Examination of the Role of Galectins in Plasma Cell Differentiation
Plasma cells are terminally differentiated B cells that develop via the stimulation of mature B cells with various agents such as antigens and mitogens. Recently, we found that plasma cell differentiation can be modulated by galectin-1 and galectin-8; these galectins appear to play additive and redundant roles in promoting the production of antibody. Here, we describe the protocols for how to investigate the roles of galectins in plasma cell differentiation. These methods include the preparation of recombinant galectins from Escherichia coli for exogenously treating primary B cells, generation of galectin_Fcm fusion proteins for determining their binding to B cells, introduction of ectopic galectins in primary B cells using retroviral vectors, and inhibition of the binding of galectins to B cells by synthetic disaccharides.
Key wordsMouse splenic B cells Plasma cell Recombinant galectin Retroviral vector Flow cytometry
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