Human natural killer (NK) cell development is a step-by-step process characterized by phenotypically identified stages. CD161 is a marker informative of the NK cell lineage commitment, whereas CD56, CD117, and CD94/NKG2A contribute to define discrete differentiation stages. In cells undergoing in vitro differentiation from CD34 ؉ umbilical cord blood (UCB) progenitors, LFA-1 expression allowed to discriminate between immature noncytolytic CD161 ؉ CD56 ؉ LFA-1 ؊ and more differentiated cytolytic CD161 ؉ CD56 ؉ LFA-1 ؉ NK cells. CD161 ؉ CD56 ؉ LFA-1 ؊ NK cells produce large amounts of CXCL8 after phorbol myristate acetate (PMA) or cytokine treatment. Remarkably, CXCL8 mRNA expression was also detected in fresh stage III immature NK cells isolated from tonsils and these cells expressed CXCL8 protein on PMA stimulation. Within in vitro UCB-derived CD161 ؉ CD56 ؉ LFA-1 ؊ NK cells, CXCL8 release was also induced on antibodymediated cross-linking of NKp44 and CD161. Such unexpected activating function of CD161 was confined to the CD161 ؉ CD56 ؉ LFA-1 ؊ subset, because it did not induce cytokine release or CD107a expression in CD161 ؉ CD56 ؉ LFA-1 ؉ cells or in mature peripheral blood NK cells. Anti-CXCL8 neutralizing antibody induced a partial inhibition of NK cell differentiation, which suggests a regulatory role of CXCL8 during early NK cell differentiation. Altogether, these data provide novel information that may offer clues to optimize NK cell maturation in hematopoietic stem cell transplantation. (Blood. 2012; 119(17):3987-3996) IntroductionNatural killer (NK) cells play a key role in the host defense because of their ability to mediate cytolytic activity and to release cytokines during the early phases of immune responses against tumor cells, viruses, and other pathogens. [1][2][3] In humans, two main cell subsets have been identified: CD3 Ϫ CD56 dim CD16 ϩ cells (CD56 dim ), which represent the majority of NK cells circulating in the peripheral blood (PB) and CD3 Ϫ CD56 bright CD16 Ϫ cells (CD56 bright ), that are a minority in PB but are largely represented in peripheral tissues. These subsets differ for the expression and the surface density of the main inhibitory receptors. Thus, killer immunoglobulin (Ig)-like receptors (KIR), specific for HLA-class I allotypes are confined to CD56 dim cells (that may also express the HLA-Especific CD94/NKG2A receptor), whereas CD56 bright cells express CD94/NKG2A but not KIRs. 2,4,5 Cytolytic activity is mostly associated with the CD56 dim subset, whereas CD56 bright NK cells are poorly cytotoxic but secrete cytokines playing an immunoregulatory role in innate and in adaptive immunity. 2,5,6 Recent data demonstrated that the "cytolytic" CD56 dim subset also releases large amounts of cytokines within a short time interval after receptor-mediated activation. Thus, CD56 dim cells may provide a prompt intervention mediating not only rapid killing, but also production of proinflammatory cytokines and chemokines. 7,8 On the other hand, CD56 bright cells would guarantee a late, b...

NK cells are innate lymphocytes characterized by the expression of nuclear factor interleukin 3 regulated (NFIL3 or E4BP4), eomesodermin (EOMES) transcription factors (TFs), and by the ability to exert cytolytic activity and release IFN-γ. In the haploidenticalhematopoietic stem cell transplantation (haplo-HSCT) setting, CD34 + donor derived NK cells play a major role in the control of leukemic relapses. Therefore, it is important to better define cytokines that influence NK-cell differentiation from CD34 + precursors. We analyzed the effects of IL-1β on NK-cell differentiation from umbilical cord blood (UCB) CD34 + cells. While IL-1β inhibited CD161 + CD56 + cell proliferation, an increased expression of LFA-1, CD94/NKG2A, KIRs, and perforin on CD56 + cells was detected. In addition, within the CD161 + CD56 + IL-1RI + LFA-1 − cell fraction (representing group 3 innate lymphoid cells, ILC3-like cells), a significant increase of EOMES, NKp46, and CD94/NKG2A receptors, cytolytic granules, and IFN-γ was detected. This increase was paralleled by a decrease of related orphan receptors (RORγt) TF, NKp44 expression, and IL-22 production. These data suggest that IL-1β inhibits ILC3-while favoring NK-cell maturation. Since in haplo-HSCT conditioning regimen, infections or residual leukemia cells may induce IL-1β production, this may influence the NK/ILC3 development from donor-derived CD34 + precursors.Keywords: IL-1β r ILC3 r Innate immunity r NK cells r NK-cell development Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionNatural killer (NK) cells are major effectors of the innate immunity. They are able to rapidly eliminate virally infected cells or tumor cells [1][2][3]. Their activity is finely regulated by a number of activating and inhibitory receptors that tune their cytolytic Correspondence: Dr. Chiara Vitale e-mail: chiara.vitale@unige.it activity and cytokine secretion [4,5]. The main activating receptors include NKp46, NKp30, and NKp44 (collectively termed natural cytotoxicity receptors), NKG2D, and DNAM-1 [6,7]. In addition, NK cells express inhibitory receptors specific for HLA class I molecules, including killer immunoglobulin-like(Ig-like) receptors (KIRs), specific for allotypic HLA class I determinants, and * These authors contributed equally to this work as senior co-authors. Eur. J. Immunol. 2015Immunol. . 45: 2061Immunol. -2071 CD94/NKG2A specific for 9]. NK cells are characterized by production of IFN-γ and the expression of cytolytic granules. Their development requires the expression of E4BP4 (NFIL3, nuclear factor interleukin 3), Tbx21 (T-bet), eomesodermin (EOMES) transcription factor (TF) [10][11][12][13][14] and occurs primarily in the BM, proceeding through a multistep process characterized by the sequential acquisition of markers/receptors (including CD117, CD161, CD56, CD94/NKG2A, LFA-1, CD16, and KIRs) and functions [10,11,15]. NK cells are developmentally related to innate lymphoid cells (ILCs) as they derive fro...

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The use of immune checkpoint inhibitors (ICIs) offers new possibilities in modern treatment of many types of cancers. Few data regarding safety and efficacy of ICIs are available, and are mainly from retrospective studies and case reports rather than from clinical trials, in the context of preexisting autoimmune disease, mainly due to the risk of severe toxicity. We present an unexpected life-threatening reactivation of systemic lupus erythematosus after one dose of chemo-immunotherapy with pembrolizumab for oligometastatic non-small-cell lung cancer. We analyze data coming from the published literature in this setting and discuss the risk–benefit balance of immunotherapy in patients with preexisting severe autoimmune disease.

BackgroundChemoimmunotherapy represents the standard of care for patients with advanced non-small cell lung cancer (NSCLC) and programmed death-ligand 1 (PD-L1) <50%. Although single-agent pembrolizumab has also demonstrated some activity in this setting, no reliable biomarkers yet exist for selecting patients likely to respond to single-agent immunotherapy. The main purpose of the study was to identify potential new biomarkers associated with progression-free-survival (PFS) within a multiomics analysis.MethodsPEOPLE (NTC03447678) was a prospective phase II trial evaluating first-line pembrolizumab in patients with advanced EGFR and ALK wild type treatment-naïve NSCLC with PD-L1 <50%. Circulating immune profiling was performed by determination of absolute cell counts with multiparametric flow cytometry on freshly isolated whole blood samples at baseline and at first radiological evaluation. Gene expression profiling was performed using nCounter PanCancer IO 360 Panel (NanoString) on baseline tissue. Gut bacterial taxonomic abundance was obtained by shotgun metagenomic sequencing of stool samples at baseline. Omics data were analyzed with sequential univariate Cox proportional hazards regression predicting PFS, with Benjamini-Hochberg multiple comparisons correction. Biological features significant with univariate analysis were analyzed with multivariate least absolute shrinkage and selection operator (LASSO).ResultsFrom May 2018 to October 2020, 65 patients were enrolled. Median follow-up and PFS were 26.4 and 2.9 months, respectively. LASSO integration analysis, with an optimal lambda of 0.28, showed that peripheral blood natural killer cells/CD56dimCD16+ (HR 0.56, 0.41–0.76, p=0.006) abundance at baseline and non-classical CD14dimCD16+monocytes (HR 0.52, 0.36–0.75, p=0.004), eosinophils (CD15+CD16−) (HR 0.62, 0.44–0.89, p=0.03) and lymphocytes (HR 0.32, 0.19–0.56, p=0.001) after first radiologic evaluation correlated with favorable PFS as well as high baseline expression levels of CD244 (HR 0.74, 0.62–0.87, p=0.05) protein tyrosine phosphatase receptor type C (HR 0.55, 0.38–0.81, p=0.098) and killer cell lectin like receptor B1 (HR 0.76, 0.66–0.89, p=0.05). Interferon-responsive factor 9 and cartilage oligomeric matrix protein genes correlated with unfavorable PFS (HR 3.03, 1.52–6.02, p 0.08 and HR 1.22, 1.08–1.37, p=0.06, corrected). No microbiome features were selected.ConclusionsThis multiomics approach was able to identify immune cell subsets and expression levels of genes associated to PFS in patients with PD-L1 <50% NSCLC treated with first-line pembrolizumab. These preliminary data will be confirmed in the larger multicentric international I3LUNG trial (NCT05537922).Trial registration number2017-002841-31.