The SW(HEL) system for high-resolution analysis of in vivo antigen-specific T-dependent B cell responses

Methods Mol Biol. 2015:1291:103-23. doi: 10.1007/978-1-4939-2498-1_9.

Abstract

T cell-dependent B cell responses generate optimal antibodies to combat foreign antigens. Naïve B cells responding to antigen undergo a complex series of differentiation events and cell fate decisions to provide long-lived memory B cells and plasma cells. Historically, B cell biologists have been challenged by the task of investigating rare antigen-specific B cells in an in vivo setting such that their interactions with antigen, regulation and migration may be accurately tracked. We have developed the SW(HEL) experimental system capable of accurately monitoring B cells that interact with a protein antigen and then subsequently undergo isotype switching, somatic hypermutation, and affinity maturation within germinal centers (GC) to generate high-affinity antibodies. Here we provide a comprehensive description of the procedures involved in establishing and using the SW(HEL) system to assess B cell responses to a foreign antigen. This system can provide a valuable measure of the functional capabilities of T follicular helper cells, whose role is ultimately to support and shape long-term humoral immunity.

MeSH terms

  • Adoptive Transfer
  • Animals
  • Antibodies / blood
  • Antibody Affinity / immunology
  • B-Lymphocytes / immunology*
  • Chickens
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / immunology*
  • Erythrocytes / metabolism
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Genotyping Techniques
  • Immunologic Techniques / methods*
  • Lymphocyte Subsets / immunology
  • Mice, Transgenic
  • Muramidase / metabolism*
  • Polymerase Chain Reaction
  • Recombinant Proteins / metabolism
  • Sheep
  • Somatic Hypermutation, Immunoglobulin
  • Spleen / cytology
  • Staining and Labeling
  • T-Lymphocytes / immunology*

Substances

  • Antibodies
  • Epitopes
  • Recombinant Proteins
  • Muramidase