Systematic evaluation of the conditions required for the generation of immature rat bone marrow-derived dendritic cells and their phenotypic and functional characterization

Muthana, Munitta; Fairburn, Barbara; Mirza, Shabana; Slack, Laura K.; Pockley, A. Graham

doi:10.1016/j.jim.2004.09.006

Cited by 16 publications

(21 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Plasmacytoid DCs are OX62 Ϫ (Hubert et al 2004). Our own studies have demonstrated that rat bone marrowderived DCs express the OX62 antigen (Muthana et al 2004).…”

Section: Effect Of Gp96 Administration On Splenic Leukocyte Populationsmentioning

confidence: 98%

Administration of the stress protein gp96 prolongs rat cardiac allograft survival, modifies rejection-associated inflammatory events, and induces a state of peripheral T-cell hyporesponsiveness

et al. 2007

Self Cite

View full text Add to dashboard Cite

High-dose gp96 has been shown to inhibit experimental autoimmune disease by a mechanism that appears to involve immunoregulatory CD4 ϩ T cells. This study tested the hypothesis that high-dose gp96 administration modifies allograft rejection and associated inflammatory events. Wistar cardiac allografts were transplanted into Lewis recipient rats and graft function was monitored daily by palpation. Intradermal administration of gp96 purified from Wistar rat livers (100 g) at the time of transplantation and 3 days later significantly prolonged allograft survival (14 vs 8 days in phosphate-buffered saline [PBS]-treated recipients; P ϭ 0.009). Rejected allografts from gp96-treated animals were significantly less enlarged than allografts from their PBS-treated counterparts (2.8 vs 4.3 g; P Ͻ 0.004). Gp96 was also effective when administered on days 1 and 8 (13 vs 7 days), but not if it was derived from recipient (Lewis) liver tissue or administered on days 0, 3, and 6. In parallel studies, CD3 ϩ T cells from gp96-treated untransplanted animals secreted less interleukin (IL)-4, IL-10, and interferon (IFN)-␥ after in vitro polyclonal stimulation than CD3 ϩ T cells from PBS-treated animals. Gp96 administration might therefore influence the induction of immunity to coencountered antigenic challenges and inflammatory events by inducing what appears to be a state of peripheral T-cell hyporesponsiveness.

show abstract

“…Plasmacytoid DCs are OX62 Ϫ (Hubert et al 2004). Our own studies have demonstrated that rat bone marrowderived DCs express the OX62 antigen (Muthana et al 2004).…”

Section: Effect Of Gp96 Administration On Splenic Leukocyte Populationsmentioning

confidence: 98%

Administration of the stress protein gp96 prolongs rat cardiac allograft survival, modifies rejection-associated inflammatory events, and induces a state of peripheral T-cell hyporesponsiveness

et al. 2007

Self Cite

View full text Add to dashboard Cite

show abstract

“…Rat BMDCs were generated as described previously (Muthana et al 2004). Briefly, bone marrow was flushed from the tibias and femurs of Wistar rats with a 25-gauge needle.…”

Section: Generation Of Rat Bmdcsmentioning

confidence: 99%

“…The OX62 ϩ BMDCs generated under these conditions exhibit an immature phenotype, as defined by low levels of CD40, CD54, CD86, and major histocompatibility complex (MHC) class II expression and a pronounced capacity for antigen uptake (Muthana et al 2004). Cells were not harvested before use in any of the assays described below because the disruption and harvesting of DCs and DC clusters induces DC maturation (Pierre et al 1997;Talmor et al 1998;Gallucci et al 1999;Delamarre et al 2003) and might therefore influence the quantitative and qualitative responsiveness of BMDCs to exogenous stimuli.…”

Section: Generation Of Rat Bmdcsmentioning

confidence: 99%

“…We have recently reported a technique for generating resting immature rat bone marrow-derived DCs (BMDCs; Muthana et al 2004), and this study evaluated the influence of gp96 on the phenotype and activities of these cells. The study also investigated the direct influence of gp96 on the activities of purified CD3 ϩ T cells in the absence and presence of costimulation via ligation of the CD28 molecule.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The stress protein gp96 is not an activator of resting rat bone marrow–derived dendritic cells, but is a costimulator and activator of CD3+ T cells

Mirza¹,

Muthana²,

Fairburn³

et al. 2006

Cell Stress Chaper

Self Cite

View full text Add to dashboard Cite

Although low doses of tumor-derived stress protein gp96 elicit protective immunity to the tumor from which it is isolated, protection is lost at high doses because of the induction of immunoregulatory CD4 ϩ T cells. This study evaluated the influence of gp96 on resting rat bone marrow-derived dendritic cells (BMDCs) and purified CD3 ϩ T cells. In contrast to previous reports, gp96 had no effect on adhesion and costimulatory molecule expression by BMDCs, nor did it influence interleukin (IL)-10 and IL-12 secretion or their allostimulatory capacity. Gp96 did not bind to BMDCs but dose-dependently bound to CD4 ϩ and CD8 ϩ T cells. At low concentrations (1 and 25 g/mL), gp96 acted as a costimulator of CD3 ϩ T cells, inducing proliferation and the secretion of interferon (IFN)-␥ and IL-10. Gp96 also increased the proliferation of CD28-costimulated CD3 ϩ T cells and their secretion of IFN-␥, IL-4, and IL-10. Gp96 had no effect at higher concentrations (50 and 100 g/mL), despite the occurrence of cell surface binding at these concentrations. These findings indicate that gp96 can act as a costimulatory molecule for CD3 ϩ T cells, and an observed increase in the IL-10:IFN-␥ secretion ratio induced by gp96 suggests that it might, at appropriate concentrations, promote a regulatory T-helper 2 (Th2)-like phenotype.

show abstract

“…We have previously reported a strategy for generating a population of immature rat bone marrow-derived DCs (BMDCs) [59,60], and demonstrated that these do not express cell surface receptors for gp96 [56]. However, these cells do internalize gp96 via non receptor-mediated endocytosis in the absence of any overt effect on their phenotype [56].…”

Section: Introductionmentioning

confidence: 99%

A non-receptor-mediated mechanism for internalization of molecular chaperones

Pockley

Fairburn

Mirza

et al. 2007

Methods

Self Cite

View full text Add to dashboard Cite

The evolving realization that stress proteins, which have for many years been considered to be exclusively intracellular molecules under normal conditions, can be released from viable cells via a number of potential routes/pathways has prompted interest into their extracellular biology and intercellular signaling properties. That the stress proteins Hsp60, Hsp70 and gp96 can elicit both proand anti-inflammatory effects suggests that these molecules play a key role in the maintenance of immunological homeostasis, and a better understanding of the immunobiology of extracellular stress proteins might reveal new and more effective approaches for controlling and managing infectious disease, inflammatory disease and cancer. A number of cell surface receptors for stress proteins have been identified, and the intracellular consequences of these cell surface receptor-ligand interactions have been characterized. To date, studies into the intercellular signaling properties of stress proteins and their interactions with antigen presenting cells have focused on specific receptor-mediated uptake, and have not considered the fact that such cells can also take up proteins via non-specific endocytosis/pinocytosis. Herein we present a methodological approach for assessing receptormediated and non-receptor mediated uptake of gp96 by rat bone marrow-derived dendritic cells.

show abstract

Systematic evaluation of the conditions required for the generation of immature rat bone marrow-derived dendritic cells and their phenotypic and functional characterization

Cited by 16 publications

References 40 publications

Administration of the stress protein gp96 prolongs rat cardiac allograft survival, modifies rejection-associated inflammatory events, and induces a state of peripheral T-cell hyporesponsiveness

Administration of the stress protein gp96 prolongs rat cardiac allograft survival, modifies rejection-associated inflammatory events, and induces a state of peripheral T-cell hyporesponsiveness

The stress protein gp96 is not an activator of resting rat bone marrow–derived dendritic cells, but is a costimulator and activator of CD3+ T cells

A non-receptor-mediated mechanism for internalization of molecular chaperones

Contact Info

Product

Resources

About