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Blood, 1 May 2001, Vol. 97, No. 9, pp. 2680-2687
IMMUNOBIOLOGY
Localization of recombination activating gene 1/green fluorescent
protein (RAG1/GFP) expression in secondary lymphoid organs after
immunization with T-dependent antigens in rag1/gfp
knockin mice
Hideya Igarashi,
Naomi Kuwata,
Kumiko Kiyota,
Kiminobu Sumita,
Toshio Suda,
Shiro Ono,
Steven R. Bauer, and
Nobuo Sakaguchi
From the Departments of Immunology, Pediatrics, and
Cell Differentiation (Institute of Molecular Embryology and Genetics),
Kumamoto University School of Medicine; the Biomedical Research Center,
Osaka University Medical School, Japan; and the Food and Drug
Administration, Center for Biologic Evaluation and Research, Division
of Cellular and Gene Therapies, Bethesda, MD.
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 or
rag1 homozygous knockout mice reconstituted with
rag1/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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