WP01 - High purity isolation of antigen specific T-cells using combinatorial MHC streptamer staining
Objectives of the workpackage
- Isolation from peripheral blood of healthy donors of immune cells (T-cells) capable of attacking viruses.
- Isolation of immune cells (T-cells) from healthy donors capable of recognizing specific target structures on tumors.
- Development of procedures to produce products of immune cell capable of recognizing viruses and tumors for the treatment of patients.
- Development of methods to monitor specific immune cells from peripheral blood of patients who are treated with these cell products.
Three viruses, adenovirus, cytomegalovirus (CMV), and Epstein Bar virus (EBV), are viruses that are dangerous to patients shortly after stem cell transplantation. Immune cells from healthy donors who have been previously exposed to these viruses are capable of recognizing these targets and these donors usually have relatively high numbers of these immune cells (T-cells) in their peripheral blood. A method will be developed to highly purified the T-cells from the peripheral blood of healthy donors to allow them to administer to the patients after stem cell transplantation. The specificity of the immune cells as well as the complexity of the product will be characterized.
Healthy donors are unlikely to have been exposed to target structures that are present on tumor cells that have to be eliminated from the patient. Therefore, the frequencies of those immune cells possibly recognizing the tumors from the patients are extraordinary low in normal peripheral blood. In this task, we will develop strategies to isolate these low frequency immune cells from normal peripheral blood of healthy donors. The isolation efficacy, and also the functionality of these isolated immune cells will be characterized, and we will determine whether it is likely that these cells are capable of eradicating the cells from the patients once they are transferred to the patients. We will identify whether a functional “immune cell product” can be generated allowing the treatment of patients with residual cancer cells after transplantation.
Development of a clinical grade product containing immune cells recognizing both viruses and tumor cells. The most optimal process to combine the isolation strategies of virus specific and tumor specific immune cells will be determined. These methods will be up-scaled and translated into procedures that are called ”standard operational procedures” allowing treatment of patients after stem cell transplantation.
Development of methods to monitor immune cells in patients treated with T cell products. We will validate laboratory methods to accurately evaluate the outcome of patients treated with the specific immune cells. Specific assays will be developed allowing the analysis of low frequency immune cells in peripheral blood of patients after treatment. We will not only develop assays to determine the presence of those cells, but also their efficacy in combating viruses and tumor cells.