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Generation of dendritic cell‐like antigen‐presenting cells in long‐term mixed leucocyte culture: phenotypic and functional studies
Author(s) -
GAO J.X.,
MADRENAS J.,
ZENG W.,
ZHONG R.,
GRANT D.
Publication year - 1997
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1046/j.1365-2567.1997.00220.x
Subject(s) - cd40 , antigen , antigen presenting cell , antigen presentation , biology , dendritic cell , immunology , population , t cell , follicular dendritic cells , spleen , cd11c , cytotoxic t cell , microbiology and biotechnology , chemistry , immune system , medicine , phenotype , in vitro , biochemistry , environmental health , gene
The mechanisms contributing to the proliferation and differentiation of antigen‐presenting cell (APC) precursors upon antigen stimulation or tissue injury are poorly understood. Herein, we report the induction of a population of dendritic cell‐like cells (DLC) with potent antigen‐presentation function from unfractionated spleen cells by means of repetitive allostimulation in long‐term mixed leucocyte cultures (LT‐MLC). Initially, only a few adherent DLC were observed. By 4–6 weeks, however, there were large numbers of DLC which survived persistently. Features of these DLC are closely related to dendritic cells (DC), including: (1) dendritic, veiled or spiny‐processed morphology; (2) expression of a wide array of leucocyte surface markers including DC‐associated or restricted antigens: 33D1, NLDC‐145, CD11c (N418), heat‐stable antigen (HSA), CD44, B7‐1 and B7‐2; (3) ability to migrate to draining lymph nodes and white pulp area of spleen; (4) expression of high level of major histocompatability complex (MHC) class II molecules and (5) more potent mixed leucocyte reaction (MLR)‐stimulating capacity than peritoneal macrophages and APC‐enriched spleen cells. DLC‐stimulated MLR was inhibited by monoclonal antibodies (mAbs) to B7‐1, B7‐2, intracellular adhesion molecule‐1 (ICAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1), leucocyte‐function associated antigen‐1 (LFA‐1) or very‐late activation antigen‐4 (VLA‐4) by 30–55%. When maintained for more than 2 months, the DLC did not lose their MLR‐stimulating activity, but many surface markers were down‐regulated except for Mac‐2 and VCAM‐1, which remained stable or were up‐regulated, respectively. In short‐term culture, the addition of granulocyte–macrophage colony‐stimulating factor (GM‐CSF) or interleukin (IL)‐2 enhanced proliferation of DLC, while tumour necrosis factor‐ α (TNF‐ α ) and IL‐4 did not. IL‐4 suppressed not only ‘spontaneous’, but also GM‐CSF‐enhanced proliferation, suggesting that cytokines play a differential role in DLC proliferation. These results confirm that professional APC can proliferate in response to repetitive antigen stimulation, and their proliferation is differentially regulated by cytokines. A comparison study of DLC with typical DC is being carried out in our laboratory.