Federica Bozzano1, Mariella Della Chiesa2, Andrea Pelosi1, Francesca Antonini3, Maria Libera Ascierto4, Genny Del Zotto3, Francesca Moretta5, Letizia Muccio2, Anna Luganini6, Giorgio Gribaudo6, Giovanni Cenderello7, Chiara Dentone8, Laura Nicolini8, Alessandro Moretta2, Lorenzo Moretta1, Andrea De Maria9. 1. Pediatric Hospital Bambino Gesù, Rome, Italy. 2. Department of Experimental Medicine and Center of Excellence for Biomedical Research, University of Genova, Genova, Italy. 3. Istituto G. Gaslini, Pediatric Hospital, Genova, Italy. 4. Medimmune, Gaithersburg, Md. 5. Istituto di Ricovero e Cura a Carattere Scientifico Sacro Cuore Don Calabria Hospital, Negrar, Verona, Italy; Department of Life Sciences and Systems Biology, University of Torino, Torino, Italy. 6. Department of Life Sciences and Systems Biology, University of Torino, Torino, Italy. 7. Unità Operativa Complessa Malattie Infettive, Ospedale Sanremo, ASL1, Italy. 8. Clinica Malattie Infettive, Ospedale Policlinico San Martino Istituto di Ricovero e Cura a Carattere Scientifico, Genova, Italy. 9. Clinica Malattie Infettive, Ospedale Policlinico San Martino Istituto di Ricovero e Cura a Carattere Scientifico, Genova, Italy; Department of Life Sciences, University of Genova, Genova, Italy. Electronic address: de-maria@unige.it.
Abstract
BACKGROUND: There is limited knowledge on the origin and development from CD34+ precursors of the ample spectrum of human natural killer (NK) cells, particularly of specialized NK subsets. OBJECTIVE: This study sought to characterize the NK-cell progeny of CD34+DNAM-1brightCXCR4+ and of other precursors circulating in the peripheral blood of patients with chronic viral infections (eg, HIV, hepatitis C virus, cytomegalovirus reactivation). METHODS: Highly purified precursors were obtained by flow cytometric sorting and cultured in standard NK-cell differentiation media (ie, SCF, FLT3, IL-7, IL-15). Phenotypic and functional analyses on progenies were performed by multiparametric cytofluorimetric assays. Transcriptional signatures of NK-cell progenies were studied by microarray analysis. Inhibition of cytomegalovirus replication was studied by PCR. RESULTS: Unlike conventional CD34+ precursors, Lin-CD34+DNAM-1brightCXCR4+ precursors from patients with chronic infection, rapidly differentiate into cytotoxic, IFN-γ-secreting CD94/NKG2C+KIR+CD57+ NK-cell progenies. An additional novel subset of common lymphocyte precursors was identified among Lin-CD34-CD56-CD16+ cells and characterized by expression of CXCR4 and lack of perforin and CD94. Lin-CD34-CD56-CD16+Perf-CD94-CXCR4+ precursors are also endowed with generation potential toward memory-like NKG2C+NK cells. Maturing NK-cell progenies mediated strong human cytomegalovirus-inhibiting activity. Microarray analysis confirmed a transcriptional signature compatible with NK-cell progenies and with maturing adaptive NK cells. CONCLUSIONS: During viral infections, precursors of adaptive NK cells are released and circulate in the peripheral blood.
BACKGROUND: There is limited knowledge on the origin and development from CD34+ precursors of the ample spectrum of human natural killer (NK) cells, particularly of specialized NK subsets. OBJECTIVE: This study sought to characterize the NK-cell progeny of CD34+DNAM-1brightCXCR4+ and of other precursors circulating in the peripheral blood of patients with chronic viral infections (eg, HIV, hepatitis C virus, cytomegalovirus reactivation). METHODS: Highly purified precursors were obtained by flow cytometric sorting and cultured in standard NK-cell differentiation media (ie, SCF, FLT3, IL-7, IL-15). Phenotypic and functional analyses on progenies were performed by multiparametric cytofluorimetric assays. Transcriptional signatures of NK-cell progenies were studied by microarray analysis. Inhibition of cytomegalovirus replication was studied by PCR. RESULTS: Unlike conventional CD34+ precursors, Lin-CD34+DNAM-1brightCXCR4+ precursors from patients with chronic infection, rapidly differentiate into cytotoxic, IFN-γ-secreting CD94/NKG2C+KIR+CD57+ NK-cell progenies. An additional novel subset of common lymphocyte precursors was identified among Lin-CD34-CD56-CD16+ cells and characterized by expression of CXCR4 and lack of perforin and CD94. Lin-CD34-CD56-CD16+Perf-CD94-CXCR4+ precursors are also endowed with generation potential toward memory-like NKG2C+NK cells. Maturing NK-cell progenies mediated strong human cytomegalovirus-inhibiting activity. Microarray analysis confirmed a transcriptional signature compatible with NK-cell progenies and with maturing adaptive NK cells. CONCLUSIONS: During viral infections, precursors of adaptive NK cells are released and circulate in the peripheral blood.
Authors: Lucia Taramasso; Federica Bozzano; Anna Casabianca; Chiara Orlandi; Francesca Bovis; Sara Mora; Mauro Giacomini; Lorenzo Moretta; Mauro Magnani; Antonio Di Biagio; Andrea De Maria Journal: Front Immunol Date: 2022-04-26 Impact factor: 8.786