Rho GTPases pp 235-246 | Cite as

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

  • Natalija Gerasimcik
  • Lisa S. Westerberg
  • Eva SeverinsonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1821)


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.

Key words

B lymphocytes In vitro responses Polyclonal activation Adhesion Spreading 

Supplementary material

Video 1

Formation of aggregates in B cells cultured with anti-CD40 + IL-4. B cells were stimulated for 2 days. Bright-field images were taken every minute. Note that big aggregates are formed by fusion of smaller ones. Magnification 10× (MOV 61702 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Natalija Gerasimcik
    • 1
  • Lisa S. Westerberg
    • 2
  • Eva Severinson
    • 3
    Email author
  1. 1.Department of Dental MedicineDivision of Periodontology, Karolinska InstitutetHuddingeSweden
  2. 2.Department of MicrobiologyTumor and Cell Biology, Karolinska InstitutetStockholmSweden
  3. 3.Department of Molecular Biosciences, The Wenner Gren InstituteStockholm UniversityStockholmSweden

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