A novel CD4-CD8α+CD205+CD11b- murine spleen dendritic cell line: establishment, characterization and functional analysis in a model of vaccination to toxoplasmosis

Ruiz, Sophie; Beauvillain, Céline; Mévélec, Marie-Noëlle; Roingeard, Philippe; Breton, Pascal; Bout, Daniel; Dimier‐Poisson, Isabelle

doi:10.1111/j.1462-5822.2005.00583.x

Cited by 27 publications

(15 citation statements)

References 31 publications

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“…Among the different examples of oncogene-based approaches to immortalize DC lines, several strategies are based on ex vivo transduction or transfection of DCs with the SV40LgT (77, 83, 86). Our approach is different because it is based on the generation of SV40LgT-transgenic mice and on the transgene-induced transformation of DCs in vivo with consequent development of DC tumors (42).…”

Section: Discussionmentioning

confidence: 99%

Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line

et al. 2018

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Dendritic cells (DCs) are the most potent antigen presenting cells and possess an incomparable ability to activate and instruct T cells, which makes them one of the cornerstones in the regulation of the cross-talk between innate and adaptive immunity. Therefore, a deep understanding of DC biology lays the foundations to describe and to harness the mechanisms that regulate the development of the adaptive response, with clear implications in a vast array of fields such as the study of autoimmune diseases and the development of new vaccines. However, the great difficulty to obtain large quantities of viable non-activated DCs for experimentation have considerably hindered the progress of DC research. Several strategies have been proposed to overcome these limitations by promoting an increase of DC abundance in vivo, by inducing DC development from DC progenitors in vitro and by generating stable DC lines. In the past years, we have described a method to derive immortalized stable DC lines, named MutuDCs, from the spleens of Mushi1 mice, a transgenic mouse strain that express the simian virus 40 Large T-oncogene in the DCs. The comparison of these DC lines with the vast variety of DC subsets described in vivo has shown that all the MutuDC lines that we have generated so far have phenotypic and functional features of type 1 conventional DCs (cDC1s). With the purpose of deriving DC lines with characteristics of type 2 conventional DCs (cDC2s), we bred a new Batf3−/− Mushi1 murine line in which the development of the cDC1 subset is severely defective. The new MutuDC line that we generated from Batf3−/− Mushi1 mice was phenotypically and functionally characterized in this work. Our results demonstrated that all the tested characteristics of this new cell line, including the expression of subset-determining transcription factors, the profile of cytokine production and the ability to present antigens, are comparable with the features of splenic CD4− cDC2s. Therefore, we concluded that our new cell line, that we named CD4− MutuDC2 line, represents a valuable model for the CD4− cDC2 subset.

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Section: Discussionmentioning

confidence: 99%

Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line

et al. 2018

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“…The resulting SVDC line represents an enhancement to GM-CSF-driven BMDC given the relatively larger number of cells available, but the requirement of culture at 33°C may represent a technical limitation. A second cell line, the SRDC line, with a CD8a + phenotype, results from transformation of splenic DC in vitro by transfection with a plasmid coding for SV40 LgT (Ruiz et al, 2005). A third cell line is the DC 2.4 cell line, based on supertransfection with myc and raf in GM-CSF-transduced BM cells (Shen et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

“…In order to obtain DC cultures of the CD8α + subset, DC are generally derived from BM using Flt3L rather than GM-CSF (Naik et al, 2005). Only one DC line with a CD8α + phenotype has been previously reported, the SRDC line (without further follow-up studies; Ruiz et al, 2005). …”

Section: Discussionmentioning

confidence: 99%

“…These include the D1 cells, a growth factor-dependent immature DC line derived from mouse spleen DC, which can be “matured” with LPS (Winzler et al, 1997; Mortellaro et al, 2009). The generation of murine DC lines based on oncogene-driven immortalization has also been reported, including the SRDC line (Ruiz et al, 2005), the SVDC line (Ebihara et al, 2004), and the DC 2.4 cell line (Shen et al, 1997). In humans, DC lines can be generated from the culture of leukemic DC found in the blood of acute myeloid leukemia patients (Mohty et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Novel Murine Dendritic Cell Lines: A Powerful Auxiliary Tool for Dendritic Cell Research

Marraco¹,

Grosjean²,

Duval³

et al. 2012

Front. Immun.

138

157

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Research in vitro facilitates discovery, screening, and pilot experiments, often preceding research in vivo. Several technical difficulties render Dendritic Cell (DC) research particularly challenging, including the low frequency of DC in vivo, thorough isolation requirements, and the vulnerability of DC ex vivo. Critically, there is not as yet a widely accepted human or murine DC line and in vitro systems of DC research are limited. In this study, we report the generation of new murine DC lines, named MutuDC, originating from cultures of splenic CD8α conventional DC (cDC) tumors. By direct comparison to normal WT splenic cDC subsets, we describe the phenotypic and functional features of the MutuDC lines and show that they have retained all the major features of their natural counterpart in vivo, the splenic CD8α cDC. These features include expression of surface markers Clec9A, DEC205, and CD24, positive response to TLR3 and TLR9 but not TLR7 stimuli, secretion of cytokines, and chemokines upon activation, as well as cross-presentation capacity. In addition to the close resemblance to normal splenic CD8α cDC, a major advantage is the ease of derivation and maintenance of the MutuDC lines, using standard culture medium and conditions, importantly without adding supplementary growth factors or maturation-inducing stimuli to the medium. Furthermore, genetically modified MutuDC lines have been successfully obtained either by lentiviral transduction or by culture of DC tumors originating from genetically modified mice. In view of the current lack of stable and functional DC lines, these novel murine DC lines have the potential to serve as an important auxiliary tool for DC research.

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“…generated an immortalized DC cell line, termed SRDCs, that have a phenotype, morphology and activity similar to CD4-CD8α+ CD205+ CD11b- DCs purified ex vivo (Ruiz et al, 2005). SRDCs were cultivated as described (Ruiz et al, 2005). MNV-1 infection of SRDCs resulted in titers similar to MNV-1 infection of RAW 264.7 cells (Fig.…”

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confidence: 99%

Murine norovirus-1 entry into permissive macrophages and dendritic cells is pH-independent

2009

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SummaryMurine norovirus (MNV) is a recently discovered mouse pathogen. Unlike the fastidious human noroviruses that cause the overwhelming majority of non-bacterial gastroenteritis worldwide, MNV readily infects cells in culture. Its replication in primary murine macrophages and dendritic cells and their derived cell lines allows the study of norovirus cell entry for the first time. In this study we determined the role of pH during MNV-1 infection since the low pH environment of endosomes often triggers uncoating of viruses. We demonstrated that MNV-1 viral titers by plaque assay and expression of the non-structural protein VPg by immunofluorescence were not affected by pH in cultured and primary macrophages and dendritic cells in the presence of two known endosome acidification inhibitors, bafilomycin A1 and chloroquine. These data indicate that MNV-1 enters permissive cells in a pH-independent manner. KeywordsRNA virus; norovirus; pH-independent entry; macrophages; dendritic cells; enteric virusNoroviruses are an understudied group of non-enveloped positive strand RNA viruses that belong to the Caliciviridae family (Green, 2007). Despite the significant impact of human noroviruses (HuNoV) on public health worldwide as the major agent of non-bacterial gastroenteritis (Green, 2007), no drug or vaccine exists to treat norovirus infections. This is partially due to the absence of a robust tissue culture system (Duizer et al., 2004;Straub et al., 2007). In contrast, murine norovirus (MNV), a highly prevalent agent in research mouse colonies (Hsu et al., 2005;Muller et al., 2007), readily infects murine macrophages and dendritic cells (DC) in culture and in vivo (Ward et al., 2006;Wobus et al., 2004;. Similar to HuNoV, MNV replicates in the gastrointestinal tract of its wild type or immunocompromised host, is shed in the feces, and is transmitted by the fecal-oral route (reviewed in: ). The ability to culture a norovirus has already led to insights into norovirus biology (Chaudhry et al., 2006;Daughenbaugh et al., 2006;Simmonds et al., 2008;Sosnovtsev et al., 2006) and inactivation (for example: (Baert et al., 2008;Belliot et al., 2008). However, no studies have yet addressed requirements for norovirus entry into cells. , email: E-mail: cwobus@umich.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. To gain access into host cells, viruses hijack cellular processes. The most commonly used endocytic pathway for virus entry is clathrin-mediated endocytosis. Viral entry can also occur via caveolin-mediated endocytosis, clathrin/caveolin-independent endocytosis, macropinocytosi...

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A novel CD4-CD8α+CD205+CD11b- murine spleen dendritic cell line: establishment, characterization and functional analysis in a model of vaccination to toxoplasmosis

Cited by 27 publications

References 31 publications

Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line

Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line

Novel Murine Dendritic Cell Lines: A Powerful Auxiliary Tool for Dendritic Cell Research

Murine norovirus-1 entry into permissive macrophages and dendritic cells is pH-independent

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