Utilizing Complexity Leadership Theory to Promote Faculty Mentorship and Resident Leadership.

OBJECTIVE: To create and evaluate a structured combined faculty mentorship/resident leadership program based on complexity leadership theory. DESIGN: In 2021, a logic model was used to create a 5 part bi-monthly resident leadership series utilizing administrative, adaptive, and enabling components of complexity leadership theory. Each of the 5 sessions had a nationally prominent senior faculty member mentor 3 junior faculty in creation of an interactive workshop that was delivered to resident physicians during scheduled didactics. Validated surveys were used to assess faculty post-mentorship experience and resident self-perception of leadership skills pre-and post-series. Descriptive statistics and 2-way ANOVA were performed; text comments underwent content analysis. SETTING: A large academic OB/GYN department at Baylor College of Medicine in Houston, Texas. PARTICIPANTS: Five faculty mentors, 15 junior faculty, and 48 residents participated in this program. All faculty mentors (5/5) and 87% (13/15) of mentees completed the post-mentorship survey. Resident response rate was 60% (29/48) pre-series and 63% (30/48) post-series. RESULTS: Both mentors and mentees rated the experience favorably (4.62 versus 5.29, p = 0.51). In open-ended comments, enabling components of mentorship process, such as approachability and expertise of the mentors, were most often noted as positive. Both mentees and mentors suggested administrative changes to the experience, such as longitudinal relationships between mentors and mentees. The mean score on the resident leadership questionnaire improved from 3.82 to 3.96 (5-point Likert scale, p = 0.30) with self-reported leadership skills improving in 8/9 domains, although none reaching statistical significance. Open-ended comments revealed that residents also most desired administrative changes in the leadership series, such as increased leadership opportunities and more interactive workshops. CONCLUSIONS: A structured combined faculty mentorship/resident leadership program formed utilizing complexity leadership theory was positively received. Participants most liked the enabling components of the series, with requested administrative changes in the future.

Defining the Effects of PKC Modulator HIV Latency-Reversing Agents on Natural Killer Cells.

Background: Latency reversing agents (LRAs) such as protein kinase C (PKC) modulators can reduce rebound-competent HIV reservoirs in small animal models. Furthermore, administration of natural killer (NK) cells following LRA treatment improves this reservoir reduction. It is currently unknown why the combination of a PKC modulator and NK cells is so potent and whether exposure to PKC modulators may augment NK cell function in some way. Methods: Primary human NK cells were treated with PKC modulators (bryostatin-1, prostratin, or the designed, synthetic bryostatin-1 analog SUW133), and evaluated by examining expression of activation markers by flow cytometry, analyzing transcriptomic profiles by RNA sequencing, measuring cytotoxicity by co-culturing with K562 cells, assessing cytokine production by Luminex assay, and examining the ability of cytokines and secreted factors to independently reverse HIV latency by co-culturing with Jurkat-Latency (J-Lat) cells. Results: PKC modulators increased expression of proteins involved in NK cell activation. Transcriptomic profiles from PKC-treated NK cells displayed signatures of cellular activation and enrichment of genes associated with the NFκB pathway. NK cell cytotoxicity was unaffected by prostratin but significantly decreased by bryostatin-1 and SUW133. Cytokines from PKC-stimulated NK cells did not induce latency reversal in J-Lat cell lines. Conclusions: Although PKC modulators have some significant effects on NK cells, their contribution in "kick and kill" strategies is likely due to upregulating HIV expression in CD4+ T cells, not directly enhancing the effector functions of NK cells. This suggests that PKC modulators are primarily augmenting the "kick" rather than the "kill" arm of this HIV cure approach.

C. elegans Presenilin Mediates Inter-Organelle Contacts and Communication that Is Required for Lysosome Activity.

Compromised lysosome function is implicated in the pathology of many neurodegenerative diseases, including Alzheimer's disease (AD). Familial Alzheimer's disease (fAD) is caused primarily by mutations in the presenilin encoding genes, but the underlying mechanism remains obscure. Loss of the conserved C. elegans presenilin orthologue SEL-12 results in increased mitochondrial calcium, which promotes neurodegeneration. Here, we find that sel-12 mutant lysosomes, independent of SEL-12 proteolytic activity, are significantly enlarged and more alkaline due to increased ER-to-mitochondrial calcium signaling and concomitant mitochondrial oxidative stress. These defects and their dependence on mitochondrial calcium are recapitulated in human fAD fibroblasts, demonstrating a conserved role for mitochondrial calcium in presenilin-mediated lysosome dysfunction. sel-12 mutants also have increased contact surface area between the ER, mitochondria, and lysosomes, suggesting sel-12 has an additional role in modulating organelle contact and communication. Overall, we demonstrate that SEL-12 maintains lysosome acidity and lysosome health by controlling ER-to-mitochondrial calcium signaling.

Drivers of men's use of intimate partner violence in conflict-affected settings: learnings from the Democratic Republic of Congo.

BACKGROUND: Intimate partner violence against women (IPVAW) is prevalent in conflict-affected settings. Yet, there is limited knowledge about the risk factors that influence men's use of IPVAW in conflict-affected settings. This paper adopts a transdisciplinary perspective to understand how experiences hypothesized to increase men's use of IPVAW relate to each other and to men's use of IPVAW. The findings may help researchers and interventionists to better select and target interventions for IPVAW in conflict-affected settings. METHODS: We used baseline data from the Tushinde Ujeuri project in the Democratic Republic of Congo. Men with at least partial data for the variables of interest were included in the analysis (n = 2080). We estimated a structural equation model that explored how five constructs - interpersonal violence, mental health, socioeconomic adversity, gender inequitable attitudes, and conflict violence - influenced men's self-reported past-year use of physical and/or sexual IPVAW. RESULTS: The model had acceptable fit (χ2 = 1576.574, p = 0.000; RMSEA = 0.041; CLI = 0.882; SRMR = 0.055). There was a statistically significant path from interpersonal violence to IPVAW (ß = 0.875; OR = 2.40). Interpersonal violence also was linked to gender inequitable attitudes (ß = 0.364), which were linked to increased use of IPVAW (ß = 0.180; OR = 1.20). Moreover, interpersonal violence was linked to trauma symptoms (ß = 0.331), which were linked to increased use of IPVAW (ß = 0.238; OR = 1.27). Use of IPVAW decreased as conflict exposures increased (ß=-0.036; OR = 0.96), and there was no path from socioeconomic adversity to IPVAW. CONCLUSIONS: Our findings suggest interpersonal violence exposures, trauma symptoms, and gender inequitable attitudes are all risk factors for the use of IPVAW in a conflict-affected setting. While continuing to focus on gender inequitable attitudes and norms, interventionists should also consider addressing men's experiences of victimization and mental wellbeing. Doing so can help to improve trauma symptoms and may hold promise to reduce IPVAW in conflict-affected settings.

Vancomycin-induced gut microbial dysbiosis alters enteric neuron-macrophage interactions during a critical period of postnatal development.

Vancomycin is a broad-spectrum antibiotic widely used in cases of suspected sepsis in premature neonates. While appropriate and potentially lifesaving in this setting, early-life antibiotic exposure alters the developing microbiome and is associated with an increased risk of deadly complications, including late-onset sepsis (LOS) and necrotizing enterocolitis (NEC). Recent studies show that neonatal vancomycin treatment disrupts postnatal enteric nervous system (ENS) development in mouse pups, which is in part dependent upon neuroimmune interactions. This suggests that early-life antibiotic exposure could disrupt these interactions in the neonatal gut. Notably, a subset of tissue-resident intestinal macrophages, muscularis macrophages, has been identified as important contributors to the development of postnatal ENS. We hypothesized that vancomycin-induced neonatal dysbiosis impacts postnatal ENS development through its effects on macrophages. Using a mouse model, we found that exposure to vancomycin in the first 10 days of life, but not in adult mice, resulted in an expansion of pro-inflammatory colonic macrophages by increasing the recruitment of bone-marrow-derived macrophages. Single-cell RNA sequencing of neonatal colonic macrophages revealed that early-life vancomycin exposure was associated with an increase in immature and inflammatory macrophages, consistent with an influx of circulating monocytes differentiating into macrophages. Lineage tracing confirmed that vancomycin significantly increased the non-yolk-sac-derived macrophage population. Consistent with these results, early-life vancomycin exposure did not expand the colonic macrophage population nor decrease enteric neuron density in CCR2-deficient mice. Collectively, these findings demonstrate that early-life vancomycin exposure alters macrophage number and phenotypes in distinct ways compared with vancomycin exposure in adult mice and results in altered ENS development.

Current Advances in Humanized Mouse Models for Studying NK Cells and HIV Infection.

Human immunodeficiency virus (HIV) has infected millions of people worldwide and continues to be a major global health problem. Scientists required a small animal model to study HIV pathogenesis and immune responses. To this end, humanized mice were created by transplanting human cells and/or tissues into immunodeficient mice to reconstitute a human immune system. Thus, humanized mice have become a critical animal model for HIV researchers, but with some limitations. Current conventional humanized mice are prone to death by graft versus host disease induced by the mouse signal regulatory protein α and CD47 signaling pathway. In addition, commonly used humanized mice generate low levels of human cytokines required for robust myeloid and natural killer cell development and function. Here, we describe recent advances in humanization procedures and transgenic and knock-in immunodeficient mice to address these limitations.

DEC-7/SUSD2, a sushi domain-containing protein, regulates an ultradian behavior mediated by intestinal epithelial Ca2+ oscillations in Caenorhabditis elegans.

In Caenorhabditis elegans, rhythmic posterior body wall muscle contractions mediate the highly regular defecation cycle. These contractions are regulated by inositol-1,4,5-trisphosphate (InsP3) receptor-dependent Ca2+ oscillations in intestinal epithelial cells. Here, we find that mutations in dec-7, which encodes the nematode ortholog of the human Sushi domain-containing 2 protein (SUSD2), lead to an increase in InsP3 receptor-dependent rhythmic posterior body wall muscle contractions. DEC-7 is highly expressed in the intestinal epithelia and localizes to the cell-cell junction. The increase in rhythmic activity caused by the loss of dec-7 is dependent on the innexin gap junction protein INX-16. Moreover, DEC-7 is required for the clustering of INX-16 to the cell-cell junction of the intestinal epithelia. We hypothesize that DEC-7/SUSD2 regulates INX-16 activity to mediate the rhythmic frequency of the defecation motor program. Thus, our data indicate a critical role of a phylogenetically conserved cell-cell junction protein in mediating an ultradian rhythm in the intestinal epithelia of C. elegans.NEW & NOTEWORTHY The conserved complement group protein DEC-7/SUSD2 acts at the apical cell-cell junction of C. elegans intestinal epithelia to mediate gap junction protein organization and function to facilitate a Ca2+ wave-regulated ultradian behavior.

Case report: Responses to the combination of gemcitabine with sirolimus in two patients with TSC-mutated sarcomas.

TSC-mutated sarcomas are rare molecular and histologic types of sarcoma. Due to the presence of their specific oncogenic driver mutation, these sarcomas are particularly sensitive to mTOR inhibitors. Recently, nab-sirolimus, an albumin-bound mTOR inhibitor, was approved by the Food and Drug Administration (FDA) for PEComas, which harbor a TSC mutation, and this drug remains the only FDA-approved systemic treatment for these tumors. We report on two cases of patients with TSC-mutated sarcomas who experienced significant responses to the combination of gemcitabine and sirolimus, after progression on prior gemcitabine-based chemotherapy and single agent mTOR inhibition with nab-sirolimus. Preclinical and clinical data support rationale for a synergistic effect of the combination. This combination may represent a valid therapeutic option after failure of nab-sirolimus in these patients, with no standard-of-care treatment options.

Secreted factors induced by PKC modulators do not indirectly cause HIV latency reversal.

HIV can establish a long-lived latent infection in cells harboring integrated non-expressing proviruses. Latency reversing agents (LRAs), including protein kinase C (PKC) modulators, can induce expression of latent HIV, thereby reducing the latent reservoir in animal models. However, PKC modulators such as bryostatin-1 also cause cytokine upregulation in peripheral blood mononuclear cells (PBMCs), including cytokines that might independently reverse HIV latency. To determine whether cytokines induced by PKC modulators contribute to latency reversal, primary human PBMCs were treated with bryostatin-1 or the bryostatin analog SUW133, a superior LRA, and supernatant was collected. As anticipated, LRA-treated cell supernatant contained increased levels of cytokines compared to untreated cell supernatant. However, exposure of latently-infected cells with this supernatant did not result in latency reactivation. These results indicate that PKC modulators do not have significant indirect effects on HIV latency reversal in vitro and thus are targeted in their latency reversing ability.

A humanized mouse model to study NK cell biology during HIV infection.

NK cells are an important subset of innate immune effectors with antiviral activity. However, NK cell development and immune responses in different tissues during acute and chronic HIV infection in vivo have been difficult to study due to the impaired development and function of NK cells in conventional humanized mouse models. In this issue of the JCI, Sangur et al. report on a transgenic MISTRG-6-15 mouse model with human IL-6 and IL-15 knocked into the previously constructed MISTRG mice. The predecessor model was deficient in Rag2 and γ chain (γc) with knock-in expression of human M-CSF, IL-3, GM-CSF, and TPO, and transgenic expression of human SIRPα. The researchers studied tissue-specific NK cell immune responses during HIV infection and clearly show that the endogenous human NK cells in the humanized mouse model suppressed HIV-1 replication in vivo. These findings provide insight into harnessing the innate immune response for clinical antiviral therapies.

Deregulation of Mitochondrial Calcium Handling Due to Presenilin Loss Disrupts Redox Homeostasis and Promotes Neuronal Dysfunction.

Mitochondrial dysfunction and oxidative stress are major contributors to the pathophysiology of neurodegenerative diseases, including Alzheimer's disease (AD). However, the mechanisms driving mitochondrial dysfunction and oxidative stress are unclear. Familial AD (fAD) is an early onset form of AD caused primarily by mutations in the presenilin-encoding genes. Previously, using Caenorhabditis elegans as a model system to study presenilin function, we found that loss of C. elegans presenilin orthologue SEL-12 results in elevated mitochondrial and cytosolic calcium levels. Here, we provide evidence that elevated neuronal mitochondrial generated reactive oxygen species (ROS) and subsequent neurodegeneration in sel-12 mutants are a consequence of the increase of mitochondrial calcium levels and not cytosolic calcium levels. We also identify mTORC1 signaling as a critical factor in sustaining high ROS in sel-12 mutants in part through its repression of the ROS scavenging system SKN-1/Nrf. Our study reveals that SEL-12/presenilin loss disrupts neuronal ROS homeostasis by increasing mitochondrial ROS generation and elevating mTORC1 signaling, which exacerbates this imbalance by suppressing SKN-1/Nrf antioxidant activity.

Violence against pregnant women and adverse maternal/ fetal outcomes in the United States: racial/ethnic disparities.

Violence against women is relatively common, and violence during pregnancy is of special concern due to potential risk of maternal and neonatal complications. Previous studies using diagnostic codes to determine prevalence and health outcomes of violence against women used ICD-9 data and lack a standard of consistency. Data from the 2002 to 2018 National Inpatient Sample (NIS) was used to analyze pregnancy-related hospitalizations of women aged 15-49 years. International Classification of Disease, Ninth Edition, Clinical Modification (ICD-9-CM) was utilized in the NIS until the third quarter of 2015, after which it transitioned to ICD-10-CM format. The exposure was violence against women whereas outcomes included preterm birth, intrauterine fetal demise, miscarriage, fetal growth restriction, hypertensive disorders of pregnancy, and gestational diabetes. Temporal trends analyses were performed using Joinpoint regression technique and adjusted survey logistic regression models were conducted to examine the association between exposure and outcomes. Certain sociodemographic characteristics including age 35-49 (2.88/1,000 hospitalizations), non-Hispanic White (2.66/1,000) and non-Hispanic Black (2.61/1,000) racial/ethnic groups, and lowest quartile income (2.91/1,000) were associated with higher prevalence of violence. There was an overall increase in hospitalizations over the study period, the most significant being among non-Hispanic White patients (AAPC 18 percent, 95 percent CI = 10.3, 26.3). When compared to those with no exposure, individuals of all ethnic groups exposed to violence had increased risk of all adverse maternal/fetal outcomes. Higher prevalence of violence was associated with certain sociodemographic characteristics. Disparities in maternal/fetal adverse outcome risk were noted between ethnic groups. Additional studies are needed to ensure accuracy of violence data using diagnostic codes.

Promising practices for the monitoring and evaluation of gender-based violence risk mitigation interventions in humanitarian response: a multi-methods study.

BACKGROUND: Risks of gender-based violence (GBV) are exacerbated in humanitarian crises. GBV risk mitigation interventions aim to reduce exposure to GBV and ensure that humanitarian response actions and services themselves do not cause harm or increase the risk of violence. The 2015 IASC Guidelines for Integrating Gender-Based Violence Interventions in Humanitarian Action ('GBV Guidelines') are a globally endorsed resource that provides comprehensive guidance for all humanitarian actors and sectors on GBV risk mitigation. While uptake of GBV risk mitigation approaches across multiple humanitarian sectors has occurred, there is limited understanding of how to monitor and evaluate GBV risk mitigation interventions. METHODS: A multi-methods study was conducted in 2019 to identify promising practices for the monitoring and evaluation (M&E) of GBV risk mitigation interventions in non-GBV sectors and to develop a set of illustrative case examples. The study included a comprehensive desk review of 145 articles, documents and resources from the published and grey literature, as well as 11 in-depth interviews and five focus group discussions with humanitarian practitioners. Using Dedoose software and a codebook developed a priori, qualitative data were transcribed and coded and a content analysis was conducted. Excerpts focusing on promising practices from the qualitative data and the desk review were analyzed together and grouped by thematic area. Similar promising practices were combined and consolidated to create a final list, and case examples were identified. RESULTS: Current promising practices for M&E of GBV risk mitigation activities in the following categories are described: (1) Coordination and collaboration, (2) Designing M&E approaches and tools for GBV risk mitigation activities, (3) Contextualization, (4) Developing and selecting indicators, (5) Data collection, (6) Data analysis and use of findings, (7) Potential safety concerns for affected populations and staff, and (8) Staff capacity and engagement. These are supplemented with seven diverse case examples to illustrate application of the promising practices using real-world examples. CONCLUSION: This paper highlights current promising practices for M&E of GBV risk mitigation interventions in humanitarian response. Further application of these practices-alongside ongoing documentation of emerging approaches-will be critical to ensuring that GBV risk mitigation interventions are more rigorously tested with the aim of building the evidence base on the effectiveness of different GBV risk mitigation interventions within specific humanitarian sectors.

Development of off-the-shelf hematopoietic stem cell-engineered invariant natural killer T cells for COVID-19 therapeutic intervention.

BACKGROUND: New COVID-19 treatments are desperately needed as case numbers continue to rise and emergent strains threaten vaccine efficacy. Cell therapy has revolutionized cancer treatment and holds much promise in combatting infectious disease, including COVID-19. Invariant natural killer T (iNKT) cells are a rare subset of T cells with potent antiviral and immunoregulatory functions and an excellent safety profile. Current iNKT cell strategies are hindered by the extremely low presence of iNKT cells, and we have developed a platform to overcome this critical limitation. METHODS: We produced allogeneic HSC-engineered iNKT (AlloHSC-iNKT) cells through TCR engineering of human cord blood CD34+ hematopoietic stem cells (HSCs) and differentiation of these HSCs into iNKT cells in an Ex Vivo HSC-Derived iNKT Cell Culture. We then established in vitro SARS-CoV-2 infection assays to assess AlloHSC-iNKT cell antiviral and anti-hyperinflammation functions. Lastly, using in vitro and in vivo preclinical models, we evaluated AlloHSC-iNKT cell safety and immunogenicity for off-the-shelf application. RESULTS: We reliably generated AlloHSC-iNKT cells at high-yield and of high-purity; these resulting cells closely resembled endogenous human iNKT cells in phenotypes and functionalities. In cell culture, AlloHSC-iNKT cells directly killed SARS-CoV-2 infected cells and also selectively eliminated SARS-CoV-2 infection-stimulated inflammatory monocytes. In an in vitro mixed lymphocyte reaction (MLR) assay and an NSG mouse xenograft model, AlloHSC-iNKT cells were resistant to T cell-mediated alloreaction and did not cause GvHD. CONCLUSIONS: Here, we report a method to robustly produce therapeutic levels of AlloHSC-iNKT cells. Preclinical studies showed that these AlloHSC-iNKT cells closely resembled endogenous human iNKT cells, could reduce SARS-CoV-2 virus infection load and mitigate virus infection-induced hyperinflammation, and meanwhile were free of GvHD-risk and resistant to T cell-mediated allorejection. These results support the development of AlloHSC-iNKT cells as a promising off-the-shelf cell product for treating COVID-19; such a cell product has the potential to target the new emerging SARS-CoV-2 variants as well as the future new emerging viruses.

Longitudinal outcomes one year following implicit bias training in medical students.

PURPOSE: Training in implicit bias is broadly recognized as important in medical education and is mandated by some accrediting bodies. This study examined medical students' retention of concepts immediately following and one-year post participation in an implicit bias workshop. METHODS: Study subjects were 272 third-year medical students who participated in workshops held between 2018-2020 that used the Implicit Associations Test (IAT) as a trigger for discussions in small groups. We developed a survey and administered it to students to capture their awareness of implicit bias pre-, post-, and one-year post-workshop attendance. Repeated Measures Analyses and independent-samples t-tests were used to examine for differences in responses on each of the seven survey items and a tabulated 7-item average of these seven items. RESULTS: Six of seven survey items and the tabulated 7-item average examined by Repeated Measures Analyses showed statistically significant increases between the pre-, post-, and one-year post-surveys (ps range: 0.01-0.07), with a small to moderate effect sizes (ƞp2s range: 0.01-0.07). Pairwise comparisons among these three surveys' results indicated statistically significant improvements between the pre- and the post-workshop surveys (ps range: 0.01-0.03) but no statistically significant differences between the post- and the one-year post-workshop surveys (ps range: 0.57-0.99). A separate sample of 17 off-cycle students who took the one-year post- workshop survey two years after the workshop did not differ statistically on the level of awareness of bias compared to those taking the same survey one year later, as examined by the two-group independent t-tests for the seven one-year post-workshop survey items (ps range: 0.56-0.99). CONCLUSIONS: The findings support one-year retention of knowledge and attitudes gained from an implicit bias workshop and suggest similar retention at two years. Future educational interventions that train learners to recognize and manage implicit and explicit behaviors in clinical practice are needed.

Latency reversal plus natural killer cells diminish HIV reservoir in vivo.

HIV is difficult to eradicate due to the persistence of a long-lived reservoir of latently infected cells. Previous studies have shown that natural killer cells are important to inhibiting HIV infection, but it is unclear whether the administration of natural killer cells can reduce rebound viremia when anti-retroviral therapy is discontinued. Here we show the administration of allogeneic human peripheral blood natural killer cells delays viral rebound following interruption of anti-retroviral therapy in humanized mice infected with HIV-1. Utilizing genetically barcoded virus technology, we show these natural killer cells efficiently reduced viral clones rebounding from latency. Moreover, a kick and kill strategy comprised of the protein kinase C modulator and latency reversing agent SUW133 and allogeneic human peripheral blood natural killer cells during anti-retroviral therapy eliminated the viral reservoir in a subset of mice. Therefore, combinations utilizing latency reversal agents with targeted cellular killing agents may be an effective approach to eradicating the viral reservoir.

A User-Friendly Computational Framework for Robust Structured Regression with the L2 Criterion.

We introduce a user-friendly computational framework for implementing robust versions of a wide variety of structured regression methods with the L2 criterion. In addition to introducing an algorithm for performing L2E regression, our framework enables robust regression with the L2 criterion for additional structural constraints, works without requiring complex tuning procedures on the precision parameter, can be used to identify heterogeneous subpopulations, and can incorporate readily available non-robust structured regression solvers. We provide convergence guarantees for the framework and demonstrate its flexibility with some examples. Supplementary materials for this article are available online.

Gender-Based Violence Risk Mitigation by Non-GBV Specialists Prior to and during COVID-19: A Global Survey of Knowledge, Attitudes and Practices of Humanitarian Practitioners.

Available evidence indicates that the COVID-19 pandemic and response measures may lead to increased risk of gender-based violence (GBV), including in humanitarian contexts. This study examined the knowledge, attitudes, and practices of humanitarian practitioners related to GBV risk mitigation approaches during COVID-19 in order to refine current guidance and inform future materials. A global, online cross-sectional survey of humanitarian practitioners was conducted between November 2020 and April 2021. We calculated descriptive statistics and used Chi-square or Fisher's exact tests to compare knowledge, attitudes, and practices among GBV specialists and non-specialists. Of 170 respondents, 58% were female and 44% were GBV specialists. Almost all (95%) of the respondents agreed or strongly agreed that they have a role to play in GBV risk mitigation. Compared to GBV specialists, a higher proportion of non-specialists reported little to no knowledge on GBV risk mitigation global guidance (38% vs. 7%, p < 0.001) and on how to respond to a disclosure of GBV (18% vs. 3%, p < 0.001). Respondents reported several barriers to integrating GBV risk mitigation into their work during COVID-19, including insufficient funding, capacity, knowledge, and guidance. Efforts to mainstream GBV risk mitigation actions should continue and intensify, leveraging the lessons and experiences generated thus far.

HLA-A∗02:01 restricted T cell receptors against the highly conserved SARS-CoV-2 polymerase cross-react with human coronaviruses.

Cross-reactivity and direct killing of target cells remain underexplored for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-specific CD8+ T cells. Isolation of T cell receptors (TCRs) and overexpression in allogeneic cells allows for extensive T cell reactivity profiling. We identify SARS-CoV-2 RNA-dependent RNA polymerase (RdRp/NSP12) as highly conserved, likely due to its critical role in the virus life cycle. We perform single-cell TCRαß sequencing in human leukocyte antigen (HLA)-A∗02:01-restricted, RdRp-specific T cells from SARS-CoV-2-unexposed individuals. Human T cells expressing these TCRαß constructs kill target cell lines engineered to express full-length RdRp. Three TCR constructs recognize homologous epitopes from common cold coronaviruses, indicating CD8+ T cells can recognize evolutionarily diverse coronaviruses. Analysis of individual TCR clones may help define vaccine epitopes that can induce long-term immunity against SARS-CoV-2 and other coronaviruses.