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Generation of DC from mouse spleen cell cultures in response to GM-CSF: Immunophenotypic and functional analyses

Lu, L and Hsieh, M and Oriss, TB and Morel, PA and Starzl, TE and Rao, AS and Thomson, AW (1995) Generation of DC from mouse spleen cell cultures in response to GM-CSF: Immunophenotypic and functional analyses. Immunology, 84 (1). 127 - 134. ISSN 0019-2805

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Abstract

In all tissues that have been studied to date, dendritic leucocytes constitute only a small proportion of total cells and are difficult both to isolate and purify. This study reports on a method for the propagation of large numbers of dendritic cells (DC) from mouse spleen using granulocyte-macrophage colony-stimulating factor (GM-CSF) and their characteristics. Within a few days of liquid culture in GM-CSF, B10 BR (H-2(k), I-E + ) mouse splenocytes formed loosely adherent myeloid cell clusters. Mononuclear progeny released from these clusters at and beyond 4 days exhibited distinct dendritic morphology and strongly expressed leucocyte common antigen (CD45), CD11b, heat-stable antigen, Pgp-1 (CD44) and intercellular adhesion molecule-1 (ICAM-1; CD54). The intensity of expression of the DC-restricted markers NLDC 145 and 33D1, the macrophage marker F4/80, and FcγRII (CDw32) was low to moderate, whereas the cells were negative for CD3, CD45RA and NK1.1. High and moderate levels, respectively, of cell surface staining for major histocompatibility complex (MHC) class II (I-E(k)) and the B7 antigens (counterreceptors of CTLA4, a structural homologue of CD28) were associated with potent stimulation of unprimed, allogeneic T-cells (B10; H-2 b , I-E - ). DC propagated in a similar fashion from DBA/2 mouse spleen proved to be strong antigen-presenting cells (APC) for MHC-restricted, syngeneic T-helper type 2 (Th2) cell clones specifically responsive to sperm whale myoglobin. Footpad or intravenous injection of GM-CSF-stimulated B10.BR spleen-derived DC into B10 (H-2 b , I-E - ) recipients resulted in homing of the allogeneic cells to T-cell-dependent areas of lymph nodes and spleen, where they strongly expressed donor MHC class II antigen 1-2 days later. These findings indicate that cells can be propagated from fresh splenocyte suspensions that exhibit distinctive features of DC, namely morphology, motility, cell-surface phenotype, potent allogeneic and syngeneic APC function and in vivo homing ability. Propagation of DC in this manner from progenitors present in lymphoid tissue provides an alternative and relatively convenient source of high numbers of these otherwise difficult to isolate but functionally important APC.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Lu, L
Hsieh, M
Oriss, TB
Morel, PA
Starzl, TEtes11@pitt.eduTES11
Rao, AS
Thomson, AW
Centers: Other Centers, Institutes, or Units > Thomas E. Starzl Transplantation Institute
Date: 1 January 1995
Date Type: Publication
Journal or Publication Title: Immunology
Volume: 84
Number: 1
Page Range: 127 - 134
Institution: University of Pittsburgh
Refereed: Yes
ISSN: 0019-2805
Other ID: uls-drl:31735062125343, Starzl CV No. 1724
Date Deposited: 08 Apr 2010 17:29
Last Modified: 13 Oct 2017 23:56
URI: http://d-scholarship.pitt.edu/id/eprint/5110

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