BackgroundTo establish a predictive model that combines radiomics, deep learning and clinical features for predicting the pathological complete response (pCR) of non-small cell lung cancer (NSCLC) patients after neoadjuvant immunochemotherapy (NIT).MethodsWe retrospectively collected patients from three centers (split into training, internal testing and external testing cohorts). In this study, tumor segmentation was performed on chest CT images before (pre-NIT) and after (post-NIT) neoadjuvant therapy. The radiomics features were extracted from pre-NIT and post-NIT images. Deep learning (DL) features were extracted from the post-NIT images. The most meaningful features were selected using the mRMR and LASSO. A logistic regression classifier was then applied to create a classification model to predict pCR or non-pCR. The predicted probabilities were referred to as the Rad-scores and Deep-scores. Finally, Rad-scores, Deep-scores, and meaningful clinical features were fused to build a combined model.ResultsA total of 178 patients were enrolled in the current study. In conventional radiomics, the efficacy of post-NIT model was better than the pre-NIT. In delta radiomics model, delta1 had the best efficacy. Subsequently, the post-NIT and delta1 features were further constructed as the combined model 1 with AUCs of 0.939 and 0.849, respectively. iRECIST was combined with the radiomics and the DL features to establish the combined model 2, which achieved the best performance among all the models, with AUCs of 0.955(training), 0.882(In-testing), and 0.839(Ex-testing).ConclusionsOur results demonstrated that combination of three dimensional features can provide complementary information to predict pCR more accurately.
Abstract Background Timely molecular surveillance of Plasmodium falciparum kelch 13 (k13) gene mutations is essential for monitoring the emergence and stemming the spread of artemisinin resistance. Widespread artemisinin resistance, as observed in Southeast Asia, would reverse significant gains that have been made against the malaria burden in Africa. The purpose of this study was to assess the prevalence of k13 polymorphisms in western Kenya and Ethiopia at sites representing varying transmission intensities between 2018 and 2022. Methods Dried blood spot samples collected through ongoing passive surveillance and malaria epidemiological studies, respectively, were investigated. The k13 gene was genotyped in P. falciparum isolates with high parasitaemia: 775 isolates from four sites in western Kenya (Homa Bay, Kakamega, Kisii, and Kombewa) and 319 isolates from five sites across Ethiopia (Arjo, Awash, Gambella, Dire Dawa, and Semera). DNA sequence variation and neutrality were analysed within each study site where mutant alleles were detected. Results Sixteen Kelch13 haplotypes were detected in this study. Prevalence of nonsynonymous k13 mutations was low in both western Kenya (25/783, 3.19%) and Ethiopia (5/319, 1.57%) across the study period. Two WHO-validated mutations were detected: A675V in three isolates from Kenya and R622I in four isolates from Ethiopia. Seventeen samples from Kenya carried synonymous mutations (2.17%). No synonymous mutations were detected in Ethiopia. Genetic variation analyses and tests of neutrality further suggest an excess of low frequency polymorphisms in each study site. Fu and Li’s F test statistic in Semera was 0.48 (P > 0.05), suggesting potential population selection of R622I, which appeared at a relatively high frequency (3/22, 13.04%). Conclusions This study presents an updated report on the low frequency of k13 mutations in western Kenya and Ethiopia. The WHO-validated R622I mutation, which has previously only been reported along the north-west border of Ethiopia, appeared in four isolates collected from eastern Ethiopia. The rapid expansion of R622I across Ethiopia signals the need for enhanced monitoring of the spread of drug-resistant P. falciparum parasites in East Africa. Although ACT remains currently efficacious in the study areas, continued surveillance is necessary to detect early indicators of artemisinin partial resistance.
Austin T. Hertel, Cynthia M. McMillen, Ryan M. Hoehl
et al.
Abstract Rift Valley fever virus (RVFV) causes high rates of spontaneous abortions and neonatal mortality in ruminants resulting in severe socioeconomic and public health consequences. Maternal vaccination may protect pregnant animals, fetuses, and neonates via transfer of maternal antibodies; however, currently available live-attenuated RVFV vaccines are generally unsafe for use during pregnancy. RVFV-delNSs/NSm is a live attenuated strain that has demonstrated favorable safety and efficacy in pregnant livestock, yet studies investigating maternal vaccination as a strategy to protect neonates from RVF are limited. Using pregnant Sprague-Dawley rats, we show that maternal vaccination with RVFV-delNSs/NSm leads to efficient transfer of anti-RVFV antibodies to offspring. These offspring were completely protected from lethal RVFV challenge. Although further investigation is required in susceptible ruminant species, our findings indicate that maternal anti-RVFV immunity is sufficient to protect offspring, highlighting maternal vaccination as a potential strategy to reduce RVF disease burden in endemic regions.
ABSTRACT Elimination of tumors is typically dependent on T cells, which require prior or ongoing activation signals. These dependencies form the basis for our understanding of “tumor‐reactive immunity” and for the successes of immunotherapies, particularly immune checkpoint blockades. Over the years, tremendous work has been done towards understanding the biology of this reactive immunity and early investigation identified dendritic cells (DC) as key contributors. Recent advances have shed more light on dendritic cell heterogeneity in tumors, revealing specialized roles for each subtype. In addition, the network of cellular interactions surrounding DCs has grown as additional cell types have been revealed to variously influence how the immune system can become most effective at eliminating malignancies. Greater understanding of intratumoral DC biology has empowered investigators to engineer dendritic cell vaccines and consider other approaches to augment this component of reactive immunity, towards the generation of anti‐tumor immune responses de novo . In this review, we will discuss the state of the field, recent advances and suggest what the near future of scientific inquiry could entail.
Xiaoqian Song, Xiaoqian Song, Jingjiao Chen
et al.
ObjectiveThis study aims to delineate the clinical features underlying the concurrent disease of neuromyelitis optica spectrum disorder (NMOSD) and myasthenia gravis (MG), and to identify efficacious therapeutic strategies.BackgroundNMOSD and MG are uncommon autoimmune diseases that infrequently co-exist. Despite previous reports, a consensus on treating NMOSD concurrent with MG is lacking.MethodsWe present the case of a 55-year-old female with both anti-aquaporin-4 (AQP4) antibody-positive NMOSD and anti-acetylcholine receptor (AChR) antibody-positive MG, who achieved stable disease control following treatment with inebilizumab without significant adverse effects. We also conducted a literature review to evaluate the clinical profile of this comorbidity.ResultsOur review identified 85 patients with concurrent NMOSD and MG. In 70 well-documented cases, MG predated NMOSD in 60 (85.8%) cases, with 42 (70%) patients having undergone thymectomy. Six (8.6%) patients were first diagnosed with NMOSD, and then thymectomy was performed in 2 (33.3%) MG patients. For NMOSD treatment, although most patients received steroid hormones and immunosuppressive agents, quite a few patients had persistent severe disability. Additionally, of 44 patients with clear records of disease recurrence, 31 patients(70.5%) experienced frequent relapses of optic neuritis and myelitis, ranging from 1 to 15 attacks, averaging five. The manifestations of MG are mainly included fatigability, diplopia, and blepharoptosis, with symptoms well-controlled in most patients. Our patient treated with inebilizumab for 1 year and no relapse was recorded to date.ConclusionsThough MG typically precedes NMOSD and thymectomy is frequently performed, it is not a prerequisite for NMOSD development but may represent a potential risk factor. MG generally follows a benign course, in contrast to the more aggressive nature of NMOSD. The utility of biological agents such as inebilizumab for patients with both NMOSD combined with MG warrants further attention.
A. A. Savchenko, D. E. Zdzitovetskiy, M. M. Adilov
et al.
The aim of our research was to study the features of activation receptor expression on various subsets of blood monocytes in patients with acute pancreatitis (AP). 69 patients aged 37-62 years with moderateand severe-grade AP were examined. The diagnosis of AP was based on the results of clinical, laboratory and instrumental examination. Phenotype and subpopulation composition of monocytes were studied by flow cytometry. Alterations in blood monocytes phenotypes and increased expression of activation receptors were noted in patients during the initial period of AP. Thus, an increased proportion of monocytes in the blood of patients with AP with co-expression of CD45RO and CD62L was detected, along with increased number of cells expressing CD25 receptor. An increased level of migratory monocyte activity in AP could be linked with CXCR4 and CCR5 receptors. Altered subset composition during the acute period of AP was linked with 2-fold increased levels of “non-classical” monocytes. The proportion of cells with expression of chemokine receptors in the subset composition of monocytes changed in AP. Thus, the number of “classical” and “nonclassical” monocytes with CXCR4 was increased within total monocyte subset in the patients. Meanwhile, the content of cell subsets with CCR5 receptor expression was almost uniformly increased. The changed expression levels of activation receptors also characterized the activation features of various monocyte subsets in patients during the initial period of AP. Elevated CCR5 was detected in AP only on “classical” monocytes, whereas increased CD64 was found only on “non-classical” monocytes. Elevated HLA-DR expression was detected on “classical” and “intermediate” monocytes of patients with AP but a high level of CXCR4 expression was found on all monocytes subsets. The registered changes in phenotype and subset composition of monocytes in patients during the initial period of the disease seem to characterize the mode of monocyte involvement into the inflammatory process in AP thus revealing not only pro-inflammatory reaction of monocytes, along with increased activity of monocyte subset with anti-inflammatory function.
Background and objectiveVenous thrombus fibrosis contributes to post-thrombotic syndrome (PTS) and chronic thromboembolic pulmonary hypertension (CTEPH). M2 macrophages promote fibrosis via TGF-β1 secretion. This study investigates whether sphingosine kinase 1 (SPHK1) promotes thrombus fibrosis by regulating M2 macrophage polarization.MethodsHistological staining and immunofluorescence (IF) were performed on thrombus tissues from patients with acute thrombosis and CTEPH. Single-cell RNA sequencing (scRNA-seq) was used to characterize immune cell heterogeneity and to identify SPHK1 expression within macrophage subsets. In vivo, a rat model of thrombus was established via inferior vena cava (IVC) ligation, and the SPHK1 inhibitor PF543 was administered to evaluate its effects on fibrosis and macrophage polarization. In vitro, bone marrow-derived macrophages (BMDMs) were subjected to M2 polarization and co-cultured with fibroblasts to assess the TGF-β1-dependent fibroblast activation.ResultsHistological analysis revealed significantly increased ECM deposition and macrophage infiltration in CTEPH thrombi compared to acute thrombi. Masson staining demonstrated extensive collagen fiber accumulation in CTEPH samples. Immunofluorescence analysis of fibrotic thrombi from a rat inferior vena cava (IVC) ligation model showed strong co-expression of SPHK1 and CD68, indicating the presence of SPHK1-expressing macrophages in thrombus remodeling. scRNA-seq analysis further revealed high SPHK1 expression in M2 macrophage subsets, particularly in the MARCO-1 cluster, and its expression was closely correlated with TGF-β1 secretion. In vivo, PF543 treatment significantly reduced collagen deposition, TGF-β1 expression, and M2 macrophage polarization in thrombus tissue. In vitro, SPHK1 knockdown markedly suppressed the expression of TGF-β1, Arg1, CD36, and FASN in BMDMs, indicating an inhibition of pro-fibrotic macrophage function. Co-culture experiments further confirmed that M2 macrophages activated fibroblasts via a TGF-β1-dependent mechanism.ConclusionThis study demonstrates that SPHK1 promotes M2 macrophage polarization and drives TGF-β1-dependent thrombus fibrosis, underscoring its critical role in the progression of CTEPH. Pharmacological inhibition of SPHK1 by PF543 effectively attenuates fibrotic remodeling and suppresses M2 macrophage polarization, suggesting that SPHK1 may serve as a promising therapeutic target for the treatment of chronic thrombus-associated fibrosis.
Megan E Amason, Cole J Beatty, Carissa K Harvest
et al.
Granulomas are defined by the presence of organized layers of immune cells that include macrophages. Granulomas are often characterized as a way for the immune system to contain an infection and prevent its dissemination. We recently established a mouse infection model where Chromobacterium violaceum induces the innate immune system to form granulomas in the liver. This response successfully eradicates the bacteria and returns the liver to homeostasis. Here, we sought to characterize the chemokines involved in directing immune cells to form the distinct layers of a granuloma. We use spatial transcriptomics to investigate the spatial and temporal expression of all CC and CXC chemokines and their receptors within this granuloma response. The expression profiles change dynamically over space and time as the granuloma matures and then resolves. To investigate the importance of monocyte-derived macrophages in this immune response, we studied the role of CCR2 during C. violaceum infection. Ccr2–/– mice had negligible numbers of macrophages, but large numbers of neutrophils, in the C. violaceum-infected lesions. In addition, lesions had abnormal architecture resulting in loss of bacterial containment. Without CCR2, bacteria disseminated and the mice succumbed to the infection. This indicates that macrophages are critical to form a successful innate granuloma in response to C. violaceum.
Maryam Amiri, Amir Kian Moaveni, Masoumeh Majidi Zolbin
et al.
Optimizing cancer treatment has become a pivotal goal in modern oncology, with advancements in immunotherapy and genetic engineering offering promising avenues. CAR-T cell therapy, a revolutionary approach that harnesses the body’s own immune cells to target and destroy cancer cells, has shown remarkable success, particularly in treating acute lymphoblastic leukemia (ALL), and in treating other hematologic malignancies. While CAR-T cell therapy has shown promise, challenges such as high cost and manufacturing complexity remain. However, its efficacy in solid tumors remains limited. The integration of CRISPR/Cas9 technology, a powerful and precise genome-editing tool, also raises safety concerns regarding unintended edits and off-target effects, offers a synergistic potential to overcome these limitations. CRISPR/Cas9 can enhance CAR-T cell therapy by improving the specificity and persistence of CAR-T cells, reducing off-target effects, and engineering resistance to tumor-induced immunosuppression. This combination can also facilitate the knockout of immune checkpoint inhibitors, boosting the anti-tumor activity of CAR-T cells. Recent studies have demonstrated that CRISPR/Cas9-edited CAR-T cells can target previously untreatable cancer types, offering new hope for patients with refractory cancers. This synergistic approach not only enhances the efficacy of cancer treatment but also paves the way for personalized therapies tailored to individual genetic profiles. This review highlights the ongoing research efforts to refine this approach and explores its potential to revolutionize cancer treatment across a broader range of malignancies. As research progresses, the integration of CAR-T cell therapy and CRISPR/Cas9 holds the promise of transforming cancer treatment, making it more effective and accessible. This review explores the current advancements, challenges, and future prospects of this innovative therapeutic strategy.
Abstract Activation of the Ca2+ release–activated Ca2+ (CRAC) channel is crucial for T cell functions. It was recently shown that naked cuticle homolog 2 (NKD2), a signaling adaptor molecule, orchestrates trafficking of ORAI1, a pore subunit of the CRAC channels, to the plasma membrane for sustained activation of the CRAC channels. However, the physiological role of sustained Ca2+ entry via ORAI1 trafficking remains poorly understood. Using NKD2 as a molecular handle, we show that ORAI1 trafficking is crucial for sustained Ca2+ entry and cytokine production, especially in inflammatory Th1 and Th17 cells. We find that murine T cells cultured under pathogenic Th17-polarizing conditions have higher Ca2+ levels that are NKD2-dependent than those under nonpathogenic conditions. In vivo, deletion of Nkd2 alleviated clinical symptoms of experimental autoimmune encephalomyelitis in mice by selectively decreasing effector T cell responses in the CNS. Furthermore, we observed a strong correlation between NKD2 expression and proinflammatory cytokine production in effector T cells. Taken together, our findings suggest that the pathogenic effector T cell response demands sustained Ca2+ entry supported by ORAI1 trafficking.
James Davies, Sofia Sirvent, Andres F. Vallejo
et al.
Human epidermal Langerhans cells (LCs) maintain immune homeostasis in the skin. To examine transcriptional programming of human primary LCs during homeostasis, we performed scRNA-seq analysis of LCs before and after migration from the epidermis, coupled with functional assessment of their regulatory T cell priming capabilities. The analysis revealed that steady-state LCs exist in a continuum of maturation states and upregulate antigen presentation genes along with an immunoregulatory module including the genes IDO1, LGALS1, LAMTOR1, IL4I, upon their migration. The migration-induced transition in genomic state is accompanied by the ability of LCs to more efficiently prime regulatory T cell responses in co-culture assays. Computational analyses of the scRNAseq datasets using SCENIC and Partial Information Decomposition in Context identified a set of migration-induced transcription factors including IRF4, KLF6 and RelB as key nodes within a immunoregulatory gene regulatory network. These findings support a model in which efficient priming of immunoregulatory responses by LCs is dependent on coordinated upregulation of a migration-coupled maturation program with a immunoregulation-promoting genomic module.
The connections between pattern recognition receptors (PRRs) and pathogen-associated molecular patterns (PAMPs) constitutes the crucial signaling pathways in the innate immune system. Cytoplasmic nucleic acid sensor melanoma differentiation-associated gene 5 (MDA5) serves as an important pattern recognition receptor in the innate immune system by recognizing viral RNA. MDA5 also plays a role in identifying the cytoplasmic RNA from damaged, dead cancer cells or autoimmune diseases. MDA5’s recognition of RNA triggers innate immune responses, induces interferon (IFN) response and a series of subsequent signaling pathways to produce immunomodulatory factors and inflammatory cytokines. Here we review the latest progress of MDA5 functions in triggering anti-tumor immunity by sensing cytoplasmic dsRNA, and recognizing SARS-CoV-2 virus infection for antiviral response, in which the virus utilizes multiple ways to evade the host defense mechanism.
Andrew H. Pham, Jennifer Mitchell, Sara Botto
et al.
Human cytomegalovirus (HCMV) microRNAs (miRNAs) significantly rewire host signaling pathways to support the viral lifecycle and regulate host cell responses. Here we show that SMAD3 expression is regulated by HCMV miR-UL22A and contributes to the IRF7-mediated induction of type I IFNs and IFN-stimulated genes (ISGs) in human fibroblasts. Addition of exogenous TGFβ interferes with the replication of a miR-UL22A mutant virus in a SMAD3-dependent manner in wild type fibroblasts, but not in cells lacking IRF7, indicating that downregulation of SMAD3 expression to limit IFN induction is important for efficient lytic replication. These findings uncover a novel interplay between SMAD3 and innate immunity during HCMV infection and highlight the role of viral miRNAs in modulating these responses.