Establishment of lymphotoxin beta receptor signaling-dependent cell lines with follicular dendritic cell phenotypes from mouse lymph nodes.
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 beta receptor mAb and TNF-alpha. 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, FcgammaRIIB, lymphotoxin beta 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.
Nishikawa
Yumiko
Y
Department of Biotechnology, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, Japan.
Hikida
Masaki
M
Magari
Masaki
M
Kanayama
Naoki
N
Mori
Masaharu
M
Kitamura
Hiroshi
H
Kurosaki
Tomohiro
T
Ohmori
Hitoshi
H
D000818:Animals; D053264:B-Cell Activating Factor / Q000235:genetics; D001402:B-Lymphocytes / Q000166:cytology; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology*; D018920:Coculture Techniques; D020566:Dendritic Cells, Follicular / Q000166:cytology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D018858:Germinal Center / Q000166:cytology; D016130:Immunophenotyping; D008198:Lymph Nodes / Q000166:cytology*; D053259:Lymphotoxin beta Receptor; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D015398:Signal Transduction
D053264:B-Cell Activating Factor; D053259:Lymphotoxin beta Receptor