Localization of recombination activating gene 1/green fluorescent protein (RAG1/GFP) expression in secondary lymphoid organs after immunization with T-dependent …

H Igarashi, N Kuwata, K Kiyota, K Sumita… - Blood, The Journal …, 2001 - ashpublications.org
H Igarashi, N Kuwata, K Kiyota, K Sumita, T Suda, S Ono, SR Bauer, N Sakaguchi
Blood, The Journal of the American Society of Hematology, 2001ashpublications.org
Secondary rearrangements of immunoglobulin gene segments that generate a new
antibody repertoire in peripheral B cells have been described as receptor revision and occur
by as yet unknown mechanisms. To determine the importance of recombination activating
gene (RAG) expression in receptor revision, heterozygous rag1/green fluorescent protein
(gfp) knockin mice were used to examine the location of RAG1 expression in the germinal
centers (GCs) of lymphoid follicles after immunization with a variety of T-cell–dependent …
Abstract
Secondary rearrangements of immunoglobulin gene segments that generate a new antibody repertoire in peripheral B cells have been described as receptor revision and occur by as yet unknown mechanisms. To determine the importance of recombination activating gene (RAG) expression in receptor revision, heterozygous rag1/green fluorescent protein (gfp) knockin mice were used to examine the location of RAG1 expression in the germinal centers (GCs) of lymphoid follicles after immunization with a variety of T-cell–dependent antigens. Immunization of rag1/gfp heterozygous mice orrag1 homozygous knockout mice reconstituted withrag1/gfp heterozygous spleen cells caused the down-regulation of RAG1/GFP signal in GCs. Although some RAG1/GFP+ cells appeared in regions surrounding the peanut agglutinin (PNA)+GL-7+ GC area, RAG1/GFP+ cells did not accumulate in the central region. In addition, the stimulation of spleen B cells with anti-μ antibody plus interleukin-4 (IL-4) or with anti-CD40 monoclonal antibody plus IL-7 did not induce GFP signals at detectable levels in vitro. These results clearly demonstrate that RAG1 re-expression either does not occur or is at extremely low levels in antigen-driven B cells in GCs of secondary lymphoid follicles, suggesting that other mechanisms may mediate the gene rearrangements observed in receptor revision.
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