Highlights
- •We detected CD20+T-cells in the blood and brains of MS patients.
- •Some CD20+T cells in the brains were positive for IL-17.
- •Blood CD20+ and CD20- T cells both produced INF-γ and IL-4.
- •Rituximab alone is a poor therapeutic at eliminating T-cells.
- •An engineered scFvRit:sFasL molecule specifically kills CD20+T-cells.
Abstract
Background
A subset of T-cells expresses the B-cell marker CD20 and in rheumatoid arthritis secretes
Interleukin (IL)-17. IL-17 secreting T-cells (Th17) have also been implicated in the
inflammatory response in the central nervous system in multiple sclerosis (MS) and
may be a potential target for elimination by biologic therapeutics. ScFvRit:sFasL
comprises of a rituximab-derived antibody fragment scFvRit genetically fused to human
soluble FasL that specifically eliminated T-cells.
Objective
To determine the presence and phenotype of CD20+T-cells in blood and brain of MS patients.
Second, to determine whether scFvRit:sFasL can selectively eliminate CD20+T-cells.
After CD20-selective binding, scFvRit:sFasL is designed to trigger FasL-mediated activation-induced
cell death of T-cells, but not B-cells.
Methods
Flow cytometry and immunohistochemistry were used to screen for CD20+inflammatory
T-cells in MS blood and brain tissue. ScFvRit:sFasL pro-apoptotic activity was evaluated
by Annexin-V/PI staining followed by flow cytometry assessment.
Results
Peripheral blood (n=11) and chronic but not active lesions of MS patient brains (n=5) contained CD20+inflammatory T-cells. Activated CD20+T-cells were predominantly
CD4+and secreted both IL-17 and INF-γ. ScFvRit:sFasL triggered CD20-restricted FasL-mediated
activation-induced cell death in peripheral blood CD20+T-cells, but not CD20+B-cells.
Conclusion
CD20+inflammatory T-cells are present in blood and chronic brain lesions of MS patients.
ScFvRit:sFasL selectively eliminated CD20+T-cells and may eliminate pathogenic T-cells
without B-cell depletion.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: June 11, 2014
Accepted:
June 2,
2014
Received in revised form:
May 9,
2014
Received:
October 14,
2013
Identification
Copyright
© 2014 Elsevier B.V. Published by Elsevier Inc. All rights reserved.
