Dendritic cells from mouse bone marrow: In vitro differentiation using low doses of recombinant granulocyte-macrophage colony-stimulating factor

Scheicher, Christoph; Mehlig, Maria; Zecher, Reinhard; Reske, Konrad

doi:10.1016/0022-1759(92)90199-4

Cited by 160 publications

(97 citation statements)

References 28 publications

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“…This, together with the morphology and phenotype of the cells, supports the notion that the numerous CD11c ϩ MHCII Ϫ cells in NOD and NOR cultures were monocytes and macrophages and not DC. The codevelopment of macrophages during in vitro culture of BM precursors with GM-CSF has been reported previously in normal non-autoimmunity-prone mice (33). Likewise, we found CD11c ϩ MHCII Ϫ monocytes/macrophages in control cultures, but the frequency of these cells was very low in controls and noteworthy in NOD and NOR.…”

Section: Discussionsupporting

confidence: 87%

Bone Marrow Precursors of Nonobese Diabetic Mice Develop into Defective Macrophage-Like Dendritic Cells In Vitro

Nikolić

Bunk

Drexhage

et al. 2004

The Journal of Immunology

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The NOD mouse spontaneously develops autoimmune diabetes. Dendritic cells (DC) play a crucial role in the autoimmune response. Previous studies have reported a defective DC generation in vitro from the NOD mouse bone marrow (BM), but a deviated development of myeloid precursors into non-DC in response to GM-CSF was not considered. In this study, we demonstrate several abnormalities during myeloid differentiation of NOD BM precursors using GM-CSF in vitro. 1) We found reduced proliferation and increased cell death in NOD cultures, which explain the previously reported low yield of DC progeny in NOD. Cell yield in NOR cultures was normal. 2) In a detailed analysis GM-CSF-stimulated cultures, we observed in both NOD and NOR mice an increased frequency of macrophages, identified as CD11c+/MHCII− cells with typical macrophage morphology, phenotype, and acid phosphatase activity. This points to a preferential maturation of BM precursors into macrophages in mice with the NOD background. 3) The few CD11c+/MHCIIhigh cells that we obtained from NOD and NOR cultures, which resembled prototypic mature DC, appeared to be defective in stimulating allogeneic T cells. These DC had also strong acid phosphatase activity and elevated expression of monocyte/macrophage markers. In conclusion, in this study we describe a deviated development of myeloid BM precursors of NOD and NOR mice into macrophages and macrophage-like DC in vitro. Potentially, these anomalies contribute to the dysfunctional regulation of tolerance in NOD mice yet are insufficient to induce autoimmune diabetes because they occurred partly in NOR mice.

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Section: Discussionsupporting

confidence: 87%

Bone Marrow Precursors of Nonobese Diabetic Mice Develop into Defective Macrophage-Like Dendritic Cells In Vitro

Nikolić

Bunk

Drexhage

et al. 2004

The Journal of Immunology

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“…For practical reasons we did not use tissue-derived LC to examine the effects of ORFV-IL-10 in cell culture, although it was possible to show that ORFV-IL-10 impaired the migration of LC in skin explant tissue. LC are difficult to isolate from skin and undergo maturation in culture (Teunissen et al, 1990) whereas BMDC are readily grown in cell culture, resemble DC morphologically and display almost all of the characteristics of DC (Inaba et al, 1992;Scheicher et al, 1992;Faulkner et al, 2000). BMDC have the ability to take up, process and present antigen, the ability to interact with and stimulate T cell responses (Inaba et al, 1992;Faulkner et al, 2000) and the ability to migrate and function as DC when injected in vivo (Inaba et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

Orf virus-encoded interleukin-10 inhibits maturation, antigen presentation and migration of murine dendritic cells

Lateef

Fleming

Halliday

et al. 2003

Journal of General Virology

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Orf virus (ORFV) belongs to the genus Parapoxvirus and induces cutaneous pustular lesions in sheep, goats and humans. ORFV is unusual in that it has the ability to reinfect its host and this suggests that the generation of immunological memory has been impaired, thus exposing the host to subsequent infection. The discovery that ORFV encodes an IL-10-like virokine raises the question of whether this factor adversely affects the cells that initiate the acquired immune response. We examined the effect of ORFV-IL-10 on immature murine bone marrow-derived dendritic cells (BMDC). Immature BMDC are activated on exposure to antigen and undergo maturation. This process is characterized by increased expression of CD80, CD86 and MHC class II and reduced antigen uptake. We found that the maturation of BMDC is impaired in cells treated with ORFV-IL-10 prior to antigen exposure and this was exemplified by the reduced expression of the cell-surface markers described above. We have also shown that the activation of a haemagglutinin peptide (HAT)-specific T cell hybridoma by dendritic cell-mediated presentation of HAT and heat-inactivated influenza virus AP8/34 was markedly reduced following exposure to ORFV-IL-10. Finally, we examined the effect of ORFV-IL-10 on Langerhans' cell (LC) migration using cultured murine skin explant tissue and showed that this virokine impaired the spontaneous migration of LC from the epidermis and induced changes in LC morphology. Our findings suggest that ORFV-IL-10 has the capacity to impair the initiation of an acquired immune response and hence inhibit the generation of immunological memory necessary for immunity on subsequent exposure. INTRODUCTIONThe initiation of an acquired immune response to a virus requires that antigen be taken up by dendritic cells (DC) in the periphery, processed and transported to the draining lymphoid tissue to be presented to T cells (reviewed in Palucka & Banchereau, 2002). For this sequence of events to occur, DC must first become activated by signals transmitted through the interaction of viral products with pattern recognition receptors expressed on the surface of these cells. DC form the vital bridge between the innate response and the acquired response. In doing so they become an attractive target for virally mediated immunosuppression.Viruses evade the immune response of the host by an array of mechanisms including the production of cytokine homologues capable of deflecting, down regulating or aborting cell-mediated immunity (Alcami & Koszinowski, 2000).Interleukin (IL)-10 homologues are produced by EpsteinBarr virus (EBV) (Moore et al., 1990), equine herpesvirus (EHV) (Rode et al., 1993), cytomegalovirus (Kotenko et al., 2000, Yaba-like disease virus (Lee et al., 2001) and orf virus (ORFV) (Fleming et al., 1997). Mammalian IL-10, a prototypic anti-inflammatory cytokine, is capable of both immunosupressive and immunostimulatory functions (reviewed in Fickenscher et al., 2002). This cytokine is known to inhibit DC function and ultimately the induction of ant...

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“…The in vitro differentiation of BMDC was adapted from previously published protocols (43,44). Myeloid DC were obtained by culture of bone marrow cells in DMEM (Life Technologies) containing 10% of heatinactivated FCS (Life Technologies), penicillin-streptomycin and 10% of X6310 derived GM-CSF as described (45).…”

Section: Differentiation Of Bone Marrow-derived DC (Bmdc)mentioning

confidence: 99%

Dendritic Cells Require STAT-1 Phosphorylated at Its Transactivating Domain for the Induction of Peptide-Specific CTL

Pilz

Kratky

Stockinger

et al. 2009

The Journal of Immunology

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Phosphorylation of transcription factor STAT-1 on Y701 regulates subcellular localization whereas phosphorylation of the transactivating domain at S727 enhances transcriptional activity. In this study, we investigate the impact of STAT-1 and the importance of transactivating domain phosphorylation on the induction of peptide-specific CTL in presence of the TLR9-dependent immune adjuvant IC31. STAT-1 deficiency completely abolished CTL induction upon immunization, which was strongly reduced in animals carrying the mutation of the S727 phospho-acceptor site. A comparable reduction of CTL was found in mice lacking the type I IFN (IFN-I) receptor, whereas IFN-γ-deficient mice behaved like wild-type controls. This finding suggests that S727-phosphorylated STAT-1 supports IFN-I-dependent induction of CTL. In adoptive transfer experiments, IFN-I- and S727-phosphorylated STAT-1 were critical for the activation and function of dendritic cells. Mice with a T cell-specific IFN-I receptor ablation did not show impaired CTL responses. Unlike the situation observed for CTL development S727-phosphorylated STAT-1 restrained proliferation of naive CD8+ T cells both in vitro and following transfer into Rag-deficient mice. In summary, our data reveal a dual role of S727-phosphorylated STAT-1 for dendritic cell maturation as a prerequisite for the induction of CTL activity and for T cell autonomous control of activation-induced or homeostatic proliferation.

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Dendritic cells from mouse bone marrow: In vitro differentiation using low doses of recombinant granulocyte-macrophage colony-stimulating factor

Cited by 160 publications

References 28 publications

Bone Marrow Precursors of Nonobese Diabetic Mice Develop into Defective Macrophage-Like Dendritic Cells In Vitro

Bone Marrow Precursors of Nonobese Diabetic Mice Develop into Defective Macrophage-Like Dendritic Cells In Vitro

Orf virus-encoded interleukin-10 inhibits maturation, antigen presentation and migration of murine dendritic cells

Dendritic Cells Require STAT-1 Phosphorylated at Its Transactivating Domain for the Induction of Peptide-Specific CTL

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