Factor VIII-pulsed dendritic cells reduce anti-factor VIII antibody formation in the hemophilia A mouse model.
OBJECTIVE
Hemophilia inhibitor formation is a T-cell-dependent immune response to infused factor VIII (F.VIII). As immature dendritic cells (DCre) regulate immune response and promote tolerance, we evaluated F.VIII-pulsed DCre, propagated from bone marrow in the presence of granulocyte-macrophage colony-stimulating factor and transforming growth factor-beta, in achieving F.VIII tolerance.


METHODS
The effects of intravenous F.VIII-pulsed DCre in C57BL/6 hemophilia A mice were determined by total F.VIII inhibitory antibodies, T-cell proliferation, thymidine uptake, cytokine profile, and surface molecule expression.


RESULTS
After tail vein injection of 2.5 U recombinant F.VIII (rF.VIII) on day 0, 2, and 4, anti-F.VIII antibody peaked on day 6, and increased further on day 17 following rF.VIII rechallenge on day 12, 14, and 16, with increased T-cell proliferative response to in vitro F.VIII. When mice were pretreated with 2 x 10(6) F.VIII-pulsed immature DCre (deficient nuclear factor-kappaB nuclear protein binding, low CD80, low CD86, high interleukin [IL]-10 phenotype) 7 days before rF.VIII challenge, anti-F.VIII was reduced on day 6 and on day 8, 0.1 +/- 0.0 (Bethesda units/mL) vs control phosphate-buffered saline-treated hemophilia A mice, 2.0 +/- 0.1 Bethesda units/mL, p < 0.01. Rechallenge with rF.VIII on day 12 produced no increase in anti-F.VIII antibody response. This was associated with high serum IL-10 and low IL-2 levels by enzyme-linked immunosorbent assay, and splenic T-cell hyporesponsiveness to F.VIII, with IL-10 production, high FoxP3 expression by quantitative polymerase chain reaction, and T regulatory cell expansion, confirmed in ovalbumin-T-cell receptor transgenic mice.


CONCLUSIONS
These findings suggest F.VIII-pulsed DCre reduce anti-F.VIII antibody formation in hemophilia A mice by induction of regulatory T-cell-mediated hyporesponsiveness of T-helper cells to F.VIII.
Ragni
Margaret V
MV
Medicine/Hematology/Oncology, University of Pittsburgh Medicine Center, Pittsburgh, PA 15213-4306, USA. ragni@pitt.edu
Wu
Wenhu
W
Liang
Xiaoyan
X
Hsieh
Ching-Chuan
CC
Cortese-Hassett
Andrea
A
Lu
Lina
L
D000818:Animals; D000906:Antibodies / Q000097:blood; D000917:Antibody Formation; D016207:Cytokines / Q000097:blood; D003713:Dendritic Cells / Q000276:immunology*; D004195:Disease Models, Animal; D005169:Factor VIII / Q000008:administration & dosage / Q000276:immunology*; D006467:Hemophilia A / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology; D013936:Thymidine / Q000493:pharmacokinetics
D000906:Antibodies; D016207:Cytokines; C078147:F8 protein, human; D005169:Factor VIII; D013936:Thymidine