Analysis of the germinal center reaction and in vivo long-lived plasma cells.

Kai-Michael Toellner, M Khan

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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 life-span of plasma cells in vivo.
Original languageEnglish
Pages (from-to)111-25
Number of pages15
JournalMethods in molecular biology
Volume271
DOIs
Publication statusPublished - 1 Jan 2004

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