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Autologous hematopoietic stem cell transplantation (autoHSCT) is a treatment option for hematological disorders and pediatric solid tumors. After an autoHSCT, natural killer (NK) cells are the first lymphocyte subset returning to normal levels. To uncover global changes during NK cell reconstitution after autoHSCT, we performed RNA-sequencing on NK cells before and after autoHSCT. Results showed profound changes in the gene expression profile of NK cells immediately after autoHSCT. Several biological processes including cell cycle, DNA replication and the mevalonate pathway were enriched. Significantly, we observed that following autoHSCT, NK cells acquired a decidual-like gene expression profile, including the expression of CD9. By using multiparametric flow cytometry, we confirmed the expansion of NK cells expressing CD9 immediately after autoHSCT, which exhibited higher granzyme B and perforin expression levels than CD9 - NK cells. These results provide insights into the physiopathology of NK cells during their reconstitution after autoHSCT., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

Although scarce in the peripheral blood of healthy people, CD56 neg NK cells are known to be expanded in some pathological conditions. However, studies on CD56 neg NK cells had revealed contradictions, probably due to the lack of a specific NK cell surface marker that helps to identify this subset. This protocol details the step-by-step procedure for the identification and functional analysis of CD56 neg NK cells, providing an improved gating strategy for the selection of this intriguing population. For complete details on the use and execution of this protocol, please refer to Orrantia et al. (2020)., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)