TY - JOUR
T1 - Inhibiting HLA-B27 homodimer-driven immune cell inflammation in spondylarthritis
AU - Payeli, Sravan K.
AU - Kollnberger, Simon
AU - Belaunzaran, Osiris Marroquin
AU - Thiel, Markus
AU - McHugh, Kirsty
AU - Giles, Joanna
AU - Shaw, Jacqueline
AU - Kleber, Sascha
AU - Ridley, Anna
AU - Wong-Baeza, Isabel
AU - Keidel, Sarah
AU - Kuroki, Kimiko
AU - Maenaka, Katsumi
AU - Wadle, Andreas
AU - Renner, Christoph
AU - Bowness, Paul
PY - 2012/10
Y1 - 2012/10
N2 - Objective. Spondylarthritides (SpA), including ankylosing spondylitis (AS), are common inflammatory rheumatic diseases that are strongly associated with positivity for the HLA class I allotype B27. HLA-B27 normally forms complexes with β2-microglobulin (β2m) and peptide to form heterotrimers. However, an unusual characteristic of HLA-B27 is its ability to form β2m-free heavy chain homodimers (HLA-B272), which, unlike classic HLA-B27, bind to killer cell immunoglobulinlike receptor 3DL2 (KIR-3DL2). Binding of HLA-B272 to KIR-3DL2-positive CD4+ T and natural killer (NK) cells stimulates cell survival and modulates cytokine production. This study was undertaken to produce an antibody to HLA-B27 2 in order to confirm its expression in SpA and to inhibit its proinflammatory properties. Methods. We generated monoclonal antibodies by screening a human phage display library positively against B272 and negatively against B27 heterotrimers. Specificity was tested by enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR) assay, and fluorescence-activated cell sorting (FACS) analysis of B272- expressing cell lines and peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) from patients with SpA. Functional inhibition of KIR-3DL2-B272 interactions was tested using cell lines and PBMCs from patients with SpA. Results. Monoclonal antibody HD6 specifically recognized recombinant HLA-B272 by ELISA and by SPR assay. HD6 bound to cell lines expressing B272. FACS revealed binding of HD6 to PBMCs and SFMCs from patients with AS but not from controls. HD6 inhibited both the binding of HLA-B272 to KIR-3DL2 and the survival and proliferation of KIR-3DL2- positive NK cells. Finally, HD6 inhibited production of the proinflammatory disease-associated cytokine interleukin-17 by PBMCs from patients with AS. Conclusion. These results demonstrate that antibody HD6 has potential for use in both the investigation and the treatment of AS and other B27-associated spondylarthritides.
AB - Objective. Spondylarthritides (SpA), including ankylosing spondylitis (AS), are common inflammatory rheumatic diseases that are strongly associated with positivity for the HLA class I allotype B27. HLA-B27 normally forms complexes with β2-microglobulin (β2m) and peptide to form heterotrimers. However, an unusual characteristic of HLA-B27 is its ability to form β2m-free heavy chain homodimers (HLA-B272), which, unlike classic HLA-B27, bind to killer cell immunoglobulinlike receptor 3DL2 (KIR-3DL2). Binding of HLA-B272 to KIR-3DL2-positive CD4+ T and natural killer (NK) cells stimulates cell survival and modulates cytokine production. This study was undertaken to produce an antibody to HLA-B27 2 in order to confirm its expression in SpA and to inhibit its proinflammatory properties. Methods. We generated monoclonal antibodies by screening a human phage display library positively against B272 and negatively against B27 heterotrimers. Specificity was tested by enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR) assay, and fluorescence-activated cell sorting (FACS) analysis of B272- expressing cell lines and peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) from patients with SpA. Functional inhibition of KIR-3DL2-B272 interactions was tested using cell lines and PBMCs from patients with SpA. Results. Monoclonal antibody HD6 specifically recognized recombinant HLA-B272 by ELISA and by SPR assay. HD6 bound to cell lines expressing B272. FACS revealed binding of HD6 to PBMCs and SFMCs from patients with AS but not from controls. HD6 inhibited both the binding of HLA-B272 to KIR-3DL2 and the survival and proliferation of KIR-3DL2- positive NK cells. Finally, HD6 inhibited production of the proinflammatory disease-associated cytokine interleukin-17 by PBMCs from patients with AS. Conclusion. These results demonstrate that antibody HD6 has potential for use in both the investigation and the treatment of AS and other B27-associated spondylarthritides.
UR - http://www.scopus.com/inward/record.url?scp=84869026015&partnerID=8YFLogxK
U2 - 10.1002/art.34538
DO - 10.1002/art.34538
M3 - Artículo
C2 - 22576154
SN - 0004-3591
VL - 64
SP - 3139
EP - 3149
JO - Arthritis and Rheumatism
JF - Arthritis and Rheumatism
IS - 10
ER -