Tissue-resident memory and circulating T cells are early responders to pre-surgical cancer immunotherapy.

Neoadjuvant immune checkpoint blockade has shown promising clinical activity. Here, we characterized early kinetics in tumor-infiltrating and circulating immune cells in oral cancer patients treated with neoadjuvant anti-PD-1 or anti-PD-1/CTLA-4 in a clinical trial (NCT02919683). Tumor-infiltrating CD8 T cells that clonally expanded during immunotherapy expressed elevated tissue-resident memory and cytotoxicity programs, which were already active prior to therapy, supporting the capacity for rapid response. Systematic target discovery revealed that treatment-expanded tumor T cell clones in responding patients recognized several self-antigens, including the cancer-specific antigen MAGEA1. Treatment also induced a systemic immune response characterized by expansion of activated T cells enriched for tumor-infiltrating T cell clonotypes, including both pre-existing and emergent clonotypes undetectable prior to therapy. The frequency of activated blood CD8 T cells, notably pre-treatment PD-1-positive KLRG1-negative T cells, was strongly associated with intra-tumoral pathological response. These results demonstrate how neoadjuvant checkpoint blockade induces local and systemic tumor immunity.

Transcriptional Atlas of Intestinal Immune Cells Reveals that Neuropeptide α-CGRP Modulates Group 2 Innate Lymphoid Cell Responses.

Signaling abnormalities in immune responses in the small intestine can trigger chronic type 2 inflammation involving interaction of multiple immune cell types. To systematically characterize this response, we analyzed 58,067 immune cells from the mouse small intestine by single-cell RNA sequencing (scRNA-seq) at steady state and after induction of a type 2 inflammatory reaction to ovalbumin (OVA). Computational analysis revealed broad shifts in both cell-type composition and cell programs in response to the inflammation, especially in group 2 innate lymphoid cells (ILC2s). Inflammation induced the expression of exon 5 of Calca, which encodes the alpha-calcitonin gene-related peptide (α-CGRP), in intestinal KLRG1+ ILC2s. α-CGRP antagonized KLRG1+ ILC2s proliferation but promoted IL-5 expression. Genetic perturbation of α-CGRP increased the proportion of intestinal KLRG1+ ILC2s. Our work highlights a model where α-CGRP-mediated neuronal signaling is critical for suppressing ILC2 expansion and maintaining homeostasis of the type 2 immune machinery.

Skin-resident innate lymphoid cells converge on a pathogenic effector state.

Tissue-resident innate lymphoid cells (ILCs) help sustain barrier function and respond to local signals. ILCs are traditionally classified as ILC1, ILC2 or ILC3 on the basis of their expression of specific transcription factors and cytokines1. In the skin, disease-specific production of ILC3-associated cytokines interleukin (IL)-17 and IL-22 in response to IL-23 signalling contributes to dermal inflammation in psoriasis. However, it is not known whether this response is initiated by pre-committed ILCs or by cell-state transitions. Here we show that the induction of psoriasis in mice by IL-23 or imiquimod reconfigures a spectrum of skin ILCs, which converge on a pathogenic ILC3-like state. Tissue-resident ILCs were necessary and sufficient, in the absence of circulatory ILCs, to drive pathology. Single-cell RNA-sequencing (scRNA-seq) profiles of skin ILCs along a time course of psoriatic inflammation formed a dense transcriptional continuum-even at steady state-reflecting fluid ILC states, including a naive or quiescent-like state and an ILC2 effector state. Upon disease induction, the continuum shifted rapidly to span a mixed, ILC3-like subset also expressing cytokines characteristic of ILC2s, which we inferred as arising through multiple trajectories. We confirmed the transition potential of quiescent-like and ILC2 states using in vitro experiments, single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq) and in vivo fate mapping. Our results highlight the range and flexibility of skin ILC responses, suggesting that immune activities primed in healthy tissues dynamically adapt to provocations and, left unchecked, drive pathological remodelling.

Integration of ζ-deficient CARs into the CD3ζ gene conveys potent cytotoxicity in T and NK cells.

ABSTRACT: Chimeric antigen receptor (CAR)-redirected immune cells hold significant therapeutic potential for oncology, autoimmune diseases, transplant medicine, and infections. All approved CAR-T therapies rely on personalized manufacturing using undirected viral gene transfer, which results in nonphysiological regulation of CAR-signaling and limits their accessibility due to logistical challenges, high costs and biosafety requirements. Random gene transfer modalities pose a risk of malignant transformation by insertional mutagenesis. Here, we propose a novel approach utilizing CRISPR-Cas gene editing to redirect T cells and natural killer (NK) cells with CARs. By transferring shorter, truncated CAR-transgenes lacking a main activation domain into the human CD3ζ (CD247) gene, functional CAR fusion-genes are generated that exploit the endogenous CD3ζ gene as the CAR's activation domain. Repurposing this T/NK-cell lineage gene facilitated physiological regulation of CAR expression and redirection of various immune cell types, including conventional T cells, TCRγ/δ T cells, regulatory T cells, and NK cells. In T cells, CD3ζ in-frame fusion eliminated TCR surface expression, reducing the risk of graft-versus-host disease in allogeneic off-the-shelf settings. CD3ζ-CD19-CAR-T cells exhibited comparable leukemia control to TCRα chain constant (TRAC)-replaced and lentivirus-transduced CAR-T cells in vivo. Tuning of CD3ζ-CAR-expression levels significantly improved the in vivo efficacy. Notably, CD3ζ gene editing enabled redirection of NK cells without impairing their canonical functions. Thus, CD3ζ gene editing is a promising platform for the development of allogeneic off-the-shelf cell therapies using redirected killer lymphocytes.

Bromhidrosis treatment modalities: A literature review.

BACKGROUND: Treatment options for Bromhidrosis include botulinum toxin therapy, microwave-based therapy, laser therapy, and surgical intervention. Limited studies compare their efficacies. OBJECTIVE: The purpose of this literature review is to compare the efficacy and safety of these treatments for bromhidrosis. METHODS: A PubMed search included terms bromhidrosis and bromhidrosis AND treatment. RESULTS: A total of 25 articles were reviewed. Botulinum toxin therapy shows consistent benefit but requires repeated therapies. Microwave therapies have shown promising results but require larger cohort sizes with bromhidrosis. Similarly, laser therapy has shown promise with biopsy-proven results, but long-lasting effects remain unknown. Surgery has the best long-term prognosis, but the ideal surgical method remains unknown. LIMITATIONS: Each study varied in their treatment interval and method of assessing bromhidrosis, making direct comparisons difficult. CONCLUSIONS: Managing bromhidrosis requires shared decision making with the patient. Mild-to-moderate symptoms may be treated initially with botulinum toxin therapy. In cases that are refractory, laser therapy should be considered, as it is better studied than microwave therapy currently. Lastly, if the condition is severe and refractory to other options, surgery can be considered, although the ideal method remains unknown.

Abametapir for the Treatment of Head Lice: A Drug Review.

OBJECTIVE: This article reviews the pharmacology, safety, efficacy, and clinical importance of abametapir 0.74% (Xeglyze) for the treatment of head lice. DATA SOURCES: From 2020 to May 2021, a systematic review of the MEDLINE and EMBASE databases was conducted using the terms abametapir, Xeglyze, Ha44, and head lice. Bibliographies, Food and Drug Administration (FDA) drug package inserts, and ClinicalTrials.gov were searched for further information. STUDY SELECTION AND DATA EXTRACTION: All relevant full-text articles in English were considered for inclusion, with a final article date range of 1999 to 2020. DATA SYNTHESIS: Abametapir chelates heavy metal cations and inhibits metalloproteinases critical to louse ova development, hatching, and adult survival. In phase II, abametapir had direct ovicidal activity inhibiting 100% of treated louse eggs from hatching, compared with 64% in the vehicle-treated group. In two identical phase III clinical trials, subjects treated with a single 10-minute application of abametapir had greater treatment success compared with vehicle-treated subjects, with 81.1% success versus 50.9% in study 1 (P = 0.001) and 81.8% versus 47.2% in study 2 (P < 0.001). Abametapir was well tolerated, with only mild adverse effects. RELEVANCE TO PATIENT CARE AND CLINICAL PRACTICE: Abametapir is a newly FDA-approved, single-application treatment for head lice in patients aged 6 months and older. This review highlights the safety and efficacy of abametapir in the treatment of head lice. CONCLUSIONS: In the wake of increasing widespread resistance to first-line treatment options, abametapir offers a safe and effective new treatment option for head lice infestations.

Review of the holistic management of pediatric atopic dermatitis.

Atopic Dermatitis (AD) is a chronic inflammatory skin disease that is broadly characterized by eczematous lesions and pruritus. This condition is detrimental in a multitude of ways, including patient quality of life (QOL), family QOL, economic burden, and psychosocial afflictions. Current management needs to incorporate a holistic approach which considers the financial, emotional, and physical limitations of both the treatments and the provider. A non-systematic search was conducted on the holistic management of pediatric AD. Various search queries were used such as the key terms of "atopic dermatitis," "pediatric," "eczema," "management," and more to encompass treatments, adherence, and comorbidities. There is an association with AD and depression in children, and its prevalence should be screened for routinely in children with AD. Collaboration with other specialties may prove to be prudent in addressing this comorbidity. Objective quality of life scores can open the door to much needed conversation with patients to get them the help they need. In expanding our scope, we find the extended consequences of AD have a ripple effect on families of pediatric patients. Lastly, we introduce a model for improving treatment adherence. CONCLUSION: Patient quality-of-life can be negatively affected by the symptoms, expense, stigma, and time commitment, and inconvenience imposed by complicated treatment regimens. To ensure proper, holistic management of pediatric AD, multiple factors must be considered; seasonal changes, lifestyle modifications, and the psychosocial impact are just a couple of factors that require monitoring. WHAT IS KNOWN: • Atopic dermatitis impacts patients and their families in quality of life, economically, and psychosocially. • Current treatment revolves largely around treating physical manifestation of disease with first line measures such as topical steroids. WHAT IS NEW: • The holistic management of AD incorporates a good physician-patient relationship, frequent follow-up, and providing structured written plans. • We introduce the house building model for improving treatment adherence. KEY POINTS: Pediatric AD can be managed in a more holistic manner which incorporates several factors from the lives of patients and their families. Pediatric patients suffer from many physical and mental comorbidities which should be screened for. Adherence with treatment may be improved by following a model which emphasizes establishing a good physician-patient relationship, frequent follow-up, and providing structured written plans.

A Case of Late-Onset Alopecia Areata.

Alopecia areata (AA) is a condition characterized by nonscarring hair loss. Cases of alopecia areata are most commonly seen in patients under age 30 and are frequently idiopathic. In this report, we discuss a woman in her 50s who developed AA shortly after receiving the Tdap vaccine and after one year of guselkumab therapy.

Unmet Needs in Psoriasis Patients.

BACKGROUND: Psoriasis can greatly impact patients' quality of life. The introduction of new treatments has improved treatment outcomes, but treatment gaps may still exist. OBJECTIVE: Identify unmet treatment needs in patients with psoriasis. METHODS: Participants aged 18 years or older, with an Amazon Mechanical Turk account, who reported diagnosis of psoriasis and correctly answered an attention check question were included. Results were analyzed using descriptive and inferential statistics. RESULTS: Of the 417 participants who met study inclusion criteria, 51.1% were female, and 39.3% were 31-40 years of age; 61.2% reported mild, 25.4% moderate, and 13.4% severe psoriasis. Most (74.8%) were currently under treatment; half (51.6%) were mostly or completely satisfied with treatment, while 24.5% were slightly or not at all satisfied. Respondents were most satisfied with topical (59.5%), followed by oral (46%), and injectable treatments (19.9%). Most (78.7%) slightly or strongly felt there should be more cost-effective options. Gaps in current psoriasis treatment options included more affordable topical and oral treatments that work faster and require less frequent use. LIMITATIONS: The use of an English survey and Amazon Mechanical Turk precludes certain populations from this study. Participants were not asked to provide their current form of psoriasis treatment. CONCLUSIONS: Despite advances in psoriasis treatment, there remains a desire for more effective, faster, longer acting, and less costly, more accessible treatments. J Drugs Dermatol. 2022;21(8):839-844. doi:10.36849/JDD.6589.

Understanding eligibility creep in psoriasis assessments: a survey study.

Psoriasis severity assessments for clinical trial entry may be unintentionally overestimated, especially if trial eligibility is chiefly dependent on rating of disease severity. When this results in patients with less severe phenotypes joining clinical trials it is referred to as eligibility creep. We investigated the potential impact of psychosocial incentives on psoriasis lesion severity grading. A survey was constructed and disseminated through Amazon Mechanical Turk. Participants completed two vignette-style questions prompted with a randomly allocated psychosocial incentive. Questions required participants to grade and select psoriasis lesion pictures for a fictional trial. Participants also decided whether or not to schedule re-evaluation of patients deemed ineligible at initial visit. There were 646 participants. There was no significant difference in number of total lesions selected for study inclusion between incentive groups (Kruskal-Wallis, P=0.30). In general, participants completing empathy and professional uncertainty incentives selected the most and least number of lesion pictures for trial inclusion, respectively. Participants prompted with empathy incentives had significantly greater rates of choosing to schedule a follow-up visit for ineligible patients compared to participants prompted with other incentives (69.7% versus 59.1%, Chi square P=0.046). Situations evoking empathy may contribute to eligibility creep.

New onset inflammatory bowel disease in patient treated with secukinumab: Case report and review of literature.

Psoriasis is a chronic autoimmune skin disorder that can vary in severity and extent of disease. While localized disease can be managed with topical medications, widespread disease often requires systemic therapy including biologics. This medication class targets different components of the immune system and thus modulates disease activity. The biologic secukinumab is a human monoclonal antibody against interleukin-17A used for the treatment of psoriasis and psoriatic arthritis; cases of inflammatory bowel disease (IBD) have been observed in clinical trials to be associated with this medication. This review aims to provide evidence for the relationships between secukinumab treatment and the development of IBD. We have examined review articles and original research papers, published between 2010 and 2020, using the following keywords: psoriasis, psoriatic arthritis, secukinumab, IBD, Crohn's disease, ulcerative colitis, interleukin-17, IL-17, IL-17 inhibitor. Case reports have noted an association between secukinumab use and IBD and have called for IBD pre-screening in patients who will be prescribed this medication. Clinical trials concluded that secukinumab was associated with IBD, while retrospective studies have had mixed results, with most studies showing no statistical significance between secukinumab and IBD but having seen patients with history of IBD or family histories experience new-onset IBD or flare-ups. Given the utilization of secukinumab as a therapy for psoriasis and psoriatic arthritis, appropriate screening and risk stratification could help limit morbidity and mortality that can be associated with secukinumab-induced IBD.

Oncolytic Adenovirus Armed with BiTE, Cytokine, and Checkpoint Inhibitor Enables CAR T Cells to Control the Growth of Heterogeneous Tumors.

No single cancer immunotherapy will likely defeat all evasion mechanisms of solid tumors, including plasticity of tumor antigen expression and active immune suppression by the tumor environment. In this study, we increase the breadth, potency, and duration of anti-tumor activity of chimeric antigen receptor (CAR) T cells using an oncolytic virus (OV) that produces cytokine, checkpoint blockade, and a bispecific tumor-targeted T cell engager (BiTE) molecule. First, we constructed a BiTE molecule specific for CD44 variant 6 (CD44v6), since CD44v6 is widely expressed on tumor but not normal tissue, and a CD44v6 antibody has been safely administered to cancer patients. We then incorporated this BiTE sequence into an oncolytic-helper binary adenovirus (CAdDuo) encoding an immunostimulatory cytokine (interleukin [IL]-12) and an immune checkpoint blocker (PD-L1Ab) to form CAdTrio. CD44v6 BiTE from CAdTrio enabled HER2-specific CAR T cells to kill multiple CD44v6+ cancer cell lines and to produce more rapid and sustained disease control of orthotopic HER2+ and HER2-/- CD44v6+ tumors than any component alone. Thus, the combination of CAdTrio with HER2.CAR T cells ensures dual targeting of two tumor antigens by engagement of distinct classes of receptor (CAR and native T cell receptor [TCR]), and significantly improves tumor control and survival.

A Distinct Transcriptional Program in Human CAR T Cells Bearing the 4-1BB Signaling Domain Revealed by scRNA-Seq.

T cells engineered to express chimeric antigen receptors (CARs) targeting CD19 have produced impressive outcomes for the treatment of B cell malignancies, but different products vary in kinetics, persistence, and toxicity profiles based on the co-stimulatory domains included in the CAR. In this study, we performed transcriptional profiling of bulk CAR T cell populations and single cells to characterize the transcriptional states of human T cells transduced with CD3ζ, 4-1BB-CD3ζ (BBζ), or CD28-CD3ζ (28ζ) co-stimulatory domains at rest and after activation by triggering their CAR or their endogenous T cell receptor (TCR). We identified a transcriptional signature common across CARs with the CD3ζ signaling domain, as well as a distinct program associated with the 4-1BB co-stimulatory domain at rest and after activation. CAR T cells bearing BBζ had increased expression of human leukocyte antigen (HLA) class II genes, ENPP2, and interleukin (IL)-21 axis genes, and decreased PD1 compared to 28ζ CAR T cells. Similar to previous studies, we also found BBζ CAR CD8 T cells to be enriched in a central memory cell phenotype and fatty acid metabolism genes. Our data uncovered transcriptional signatures related to costimulatory domains and demonstrated that signaling domains included in CARs uniquely shape the transcriptional programs of T cells.

How should medical students prepare for a clinical dermatology rotation?

Skin diseases are commonly encountered in medical practice, yet medical students often receive little dermatology training. There is little research on what self-study materials best prepare students. We aim to identify which resources dermatology residents have found to be most useful in preparing for clinical dermatology rotations and dermatology residency. Forty current dermatology residents and fellows responded to our REDCap-generated survey. Data was analyzed using descriptive statistics. Most respondents (N=36, 90%) reported using outside resources to prepare for clinical dermatology rotations and dermatology residency. American Academy of Dermatology (AAD) modules and other online resources were most used (N=31, 77.5%) and most recommended (N=32, 80%). However, 67.5% of all respondents also used printed textbooks in some capacity, but low-to-no cost, usefulness, and easy accessibility of online resources made them more favorable among study participants. Multiple clinical dermatology rotations were recommended for preparing for dermatology residency (N= 34, 85%), as were other rotations, including internal medicine (N=22, 55%) and rheumatology (N=17, 42.5%). Overall, the AAD modules and online resources are most useful when preparing for clinical dermatology rotations because of favorable cost and accessibility. Compared to clinical rotations in other specialties, multiple rotations in dermatology may be most helpful for dermatology residency.

The effect of physician appearance on patient perceptions of treatment plans.

The presentation of a physician's clothing and themselves is an important part of the physician-patient interaction. Physician attire can impact patient satisfaction and trust in their physician. We sought to discover the influence physician attire may have on patients' comfort level with proposed treatment plans, which could influence treatment adherence. We surveyed 495 subjects to better understand the relationship between physician attire and patients' perceptions of treatment plans. We found subjects' comfort level in proposed treatment plans was not significantly affected by physician attire. This finding suggests physician attire may not be as important when considering the potential effect on treatment adherence.

The effect of the COVID-19 pandemic on perceptions of teledermatology.

There is limited information of the effect of the COVID-19 pandemic on the general population's perceptions towards teledermatology. This study aims to assess the pandemic's impact on people's willingness to use teledermatology as well as to investigate influencing factors. We recruited 544 participants through Amazon Mechanical Turk (MTurk) and surveyed them using REDCap. Participants' willingness to use teledermatology before, during, and after the COVID-19 pandemic was measured via a 5-point Likert scale. The survey also included questions regarding factors influencing participants' attitudes towards teledermatology and their sociodemographic characteristics. Of the 185 participants who reported unwillingness to use teledermatology prior to the COVID-19 pandemic, 79.2% and 66.5% became either neutral or willing to use teledermatology during and after the pandemic, respectively. Less than half of prior satisfactory telemedicine users reported willingness to use teledermatology before the pandemic; willingness to use teledermatology increased to 80.1% and 63.8% during and after the COVID-19 pandemic, respectively. The top reason for lack of interest in teledermatology was concern for security and privacy (24.4%). Although a useful tool, teledermatology has been met with reluctance by the public. However, the unique circumstances of the COVID-19 pandemic have improved the public's perceptions and readiness to use teledermatology.

Development of the NEC-Zero Toolkit: Supporting Reliable Implementation of Necrotizing Enterocolitis Prevention and Recognition.

The goal of the NEC-Zero project is to reduce the burden of necrotizing enterocolitis (NEC) by increasing access to evidence-based tools to help clinicians and parents integrate evidence into daily care. It involves (a) human milk feeding with prioritized mother's own milk; (b) use of a unit-adopted standardized feeding protocol; (c) a unit-adopted strategy for timely recognition that integrates risk awareness and a structured communication tool when symptoms develop; and (d) stewardship of empiric antibiotics and avoidance of antacids. A toolkit for caregivers and parents was developed to make implementation consistent. For clinicians the toolkit includes: the GutCheckNEC risk score, a structured communication tool, the "Avoiding NEC" checklist, and the NEC-Zero website. For parents, NEC-Zero tools include the website, three educational brochures in English and Spanish, and a collaborative care video produced with the NEC Society. This article describes the toolkit and how it has been accessed and used.

Adenovirotherapy Delivering Cytokine and Checkpoint Inhibitor Augments CAR T Cells against Metastatic Head and Neck Cancer.

In solid tumors, chimeric antigen receptor (CAR)-modified T cells must overcome the challenges of the immunosuppressive tumor microenvironment. We hypothesized that pre-treating tumors with our binary oncolytic adenovirus (CAd), which produces local oncolysis and expresses immunostimulatory molecules, would enhance the antitumor activity of HER2-specific CAR T cells, which alone are insufficient to cure solid tumors. We tested multiple cytokines in conjunction with PD-L1-blocking antibody and found that Ad-derived IL-12p70 prevents the loss of HER2.CAR-expressing T cells at the tumor site. Accordingly, we created a construct encoding the PD-L1-blocking antibody and IL-12p70 (CAd12_PDL1). In head and neck squamous cell carcinoma (HNSCC) xenograft models, combining local treatment with CAd12_PDL1 and systemic HER2.CAR T cell infusion improved survival to >100 days compared with approximately 25 days with either approach alone. This combination also controlled both primary and metastasized tumors in an orthotopic model of HNSCC. Overall, our data show that CAd12_PDL1 augments the anti-tumor effects of HER2.CAR T cells, thus controlling the growth of both primary and metastasized tumors.

Antibiotic efficacy is linked to bacterial cellular respiration.

Bacteriostatic and bactericidal antibiotic treatments result in two fundamentally different phenotypic outcomes--the inhibition of bacterial growth or, alternatively, cell death. Most antibiotics inhibit processes that are major consumers of cellular energy output, suggesting that antibiotic treatment may have important downstream consequences on bacterial metabolism. We hypothesized that the specific metabolic effects of bacteriostatic and bactericidal antibiotics contribute to their overall efficacy. We leveraged the opposing phenotypes of bacteriostatic and bactericidal drugs in combination to investigate their activity. Growth inhibition from bacteriostatic antibiotics was associated with suppressed cellular respiration whereas cell death from most bactericidal antibiotics was associated with accelerated respiration. In combination, suppression of cellular respiration by the bacteriostatic antibiotic was the dominant effect, blocking bactericidal killing. Global metabolic profiling of bacteriostatic antibiotic treatment revealed that accumulation of metabolites involved in specific drug target activity was linked to the buildup of energy metabolites that feed the electron transport chain. Inhibition of cellular respiration by knockout of the cytochrome oxidases was sufficient to attenuate bactericidal lethality whereas acceleration of basal respiration by genetically uncoupling ATP synthesis from electron transport resulted in potentiation of the killing effect of bactericidal antibiotics. This work identifies a link between antibiotic-induced cellular respiration and bactericidal lethality and demonstrates that bactericidal activity can be arrested by attenuated respiration and potentiated by accelerated respiration. Our data collectively show that antibiotics perturb the metabolic state of bacteria and that the metabolic state of bacteria impacts antibiotic efficacy.