The Expression of HLA-DR, IL-2R, ICAM-1 and TNF Alpha Molecules as Possible Prognostic Elements of Hypertrophic Scarring in Burns

Granulocyte/macrophage CSF (GM-CSF) has recently been identified in rheumatoid arthritis (RA) synovial effusions. To study a potential role for GM-CSF and other cytokines on the induction of HLA-DR expression on monocytes and synovial macrophages, we analyzed the relative ability of recombinant human cytokines to induce the surface expression of class II MHC antigens on normal peripheral blood monocytes by FACS analysis. GM-CSF (800 U/ml) (mean fluorescence channel 2.54 +/- 0.33 times the control, p less than 0.001) and IFN-gamma (100 U/ml) (5.14 +/- 0.60, p less than 0.001) were the most potent inducers of HLA-DR. TNF-alpha and IL-4 also increased HLA-DR expression, although to a lesser degree [1.31 +/- 0.06 (p less than 0.02) and 1.20 +/- 0.03 (p less than 0.01), respectively]. IL-1 (40 U/ml), IL-2 (10 ng/ml), IL-3 (50 U/ml), IL-6 (100 U/ml), and CSF-1 (1,000 U/ml) did not affect surface HLA-DR density. GM-CSF also increased HLA-DR mRNA expression and surface HLA-DQ expression, but decreased CD14 (a monocyte/macrophage antigen) expression. The effect of GM-CSF on HLA-DR was not mediated by the generation of IFN-gamma in vitro because it was not blocked by anti-IFN-gamma mAb. GM-CSF was additive with IL-4 and low amounts (less than 3 U/ml) of IFN-gamma and synergistic with TNF-alpha. Because we have recently reported that supernatants of cultured RA synovial cells produce a non-IFN-gamma factor that induces HLA-DR on monocytes, we then attempted to neutralize this factor with specific anti-GM-CSF mAb. Four separate synovial tissue supernatants were studied, and the antibody neutralized the HLA-DR-inducing factor in each (p less than 0.01).

Download Full-text

Bicycle exercise enhances plasma IL-6 but does not change IL-1 alpha, IL-1 beta, IL-6, or TNF-alpha pre-mRNA in BMNC

Journal of Applied Physiology ◽

10.1152/jappl.1994.77.1.93 ◽

1994 ◽

Vol 77 (1) ◽

pp. 93-97 ◽

Cited By ~ 125

Author(s):

H. Ullum ◽

P. M. Haahr ◽

M. Diamant ◽

J. Palmo ◽

J. Halkjaer-Kristensen ◽

...

Keyword(s):

Plasma Levels ◽

Mononuclear Cells ◽

Absolute Number ◽

Tnf Alpha ◽

Bicycle Exercise ◽

Necrosis Factor Alpha ◽

Concentric Exercise ◽

Hla Dr ◽

Cytokine Plasma ◽

Activated Monocytes

The present study investigated the effect of concentric exercise on cytokine plasma levels and cytokine pre-mRNA in blood mononuclear cells (BMNCs). Healthy young moderately trained men performed ergometer bicycle exercise for 1 h at 75% of maximal oxygen uptake. The levels of plasma interleukin (IL)-6 increased significantly during exercise, but plasma levels of IL-1 alpha, IL-1 beta, and tumor necrosis factor-alpha (TNF-alpha) were below the detection limit in most subjects. Pre-mRNA for IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha could be detected in BMNCs, but the amounts did not change in relation to exercise. These results indicate that, although the absolute number of monocytes increases during exercise and the percentage of CD14+/HLA-DR+ and CD14+/HLA-DR- monocytes increases after exercise, the increased plasma levels of IL-6 during exercise is not likely to be a result of activated monocytes in peripheral blood.

Download Full-text

Beneficial Effect of Pentoxifylline on Thyroid Associated Ophthalmopathy (TAO)*: a pilot study

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.82.6.9995 ◽

1997 ◽

Vol 82 (6) ◽

pp. 1999-2002 ◽

Cited By ~ 37

Author(s):

Cs. Balazs ◽

E. Kiss ◽

A. Vamos ◽

I. Molnar ◽

N.R. Farid

Keyword(s):

Pilot Study ◽

Soft Tissue ◽

Beneficial Effect ◽

Tnf Alpha ◽

Thyroid Peroxidase ◽

Thyroid Antibodies ◽

Muscle Involvement ◽

Eye Muscle ◽

Hla Dr ◽

Thyroid Associated Ophthalmopathy

Abstract We have previously found that pentoxifylline (Ptx) inhibited cytokine induced HLA-DR expression and glycosaminoglycan (GAG) synthesis by retroorbital fibroblasts. We have now tested the clinical efficacy of Ptx in treating TAO. Ten patients with moderately severe ophthalmopathy were selected for study. All patients were euthyroid before and during the 12 weeks of the Ptx therapy. Serum GAG, TNF-alpha, anti-TSH-receptor, anti-eye muscle, anti-thyroglobulin and anti-thyroid peroxidase antibodies were determined sequentially. At the end of 12 weeks eight of the ten patients showed improvement in soft tissue but not in proptosis or extraocular muscle involvement. At baseline the levels of GAG (5.2±0.92 mg/dl v.s. 0.7±0.14 mg/dl, p<0.001) and TNF-alpha (33.6±6.6 pg/ml v.s. 5.4±1.3 pg/ml, p<0.001) were increased in patients compared to controls. They gradually decreased in the eight patients who responded to Ptx: after 4, 8 and 12 weeks of therapy serum GAG was 3.4±0.42 mg/dl, 2.5±0.77 mg/dl (p<0.01) and 1.1±0.2 mg/dl (p<0.001), respectively and serum TNF-alpha was 20.9±4.8 pg/ml, 14.9±2.2 pg/ml (p<0.05) and9.7±1.8 pg/ml (p<0.01), respectively. Serum GAG and TNF alpha did not fall in the two patients who did not respond. The titre of anti-eye muscle antibodies but not anti-thyroid antibodies were lower at 12 weeks. Ptx has a beneficial effect on inflammatory symptoms of TAO and associated laboratory parameters in the majority of patients.

Download Full-text

Dendritic cells and macrophages can mature independently from a human bone marrow-derived, post-colony-forming unit intermediate

Blood ◽

10.1182/blood.v87.11.4520.bloodjournal87114520 ◽

1996 ◽

Vol 87 (11) ◽

pp. 4520-4530 ◽

Cited By ~ 160

Author(s):

P Szabolcs ◽

D Avigan ◽

S Gezelter ◽

DH Ciocon ◽

MA Moore ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Tnf Alpha ◽

Intermediate Cell ◽

Cell Type ◽

Gm Csf ◽

Kit Ligand ◽

Hla Dr

CD34+ precursors in normal human bone marrow (BM) generate large numbers of dendritic cells alongside macrophages and granulocytic precursors when cultured for 12 to 14 days in c-kit ligand, granulocyte- macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). This study reports an intermediate cell type that develops by day 6, and has the potential to differentiate into either macrophages or dendritic cells. When the d6 progeny are depleted of mature macrophages and residual CD34+ precursors, a discrete CD14+ HLA-DR+ population persists in addition to immunostimulatory CD14- HLA- DR() dendritic cells. Half of the CD14+ HLA-DR+ population is in cell cycle (Ki-67+), but colony-forming units (CFUs) are no longer detectable. The calls are c-fms+, but lack myeloperoxidase and nonspecific esterase. They also possess substantial phagocytic and allostimulatory activity. These post-CFU, CD14+ HLA-DR+ intermediates develop into typical macrophages when recultured in the absence of exogenous cytokines. M-CSF supports up to approximately 2.5-fold expansion of macrophage progeny. In contrast, the combination of GM-CSF and TNF-alpha supports quantitative differentiation into dendritic cells, lacking c-fms, CD14, and other macrophage properties, and expressing HLA-DR, CD1a, CD83, CD80, CD86, and potent allostimulatory activity. Therefore, normal CD34+ BM precursors can generate a post-CFU bipotential intermediate in the presence of c-kit ligand, GM-CSF, and TNF-alpha. This intermediate cell type will develop along the dendritic cell pathway when macrophages are removed and GM-CSF and TNF-alpha are provided. Alternatively, it can differentiate along a macrophage pathway when recultured with or without M-CSF.

Download Full-text

Human BDCA-1 Positive Blood Dendritic Cells in Immunosuppressed Patients: Phenotype, Function and Maturation.

Blood ◽

10.1182/blood.v106.11.2231.2231 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2231-2231

Author(s):

Kathrin Sebelin ◽

Antje Meier ◽

Carola Beier ◽

Bernd Dörken ◽

Antonio Pezzutto ◽

...

Keyword(s):

T Cells ◽

Ex Vivo ◽

Immunosuppressive Drugs ◽

Tnf Alpha ◽

Hla Dr ◽

And Function ◽

Cellular Immune ◽

Vitro Data ◽

In Vitro Data

Abstract Immunosuppressive drugs used in patients (pts) after stem cell / organ transplantation (Tx) as well as in pts with autoimmune disease are known to impair the cellular immune response. This results in an increased incidence of viral infections and viral associated malignancies which has been ascribed to the effect of immunosuppressive drugs on lymphocytes. However, in vitro data indicate that immunosuppressive drugs also target Dendritic Cells (DCs), the most potent antigen-presenting cells and initiators of lymphocyte responses. So far, most studies are based on in vitro data obtained with DC culture in the presence of different concentrations of single immunosuppressive drugs. To investigate the effect of immunosuppression on DC phenotype and function in vivo, we quantitatively and qualitatively analyzed freshly isolated human BDCA-1(CD1c) positive DCs from 15 solid organ transplant (SOT) recipients under immunosuppressive treatment. The percentage of BDCA-1 positive cells among total PBMCs was not statistically different in pts vs ctrls (0,52 vs 0,65, p<0,18). BDCA-1 positive DCs were analyzed for expression of HLA class I and II, CD14, costimmulatory molecules and chemokine expression. Interestingly, CD14 was found to be significantly higher expressed on pt-DCs vs ctrl-DCs suggesting a more immature DC-phenotype. We observed a trend toward a reduced expression of HLA-DR and CD86 on pts-DCs as compared to ctrls-DCs (p=0,059). Surface profile of BDCA-1 positive DCs was also analyzed after 48h of LPS and CD40L stimulation. Here we found a marked upregulation of HLA-DR and CD86 in pts- DCs as well as ctrl-DCs. Supernatant of stimulated DCs was analyzed with cytokine capture beads for secretion of inflammatory cytokines. High secretion of IL-6, IL-1 beta and partially of TNF-alpha by stimulated DCs was observed in both groups. Other Th2 type cytokines (IL-10, IL-4, IL-5) and Th1 type cytokines like IFN-gamma and Il-2 were not significantly secreted. We additionally addressed the question if mature and functionally competent DCs could be generated ex vivo from this pts cohort. After 9 days of culture with GM-CSF, IL-4, IL-1, IL-6, TNF-alpha and PGE2 fully mature DCs could be generated. Co-culture of EBV-peptide-pulsed DCs with autologous T-cells resulted in significant expansion of EBV-specific T cells in pts and ctrls. These T cells were fully functional as shown by IFN-γ secretion detected by ELISPOT. In summary, this is the first analysis of freshly isolated BDCA-1 positive DCs from immunosuppressed pts. Our data support the notion that immunosuppressive drugs target DCs and contribute to a maturation defect of circulating blood DCs which may help to understand the mechanism of impaired cellular immune responses in immunosuppressed pts. However, ex vivo generated DCs from immunosuppressed pts do not show an impairment in phenotype and function, suggesting that they could be efficiently be used in immunotherapeutic strategies.

Download Full-text