Effector Alloreactive CTL Engineered as Replication Competent Retrovirus Vector Producer Cells to Deliver Payload to BC Metastatic to the Brain
Abstract
This Idea Proposal was to explore multi-modal immunogene treatment for breast cancer metastatic to brain. The treatments involve using 1) alloreactive cytotoxic T lymphocytes (alloCTL) effector cells that are trained to recognize major histocompatibility antigens (HLA) with 2) replication competent retroviral vectors that code for suicide genes such that infected cells become susceptible to antiviral agents like ganciclovir. We demonstrated that HLA was expressed quite well by several breast cancer cell lines, two of which had a known propensity to metastasize to the brain. We also showed that alloCTL generated by one-way mixed tumor lymphocyte reaction (MLTR) were cytotoxic towards the breast cancer cells and that interferon-gamma is produced upon their contact with relevant target antigens. We know that alloreactivity is involved in this process since antibody to HLA class I inhibits the percentage lysis obtained. We have attempted the knock-down of HLA in the breast cancer cells using siRNA to obtain targets with low, intermediate and high HLA class I expression such that sensitivity to alloCTL can be determined. alloCTL transduced with replication competent retroviral vector (RCR) by spinoculation was most effective and with this technique we achieved up to 35% gene transfer to tumor cells. It was harder to get higher transduction percentages over time as apoptosis was also turned on in the transduced cell population. Finally, we performed a small pilot bioluminescence animal experiment and administered tumor cells with a luciferase marker gene into the brain. After five days, alloCTL was injected right into the center of the tumor or several millimeters away. Histological examination of the brain was performed, and as well, we began working out the technique for detection of CD3+ cells on frozen and fixed brain tissue.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2009
- Accession Number
- ADA542569
Entities
People
- Carol A. Kruse
Organizations
- Sanford Burnham Prebys Medical Discovery Institute