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HLA-B27-Homodimer-Specific Antibody Modulates the Expansion of Pro-Inflammatory T-Cells in HLA-B27 Transgenic Rats.

Marroquin Belaunzaran O, Kleber S, Schauer S, Hausmann M, Nicholls F, Van den Broek M, Payeli S, Ciurea A, Milling S, Stenner F, Shaw J, Kollnberger S, Bowness P, Petrausch U, Renner C - PLoS ONE (2015)

Bottom Line: HD5 bound B272 with high specificity and affinity (Kd = 0.32 nM).HD5 blocked cell-surface interaction of B272 with immune regulatory receptors KIR3DL2, LILRB2 and Pirb.In addition, HD5 modulated the production of TNF from CD4+ T-cells by limiting B272 interactions in vitro.

View Article: PubMed Central - PubMed

Affiliation: Division of Oncology, University Hospital Zurich, Zurich, Switzerland.

ABSTRACT

Objectives: HLA-B27 is a common genetic risk factor for the development of Spondyloarthritides (SpA). HLA-B27 can misfold to form cell-surface heavy chain homodimers (B272) and induce pro-inflammatory responses that may lead to SpA pathogenesis. The presence of B272 can be detected on leukocytes of HLA-B27+ Ankylosing spondylitis (AS) patients and HLA-B27 transgenic rats. We characterized a novel B272-specific monoclonal antibody to study its therapeutic use in HLA-B27 associated disorders.

Methods: The monoclonal HD5 antibody was selected from a phage library to target cell-surface B272 homodimers and characterized for affinity, specificity and ligand binding. The immune modulating effect of HD5 was tested in HLA-B27 transgenic rats. Onset and progression of disease profiles were monitored during therapy. Cell-surface B272 and expansion of pro-inflammatory cells from blood, spleen and draining lymph nodes were assessed by flow cytometry.

Results: HD5 bound B272 with high specificity and affinity (Kd = 0.32 nM). HD5 blocked cell-surface interaction of B272 with immune regulatory receptors KIR3DL2, LILRB2 and Pirb. In addition, HD5 modulated the production of TNF from CD4+ T-cells by limiting B272 interactions in vitro. In an HLA-B27 transgenic rat model repetitive dosing of HD5 reduced the expansion of pro-inflammatory CD4+ T-cells, and decreased the levels of soluble TNF and number of cell-surface B272 molecules.

Conclusion: HD5 predominantly inhibits early TNF production and expansion of pro-inflammatory CD4+ T-cells in HLA-B27 transgenic rats. Monoclonal antibodies targeting cell-surface B272 propose a new concept for the modulation of inflammatory responses in HLA-B27 related disorders.

No MeSH data available.


Related in: MedlinePlus

Reduced amount of soluble TNF in Tg-HD5 rats.Serum obtained from the blood after euthanasia was assessed for the presence of soluble TNF. (A-B) TNF serum ELISA from WT, Tg-HD5 and Tg-ctrl (n = 5 per group) at 15 weeks (A) and at 23 weeks (B).
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pone.0130811.g007: Reduced amount of soluble TNF in Tg-HD5 rats.Serum obtained from the blood after euthanasia was assessed for the presence of soluble TNF. (A-B) TNF serum ELISA from WT, Tg-HD5 and Tg-ctrl (n = 5 per group) at 15 weeks (A) and at 23 weeks (B).

Mentions: Serum samples were analyzed for the presence of soluble TNF. TNF levels were significantly reduced in Tg-HD5 rats at early treatment stages when compared to Tg-ctrl (13.2 pg/mL vs. 22.13 pg/mL, respectively, Fig 7A), but not at later time points (14.3 pg/mL vs. 18.6 pg/mL, respectively, Fig 7B). Serum IL-17 levels measured by ELISA were below detection limit and, therefore, not determined.


HLA-B27-Homodimer-Specific Antibody Modulates the Expansion of Pro-Inflammatory T-Cells in HLA-B27 Transgenic Rats.

Marroquin Belaunzaran O, Kleber S, Schauer S, Hausmann M, Nicholls F, Van den Broek M, Payeli S, Ciurea A, Milling S, Stenner F, Shaw J, Kollnberger S, Bowness P, Petrausch U, Renner C - PLoS ONE (2015)

Reduced amount of soluble TNF in Tg-HD5 rats.Serum obtained from the blood after euthanasia was assessed for the presence of soluble TNF. (A-B) TNF serum ELISA from WT, Tg-HD5 and Tg-ctrl (n = 5 per group) at 15 weeks (A) and at 23 weeks (B).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488392&req=5

pone.0130811.g007: Reduced amount of soluble TNF in Tg-HD5 rats.Serum obtained from the blood after euthanasia was assessed for the presence of soluble TNF. (A-B) TNF serum ELISA from WT, Tg-HD5 and Tg-ctrl (n = 5 per group) at 15 weeks (A) and at 23 weeks (B).
Mentions: Serum samples were analyzed for the presence of soluble TNF. TNF levels were significantly reduced in Tg-HD5 rats at early treatment stages when compared to Tg-ctrl (13.2 pg/mL vs. 22.13 pg/mL, respectively, Fig 7A), but not at later time points (14.3 pg/mL vs. 18.6 pg/mL, respectively, Fig 7B). Serum IL-17 levels measured by ELISA were below detection limit and, therefore, not determined.

Bottom Line: HD5 bound B272 with high specificity and affinity (Kd = 0.32 nM).HD5 blocked cell-surface interaction of B272 with immune regulatory receptors KIR3DL2, LILRB2 and Pirb.In addition, HD5 modulated the production of TNF from CD4+ T-cells by limiting B272 interactions in vitro.

View Article: PubMed Central - PubMed

Affiliation: Division of Oncology, University Hospital Zurich, Zurich, Switzerland.

ABSTRACT

Objectives: HLA-B27 is a common genetic risk factor for the development of Spondyloarthritides (SpA). HLA-B27 can misfold to form cell-surface heavy chain homodimers (B272) and induce pro-inflammatory responses that may lead to SpA pathogenesis. The presence of B272 can be detected on leukocytes of HLA-B27+ Ankylosing spondylitis (AS) patients and HLA-B27 transgenic rats. We characterized a novel B272-specific monoclonal antibody to study its therapeutic use in HLA-B27 associated disorders.

Methods: The monoclonal HD5 antibody was selected from a phage library to target cell-surface B272 homodimers and characterized for affinity, specificity and ligand binding. The immune modulating effect of HD5 was tested in HLA-B27 transgenic rats. Onset and progression of disease profiles were monitored during therapy. Cell-surface B272 and expansion of pro-inflammatory cells from blood, spleen and draining lymph nodes were assessed by flow cytometry.

Results: HD5 bound B272 with high specificity and affinity (Kd = 0.32 nM). HD5 blocked cell-surface interaction of B272 with immune regulatory receptors KIR3DL2, LILRB2 and Pirb. In addition, HD5 modulated the production of TNF from CD4+ T-cells by limiting B272 interactions in vitro. In an HLA-B27 transgenic rat model repetitive dosing of HD5 reduced the expansion of pro-inflammatory CD4+ T-cells, and decreased the levels of soluble TNF and number of cell-surface B272 molecules.

Conclusion: HD5 predominantly inhibits early TNF production and expansion of pro-inflammatory CD4+ T-cells in HLA-B27 transgenic rats. Monoclonal antibodies targeting cell-surface B272 propose a new concept for the modulation of inflammatory responses in HLA-B27 related disorders.

No MeSH data available.


Related in: MedlinePlus