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, Nobuo Sakaguchi

Research output: Contribution to journalArticle

37 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)2680-2687
Number of pages8
JournalBlood
Volume97
Issue number9
DOIs
Publication statusPublished - 2001 May 1

Fingerprint

RAG-1 Genes
Immunization
Viral Tumor Antigens
Green Fluorescent Proteins
Germinal Center
Genes
Antigens
B-Lymphocytes
Cells
Blastodisc
Spleen
Peanut Agglutinin
Interleukin-7
Immunoglobulin Genes
T-cells
Gene Rearrangement
Gene expression
Knockout Mice
Interleukin-4
Genetic Recombination

ASJC Scopus subject areas

  • Hematology

Cite this

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. / Igarashi, Hideya; Kuwata, Naomi; Kiyota, Kumiko; Sumita, Kiminobu; Suda, Toshio; Ono, Shiro; Bauer, Steven R.; Sakaguchi, Nobuo.

In: Blood, Vol. 97, No. 9, 01.05.2001, p. 2680-2687.

Research output: Contribution to journalArticle

Igarashi, Hideya ; Kuwata, Naomi ; Kiyota, Kumiko ; Sumita, Kiminobu ; Suda, Toshio ; Ono, Shiro ; Bauer, Steven R. ; Sakaguchi, Nobuo. / 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. In: Blood. 2001 ; Vol. 97, No. 9. pp. 2680-2687.
@article{a310c7b0b20549fe8f9e1f13accbfc2d,
title = "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",
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 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.",
author = "Hideya Igarashi and Naomi Kuwata and Kumiko Kiyota and Kiminobu Sumita and Toshio Suda and Shiro Ono and Bauer, {Steven R.} and Nobuo Sakaguchi",
year = "2001",
month = "5",
day = "1",
doi = "10.1182/blood.V97.9.2680",
language = "English",
volume = "97",
pages = "2680--2687",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "9",

}

TY - JOUR

T1 - 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

AU - Igarashi, Hideya

AU - Kuwata, Naomi

AU - Kiyota, Kumiko

AU - Sumita, Kiminobu

AU - Suda, Toshio

AU - Ono, Shiro

AU - Bauer, Steven R.

AU - Sakaguchi, Nobuo

PY - 2001/5/1

Y1 - 2001/5/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0035353180&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035353180&partnerID=8YFLogxK

U2 - 10.1182/blood.V97.9.2680

DO - 10.1182/blood.V97.9.2680

M3 - Article

C2 - 11313258

AN - SCOPUS:0035353180

VL - 97

SP - 2680

EP - 2687

JO - Blood

JF - Blood

SN - 0006-4971

IS - 9

ER -