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Preclinical Evaluation of Recombinant T Cell Receptor Ligand RTL1000 as a Therapeutic Agent in Ischemic Stroke

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Abstract

Recombinant T cell Receptor Ligand 1000 (RTL1000), a partial human major histocompatibility complex (MHC) molecule coupled to a human myelin peptide, reduces infarct size after experimental stroke in HLA-DRB1*1502 transgenic (DR2-Tg) mice. In this study, we characterized the therapeutic time window of opportunity for RTL1000; we explored the efficacy of a single dose of RTL1000 administration and determined if RTL1000 affords long-term neurobehavioral functional improvement after ischemic stroke. Male DR2-Tg mice underwent 60 min of intraluminal reversible middle cerebral artery occlusion (MCAO). RTL1000 or vehicle was injected 4, 6, or 8 h after MCAO, followed by three daily injections. In the single-dose study, one-time injection of RTL1000 was applied 4 h after MCAO. Cortical, striatal, and hemispheric infarct sizes were measured 24 or 96 h after stroke. Behavioral testing, including neuroscore evaluation, open field, paw preference, and novel object recognition, was performed up to 28 days after stroke. Our data showed that RTL1000 significantly reduced the infarct size 96 h after MCAO when the first injection was given at 4 and 6 h, but not 8 h, after the onset of stroke. A single dose of 400 or 100 μg RTL1000 also significantly reduced the infarct size 24 h after MCAO. Behavioral testing showed that RTL1000 treatment used 4 h after MCAO improved long-term cognitive outcome 28 days after stroke. Taken together, RTL1000 protects against acute injury if applied within a 6-h time window and improves long-term functional recovery after experimental stroke in DR2-Tg mice.

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Acknowledgments

This work was supported by NIH Grants #NS076013 (STTR) and by the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development.

Conflict of Interest

Dr. Offner, Dr. Alkayed and OHSU have a significant financial interest in ArtielleImmuno Therapeutics, Inc., a company that may have a commercial interest in the results of this research and technology. This potential conflict of interest has been reviewed and managed by the OHSU and VAMC Conflict of Interest in Research Committees. Wenbin Zhu declares that he has no conflict of interest. Amanda Casper declares that she has no conflict of interest. Nicole L. Libal declares that she has no conflict of interest. Stephanie J. Murphy declares that she has no conflict of interest. Sheetal Bodhankar declares that she has no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed. This article does not contain any studies with human subjects.

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Authors and Affiliations

  1. Department of Anesthesiology & Perioperative Medicine, Oregon Health & Science University, Portland, OR, 97239, USA

    Wenbin Zhu, Amanda Casper, Nicole L. Libal, Stephanie J. Murphy, Halina Offner & Nabil J. Alkayed

  2. Department of Neurology, Oregon Health & Science University, Portland, OR, 97239, USA

    Sheetal Bodhankar, Halina Offner & Nabil J. Alkayed

  3. Department of Behavioral Neuroscience, Oregon Health & Science University, Portland, OR, 97239, USA

    Stephanie J. Murphy

  4. The Knight Cardiovascular Institute, Oregon Health & Science University, Portland, OR, 97239, USA

    Nabil J. Alkayed

  5. Neuroimmunology Research, Portland VA Medical Center, Portland, OR, 97239, USA

    Sheetal Bodhankar & Halina Offner

Authors
  1. Wenbin Zhu
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  2. Amanda Casper
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  3. Nicole L. Libal
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  6. Halina Offner
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  7. Nabil J. Alkayed
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Correspondence to Nabil J. Alkayed.

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Zhu, W., Casper, A., Libal, N.L. et al. Preclinical Evaluation of Recombinant T Cell Receptor Ligand RTL1000 as a Therapeutic Agent in Ischemic Stroke. Transl. Stroke Res. 6, 60–68 (2015). https://doi.org/10.1007/s12975-014-0373-7

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  • DOI: https://doi.org/10.1007/s12975-014-0373-7

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