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Receptor Signaling-Dependent Cell Lines with Follicular Dendritic Cell Phenotypes from Mouse Lymph Nodes1
* Department of Biotechnology, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, Japan;
Laboratory for Lymphocyte Differentiation, RIKEN Research Center for Allergy and Immunology, Tsurumi-ku, Yokohama, Kanagawa, Japan;
Laboratory of Immunogenomics, RIKEN Research Center for Allergy and Immunology, Tsurumi-ku, Yokohama, Kanagawa, Japan; and
Faculty of Health and Welfare Sciences, Okayama Prefectural University, Soja, Okayama, Japan
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 
receptor mAb and TNF-
. 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, Fc
RIIB, lymphotoxin 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.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This work was supported by a grant-in-aid from the Ministry of Education, Science, Culture, and Sports of Japan (16360414; to H.O.).
2 Address correspondence and reprint requests to Dr. Hitoshi Ohmori, Department of Biotechnology, Okayama University, 3-1-1 Tsushima-Naka, Okayama 700-8530, Japan. E-mail address: hit2224{at}cc.okayama-u.ac.jp
3 Abbreviations used in this paper: GC, germinal center; FDC, follicular dendritic cell; BAFF, B cell-activating factor; BMDC, bone marrow-derived DC; CGG, chicken -globulin; Con A-sup, Con A-stimulated rat spleen cell culture; HS, horse serum; IC, immune complex; KLH, keyhole limpet hemocyanin; LN, lymph node; LT, lymphotoxin; NIP, 4-hydroxy-3-iodo-5-nitrophenylacetyl; NP, 4-hydroxy-3-nitrophenylacetyl; TNP, 2,4,6-trinitrophenyl.
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