To the top

Page Manager: Webmaster
Last update: 9/11/2012 3:13 PM

Tell a friend about this page
Print version

Flow-Cytometric Method Me… - University of Gothenburg, Sweden Till startsida
Sitemap
To content Read more about how we use cookies on gu.se

Flow-Cytometric Method Measuring B Cell Surface Immunoglobulin Avidity.

Journal article
Authors Davide Angeletti
Gregory M Frank
Jonathan W Yewdell
Published in Methods in molecular biology (Clifton, N.J.)
Volume 1623
Pages 181-189
ISSN 1940-6029
Publication year 2017
Published at
Pages 181-189
Language en
Links dx.doi.org/10.1007/978-1-4939-7095-...
www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Animals, Antibody Affinity, Antigens, immunology, metabolism, B-Lymphocytes, immunology, metabolism, Flow Cytometry, methods, Germinal Center, cytology, immunology, metabolism, Lymphoid Tissue, cytology, immunology, metabolism, Mice, Protein Binding, immunology, Receptors, Antigen, B-Cell, metabolism
Subject categories Immunology in the medical area

Abstract

The affinity of antibodies for their cognate antigens is a critical aspect of humoral immunity. The immune system has gone to great lengths to evolve a mechanism that enables real time increases in antibody affinity during the course of an immune response. This occurs in germinal centers (GC), which form in spleen and lymph nodes following immunization. GC B cell competition for limiting amount of antigen drives the selection of B cells expressing higher affinity Abs. Remarkably little is known about affinity maturation of B cells in immune responses to all but a handful of small model antigens. It has proven challenging to measure the avidity of specific Abs in polyclonal sera to more complex antigens, including viruses. In this chapter we present a simple, flow cytometry based, method that determines the average avidity of GC B cells for the influenza A virus hemagglutinin glycoprotein, the target antigen of traditional influenza vaccines. Flow cytometry using fluorescent hemagglutinin and B cell marker specific Abs enables high throughput qualitative and quantitative detection of individual B cells. By using a graded amount of antigen and gating on GC B cells we define the AC50 the amount of antigen required to stain 50% of hemagglutinin specific B cells. This number is in remarkable agreement with the avidity of the B cell population. This method can be generally employed to include antibody avidity measurements basic and clinical studies of immunity to viruses and other medically relevant immunogens.

Page Manager: Webmaster|Last update: 9/11/2012
Share:

The University of Gothenburg uses cookies to provide you with the best possible user experience. By continuing on this website, you approve of our use of cookies.  What are cookies?