Up-regulation of the dendritic cell marker CD83 on polymorphonuclear neutrophils (PMN): divergent expression in acute bacterial infections and chronic inflammatory disease - PubMed (original) (raw)

Comparative Study

Up-regulation of the dendritic cell marker CD83 on polymorphonuclear neutrophils (PMN): divergent expression in acute bacterial infections and chronic inflammatory disease

C Iking-Konert et al. Clin Exp Immunol. 2002 Dec.

Abstract

Upon cultivation with interferon-gamma (IFN-gamma ) and granulocyte/macrophage-colony stimulating factor (GM-CSF) polymorphonuclear neutrophils (PMN) acquire characteristics of dendritic cells, including expression of major histocompatibility complex (MHC) class II antigens, of the co-stimulatory antigens CD80, CD86 and of CD83, the latter considered to be specific for dendritic cells. Dendritic-like PMN were also able to present to T cells antigens in a MHC class II-restricted manner. To assess whether dendritic-like PMN are also generated in vivo, cells of patients with acute bacterial infections and of patients with chronic inflammatory diseases (primary vasculitis) were tested. During acute infection up to 80% of PMN acquired CD83, but remained negative for MHC class II, CD80 or CD86. PMN of patients with primary vasculitis expressed MHC class II antigens, CD80 and CD86, but not CD83, indicating that up-regulation of MHC class II and of CD83 are not necessarily linked to each other. Indeed, parallel studies with PMN of healthy donors showed that while IFN-gamma and granulocyte/macrophage colony stimulating factor (GM-CSF) induced both, MHC class II and CD83, tumour necrosis factor (TNF)-alpha selectively induced de novo synthesis of CD83. The function of CD83 on PMN is still elusive. A participation in the MHC class II-restricted antigen presentation could be ruled out, consistent with the segregation of MHC class II and CD83 expression. Regardless, however, of its function, CD83 expression could serve as a marker to differentiate between acute and chronic inflammation.

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Figures

Fig. 1

Expression of CD83 on PMN of patients with acute bacterial infection. (a) PMN in whole blood were examined by flow cytofluorometry; cells were labelled with CD66b-FITC as a specific marker for PMN and CD83 PE (clone HB15A) or the isotypic-negative control, respectively. The upper panel shows PMN of a patient with infection; here 33·4% of PMN are double-positive for CD66b/CD83 (right upper quadrant). The right panel shows CD64 expression as a ‘positive control’ for PMN activation. The lower panel shows cells of a patient with active Wegener's granulomatosis. (b) Expression of CD83 on PMN of patients with either bacterial infections, or Wegener's granulomatosis, or of healthy donors. Data are shown for patients with acute bacterial infections (n = 26), patients with Wegener's granulomatosis with active disease (BVAS > 2) (n = 7) or inactive disease (n = 12), and healthy donors (n = 22) and is expressed as percentage of CD83-positive PMN. Each dot represents one patient or donor, respectively, The values of the group ‘bacterial infection’ differed from all the others (largest _P_-value: 0·05).

Fig. 1

Fig. 2

Identification of CD83 on PMN of a patient with acute bacterial infection: of PMN derived from a patient with staphylococcus-induced soft tissue infection, expressing CD83 (cytofluorometry using anti-CD83 clone clone HB15E, panel (a), membranes were prepared and CD83 was precipitated using anti-CD83 (clone HB15A). (b) Silver-stained gel; a band with an apparent molecular weight of 45 kDa (lane 4) is seen; for comparison, immunoprecipitates from in vitro differentiated monocytes are shown (lane 1), freshly isolated PMN (lane 2) or PMN stimulated with IFN-γ+ GM-CSF for 48 h (lane 3). (c) CD83-specific RT-PCR products with the expected size of 445 bp derived from RNA from: PMN cultivated for 8 h in AIM V/NHS (lane 1); IFN-γ (lane 2); TNF-α (lane 3), freshly isolated PMN (lane 4) and patient's PMN (lane 5). The right panel shows the respective amplification products of β-actin.

Fig. 3

Expression of CD83 on isolated PMN: highly purified PMN were cultivated for 48 h under different conditions. From left to right: AIM V containing 2·5% NHS (extreme left), 100 U/ml IFN-γ, 100 U/ml IFN-γ/ 50 U/ml GM-CSF or (extreme right) 2 ng/ml TNF-α.

Fig. 4

Coincidence of expression of CD83 on PMN and presence of TNF-α in the plasma: of 25 patients, CD83 expression on PMN was determined by cytofluorometry; in parallel TNF-α levels in the plasma were determined. The scattergram (a) shows that there is no linear correlation; TNF-α levels higher than 10 pg/ml, however, were found more frequently in patients with PMN expressing CD83 (b).

Fig. 5

Induction of T cell proliferation by PMN: (a) Of a healthy donor TT-specific T cell clones was established. PMN were induced to acquire MHC class II and CD83 by culturing the cells with either IFN-γ (100 U/ml) and GM-CSF (50 U/ml) or additionally TNF-α (2 ng/ml) to further up-regulate CD83 for 24 h. After 5 days of co-culture of T cells with PMN and TT proliferation of T cells was measured by incorporation of [3H]-thymidine. (b) Antibodies (2 µg/ml) to MHC class II or CD83 were added during PMN/T cell/TT co-culture, as was mouse IgG as a negative control. Proliferation was measured after 5 days.

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Up-regulation of the dendritic cell marker CD83 on polymorphonuclear neutrophils (PMN): divergent expression in acute bacterial infections and chronic inflammatory disease - PubMed (original) (raw)