Cancer immunotherapy is a clinically validated therapeutic modality for cancer and has been rapidly advancing in recent years. Adoptive transfer of immune cells such as T cells and natural killer (NK) cells has emerged as a viable method of controlling the immune system against cancer. Recent evidence indicates that even immune-cell-released vesicles such as NK-cell-derived exosomes also exert anticancer effect. Nevertheless, the underlying mechanisms remain elusive. In the present study, the anticancer potential of isolated extracellular vesicles (EVs) from expanded and activated NK-cell-enriched lymphocytes (NKLs) prepared by house-developed protocol was evaluated both in vitro and in vivo. Moreover, isolated EVs were characterized by using two-dimensional electrophoresis (2-DE)-based proteome and network analysis, and functional study using identified factors was performed. Our data indicated that the EVs from expanded and active NKLs had anticancer properties, and a number of molecules, such as Fas ligand, TRAIL, NKG2D, β-actin, and fibrinogen, were identified as effector candidates based on the proteome analysis and functional study. The results of the present study suggest the possibility of NK-cell-derived EVs as a viable immunotherapeutic strategy for cancer.

Adoptive transfer of immune cells such as T cells and natural killer (NK) cells has emerged as a targeted method of controlling the immune system against cancer. Despite their significant therapeutic potential, efficient methods to generate adequate numbers of NK cells are lacking and ex vivo-expansion and activation of NK cells is currently under intensive investigation. The primary purpose of this study was to develop an effective method for expansion and activation of the effector cells with high proportion of NK cells and increasing cytotoxicity against liver cancer in a short time period. Expanded NK cell-enriched lymphocytes (NKL) designated as “MYJ1633” were prepared by using autologous human plasma, cytokines (IL-2, IL-12 and IL-18) and agonistic antibodies (CD16, CD56 and NKp46) without an NK cell-sorting step. The characteristics of NKL were compared to those of freshly isolated PBMCs. In addition, the cytotoxic effect of the NKL on liver cancer cell was examined in vitro and in vivo. The total cell number after ex vivo-expansion increased about 140-fold compared to that of freshly isolated PBMC within 2 weeks. Approximately 78% of the expanded and activated NKL using the house-developed protocol was NK cell and NKT cells even without a NK cell-sorting step. In addition, the expanded and activated NKL demonstrated potent cytotoxicity against liver cancer in vitro and in vivo. The house-developed method can be a new and effective strategy to prepare clinically applicable NKL for autologous NK cell-based anti-tumor immunotherapy.

Purpose: Regulatory T (Treg) cells, a type of immune cell, play a very important role in the immune response as a subpopulation of T cells. In this study, we investigated the effects of Treg cells conditioned media (CM) on cell migration. Various cytokines and growth factors of Treg cells CM can effect on re-epithelialization stage during the wound healing. Methods: Isolated CD4+CD25+ Treg cells from Peripheral Blood Mononuclear Cells (PBMCs) were cultured and CM obtained. HaCaT keratinocytes were treated with various concentration of Treg cells CM. Cell migration, proliferation and expression of proteins that are related to the Epithelial-Mesenchymal Transition (EMT) process, matrix metalloproteinase-1 (MMP-1) were analyzed. Results: Above 90% CD4+CD25+ Treg cells were obtained from CD8+ depleted PBMCs and the CM have various cytokines and growth factors.One percent and 5% concentration of Treg cells CM increased HaCaT keratinocytes migration. The Treg cells CM stimulated EMT, which led to the down-regulation of E-cadherin in the HaCaT keratinocytes at the wound edge. The Treg cells CM increased MMP-1, which is involved in tissue remodeling. Conclusion: Our results suggest that Treg cells CM which has various cytokines and growth factors promote wound healing by stimulating HaCaT keratinocytes migration.

Various vaccines were rapidly developed during the COVID-19 pandemic to prevent and treat infections but global infections continue, and concerns about new mutations and infectious diseases persist. Thus, active research focuses on developing, producing, and supplying vaccines and treatments for various infectious diseases and potential pandemics.Natural killer(NK) cells, as innate immune cells, can recognize and eliminate abnormal cells like virus-infected and cancer cells. Hence, their development as anticancer and antiviral treatments is rapidly advancing. In this study, optimal short-term culture conditions were identified for allogeneic NK cells by simplifying the culture process through the isolation of NK cells(referred to as NKi cells) and eliminating CD3+ cells(referred to as CD3-cells). NK cells demonstrated reduced viral titer in injection of NK cells into SARS-CoV-2 infected ACE-tg mice increased survival.The study's findings could form the basis for an antiviral treatment platform that swiftly responds to new viral disease pandemics.