Rapid flow cytometric identification of putative CD14? and CD64? dendritic cells in whole blood

Macey, Marion G.; McCarthy, D.; Vogiatzi, D.; Brown, Kenneth A.; Newland, Adrian C.

doi:10.1002/(sici)1097-0320(19980301)31:3<199::aid-cyto7>3.0.co;2-g

Cited by 19 publications

(8 citation statements)

References 36 publications

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“…Human blood DC have been described to express Fc␥RI and Fc␥RII, whereas mouse DC express all Fc␥Rs: Fc␥RI, RII, and RIII (13)(14)(15). The expression of Fc␥RI on a subset of blood-derived DC has been observed (24), whereas other groups could not detect this surface protein (25). Because the level of Fc␥RI expression on monocytes can be reduced by IL-4 (26), it was suggested that Fc␥RI expression is down-regulated on MDC due to the presence of this cytokine during the culture (15).…”

Section: Discussionmentioning

confidence: 99%

Cross-Linking of CD32 Induces Maturation of Human Monocyte-Derived Dendritic Cells Via NF-κB Signaling Pathway

Bánki

Kacani

Müllauer

et al. 2003

The Journal of Immunology

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Dendritic cells (DC) represent a unique set of APCs that initiate immune responses through priming of naive T cells. Maturation of DC is a crucial step during Ag presentation and can be induced by triggering a broad spectrum of DC surface receptors. Although human DC express several receptors for the Fc portion of IgG which were described to play an important role in Ag internalization, little is known about the effects of IgG or immune complexes on DC maturation. In this study, we show that cross-linking of FcγR-type II (CD32) with immobilized IgG (imIgG) can induce maturation of human monocyte-derived DC via the NF-κB signaling pathway. IgG-mediated maturation was accompanied by a moderate increase of IL-10 secretion, whereas no IL-12 production was observed. Involvement of CD32 was further supported by experiments with the anti-CD32 mAb, which blocked IgG-triggered DC maturation and cytokine secretion significantly. Furthermore, DC cultivated in the presence of imIgG induced allogeneic T cell proliferation. Because this imIgG-induced maturation was considerably impaired in monocyte-derived DC from systemic lupus erythematosus patients, we suggest that DC, which matured in the presence of immune complexes, may contribute to prevention of pathological immune responses.

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Section: Discussionmentioning

confidence: 99%

Cross-Linking of CD32 Induces Maturation of Human Monocyte-Derived Dendritic Cells Via NF-κB Signaling Pathway

Bánki

Kacani

Müllauer

et al. 2003

The Journal of Immunology

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“…Flow cytometric sorting was performed on a FACS Vantage (Becton Dickinson). Only CD64 Ϫ cells (20) were included in the acquisition. OspA-tetramer ϩ CD4 ϩ CD64 Ϫ cells were sorted by single cells into 96-well U-bottom plates, containing 200 l of complete medium and 150,000 irradiated human PBMC (5,000 rads) plus 2 g͞ml phytohemagglutinin, and incubated at 37°C.…”

Section: )mentioning

confidence: 93%

“…To include both small resting and large activated antigen-specific T cells, the analysis was not restricted with respect to size and granularity of cells. Contaminating monocytes, which nonspecifically bind tetramer, were excluded from analysis by selecting the CD64 Ϫ [Fc␥IR Ϫ (20)] population (Fig. 1B, gate R1).…”

Section: Visualization and Frequency Of Ospa-specific T Cells In Lymementioning

confidence: 99%

Direct enumeration ofBorrelia-reactive CD4 T cellsex vivoby using MHC class II tetramers

Meyer

Trollmo

Crawford

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

138

107

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We characterized antigen-specific CD4 ؉ T cells in six patients with treatment-resistant Lyme arthritis, using an HLA-DRB1*0401 major histocompatibility complex (MHC) class II tetramer covalently loaded with OspA164-175, an immunodominant epitope of Borrelia burgdorferi. Direct analysis of OspA-tetramer binding CD4 ؉ cells in patients expressing the HLA-DRB1*0401 allele revealed frequencies of between <0.005 and 0.1% in peripheral blood (n ‫؍‬ 6), and between <0.005 and 3.1% in synovial fluid (n ‫؍‬ 3). OspAtetramer ؉ CD4 ؉ cells were directly cloned at 1 cell per well and expanded by mitogen and IL-2 on allogeneic feeder cells. As measured by [ 3 H]thymidine incorporation, 95% of 168 T cell clones from synovial fluid binding the OspA-tetramer were antigenreactive. Clones generated from peripheral blood revealed a different pattern of responsiveness when compared with clones generated from synovial fluid, as measured by proliferation, IFN-␥, and IL-13 secretion. These clones, selected on the basis of their peptide binding, also responded to whole protein, but with a different cytokine profile. Our studies demonstrate that MHC class II tetramers can be used in humans to directly identify, isolate, and characterize antigen-reactive T cells from an inflammatory compartment.

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“…However, patients' MO surface EP expression levels during PGE 2 insensitivity or EP receptor level relationships to APC defects are still uncharacterized (24). Similarly, the patient MO levels of the two bestdescribed TLR receptors, TLR2 for gram-positive organisms and TLR4 for gram-negative bacteria, during PGE 2 insensitivity, are poorly defined (25,26).…”

Section: Introductionmentioning

confidence: 99%

Abnormal Pge2 Regulation of Monocyte TNF-?? Levels in Trauma Patients Parallels Development of a More Macrophage-Like Phenotype

Laudański

Brouxhon

et al. 2004

Shock

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Some trauma patients' monocytes (MO) increase TNF-alpha levels concomitant to augmenting production of the TNF-alpha inhibitor prostaglandin E2 (PGE2), suggesting posttrauma MO insensitivity to PGE2 effects. This study assesses additional posttrauma MO PGE2 insensitivity effects on altering TNF-alpha form (membrane versus secreted), down-regulating MO receptor expression, and depressing MO APC function. Posttrauma MO TNF-alpha insensitivity to exogenous and autocrine PGE2 correlated to accumulation of TNF-alpha primarily as a membrane-bound cytokine (mTNF-alpha). MO retention of mTNF-alpha correlated with unfavorable clinical outcomes and loss of antigen-presenting cell (APC) function as assessed by depressed MLR and dendritic cell (DC) differentiation. MO TNF-alpha sensitivity to down-regulation by IL-10 was retained, suggesting that PGE2-related functions are specifically altered in these patients' MO. Freshly isolated MO from all trauma patients had decreased expression of Toll-like receptor 4 (TLR4) for gram-negative bacteria. Exogenous PGE2 at high (10 (-6) M) or low (10 (-8) M) concentrations decreased normals' and further decreased APC-competent patients' MO TLR4 expression but had no effect on TLR2. Patients' APC-dysfunctional MO failed to further down-regulate their TLR4 expression in response to additional PGE2, demonstrating another form of PGE2 insensitivity. One of the primary MO prostaglandin receptors, eicosanoid receptor 4 (EP4), was decreased on patients' APC dysfunctional MO, suggesting that depressed EP4 expression could contribute to PGE2 insensitivity in patients' MO. The APC dysfunctional MO's dysregulation of TLR4 expression paralleled increased macrophage-like characteristics such as increased CD64 expression density, elevated mTNF-alpha production, and increased PGE2 levels. Increased PGE2 levels still decreased patients' MO APC functions but failed to depress either MO TLR4 expression or mTNF-alpha levels, suggesting differential involvement of EP receptors in postinjury PGE2-mediated effects.

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Rapid flow cytometric identification of putative CD14? and CD64? dendritic cells in whole blood

Cited by 19 publications

References 36 publications

Cross-Linking of CD32 Induces Maturation of Human Monocyte-Derived Dendritic Cells Via NF-κB Signaling Pathway

Cross-Linking of CD32 Induces Maturation of Human Monocyte-Derived Dendritic Cells Via NF-κB Signaling Pathway

Direct enumeration ofBorrelia-reactive CD4 T cellsex vivoby using MHC class II tetramers

Abnormal Pge2 Regulation of Monocyte TNF-?? Levels in Trauma Patients Parallels Development of a More Macrophage-Like Phenotype

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