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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

Increased body weight and decreased expansion of pro-inflammatory CD4+ T-cells from blood in Tg-HD5 rats.(A) Body weight gain from the 6th week of age where the first antibody injections were performed. (B) Stool score from Tg-HD5 and Tg-ctrl groups. (C-E)In vitro stimulated cells obtained at different time points from the blood of Tg-HD5, Tg-ctrl and WT-littermates were assessed by ICS for the presence of pro-inflammatory cells expressing TNF (C), IL-17 (D) and IFN-γ (E). Results are expressed as percentages of CD4+ T gated cells. Values are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.005 as determined by one-way ANOVA followed by Bonferroni post-hoc analysis.
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pone.0130811.g005: Increased body weight and decreased expansion of pro-inflammatory CD4+ T-cells from blood in Tg-HD5 rats.(A) Body weight gain from the 6th week of age where the first antibody injections were performed. (B) Stool score from Tg-HD5 and Tg-ctrl groups. (C-E)In vitro stimulated cells obtained at different time points from the blood of Tg-HD5, Tg-ctrl and WT-littermates were assessed by ICS for the presence of pro-inflammatory cells expressing TNF (C), IL-17 (D) and IFN-γ (E). Results are expressed as percentages of CD4+ T gated cells. Values are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.005 as determined by one-way ANOVA followed by Bonferroni post-hoc analysis.

Mentions: Based on our data in vitro we hypothesized that HD5 could modulate B272-dependent pro-inflammatory responses in vivo. Therefore, treatment with repetitive injections of HD5 in Tg rats was evaluated in a preventive setting (6 weeks of age). Body weight gain (Fig 5A) and presence of diarrhea (Fig 5B) was measured as an in vivo surrogate marker of B27-dependent colitis activity. HLA-B27/hβ2m transgenic (33–3 line) rats in our animal facility did not develop any other clinical symptoms described for this animal model [13, 16].


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)

Increased body weight and decreased expansion of pro-inflammatory CD4+ T-cells from blood in Tg-HD5 rats.(A) Body weight gain from the 6th week of age where the first antibody injections were performed. (B) Stool score from Tg-HD5 and Tg-ctrl groups. (C-E)In vitro stimulated cells obtained at different time points from the blood of Tg-HD5, Tg-ctrl and WT-littermates were assessed by ICS for the presence of pro-inflammatory cells expressing TNF (C), IL-17 (D) and IFN-γ (E). Results are expressed as percentages of CD4+ T gated cells. Values are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.005 as determined by one-way ANOVA followed by Bonferroni post-hoc analysis.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0130811.g005: Increased body weight and decreased expansion of pro-inflammatory CD4+ T-cells from blood in Tg-HD5 rats.(A) Body weight gain from the 6th week of age where the first antibody injections were performed. (B) Stool score from Tg-HD5 and Tg-ctrl groups. (C-E)In vitro stimulated cells obtained at different time points from the blood of Tg-HD5, Tg-ctrl and WT-littermates were assessed by ICS for the presence of pro-inflammatory cells expressing TNF (C), IL-17 (D) and IFN-γ (E). Results are expressed as percentages of CD4+ T gated cells. Values are expressed as mean±SEM. *p<0.05, **p<0.01, ***p<0.005 as determined by one-way ANOVA followed by Bonferroni post-hoc analysis.
Mentions: Based on our data in vitro we hypothesized that HD5 could modulate B272-dependent pro-inflammatory responses in vivo. Therefore, treatment with repetitive injections of HD5 in Tg rats was evaluated in a preventive setting (6 weeks of age). Body weight gain (Fig 5A) and presence of diarrhea (Fig 5B) was measured as an in vivo surrogate marker of B27-dependent colitis activity. HLA-B27/hβ2m transgenic (33–3 line) rats in our animal facility did not develop any other clinical symptoms described for this animal model [13, 16].

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