection. This novel approach using FLVs in a FluoroSpot assay can be applied to other diseases such as influenza in which prior immunity with homosubtype- or heterosubtype-specific MBCs may influence subsequent infections. The Journ Al of Immunology, 2018, 201: 3804-3814.

Publication Title, e.g., Journal

Journal of Immunology

Volume

201

Issue

12

Citation/Publisher Attribution

Adam, Awadalkareem, Marcia Woda, Sonia Kounlavouth, Alan L. Rothman, Richard G. Jarman, Josephine P. Cox, Julie E. Ledgerwood, Gregory D. Gromowski, Jeffrey R. Currier, Heather Friberg, and Anuja Mathew. "Multiplexed fluorospot for the analysis of dengue virus- And zika virus-specific and cross-reactive memory B cells." Journal of Immunology 201, 12 (2018). doi: 10.4049/jimmunol.1800892.