Establishment of lymphotoxin β receptor signaling-dependent cell lines with follicular dendritic cell phenotypes from mouse lymph nodes

Yumiko Nishikawa, Masaki Hikida, Masaki Magari, Naoki Kanayama, Masaharu Mori, Hiroshi Kitamura, Tomohiro Kurosaki, Hitoshi Ohmori

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Follicular dendritic cells (FDCs) have been shown to play a crucial role in the positive selection of high-affinity B cells that are generated by somatic hypermutation in germinal center (GC). Because of technical difficulties in preparing and maintaining pure FDCs, a role for FDCs in this complicated process has not been fully elucidated. In this study, we established a cell line designated as pFL that retained major FDC phenotypes from a three-dimensional culture of mouse lymph node cells. pFL cells proliferated slowly in response to an agonistic anti-lymphotoxin β receptor mAb and TNF-α. A more rapidly growing clone, named FL-Y, with similar requirements for growth was isolated from a long-term culture of pFL. Analysis of surface markers in these two cell lines by immunostaining, flow cytometry, and DNA microarray revealed the expression of genes, including those of CD21, FcγRIIB, lymphotoxin β receptor, ICAM-1, VCAM-1, IL-6, and C4, which have been shown to be characteristic of FDCs. In addition, B cell-activating factor was expressed in these two cell lines. At the pFL or FL-Y:B cell ratio of 1:100, the cell lines markedly sustained B cell survival and Ab production during 2 wk of culture, while most B cells collapsed within 1 wk in the absence of the FDC-like cells. Interestingly, expression of typical GC markers, Fas and GL-7, was notably augmented in B cells that were cocultured with Th cells on these two cell lines. Thus, pFL and FL-Y cells may be useful for providing insight into the functional role for FDCs in GC.

Original languageEnglish
Pages (from-to)5204-5214
Number of pages11
JournalJournal of Immunology
Volume177
Issue number8
Publication statusPublished - Oct 15 2006

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Follicular Dendritic Cells
Lymphotoxin-alpha
Lymph Nodes
Phenotype
Cell Line
B-Lymphocytes
Germinal Center
B-Cell Activating Factor
Vascular Cell Adhesion Molecule-1
Tumor Necrosis Factor Receptors
Intercellular Adhesion Molecule-1
Oligonucleotide Array Sequence Analysis
Interleukin-6
Cell Survival
Flow Cytometry
Clone Cells
Gene Expression
Growth

ASJC Scopus subject areas

  • Immunology

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Establishment of lymphotoxin β receptor signaling-dependent cell lines with follicular dendritic cell phenotypes from mouse lymph nodes. / Nishikawa, Yumiko; Hikida, Masaki; Magari, Masaki; Kanayama, Naoki; Mori, Masaharu; Kitamura, Hiroshi; Kurosaki, Tomohiro; Ohmori, Hitoshi.

In: Journal of Immunology, Vol. 177, No. 8, 15.10.2006, p. 5204-5214.

Research output: Contribution to journalArticle

Nishikawa, Yumiko ; Hikida, Masaki ; Magari, Masaki ; Kanayama, Naoki ; Mori, Masaharu ; Kitamura, Hiroshi ; Kurosaki, Tomohiro ; Ohmori, Hitoshi. / Establishment of lymphotoxin β receptor signaling-dependent cell lines with follicular dendritic cell phenotypes from mouse lymph nodes. In: Journal of Immunology. 2006 ; Vol. 177, No. 8. pp. 5204-5214.
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