Methods to Study the Role of Cdc42, Rac1, and Rac2 in B-Cell Cytoskeletal Responses

Methods Mol Biol. 2018:1821:235-246. doi: 10.1007/978-1-4939-8612-5_16.

Abstract

B-cell migration and adhesion are critical to form a germinal center response, the site for B-cell production of high-affinity antibodies. Here, we describe two assays that can be used to examine B-cell cytoskeletal responses needed during the germinal center response: B-cell spreading and homotypic adhesion. Spreading of B cells is dependent on Cdc42, while Rac1 and Rac2 are necessary for homotypic adhesion. These in vitro assays can be used to examine functional responses of B cells mediated by the cell cytoskeleton, for example when comparing B cells from different gene knockout animals.

Keywords: Adhesion; B lymphocytes; In vitro responses; Polyclonal activation; Spreading.

MeSH terms

  • Animals
  • B-Lymphocytes / cytology
  • B-Lymphocytes / enzymology*
  • Cell Adhesion / physiology
  • Cytoskeleton / enzymology*
  • Cytoskeleton / genetics
  • Germinal Center / cytology
  • Germinal Center / enzymology*
  • Mice
  • Mice, Knockout
  • Neuropeptides / genetics
  • Neuropeptides / metabolism*
  • RAC2 GTP-Binding Protein
  • cdc42 GTP-Binding Protein / genetics
  • cdc42 GTP-Binding Protein / metabolism*
  • rac GTP-Binding Proteins / genetics
  • rac GTP-Binding Proteins / metabolism*
  • rac1 GTP-Binding Protein / genetics
  • rac1 GTP-Binding Protein / metabolism*

Substances

  • Cdc42 protein, mouse
  • Neuropeptides
  • Rac1 protein, mouse
  • cdc42 GTP-Binding Protein
  • rac GTP-Binding Proteins
  • rac1 GTP-Binding Protein