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Multiplex analysis ...
Multiplex analysis of antigen-specific memory B cells in humans using reversed B-cell FluoroSpot
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- Jahnmatz, Peter (author)
- Karolinska Institutet
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- Sundling, Christopher (author)
- Karolinska Institutet
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Makower, Bartek (author)
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- Sondén, Klara (author)
- Karolinska Institutet
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- Färnert, Anna (author)
- Karolinska Institutet
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- Ahlborg, Niklas (author)
- Stockholms universitet,Institutionen för molekylär biovetenskap, Wenner-Grens institut,Mabtech AB, Sweden
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(creator_code:org_t)
- Elsevier BV, 2020
- 2020
- English.
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In: JIM - Journal of Immunological Methods. - : Elsevier BV. - 0022-1759 .- 1872-7905. ; 478
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https://doi.org/10.1...
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Abstract
Subject headings
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- Analysis of B-cell specificities at the single cell level provides important information on how the B-cell compartment responds when challenged by infection or vaccination. We recently developed a reversed B-cell FluoroSpot assay and showed that it could be used to detect B cells specific for different antigens simultaneously in a mouse model. The aim of this study was to further develop the method to detect and quantify antigen-specific memory B cells (MBCs) in humans where circulating antigen-specific cells are less frequent. We show that MBCs specific for three antigens, tetanus toxoid, hepatitis B surface antigen and cytomegalovirus pp65, could be detected simultaneously in one well. In addition to enumerating antigen-specific MBCs, we also assessed the spot volume to estimate the intensity of the response in individual cells and found this to be a new and sensitive approach to study MBC responses after vaccination. This unique B-cell FluoroSpot approach provides a simple and sensitive multiplex analysis of MBCs and can be adapted to most antigens and host species.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Immunologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Immunology in the medical area (hsv//eng)
Keyword
- Antibody
- Antigen
- B cell ELISpot
- FluoroSpot
- Vaccine
Publication and Content Type
- ref (subject category)
- art (subject category)
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