This invention relates to a genetically engineered bacterial outer membrane vesicle and its preparation method. By using in-cell peptides to attach exogenous proteins to the surface of OMVs via peptide bonds, a variety of desired antigens can be conveniently delivered. The reaction conditions are mild, less affected by antigen size, and unlike the SpyCatcher / SpyTag system, the in-cell peptides do not remain in the mature protein after attaching the exogenous protein to the OMV surface. This invention can serve as a universal tool for simultaneously or separately displaying different antigens on the surface of OMVs.
Various embodiments of the invention provide compositions of tumor infiltrating lymphocytes (TILs) enriched in tumor reactive cells, methods for manufacturing TILs enriched in tumor reactive cells and methods and uses of the provided enriched tumor reactive TILs for treating cancer in a human or other subject. Among the provided embodiments of TIL compositions may include those that exhibit substantial tumor reactivity activity, including degranulation and the ability to express one or more of IFN-gamma and TNF-alpha, in response to antigen presenting cells presenting neo antigenic peptides.