NK cells play a decisive role in controlling hCMV infection by combining innate and adaptive-like immune reactions. The hCMV-derived VMAPRTLFL (LFL) peptide is a potent activator of NKG2C+ NK cells. Proposed here is an autologous system of LFL stimulation without T lymphocytes and exogenous cytokines that allows us to evaluate NK-cell hCMV-specific responses in more native settings. In this model, we evaluated LFL-induced IFNγ production, focusing on signaling pathways and the degranulation and proliferation of NK cells orchestrated by microenvironment cytokine production and analyzed the transcriptome of expanded NK cells. NK cells of individuals having high anti-hCMV-IgG levels, in contrast to NK cells of hCMV-seronegative and low-positive donors, displayed increased IFNγ production and degranulation and activation levels and enhanced proliferation upon LFL stimulation. Cytokine profiles of these LFL-stimulated cultures demonstrated a proinflammatory shift. LFL-induced NK-cell IFNγ production was dependent on the PI3K and Ras/Raf/Mek signaling pathways, independently of cytokines. In hCMV-seropositive individuals, this model allowed obtaining NK-cell antigen-specific populations proliferating in response to LFL. The transcriptomic profile of these expanded NK cells showed increased adaptive gene expression and metabolic activation. The results complement the existing knowledge about hCMV-specific NK-cell response. This model may be further exploited for the identification and characterization of antigen-specific NK cells.

• Klíčová slova
• ERK1/2, HLA-E, IFNγ, NKG2C, RNAseq, cytokines, hCMV, memory NK cells,
• MeSH
• buňky NK MeSH
• cytokiny metabolismus   MeSH
• cytomegalovirové infekce * MeSH
• Cytomegalovirus MeSH
• lidé MeSH
• prezentace antigenu * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH

The genomic characteristics of human cytomegalovirus (HCMV) strains sequenced directly from clinical pathology samples were investigated, focusing on variation, multiple-strain infection, recombination, and gene loss. A total of 207 datasets generated in this and previous studies using target enrichment and high-throughput sequencing were analyzed, in the process enabling the determination of genome sequences for 91 strains. Key findings were that (i) it is important to monitor the quality of sequencing libraries in investigating variation; (ii) many recombinant strains have been transmitted during HCMV evolution, and some have apparently survived for thousands of years without further recombination; (iii) mutants with nonfunctional genes (pseudogenes) have been circulating and recombining for long periods and can cause congenital infection and resulting clinical sequelae; and (iv) intrahost variation in single-strain infections is much less than that in multiple-strain infections. Future population-based studies are likely to continue illuminating the evolution, epidemiology, and pathogenesis of HCMV.

• Klíčová slova
• gene loss, genome sequence, genotype, human cytomegalovirus, multiple-strain infection, mutation, recombination, target enrichment, variation,
• MeSH
• cytomegalovirové infekce virologie   MeSH
• Cytomegalovirus genetika   MeSH
• databáze nukleových kyselin MeSH
• datové soubory jako téma MeSH
• DNA virů genetika   MeSH
• genetická variace MeSH
• genom virový * genetika   MeSH
• genotyp MeSH
• lidé MeSH
• molekulární evoluce MeSH
• mutace MeSH
• rekombinace genetická * MeSH
• sekvence nukleotidů * MeSH
• sekvenční analýza DNA MeSH
• sekvenování celého genomu MeSH
• virové geny MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA virů MeSH

The human cytomegalovirus (HCMV) US12 family consists of ten sequentially arranged genes (US12-21) with poorly characterized function. We now identify novel natural killer (NK) cell evasion functions for four members: US12, US14, US18 and US20. Using a systematic multiplexed proteomics approach to quantify ~1300 cell surface and ~7200 whole cell proteins, we demonstrate that the US12 family selectively targets plasma membrane proteins and plays key roles in regulating NK ligands, adhesion molecules and cytokine receptors. US18 and US20 work in concert to suppress cell surface expression of the critical NKp30 ligand B7-H6 thus inhibiting NK cell activation. The US12 family is therefore identified as a major new hub of immune regulation.

• Klíčová slova
• cytomegalovirus, human, immune evasion, immunology, infectious disease, microbiology, natural killer cell, virus,
• MeSH
• buňky NK imunologie   MeSH
• Cytomegalovirus imunologie   patogenita   MeSH
• imunitní únik MeSH
• imunologické faktory antagonisté a inhibitory   MeSH
• interakce hostitele a patogenu * MeSH
• lidé MeSH
• membránové proteiny antagonisté a inhibitory   MeSH
• proteomika MeSH
• virové proteiny metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• imunologické faktory MeSH
• membránové proteiny MeSH
• virové proteiny MeSH