In the pathogenesis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, epithelial populations in the distal lung expressing Angiotensin-converting enzyme 2 (ACE2) are infrequent, and therefore, the model of viral expansion and immune cell engagement remains incompletely understood. Using human lungs to investigate early host-viral pathogenesis, we found that SARS-CoV-2 had a rapid and specific tropism for myeloid populations. Human alveolar macrophages (AMs) reliably expressed ACE2 allowing both spike-ACE2–dependent viral entry and infection. In contrast to Influenza A virus, SARS-CoV-2 infection of AMs was productive, amplifying viral titers. While AMs generated new viruses, the interferon responses to SARS-CoV-2 were muted, hiding the viral dissemination from specific antiviral immune responses. The reliable and veiled viral depot in myeloid cells in the very early phases of SARS-CoV-2 infection of human lungs enables viral expansion in the distal lung and potentially licenses subsequent immune pathologies.

Pre-metastatic niche formation is a critical step during the metastatic spread of cancer. One way by which primary tumors prime host cells at future metastatic sites is through the shedding of tumor-derived micropar-ticles as a consequence of vascular sheer flow. However, it remains unclear how the uptake of such particles by resident immune cells affects their phenotype and function. Here, we show that ingestion of tumor-derived microparticles by macrophages induces a rapid metabolic and phenotypic switch that is characterized by enhanced mitochondrial mass and function, increased oxidative phosphorylation, and upregulation of adhesion molecules, resulting in reduced motility in the early metastatic lung. This reprogramming event is dependent on signaling through the mTORC1, but not the mTORC2, pathway and is induced by uptake of tumor-derived microparticles. Together, these data support a mechanism by which uptake of tumor-derived microparticles induces reprogramming of macrophages to shape their fate and function in the early metasta-tic lung.

Supplementary Data from Visualizing Spatial and Stoichiometric Barriers to Bispecific T-Cell Engager Efficacy

Immune-checkpoint inhibitor (ICI) combined with antiangiogenic therapy have the potential for synergistic activity through modulation of the microenvironment. We report the activity and safety of camrelizumab plus apatinib in patients with recurrent or metastatic nasopharyngeal carcinoma (NPC) who were refractory to at least one line of systemic therapy. This single-arm, phase II study enrolled patients with recurrent or metastatic NPC (nonkeratinizing carcinoma) who were refractory to at least one line of systemic therapy and treatment-naïve to ICI. Patients received camrelizumab 200 mg every 3 weeks and apatinib 250 mg once per day. The primary end point was objective response rate (ORR) assessed by independent radiologists per RECIST version 1.1. Key secondary end points included progression-free survival (PFS), duration of response (DoR), disease control rate (DCR), and safety. Between Oct 14, 2020, and Dec 23, 2021, 71 patients were assessed for eligibility, of whom 58 patients were enrolled. All patients (mean [SD] age: 48 [9.59] years; 79.3% male) were included in the efficacy and safety analysis. The ORR was 65.5% (95% CI, 51.9-77.5) and the DCR was 86.2% (95% CI, 74.6-93.9). The median DoR was not reached and a median PFS was 10.4 months (95% CI, 7.2-13.6), with a median follow-up duration of 12.4 months (IQR, 5.6-13.2). Treatment-related adverse events (TRAEs) of grade 3 or higher were reported in 34 (58.6%) patients, mainly hypertension (19.0%), nasopharyngeal necrosis (15.5%), headache (12.1%), and creatine phosphokinase elevation (10.3%). 16 (27.6%) patients discontinued apatinib treatment ahead of progression because of unbearable TRAEs, and the most common one was nasopharyngeal necrosis (9/16, 56.3%). Nasopharyngeal recurrent lesion (odd ratio = 5.3, p=0.014) and courses of nasopharyngeal radiotherapy (p=0.015) were significantly positively correlated with nasopharyngeal necrosi. Camrelizumab plus apatinib had promising antitumor activity and manageable toxicities in recurrent or metastatic NPC. Larger randomized controlled trials are warranted to validate our findings.

Chronic exposure to airborne carbon black ultrafine (nCB) particles generated from incomplete combustion of organic matter drives IL-17A–dependent emphysema. However, whether and how they alter the immune responses to lung cancer remains unknown. Here, we show that exposure to nCB particles increased PD-L1+ PD-L2+ CD206+ antigen-presenting cells (APCs), exhausted T cells, and Treg cells. Lung macrophages that harbored nCB particles showed selective mitochondrial structure damage and decreased oxidative respiration. Lung macrophages sustained the HIF1α axis that increased glycolysis and lactate production, culminating in an immunosuppressive microenvironment in multiple mouse models of non–small cell lung cancers. Adoptive transfer of lung APCs from nCB-exposed wild type to susceptible mice increased tumor incidence and caused early metastasis. Our findings show that nCB exposure metabolically rewires lung macrophages to promote immunosuppression and accelerates the development of lung cancer.

In the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic1, considerable focus has been placed on a model of viral entry into host epithelial populations, with a separate focus upon the responding immune system dysfunction that exacerbates or causes disease. We developed a precision-cut lung slice model2,3 to investigate very early host-viral pathogenesis and found that SARS-CoV-2 had a rapid and specific tropism for myeloid populations in the human lung. Infection of alveolar macrophages was partially dependent upon their expression of ACE2, and the infections were productive for amplifying virus, both findings which were in contrast with their neutralization of another pandemic virus, Influenza A virus (IAV). Compared to IAV, SARS-CoV-2 was extremely poor at inducing interferon-stimulated genes in infected myeloid cells, providing a window of opportunity for modest titers to amplify within these cells. Endotracheal aspirate samples from humans with the acute respiratory distress syndrome (ARDS) from COVID-19 confirmed the lung slice findings, revealing a persistent myeloid depot. In the early phase of SARS-CoV-2 infection, myeloid cells may provide a safe harbor for the virus with minimal immune stimulatory cues being generated, resulting in effective viral colonization and quenching of the immune system.

Although infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has pleiotropic and systemic effects in some individuals1-3, many others experience milder symptoms. Here, to gain a more comprehensive understanding of the distinction between severe and mild phenotypes in the pathology of coronavirus disease 2019 (COVID-19) and its origins, we performed a whole-blood-preserving single-cell analysis protocol to integrate contributions from all major immune cell types of the blood-including neutrophils, monocytes, platelets, lymphocytes and the contents of the serum. Patients with mild COVID-19 exhibit a coordinated pattern of expression of interferon-stimulated genes (ISGs)3 across every cell population, whereas these ISG-expressing cells are systemically absent in patients with severe disease. Paradoxically, individuals with severe COVID-19 produce very high titres of anti-SARS-CoV-2 antibodies and have a lower viral load compared to individuals with mild disease. Examination of the serum from patients with severe COVID-19 shows that these patients uniquely produce antibodies that functionally block the production of the ISG-expressing cells associated with mild disease, by activating conserved signalling circuits that dampen cellular responses to interferons. Overzealous antibody responses pit the immune system against itself in many patients with COVID-19, and perhaps also in individuals with other viral infections. Our findings reveal potential targets for immunotherapies in patients with severe COVID-19 to re-engage viral defence.

Intratumoral T cells that might otherwise control tumors are often identified in an "exhausted" state, defined by specific epigenetic modifications and upregulation of genes such as CD38, cytotoxic T-lymphocyte-associated protein 4 (CTLA4), and programmed cell death 1 (PD1). Although the term might imply inactivity, there has been little study of this state at the phenotypic level in tumors to understand the extent of their incapacitation. Starting with the observation that T cells move more quickly through mouse tumors the longer they reside there and progress toward exhaustion, we developed a nonstimulatory, live-biopsy method for the real-time study of T cell behavior within individual patient tumors. Using 2-photon microscopy, we studied native CD8+ T cell interaction with antigen-presenting cells (APCs) and cancer cells in different microniches of human tumors and found that T cell speed was variable by region and by patient and was inversely correlated with local tumor density. Across a range of tumor types, we found a strong relationship between CD8+ T cell motility and the exhausted T cell state that corresponded with our observations made in mouse models in which exhausted T cells moved faster. Our study demonstrates T cell dynamic states in individual human tumors and supports the existence of an active program in "exhausted" T cells that extends beyond incapacitating them.

外科在初治鼻咽癌中的价值一直以来存在争议。初治鼻咽癌的外科发展经历了3个阶段：鼻外径路外科辅助治疗、微创外科辅助治疗、微创外科根治治疗。前2个阶段是在放化疗之前，先进行鼻咽原发灶切除术，目的是提高疗效或通过术后减量放射治疗降低放射毒性。第3阶段指针对拒绝放射治疗的“极早期”Ⅰ期鼻咽癌患者施行微创手术，彻底切除鼻咽原发灶，术后一般不进行放化疗，避免了放射性损伤，使“极早期”鼻咽癌多一个治疗选择。.

Electronic nicotine delivery systems (ENDS) or e-cigarettes have emerged as a popular recreational tool among adolescents and adults. Although the use of ENDS is often promoted as a safer alternative to conventional cigarettes, few comprehensive studies have assessed the long-term effects of vaporized nicotine and its associated solvents, propylene glycol (PG) and vegetable glycerin (VG). Here, we show that compared with smoke exposure, mice receiving ENDS vapor for 4 months failed to develop pulmonary inflammation or emphysema. However, ENDS exposure, independent of nicotine, altered lung lipid homeostasis in alveolar macrophages and epithelial cells. Comprehensive lipidomic and structural analyses of the lungs revealed aberrant phospholipids in alveolar macrophages and increased surfactant-associated phospholipids in the airway. In addition to ENDS-induced lipid deposition, chronic ENDS vapor exposure downregulated innate immunity against viral pathogens in resident macrophages. Moreover, independent of nicotine, ENDS-exposed mice infected with influenza demonstrated enhanced lung inflammation and tissue damage. Together, our findings reveal that chronic e-cigarette vapor aberrantly alters the physiology of lung epithelial cells and resident immune cells and promotes poor response to infectious challenge. Notably, alterations in lipid homeostasis and immune impairment are independent of nicotine, thereby warranting more extensive investigations of the vehicle solvents used in e-cigarettes.

Alteration of innate immune cells in the lungs can promote loss of peripheral tolerance that leads to autoimmune responses in cigarette smokers. Development of autoimmunity in smokers with emphysema is also strongly linked to the expansion of autoreactive T helper (Th) cells expressing interferon gamma (Th1), and interleukin 17A (Th17). However, the mechanisms responsible for enhanced self-recognition and reduced immune tolerance in smoker with emphysema remain less clear. Here we show that C1q, a component of the complement protein 1 complex (C1), is downregulated in lung CD1a+ antigen presenting cells (APCs) isolated from emphysematous human, and mouse lung APCs after chronic cigarette smoke exposure. C1q potentiated the function of APCs to differentiate CD4+ T cells to Tregs, while it inhibited Th17 cell development and proliferation. Mice deficient in C1q that were exposed to chronic smoke exhibited exaggerated lung inflammation marked by increased Th17 cells, while reconstitution of C1q in the lungs enhanced Tregs abundance, dampened smoke-induced lung inflammation, and reversed established emphysema. Our findings demonstrate that cigarette smoke-mediated loss of C1q could play a key role in reduced peripheral tolerance, which could be explored to treat emphysema.

Chronic inflammation induced by inhalation of airborne contaminants can contribute to cancer development. We have found that the anthracotic pigment in lungs of smokers is composed of nano-particulate carbon black (nCB), which accumulates in the antigen presenting cells (APCs) and promotes inflammation. Other occupational nCB exposures, such as coal-mining and carbon-black manufacturing, also raise the risk of chronic inflammatory lung diseases. No direct evidence establishes that nCB is a carcinogen; however, several cohort studies have shown a strong correlation between lung inflammation in carbon black industry workers and cancer. We examined whether nCB can promote non-small cell lung cancer (NSCLC) development. Using a novel non-Kras model of NSCLC, in which mice have airway epithelia-specific deletion of tumor suppressors Pten and Smad4 (Ptsd/d), we have found that intranasal challenge with nCB accelerates NSCLC progression. nCB exposure results in myeloid-derived suppressor recruitment and lactate accumulation to paralyze anti-tumor effects. Further, adoptive transfer of APCs from nCB exposed wild type mice (nCB-APC) to Ptsd/d mice showed increased incidences of lung tumor and metastasis, indicating a critical role for APCs in NSCLC progression. nCB-APC-induced immune suppression is in part mediated by expression of immunosuppressive enzymes and lactate production. Additionally, nCB-APCs express both pro-inflammatory and immunosuppressive cytokines to promote tumor growth. These findings suggest that nCB-APCs foster an immunosuppressive microenvironment, and indicate a mechanism responsible for environmental and occupational exposure to nCB in accelerating NSCLC progression.

Background: The role of locoregional radiotherapy in patients with primary metastatic nasopharyngeal carcinoma (mNPC) is unclear.Methods: In our open-label, phase 3, multi-centre randomized controlled trial, patients with primary mNPC, staged at IVc at the diagnosis of NPC were enrolled.Key inclusion criteria were CR or PR evaluated by imaging study after three cycles of chemotherapy according to the RECST v1.1; a KPS of at least 70.Eligible patients were randomly assigned in a 1:1 ratio to receive either chemotherapy plus radiotherapy or chemotherapy alone.Chemotherapy regimens were fluorouracil at 5 g/m 2 over 120 h and cisplatin at 100 mg/m 2 on day 1 once every 3 weeks for a maximum of six cycles.The primary endpoint was OS.We did efficacy analyses in ITT population.Safety analyses were done in patients receiving allocated treatment.This study is registered with ClinicalTrials.gov,number NCT02111460, and is ongoing.Results: Between April 2014 and August 2018, 126 eligible patients were randomly assigned to receive chemotherapy plus radiotherapy (n ¼ 63), or chemotherapy alone (n ¼ 63).In August 2018, the randomization was temporarily suspended due to an imbalance in deaths between the two groups and the ad hoc IDMC and the ethics committee of SYSUCC both recommended that the trial be permanently closed to new patient enrollment after IDMC confirmed the previously identified imbalance with this additional follow-up data in February 2019.The median follow-up time for OS was 25.2 months.The median OS was 40.2 months (95%CI 25.7-54.7) in the chemotherapy plus radiotherapy group and 24.5 months (95%CI 15.3-33.7) in the chemotherapy alone (HR 0.45 95% CI 0.25-0.80;P ¼ 0.007).No significant differences between the two treatment groups were observed in terms of hematological toxicity and gastrointestinal reaction.The frequency of grade 2-3 skin reaction and grade 3-4 mucositis in chemotherapy plus radiotherapy was significantly higher than those in chemotherapy alone groups (P < 0.05).Conclusions: Chemotherapy plus radiotherapy significantly improved overall survival in primary metastatic nasopharyngeal carcinoma with acceptable toxicity and tolerability.

Dendritic cells (DCs) must integrate a broad array of environmental cues to exact control over downstream immune responses including TH polarization. The multienzyme aminoacyl- tRNA synthetase complex component AIMp1/p43 responds to cellular stress and exerts pro-inflammatory functions; however, a role for DC-expressed AIMp1 in TH polarization has not previously been shown. Here, we demonstrate that the absence of AIMp1 in bone marrow-derived DC (BMDC) significantly impairs cytokine and costimulatory molecule expression, p38 MAPK signaling, and TH1 polarization of cocultured T-cells while significantly dysregulating immune-related gene expression. These deficits resulted in significantly compromised BMDC vaccine-mediated protection against melanoma. AIMp1 within the host was also critical for innate and adaptive antiviral immunity against influenza virus infection in vivo. Cancer patients with AIMp1 expression levels in the highest tertiles exhibited a 70% survival advantage at 15-year postdiagnosis as determined by bioinformatics analysis of nearly 9,000 primary human tumor samples in The Cancer Genome Atlas database. These data establish the importance of AIMp1 for the effective governance of antitumor and antiviral immune responses.

Background: To investigate and compare circulating tumor cells (CTCs) with Epstein-Barr virus DNA (EBV DNA) in noninvasive diagnosis, monitoring, and outcome prognosis of patients with metastatic nasopharyngeal carcinoma (mNPC). Methods: We prospectively enrolled 148 mNPC patients and 122 non-metastatic NPC patients, between December 2014 and August 2016. The levels of CTCs and EBV DNA were measured before the patients were due to start established chemotherapy and at the first follow-up visit. 281 mNPC patients recruited from January 2011 to December 2014 formed independent validation cohort for EBV DNA. The progression of the disease and the response to treatment were determined by standard imaging studies at participating centers. Results: In 270 prospectively recruited NPC patients, we observed improved specificity (86.0% vs 51.0%) of CTC levels as compared with EBV DNA copy number. 148 mNPC patients were stratified into two categories based on cutoff points of 12 in CTCs and 10000 in EBV DNA copy number. The cutoff point of 10000 in EBV DNA was confirmed in independent validation cohort. Baseline CTC level of 12 remained an independent prognostic factor of progression-free survival (PFS), but not EBV DNA. The median PFS was significantly longer for patients with CTC level of 0 at first follow-up compared with that of patients with CTC levels of > 1 at first follow-up regardless of CTC levels at baseline (P < 0.05). Among non-progressing patients, the PFS with complete/partial response and CTC level of 0 at first follow-up was significantly longer compared with rest of the patients (P < 0.05). Conclusions: Our data demonstrated an important role of CTC determination in mNPC patients. Clinical trial identification: NCT02505139. Legal entity responsible for the study: Sun Yat-sen University Cancer Center. Funding: The National Natural Science Foundation of China (No.81572912, 81772895, 81572848, and 81772877), Guangdong Public Welfare Research and Capacity Building Projects (2014B020212005), the Program of Sun Yat-sen University for Clinical Research 5010 Program (No.201310), the Major Project of Sun Yat-sen University for the New Cross Subject, the Special Support Program for High-level Talents in Sun Yat-sen University Cancer Center (to M.Y. Chen), and the National Key Research and Development Program of China (2016YFC0905000), Guangdong Province Science and Technology Development Special Funds (Frontier and Key Technology Innovation Direction – Major Science and Technology Project), Guangzhou Science and Technology Planning Project - Production and Research Collaborative Innovation Major Project. Disclosure: All authors have declared no conflicts of interest.

Somatic mutations can promote malignant transformation of airway epithelial cells and induce inflammatory responses directed against resultant tumors. Tumor-infiltrating T lymphocytes (TIL) in early-stage non-small cell lung cancer (NSCLC) secrete distinct proinflammatory cytokines, but the contribution of these TILs to tumor development and metastasis remains unknown. We show here that TILs in early-stage NSCLC are biased toward IL17A expression (Th17) when compared with adjacent tumor-free tissue, whereas Th17 cells are decreased in tumor infiltrating locoregional lymph nodes in advanced NSCLC. Mice in which Pten and Smad4 (Pts4(d/d)) are deleted from airway epithelial cells develop spontaneous tumors, that share genetic signatures with squamous- (SQ.2b), and adeno- (AD.1) subtypes of human NSCLC. Pts4(d/d) mice globally lacking in IL17a (Pts4(d/d)Il17a(-/-)) showed decreased tumor latency and increased metastasis. Th17 cells were required for recruitment of CD103(+) dendritic cells, and adoptive transfer of IL17a-sufficient CD4(+) T cells reversed early tumor development and metastasis in Pts4(d/d)Il17a(-/-) mice. Together, these findings support a key role for Th17 cells in TILs associated with the Pts4(d/d) model of NSCLC and suggest therapeutic and biomarker strategies for human SQ2b and AD1 lung cancer. (C) 2018 AACR.

Airborne pathogens commonly trigger severe respiratory failure or death in smokers with lung disease. Cigarette smoking compromises the effectiveness of innate immunity against infections but the underlying mechanisms responsible for defective acquired immune responses in smokers remains less clear. We found that mice exposed to chronic cigarette smoke recovered poorly from primary Influenza A pneumonia with reduced type I and II interferons (IFNs) and viral-specific immunoglobulins, but recruited γδ T cells to the lungs that predominantly expressed interleukin 17A (IL-17A). Il-17a−/− mice exposed to smoke and infected with Influenza A also recruited γδ T cells to the lungs, but in contrast to wild-type mice, expressed increased IFNs, made protective influenza-specific antibodies, and recovered from infection. Depletion of IL-17A with blocking antibodies significantly increased T-bet expression in γδ T cells and improved recovery from acute Influenza A infection in air, but not smoke-exposed mice. In contrast, when exposed to smoke, γδ T cell deficient mice failed to mount an effective immune response to Influenza A and showed increased mortality. Our findings demonstrate a protective role for γδ T cells in smokers and suggest that smoke-induced increase in IL-17A inhibits the transcriptional programs required for their optimal anti-viral responses. Cigarette smoke induces IL-17A expression in the lungs and inhibits γδ T-cell-mediated protective anti-viral immune responses.

Smoking-related lung diseases are among the most preventable and incurable ailments in the world. Smokers are at increased risk of developing chronic obstructive pulmonary disease that can be further complicated by emphysema and lung cancer. A subset of former smokers shows persistent lung inflammation and progressive loss of lung function, indicating a role for activation of acquired immunity in smoking-induced lung diseases. In addition to the well-established noxious effects of volatile compounds in cigarette smoke, incomplete combustion of tobacco generates nano-sized carbon black (nCB) that accumulate in lung myeloid dendritic cells and macrophages. Experimentally, intra-nasal instillation nCB can cause airway inflammation and emphysema in mice, underscoring their pathogenic role in inflammatory lung diseases. High throughput analyses of macrophages that have engulfed nCB reveal de novo activation of DNA repair enzymes, and histological studies provide evidence for DNA double-stranded breaks. Emphysematous lung myeloid dendritic cells that contain nCB express pro-inflammatory cytokines, and can efficiently differentiate naive CD4 T cells to interferon-g-secreting T helper 1 and interleukin 17A expressing cell subsets. Together these findings indicate that nCB accumulation in lung innate immune cells can initiate and sustain lung inflammation and promote emphysema development.