A new T cell therapy method for cancer: polyclonal effector tumor antigen-specific T cells isolated from autologous blood or lymph nodes by nanoparticles loading whole tumor antigens.

Most immunotherapies work through activating new effector tumor antigen-specific T cells (ETASTs) or re-activating pre-existing ETASTs repertoire. Increasing ETASTs can treat cancer. Expanding autologous ETASTs is an ideal source to increase ETASTs. However, to get better therapeutic efficacy, isolating polyclonal ETASTs is needed and unachievable currently. No distinct structure characteristics exist between ETASTs and other T cells. Therefore, nanoparticles loading whole-tumor antigens are utilized to activate pre-existing pan-clones ETASTs to transform the differences between ETASTs and other T cells to the differences between activated and non-activated status, followed by isolating ETASTs by using proper activation markers. Herein, by optimizing 15 activation markers, CD137, CD134 and CD39 were identified to be ideal markers to isolate ETASTs simultaneously possessing both activation and cytotoxic properties. CD137+, CD134+ and CD39+ ETASTs are better than CD25+ and CD95L+ ETASTs, due to Treg exist in CD25+ETASTs and the overlapping of surface markers with IFN-γ is more important than that with granzyme B. In melanoma and lung cancer mouse model, ETASTs activated by nanoparticles loading whole-tumor antigens and sorted by selected markers can cure ≥75 % tumor-bearing mice, either ETASTs are from blood, lymph node, or splenocytes. Furthermore, ETASTs isolated from PBMC of lung cancer patients can efficiently kill autologous cancer cells. In summary, herein provides a new method to isolate pan-clones pre-existing ETASTs in tumor-bearing bodies using nanoparticles and proves that CD137/CD134/CD39 are ideal biomarkers to isolate activated ETASTs. Such isolated pan-clones ETASTs are more efficient, highly specific, more diverse, autologous, low cost and gene-editing free etc.