NK Cell-Monocyte Cross-talk Underlies NK Cell Activation in Severe COVID-19.

NK cells in the peripheral blood of severe COVID-19 patients exhibit a unique profile characterized by activation and dysfunction. Previous studies have identified soluble factors, including type I IFN and TGF-ß, that underlie this dysregulation. However, the role of cell-cell interactions in modulating NK cell function during COVID-19 remains unclear. To address this question, we combined cell-cell communication analysis on existing single-cell RNA sequencing data with in vitro primary cell coculture experiments to dissect the mechanisms underlying NK cell dysfunction in COVID-19. We found that NK cells are predicted to interact most strongly with monocytes and that this occurs via both soluble factors and direct interactions. To validate these findings, we performed in vitro cocultures in which NK cells from healthy human donors were incubated with monocytes from COVID-19+ or healthy donors. Coculture of healthy NK cells with monocytes from COVID-19 patients recapitulated aspects of the NK cell phenotype observed in severe COVID-19, including decreased expression of NKG2D, increased expression of activation markers, and increased proliferation. When these experiments were performed in a Transwell setting, we found that only CD56bright CD16- NK cells were activated in the presence of severe COVID-19 patient monocytes. O-link analysis of supernatants from Transwell cocultures revealed that cultures containing severe COVID-19 patient monocytes had significantly elevated levels of proinflammatory cytokines and chemokines, as well as TGF-ß. Collectively, these results demonstrate that interactions between NK cells and monocytes in the peripheral blood of COVID-19 patients contribute to NK cell activation and dysfunction in severe COVID-19.

Defining the role of natural killer cells in COVID-19.

Natural killer (NK) cells are critical effectors of antiviral immunity. Researchers have therefore sought to characterize the NK cell response to coronavirus disease 2019 (COVID-19) and the virus that causes it, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The NK cells of patients with severe COVID-19 undergo extensive phenotypic and functional changes. For example, the NK cells from critically ill patients with COVID-19 are highly activated and exhausted, with poor cytotoxic function and cytokine production upon stimulation. The NK cell response to SARS-CoV-2 is also modulated by changes induced in virally infected cells, including the ability of a viral peptide to bind HLA-E, preventing NK cells from receiving inhibitory signals, and the downregulation of major histocompatibility complex class I and ligands for the activating receptor NKG2D. These changes have important implications for the ability of infected cells to escape NK cell killing. The implications of these findings for antibody-dependent NK cell activity in COVID-19 are also reviewed. Despite these advances in the understanding of the NK cell response to SARS-CoV-2, there remain critical gaps in our current understanding and a wealth of avenues for future research on this topic.

Single-cell RNA-seq methods to interrogate virus-host interactions.

The twenty-first century has seen the emergence of many epidemic and pandemic viruses, with the most recent being the SARS-CoV-2-driven COVID-19 pandemic. As obligate intracellular parasites, viruses rely on host cells to replicate and produce progeny, resulting in complex virus and host dynamics during an infection. Single-cell RNA sequencing (scRNA-seq), by enabling broad and simultaneous profiling of both host and virus transcripts, represents a powerful technology to unravel the delicate balance between host and virus. In this review, we summarize technological and methodological advances in scRNA-seq and their applications to antiviral immunity. We highlight key scRNA-seq applications that have enabled the understanding of viral genomic and host response heterogeneity, differential responses of infected versus bystander cells, and intercellular communication networks. We expect further development of scRNA-seq technologies and analytical methods, combined with measurements of additional multi-omic modalities and increased availability of publicly accessible scRNA-seq datasets, to enable a better understanding of viral pathogenesis and enhance the development of antiviral therapeutics strategies.

SARS-CoV-2 escapes direct NK cell killing through Nsp1-mediated downregulation of ligands for NKG2D.

Natural killer (NK) cells are cytotoxic effector cells that target and lyse virally infected cells; many viruses therefore encode mechanisms to escape such NK cell killing. Here, we interrogate the ability of SARS-CoV-2 to modulate NK cell recognition and lysis of infected cells. We find that NK cells exhibit poor cytotoxic responses against SARS-CoV-2-infected targets, preferentially killing uninfected bystander cells. We demonstrate that this escape is driven by downregulation of ligands for the activating receptor NKG2D (NKG2D-L). Indeed, early in viral infection, prior to NKG2D-L downregulation, NK cells are able to target and kill infected cells; however, this ability is lost as viral proteins are expressed. Finally, we find that SARS-CoV-2 non-structural protein 1 (Nsp1) mediates downregulation of NKG2D-L and that Nsp1 alone is sufficient to confer resistance to NK cell killing. Collectively, our work demonstrates that SARS-CoV-2 evades direct NK cell cytotoxicity and describes a mechanism by which this occurs.

Deep Phenotypic Analysis of Blood and Lymphoid T and NK Cells From HIV+ Controllers and ART-Suppressed Individuals.

T and natural killer (NK) cells are effector cells with key roles in anti-HIV immunity, including in lymphoid tissues, the major site of HIV persistence. However, little is known about the features of these effector cells from people living with HIV (PLWH), particularly from those who initiated antiretroviral therapy (ART) during acute infection. Our study design was to use 42-parameter CyTOF to conduct deep phenotyping of paired blood- and lymph node (LN)-derived T and NK cells from three groups of HIV+ aviremic individuals: elite controllers (N = 5), and ART-suppressed individuals who had started therapy during chronic (N = 6) vs. acute infection (N = 8), the latter of which is associated with better outcomes. We found that acute-treated individuals are enriched for specific subsets of T and NK cells, including blood-derived CD56-CD16+ NK cells previously associated with HIV control, and LN-derived CD4+ T follicular helper cells with heightened expansion potential. An in-depth comparison of the features of the cells from blood vs. LNs of individuals from our cohort revealed that T cells from blood were more activated than those from LNs. By contrast, LNs were enriched for follicle-homing CXCR5+ CD8+ T cells, which expressed increased levels of inhibitory receptors and markers of survival and proliferation as compared to their CXCR5- counterparts. In addition, a subset of memory-like CD56brightTCF1+ NK cells was enriched in LNs relative to blood. These results together suggest unique T and NK cell features in acute-treated individuals, and highlight the importance of examining effector cells not only in blood but also the lymphoid tissue compartment, where the reservoir mostly persists, and where these cells take on distinct phenotypic features.

Multi-omic profiling reveals widespread dysregulation of innate immunity and hematopoiesis in COVID-19.

Our understanding of protective versus pathological immune responses to SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), is limited by inadequate profiling of patients at the extremes of the disease severity spectrum. Here, we performed multi-omic single-cell immune profiling of 64 COVID-19 patients across the full range of disease severity, from outpatients with mild disease to fatal cases. Our transcriptomic, epigenomic, and proteomic analyses revealed widespread dysfunction of peripheral innate immunity in severe and fatal COVID-19, including prominent hyperactivation signatures in neutrophils and NK cells. We also identified chromatin accessibility changes at NF-κB binding sites within cytokine gene loci as a potential mechanism for the striking lack of pro-inflammatory cytokine production observed in monocytes in severe and fatal COVID-19. We further demonstrated that emergency myelopoiesis is a prominent feature of fatal COVID-19. Collectively, our results reveal disease severity-associated immune phenotypes in COVID-19 and identify pathogenesis-associated pathways that are potential targets for therapeutic intervention.

Transforming Lives of Former Foster Youths: A Campus-Based Support Program Grounded in Social Work Values.

Support programs to improve higher education outcomes for former foster youths have emerged in the past 20 years, but literature to guide their development and implementation is limited. This conceptual article presents the program logic model for ACE [Achieving College Excellence] Scholars Services, the comprehensive, campus-based program supporting former foster youths grounded in social work values at California State University San Marcos (CSUSM). ACE has a noncompetitive application process that accepts an unrestricted number of students, known as ACE Scholars. To focus on the unique needs of former foster youths, students who are on probation, experiencing homeless, and unaccompanied minors are included if they have also experienced foster care. Funded by private donors, the Center for ACE Scholars on CSUSM's campus provides an inviting, welcoming space that reflects the inclusive support ACE Scholars receive. By increasing educational attainment and opportunities for this vulnerable population, ACE aims to improve outcomes for former foster youths and their next generation. Articulating this program logic model provides implications for developing and implementing support programs for former foster youths on college campuses and sets the foundation for evaluating the impact of ACE Scholars Services.

Geocorrection of Airborne Mid-Wave Infrared Imagery for Mapping Wildfires without GPS or IMU.

The increase in annual wildfires in many areas of the world has triggered international efforts to deploy sensors on airborne and space platforms to map these events and understand their behaviour. During the summer of 2017, an airborne flight campaign acquired mid-wave infrared imagery over active wildfires in Northern Ontario, Canada. However, it suffered multiple position-based equipment issues, thus requiring a non-standard geocorrection methodology. This study presents the approach, which utilizes a two-step semi-automatic geocorrection process that outputs image mosaics from airborne infrared video input. The first step extracts individual video frames that are combined into orthoimages using an automatic image registration method. The second step involves the georeferencing of the imagery using pseudo-ground control points to a fixed coordinate systems. The output geocorrected datasets in units of radiance can then be used to derive fire products such as fire radiative power density (FRPD). Prior to the georeferencing process, the Root Mean Square Error (RMSE) associated with the imagery was greater than 200 m. After the georeferencing process was applied, an RMSE below 30 m was reported, and the computed FRPD estimations are within expected values across the literature. As such, this alternative geocorrection methodology successfully salvages an otherwise unusable dataset and can be adapted by other researchers that do not have access to accurate positional information for airborne infrared flight campaigns over wildfires.

Stage-Specific Requirement for Eomes in Mature NK Cell Homeostasis and Cytotoxicity.

Natural killer (NK) cells are cytotoxic innate lymphoid cells (ILCs) that mediate antiviral and antitumor responses and require the transcriptional regulator Eomesodermin (Eomes) for early development. However, the role of Eomes and its molecular program in mature NK cell biology is unclear. To address this, we develop a tamoxifen-inducible, type-1-ILC-specific (Ncr1-targeted) cre mouse and combine this with Eomes-floxed mice. Eomes deletion after normal NK cell ontogeny results in a rapid loss of NK cells (but not ILC1s), with a particularly profound effect on penultimately mature stage III NK cells. Mechanisms responsible for stage III reduction include increased apoptosis and impaired maturation from stage II precursors. Induced Eomes deletion also decreases NK cell cytotoxicity and abrogates in vivo rejection of major histocompatibility complex (MHC)-class-I-deficient cells. However, other NK cell functional responses, and stage IV NK cells, are largely preserved. These data indicate that mature NK cells have distinct Eomes-dependent and -independent stages.

Current Status of Septic Cardiomyopathy: Basic Science and Clinical Progress.

Septic cardiomyopathy (SCM) is a complication that is sepsis-associated cardiovascular failure. In the last few decades, there is progress in diagnosis and treatment despite the lack of consistent diagnostic criteria. According to current studies, several hypotheses about pathogenic mechanisms have been revealed to elucidate the pathophysiological characteristics of SCM. The objective of this manuscript is to review literature from the past 5 years to provide an overview of current knowledge on pathogenesis, diagnosis and treatment in SCM.

The Cost of Quality: Organizational Financial Health and Program Quality.

Purpose: Nonprofit health and human service organizations (NPHSOs) encounter pressure to improve program quality to continue to secure financial resources. Yet, organizations also struggle to innovate, evaluate, and monitor quality absent sufficient resources. This study explores the relationship between NPHSOs' financial health and quality indicators. Methods: Quality indicators were measured in an online survey with leaders from 32 nonprofit children's behavioral health organizations within a Midwestern urban region. Survey responses were matched with financial health metrics extracted from publicly available IRS 990 forms from 2010 to 2014. Results: Organizations struggled to maintain adequate reserve levels, although those with more quality capacities had greater assets and growth in those assets. Discussion: Although financial health appeared to be related to quality, it is unclear whether quality attracts financial resources, or if financial reserves support quality. Conclusion: Continuing research and dialogue are needed to understand the financial implications of an increasing emphasis on quality.

A National Perspective on Exploring Correlates of Accreditation in Children's Mental Health Care.

This study is the first to explore national accreditation rates and the relationship between accreditation status and organizational characteristics and quality indicators in children's mental health. Data from the Substance Abuse and Mental Health Services Administration's (SAMHSA's) National Survey of Mental Health Treatment Facilities (NSMHTF) were used from 8,247 facilities that serve children and/or adolescents. Nearly 60% (n=4,925) of the facilities were accredited by the Council on Accreditation (COA), the Commission on Accreditation of Rehabilitation Facilities (CARF), or The Joint Commission (TJC). Chi-square analyses were conducted to explore relationships. Compared to non-accredited facilities, more accredited facilities reported greater number of admissions, acceptance of government funding and client funds, and implementation of several quality indicators. Policies with incentives for accreditation could influence accreditation rates, and accreditation could influence quality indicators. These results set the foundation for future research about the drivers of the accreditation phenomenon and its impact on children's mental health outcomes.

Challenges to quality assurance and improvement efforts in behavioral health organizations: a qualitative assessment.

Behavioral health organizations have been increasingly required to implement plans to monitor and improve service quality. This qualitative study explores challenges that quality assurance and improvement (QA/I) personnel experience in performing their job in those practice settings. Sixteen QA/I personnel from different agencies in St. Louis, Missouri, U.S.A., were interviewed face-to-face using a semi-structured instrument to capture challenges and a questionnaire to capture participant and agency characteristics. Data analysis followed a grounded theory approach. Challenges involved agency resources, agency buy-in, personnel training, competing demands, shifting standards, authority, and research capacity. Further research is needed to assess these challenges given expected outcomes.

Needs and outcomes for low income youth in special education: Variations by emotional disturbance diagnosis and child welfare contact.

Despite the high rates of service for emotional disturbance among child welfare involved youth, much remains to be understood about this population. This study is the first to use longitudinal data to examine the needs and outcomes of children in special education (comparing those with emotional disturbance (ED) and those without) according to child welfare involvement (none, child abuse and neglect report but no services, in-home child welfare services, and foster care). Administrative data linked with special education case file data on 471 youth found that those involved with child welfare were most likely to have an ED diagnosis. Special education assessments revealed that children with in-home services or reports of maltreatment without services generally had equal or greater levels of needs indicated than those placed in foster care. Youth with an ED diagnosis were more likely to experience a negative outcome, such as emergency room treatment for mental health, school problems, or juvenile delinquency. Almost all of the ED youth had child welfare contact by the end of the study period. These findings underline the unmet needs of this population and the need for system coordination to improve their outcomes.

Quality assurance and improvement practice in mental health agencies: roles, activities, targets and contributions.

Accompanying the rise in the number of mental health agency personnel tasked with quality assurance and improvement (QA/I) responsibilities is an increased need to understand the nature of the work these professionals undertake. Four aspects of the work of quality assurance and improvement (QA/I) professionals in mental health were explored in this qualitative study: their perceived roles, their major activities, their QA/I targets, and their contributions. In-person interviews were conducted with QA/I professionals at 16 mental health agencies. Respondents perceived their roles at varying levels of complexity, focused on different targets, and used different methods to conduct their work. Few targets of QA/I work served as indicators of high quality care. Most QA/I professionals provided concrete descriptions of how they had improved agency services, while others could describe none. Accreditation framed much of agency QA/I work, perhaps to its detriment.