Early transcriptional and epigenetic divergence of CD8+ T cells responding to acute versus chronic infection.

During a microbial infection, responding CD8+ T cells give rise to effector cells that provide acute host defense and memory cells that provide sustained protection. An alternative outcome is exhaustion, a state of T cell dysfunction that occurs in the context of chronic infections and cancer. Although it is evident that exhausted CD8+ T (TEX) cells are phenotypically and molecularly distinct from effector and memory CD8+ T cells, the factors regulating the earliest events in the differentiation process of TEX cells remain incompletely understood. Here, we performed single-cell RNA-sequencing and single-cell ATAC-sequencing of CD8+ T cells responding to LCMV-Armstrong (LCMV-Arm) or LCMV-Clone 13 (LCMV-Cl13), which result in acute or chronic infections, respectively. Compared to CD8+ T cells that had undergone their first division in response to LCMV-Arm (Div1ARM) cells, CD8+ T cells that had undergone their first division in response to LCMV-Cl13 (Div1CL13) expressed higher levels of genes encoding transcription factors previously associated with exhaustion, along with higher levels of Ezh2, the catalytic component of the Polycomb Repressive Complex 2 (PRC2) complex, which mediates epigenetic silencing. Modulation of Ezh2 resulted in altered expression of exhaustion-associated molecules by CD8+ T cells responding to LCMV-Cl13, though the specific cellular and infectious contexts, rather than simply the level of Ezh2 expression, likely determine the eventual outcome. Taken together, these findings suggest that the differentiation paths of CD8+ T cells responding to acute versus chronic infections may diverge earlier than previously appreciated.

Small intestine and colon tissue-resident memory CD8+ T cells exhibit molecular heterogeneity and differential dependence on Eomes.

Tissue-resident memory CD8+ T (TRM) cells are a subset of memory T cells that play a critical role in limiting early pathogen spread and controlling infection. TRM cells exhibit differences across tissues, but their potential heterogeneity among distinct anatomic compartments within the small intestine and colon has not been well recognized. Here, by analyzing TRM cells from the lamina propria and epithelial compartments of the small intestine and colon, we showed that intestinal TRM cells exhibited distinctive patterns of cytokine and granzyme expression along with substantial transcriptional, epigenetic, and functional heterogeneity. The T-box transcription factor Eomes, which represses TRM cell formation in some tissues, exhibited unexpected context-specific regulatory roles in supporting the maintenance of established TRM cells in the small intestine, but not in the colon. Taken together, these data provide previously unappreciated insights into the heterogeneity and differential requirements for the formation vs. maintenance of intestinal TRM cells.

The long noncoding RNA Malat1 regulates CD8+ T cell differentiation by mediating epigenetic repression.

During an immune response to microbial infection, CD8+ T cells give rise to short-lived effector cells and memory cells that provide sustained protection. Although the transcriptional programs regulating CD8+ T cell differentiation have been extensively characterized, the role of long noncoding RNAs (lncRNAs) in this process remains poorly understood. Using a functional genetic knockdown screen, we identified the lncRNA Malat1 as a regulator of terminal effector cells and the terminal effector memory (t-TEM) circulating memory subset. Evaluation of chromatin-enriched lncRNAs revealed that Malat1 grouped with trans lncRNAs that exhibit increased RNA interactions at gene promoters and gene bodies. Moreover, we observed that Malat1 was associated with increased H3K27me3 deposition at a number of memory cell-associated genes through a direct interaction with Ezh2, thereby promoting terminal effector and t-TEM cell differentiation. Our findings suggest an important functional role of Malat1 in regulating CD8+ T cell differentiation and broaden the knowledge base of lncRNAs in CD8+ T cell biology.

Checkpoint Inhibitor Immune-Related Adverse Events: A Multimodality Pictorial Review.

Cancer immunotherapies are drugs that modulate the body's own immune system as an anticancer strategy. Checkpoint inhibitor immunotherapies interfere with cell surface binding proteins that function to promote self-recognition and tolerance, ultimately leading to upregulation of the immune response. Given the striking success of these agents in early trials in melanoma and lung cancer, they have now been studied in many types of cancer and have become a pillar of anticancer therapy for many tumor types. However, abundant upregulation results in a new class of side effects, known as immune-related adverse events (IRAEs). It is critical for the practicing radiologist to be able to recognize these events to best contribute to care for patients on checkpoint inhibitor immunotherapy. Here, we provide a comprehensive system-based review of immune-related adverse events and associated imaging findings. Further, we detail the best imaging modalities for each as well as describe problem solving modalities. Given that IRAEs can be subclinical before becoming clinically apparent, radiologists may be the first provider to recognize them, providing an opportunity for early treatment. Awareness of IRAEs and how to best image them will prepare radiologists to make a meaningful contribution to patient care as part of the clinical team.

The Role of Cost-Effectiveness Analysis in Patient-Centered Cancer Care in the Era of Precision Medicine.

Over the last few decades, changes in diagnostic and treatment paradigms have greatly advanced cancer care and improved outcomes [...].

Heterogeneity and clonal relationships of adaptive immune cells in ulcerative colitis revealed by single-cell analyses.

Inflammatory bowel disease (IBD) encompasses a spectrum of gastrointestinal disorders driven by dysregulated immune responses against gut microbiota. We integrated single-cell RNA and antigen receptor sequencing to elucidate key components, cellular states, and clonal relationships of the peripheral and gastrointestinal mucosal immune systems in health and ulcerative colitis (UC). UC was associated with an increase in IgG1+ plasma cells in colonic tissue, increased colonic regulatory T cells characterized by elevated expression of the transcription factor ZEB2, and an enrichment of a γδ T cell subset in the peripheral blood. Moreover, we observed heterogeneity in CD8+ tissue-resident memory T (TRM) cells in colonic tissue, with four transcriptionally distinct states of differentiation observed across health and disease. In the setting of UC, there was a marked shift of clonally related CD8+ TRM cells toward an inflammatory state, mediated, in part, by increased expression of the T-box transcription factor Eomesodermin. Together, these results provide a detailed atlas of transcriptional changes occurring in adaptive immune cells in the context of UC and suggest a role for CD8+ TRM cells in IBD.

OCT Circle Scans Can Be Used to Study Many Eyes with Advanced Glaucoma.

Purpose: To examine the utility of optical coherence tomography (OCT) for studying eyes with advanced glaucoma [i.e., eyes with a 24-2 visual field (VF) mean deviation (MD) worse than -15 dB], we tested the hypothesis that if these eyes had a 10-2 total deviation (TD) map with points better than -8 dB, then the topographically corresponding regions on the circumpapillary retinal nerve fiber layer (cpRNFL) should show a preserved region. Design: Evaluation of technology study. Participants: 39 eyes from 33 patients (mean: 68.8 ± 9.2 years) with a diagnosis of glaucoma had a 24-2 VF with a MD ≤ -15 dB (mean: -18.94 ± 2.95 dB). All eyes additionally had a 10-2 VF and an averaged OCT circle scan. Methods: Each scan was inspected, and preserved cpRNFL regions of the disc associated with the macula (central ±8° were delin eated. Main Outcome Measures: The number of eyes with preserved cpRNFL regions and their association with preserved VF locations (i.e. better than -8 dB) shown in the 10-2 VF TD map. Results: 38 of the 39 eyes had one or more points on the 10-2 VF with TD values that were better than -8 dB (mean: 25.7 ± 12.6 points). For all 39 eyes, there was a preserved portion of the cpRNFL on the circle scan within the disc region associated with the macula. However, for 3 of these eyes, this region was hypodense and could be a challenge for the clinician to identify. Conclusion: OCT scans can be used to assess and potentially follow the preserved regions of cpRNFL associated with the macula in eyes with advanced glaucoma if there is a preserved region on the 10-2 VF better than -8 dB.

Galleria mellonella as an insect model for P. destructans, the cause of White-nose Syndrome in bats.

Pseudogymnoascus destructans is the fungal pathogen responsible for White-nose Syndrome (WNS), a disease that has killed millions of bats in North America over the last decade. A major obstacle to research on P. destructans has been the lack of a tractable infection model for monitoring virulence. Here, we establish a high-throughput model of infection using larvae of Galleria mellonella, an invertebrate used to study host-pathogen interactions for a wide range of microbial species. We demonstrate that P. destructans can kill G. mellonella larvae in an inoculum-dependent manner when infected larvae are housed at 13°C or 18°C. Larval killing is an active process, as heat-killed P. destructans spores caused significantly decreased levels of larval death compared to live spores. We also show that fungal spores that were germinated prior to inoculation were able to kill larvae 3-4 times faster than non-germinated spores. Lastly, we identified chemical inhibitors of P. destructans and used G. mellonella to evaluate these inhibitors for their ability to reduce virulence. We demonstrate that amphotericin B can effectively block larval killing by P. destructans and thereby establish that this infection model can be used to screen biocontrol agents against this fungal pathogen.

Public health professionals' perceptions of mental health services in Equatorial Guinea, Central-West Africa.

INTRODUCTION: Mental health disorders constitute 13% of global disease burden, the impacts of which are disproportionality felt in sub-Saharan Africa. Equatorial Guinea, located in Central-West Africa, has the highest per-capita investment in healthcare on the African continent, but only two studies have discussed mental health issues in the country and none of have examined the perspective of professionals working in the field. The purpose of this study was to gain a preliminary understanding of Equatoguinean health care professionals' perspectives on the mental health care system. METHODS: Nine adult participants (directors or program managers) were interviewed in July 2013 in Malabo, Equatorial Guinea from government agencies, aid organizations, hospitals, and pharmacies. Interviews were designed to collect broad information about the mental healthcare system in Equatorial Guinea including the professionals' perspectives and access to resources. This research was reviewed and approved by an ethical oversight committee. RESULTS: All individuals interviewed indicated that the mental health system does not currently meet the needs of the community. Professionals cited infrastructural capacity, stigmatization, and a lack of other resources (training programs, knowledgeable staff, medications, data) as key factors that limit the effectiveness of mental healthcare. CONCLUSION: This study provides a preliminary understanding of the existing mental health care needs in the country, highlighting opportunities for enhanced healthcare services.

Expression of genes of the cardiac and renal renin-angiotensin systems in preterm piglets: is this system a suitable target for therapeutic intervention?

OBJECTIVES: The newborn circulating, cardiac and renal renin-angiotensin systems (RASs) are essential for blood pressure control, and for cardiac and renal development. If cardiac and renal RASs are immature this may contribute to cardiovascular compromise in preterm infants. This study measured mRNA expression of cardiac and renal RAS components in preterm, glucocorticoid (GC) exposed preterm, and term piglets. METHODS: Renal and cardiac RAS mRNA levels were measured using real-time polymerase chain reaction (PCR). Genes studied were: (pro)renin receptor, renin, angiotensinogen, angiotensin converting enzyme (ACE), ACE2, angiotensin type 1 receptor (AT1R) and angiotensin type 2 receptor (AT2R). RESULTS: All the genes studied were expressed in the kidney; neither renin nor AT2R mRNA were detected in the heart. There were no gestational changes in (pro)renin receptor, renin, ACE or AT1R mRNA levels. Right ventricular angiotensinogen mRNA levels in females were lower in preterm animals than at term, and GC exposure increased levels in male piglets. Renal angiotensinogen mRNA levels in female term piglets were lower than females from both preterm groups, and lower than male term piglets. Left ventricular ACE2 mRNA expression was lower in GC treated preterm piglets. Renal AT2R mRNA abundance was highest in GC treated preterm piglets, and the AT1R/AT2R ratio was increased at term. CONCLUSIONS: Preterm cardiac and renal RAS mRNA levels were similar to term piglets, suggesting that immaturity of these RASs does not contribute to preterm cardiovascular compromise. Since preterm expression of both renal and cardiac angiotensin II-AT1R is similar to term animals, cardiovascular dysfunction in the sick preterm human neonate might be effectively treated by agents acting on their RASs.

Healthcare in Equatorial Guinea, West Africa: obstacles and barriers to care.

INTRODUCTION: The provision of healthcare services in developing countries has received increasing attention, but inequalities persist. One nation with potential inequalities in healthcare services is Equatorial Guinea (Central-West Africa). Mitigating these inequalities is difficult, as the Equatoguinean healthcare system remains relatively understudied. METHODS: In this study, we interviewed members of the healthcare community in order to: 1) learn which diseases are most common and the most common cause of death from the perspective of healthcare workers; and 2) gain an understanding of the healthcare community in Equatorial Guinea by describing how: a) healthcare workers gain their professional knowledge; b) summarizing ongoing healthcare programs aimed at the general public; c) discussing conflicts within the healthcare community and between the public and healthcare providers; d) and addressing opportunities to improve healthcare delivery. RESULTS: We found that some causes of death, such as serious injuries, may not be currently treatable in country, potentially due to a lack of resources and trauma care facilities. In addition, training and informational programs for both healthcare workers and the general public may not be effectively transmitting information to the intended recipients. This presents hurdles to the healthcare community, both in terms of having professional competence in healthcare delivery and in having a community that is receptive to medical care. CONCLUSION: Our data also highlight government-facility communication as an opportunity for improvement. Our research is an important first step in understanding the context of healthcare delivery in Equatorial Guinea, a country that is relatively data poor.

A randomized trial of a brief primary-care-based intervention for reducing at-risk drinking practices.

This randomized trial evaluated an intervention for reducing at-risk drinking practices in a sample of 307 patients. Eligible drinking patterns included chronic drinking (> or = 2 drinks per day in the past month), binge drinking (> or = 5 drinks per occasion at least twice in the past month), and drinking and driving (driving after > 2 drinks in the past month). Members of the intervention group received a message from their physician during their regularly scheduled visit, a self-help manual, written personalized feedback, and up to 3 telephone counseling calls. Dropout was significantly higher in the intervention than control group.