Introduction of genes into primary murine splenic B cells using retrovirus vectors.
Primary murine splenic B cells can be cultured ex vivo and, following treatment with LPS, cytokines and/or CD40L proliferate, and undergo class switch recombination and terminal differentiation to become immunoglobulin-secreting plasmacytic cells. Methods are described here for introducing genes, encoding either normal or blocking forms of experimental proteins, into murine splenic B cells using retrovirus vectors. This makes it possible to study the effects of these proteins on late stages of B-cell development, including proliferation, class switch recombination, and plasmacytic differentiation.
Lin
Kuo-I
KI
Department of Microbiology, Columbia University, New York, NY, USA.
Calame
Kathryn
K
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000821:virology; D023201:CD40 Ligand / Q000494:pharmacology; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006801:Humans; D007075:Immunoglobulin M / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D012327:RNA Virus Infections / Q000523:psychology; D012190:Retroviridae / Q000502:physiology*; D013154:Spleen / Q000166:cytology* / Q000276:immunology; D019065:Virus Assembly / Q000502:physiology
D016207:Cytokines; D007075:Immunoglobulin M; D008070:Lipopolysaccharides; D008164:Luminescent Proteins; D023201:CD40 Ligand; D049452:Green Fluorescent Proteins