Capsid antigen presentation flags human hepatocytes for destruction after transduction by adeno-associated viral vectors
| Autor: | Nicole C. Hasbrouck, Samuel J.H. Murphy, Jordan S. Orange, Federico Mingozzi, Gary C. Pien, Katherine A. High, Daniel J. Hui, Jonathan D. Finn, Etiena Basner-Tschakarjan, Shangzhen Zhou, Ashley N. Mentlik, Marcela V. Maus |
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| Rok vydání: | 2008 |
| Předmět: |
Cytotoxicity
Immunologic viruses Antigen presentation Genetic Vectors Receptors Antigen T-Cell chemical and pharmacologic phenomena Gene delivery Biology Viral vector Cell Line Substrate Specificity Transduction (genetics) Antigen Histocompatibility Antigens MHC class I Humans Antigen Presentation T-cell receptor General Medicine Dependovirus Molecular biology CTL Solubility biology.protein Hepatocytes Capsid Proteins Protein Multimerization T-Lymphocytes Cytotoxic Research Article |
| Zdroj: | The Journal of clinical investigation. 119(6) |
| ISSN: | 1558-8238 |
| Popis: | Adeno-associated virus (AAV) vectors are effective gene delivery vehicles mediating long-lasting transgene expression. Data from a clinical trial of AAV2-mediated hepatic transfer of the Factor IX gene (F9) into hemophilia B subjects suggests that CTL responses against AAV capsid can eliminate transduced hepatocytes and prevent long-term F9 expression. However, the capacity of hepatocytes to present AAV capsid-derived antigens has not been formally demonstrated, nor whether transduction by AAV sensitizes hepatocytes for CTL-mediated destruction. To investigate the fate of capsids after transduction, we engineered a soluble TCR for the detection of capsid-derived peptide:MHC I (pMHC) complexes. TCR multimers exhibited antigen and HLA specificity and possessed high binding affinity for cognate pMHC complexes. With this reagent, capsid pMHC complexes were detectable by confocal microscopy following AAV-mediated transduction of human hepatocytes. Although antigen presentation was modest, it was sufficient to flag transduced cells for CTL-mediated lysis in an in vitro killing assay. Destruction of hepatocytes was inhibited by soluble TCR, demonstrating a possible application for this reagent in blocking undesirable CTL responses. Together, these studies provide a mechanism for the loss of transgene expression and transient elevations in aminotransferases following AAV-mediated hepatic gene transfer in humans and a potential therapeutic intervention to abrogate these limitations imposed by the host T cell response. |
| Databáze: | OpenAIRE |
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