Analysis of the Germinal Center Reaction and In Vivo Long-Lived Plasma Cells

This chapter describes the analysis of long- and short-lived plasma cells on tissue sections. Mice are immunized with 4-hydroxy-3-nitrophenyl acetyl (NP) coupled to a T-cell-dependent carrier. Antigen-specific germinal center cells and extrafollicular plasma cells are identified by immunohistology on tissue sections. Plasma cells labeled with 5-bromo-2′-deoxyuridine (BrdU) pulses given during different phases of B-cell response are identified on spleen sections. To identify mutated cells originating from cells in germinal centers, antigen-specific cells from spleen sections are isolated by microdissection. From these NP-specific recombined VDJ genes are amplified with family-specific primers by polymerase chain reaction (PCR) for deoxyribonucleic acid (DNA) sequencing. The methods described can be used to characterize origins and lifespan of plasma cells in vivo.