Neutrophils are essential components of the innate immune system, playing a pivotal role in immune responses. These cells rapidly migrate to sites of inflammation or tissue injury, facilitating pathogen clearance and tissue repair. The chemotactic signaling network regulating neutrophil recruitment is complex and not fully understood, particularly regarding damage-associated molecular patterns (DAMPs). In our previous research, we identified NMI as a DAMP that activates dendritic cells and macrophages, amplifying inflammatory responses and contributing to both acute and chronic inflammation. In this study, we investigated the role of NMI in neutrophil recruitment. We purified and characterized a recombinant murine NMI protein, ensuring endotoxin removal while preserving biological activity. In vivo experiments demonstrated that NMI enhances neutrophil recruitment in both a murine air pouch model and an acute peritonitis model, mediated by macrophage-derived chemokines. In vitro assays revealed a concentration-dependent increase in neutrophil migration induced by NMI, facilitated by chemokine secretion and subsequent migration through the CXCR2 receptor. Importantly, we established that NMI activates chemokine expression via the NF-kappaB signaling pathway. These findings provide insights into the mechanisms of NMI-induced neutrophil migration, enhancing our understanding of neutrophil recruitment during inflammation.
{"title":"NMI induces chemokine release and recruits neutrophils through the activation of NF-kappaB pathway","authors":"Zhenxing Chen, Yongjie Yao, Yuzhou Peng, Zhuangfeng Weng, Yingfang Liu, Na Xu","doi":"10.1101/2024.09.13.612823","DOIUrl":"https://doi.org/10.1101/2024.09.13.612823","url":null,"abstract":"Neutrophils are essential components of the innate immune system, playing a pivotal role in immune responses. These cells rapidly migrate to sites of inflammation or tissue injury, facilitating pathogen clearance and tissue repair. The chemotactic signaling network regulating neutrophil recruitment is complex and not fully understood, particularly regarding damage-associated molecular patterns (DAMPs). In our previous research, we identified NMI as a DAMP that activates dendritic cells and macrophages, amplifying inflammatory responses and contributing to both acute and chronic inflammation. In this study, we investigated the role of NMI in neutrophil recruitment. We purified and characterized a recombinant murine NMI protein, ensuring endotoxin removal while preserving biological activity. In vivo experiments demonstrated that NMI enhances neutrophil recruitment in both a murine air pouch model and an acute peritonitis model, mediated by macrophage-derived chemokines. In vitro assays revealed a concentration-dependent increase in neutrophil migration induced by NMI, facilitated by chemokine secretion and subsequent migration through the CXCR2 receptor. Importantly, we established that NMI activates chemokine expression via the NF-kappaB signaling pathway. These findings provide insights into the mechanisms of NMI-induced neutrophil migration, enhancing our understanding of neutrophil recruitment during inflammation.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"54 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-18DOI: 10.1101/2024.09.13.612684
Caio Foulkes, Nikolas Friedrich, Branislav Ivan, Emanuel Stiegeler, Carsten Magnus, Daniel Schmidt, Umut Karakus, Jacqueline Weber, Huldrych F. Günthard, Chloé Pasin, Peter Rusert, Alexandra Trkola
The ability of broadly neutralizing antibodies (bnAbs) to interact with the closed, pre-fusion HIV-1 envelope (Env) trimer distinguishes them from weakly neutralizing antibodies (weak-nAbs) that depend on trimer opening to bind. Comparative analysis of neutralization data from the CATNAP database revealed a nuanced relationship between bnAb activity and Env conformational plasticity, with substantial epitope-specific variation of bnAb potency ranging from increased to decreased activity against open, neutralization-sensitive Env. To systematically investigate the impact of Env conformational dynamics on bnAb potency we screened 126 JR-CSF point mutants for generalized neutralization sensitivity to weak-nAbs and plasma from people with chronic HIV-1 infection. 23 mutations at highly conserved sites resulted in neutralization phenotype with high Tier 1 sensitivity, which was associated with de-stabilization of the closed, prefusion conformation. Including 19 of these mutants into a Sensitivity Env mutant panel (SENSE-19), we classified bnAbs according to potency variations in response to trimer opening. To verify that these sensitivity patterns are independent of the in vitro assay system, replication-competent SENSE-19 mutant viruses were tested on primary CD4 T cells. While loss of potency on SENSE-19 was registered for bnAbs recognizing quaternary epitopes on pre-triggered Env, structural destabilization benefitted MPER bnAbs and other inhibitors known to have post-CD4 attachment neutralization activity. Importantly, for certain bnAbs targeting CD4bs, V3-glycan and interface epitopes, particularly low potency variation was noted, suggesting that Env conformational tolerance can be achieved but is not the rule. In summary, SENSE-19 screens revealed distinct Env flexibility tolerance levels between bnAb types that provide mechanistic insights in their function and broaden current neutralization breadth assessments.
{"title":"Assessing bnAb potency in the context of HIV-1 Envelope conformational plasticity","authors":"Caio Foulkes, Nikolas Friedrich, Branislav Ivan, Emanuel Stiegeler, Carsten Magnus, Daniel Schmidt, Umut Karakus, Jacqueline Weber, Huldrych F. Günthard, Chloé Pasin, Peter Rusert, Alexandra Trkola","doi":"10.1101/2024.09.13.612684","DOIUrl":"https://doi.org/10.1101/2024.09.13.612684","url":null,"abstract":"The ability of broadly neutralizing antibodies (bnAbs) to interact with the closed, pre-fusion HIV-1 envelope (Env) trimer distinguishes them from weakly neutralizing antibodies (weak-nAbs) that depend on trimer opening to bind. Comparative analysis of neutralization data from the CATNAP database revealed a nuanced relationship between bnAb activity and Env conformational plasticity, with substantial epitope-specific variation of bnAb potency ranging from increased to decreased activity against open, neutralization-sensitive Env. To systematically investigate the impact of Env conformational dynamics on bnAb potency we screened 126 JR-CSF point mutants for generalized neutralization sensitivity to weak-nAbs and plasma from people with chronic HIV-1 infection. 23 mutations at highly conserved sites resulted in neutralization phenotype with high Tier 1 sensitivity, which was associated with de-stabilization of the closed, prefusion conformation. Including 19 of these mutants into a Sensitivity Env mutant panel (SENSE-19), we classified bnAbs according to potency variations in response to trimer opening. To verify that these sensitivity patterns are independent of the in vitro assay system, replication-competent SENSE-19 mutant viruses were tested on primary CD4 T cells. While loss of potency on SENSE-19 was registered for bnAbs recognizing quaternary epitopes on pre-triggered Env, structural destabilization benefitted MPER bnAbs and other inhibitors known to have post-CD4 attachment neutralization activity. Importantly, for certain bnAbs targeting CD4bs, V3-glycan and interface epitopes, particularly low potency variation was noted, suggesting that Env conformational tolerance can be achieved but is not the rule. In summary, SENSE-19 screens revealed distinct Env flexibility tolerance levels between bnAb types that provide mechanistic insights in their function and broaden current neutralization breadth assessments.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"192 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objectives Tumor-associated macrophages (TAMs) accumulate in the majority of solid tumors, producing inflammatory cytokines and growth factors involved in tumor maintenance. They have recently emerged as targets for restoring an effective antitumor response and limiting tumor growth. In the present study, we investigated the potential of IL-27 neutralization to modify macrophage polarization and thus the tumor immune microenvironment. Methods and Analysis We monitored the effect of IL-27 neutralization on human macrophages. Flow cytometry, quantitative reverse transcription-PCR, ELISA and western blot were performed to validate, in vitro, the role of IL-27 on the acquisition of macrophage immunoregulatory functions. A murine Colon Adenocarcinoma model (MC38) was used to assess IL-27 neutralization in the microenvironment in vivo. Results In this study, we demonstrated the importance of IL-27 in the generation of human immunoregulatory macrophages. Mechanistically, IL-27 neutralization reduced the immunosuppressive properties of macrophages, such as cytokine secretion and membrane expression of immunosuppressive molecules. These modifications led to a reduction in the ability of macrophages to inhibit the function of CD4+ and CD8+ T cells. Furthermore, in vivo neutralization of IL-27 reduced MC38 tumor growth and could improve immune checkpoint inhibitor efficacy. Conclusions Collectively, we uncovered the role of IL-27 in the immunosuppressive tumor microenvironment (TME). We showed that IL-27 could be a target for macrophage repolarization and boosting of CD4+ and CD8+ T cell responses. IL-27 neutralization thus appears as a promising strategy to target macrophages in immunosuppressive TME and improve the clinical efficacy of immunotherapy protocols.
{"title":"IL-27 neutralization to modulate the tumor microenvironment and increase immune checkpoint immunotherapy efficacy","authors":"Loukas Papargyris, Quentin Glaziou, Laetitia Basset, Senan d'Almeida, Pascale Pignon, Nabila Jabrane-Ferrat, Christophe Blanquart, Yves Delneste, Julie TABIASCO","doi":"10.1101/2024.09.13.612803","DOIUrl":"https://doi.org/10.1101/2024.09.13.612803","url":null,"abstract":"Objectives\u0000Tumor-associated macrophages (TAMs) accumulate in the majority of solid tumors, producing inflammatory cytokines and growth factors involved in tumor maintenance. They have recently emerged as targets for restoring an effective antitumor response and limiting tumor growth. In the present study, we investigated the potential of IL-27 neutralization to modify macrophage polarization and thus the tumor immune microenvironment.\u0000Methods and Analysis\u0000We monitored the effect of IL-27 neutralization on human macrophages. Flow cytometry, quantitative reverse transcription-PCR, ELISA and western blot were performed to validate, in vitro, the role of IL-27 on the acquisition of macrophage immunoregulatory functions. A murine Colon Adenocarcinoma model (MC38) was used to assess IL-27 neutralization in the microenvironment in vivo.\u0000Results\u0000In this study, we demonstrated the importance of IL-27 in the generation of human immunoregulatory macrophages. Mechanistically, IL-27 neutralization reduced the immunosuppressive properties of macrophages, such as cytokine secretion and membrane expression of immunosuppressive molecules. These modifications led to a reduction in the ability of macrophages to inhibit the function of CD4+ and CD8+ T cells. Furthermore, in vivo neutralization of IL-27 reduced MC38 tumor growth and could improve immune checkpoint inhibitor efficacy.\u0000Conclusions\u0000Collectively, we uncovered the role of IL-27 in the immunosuppressive tumor microenvironment (TME). We showed that IL-27 could be a target for macrophage repolarization and boosting of CD4+ and CD8+ T cell responses. IL-27 neutralization thus appears as a promising strategy to target macrophages in immunosuppressive TME and improve the clinical efficacy of immunotherapy protocols.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"20 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-18DOI: 10.1101/2024.09.13.612848
Asha Bhardwaj, Leena Sapra, Divya Madan, Vineet Ahuja, Pradyumna Kumar Mishra, Rupesh K. Srivastava
Osteoporosis is a skeletal condition characterized by the deterioration of bone tissue. The immune system plays a crucial role in maintaining bone homeostasis and combating the development of osteoporosis. Immunoporosis is the term used to describe the recent convergence of research on the immune system's role in osteoporosis. Gut harbors the largest component of the immune system and there is growing evidence that intestinal immunity plays a vital role in regulating bone health. Gut-resident regulatory T cells (GTregs) play an essential role in inhibiting immune responses and preventing various inflammatory manifestations. Our findings show that GTregs have a pivotal role in the pathophysiology of post-menopausal osteoporosis (PMO). We investigated the potential of GTregs in regulating the development of bone cells in vitro. We observed that GTregs significantly enhance osteoblastogenesis with concomitant inhibition of osteoclastogenesis in a cell-ratio-dependent manner. We further report that the deficiency of short-chain fatty acids (SCFAs) in osteoporotic conditions substantially disrupts the composition of GTregs, leading to a loss of peripherally derived Tregs (pTregs) and an expansion of thymus-derived Tregs (tTregs). Moreover, the administration of probiotics Lactobacillus rhamnosus and Bifidobacterium longum modulated the GTregs compartment in an SCFA-dependent manner to mitigate inflammatory bone loss in PMO. Notably, SCFAs-primed GTregs were found to be significantly more effective in inhibiting osteoclastogenesis compared to unprimed GTregs. Altogether our results, for the first time, highlight the crucial role of GTregs in the pathophysiology of PMO, with potential clinical implications in the near future.
{"title":"Homeostatic balance of Gut-resident Tregs (GTregs) plays a pivotal role in maintaining bone health under post-menopausal osteoporotic conditions","authors":"Asha Bhardwaj, Leena Sapra, Divya Madan, Vineet Ahuja, Pradyumna Kumar Mishra, Rupesh K. Srivastava","doi":"10.1101/2024.09.13.612848","DOIUrl":"https://doi.org/10.1101/2024.09.13.612848","url":null,"abstract":"Osteoporosis is a skeletal condition characterized by the deterioration of bone tissue. The immune system plays a crucial role in maintaining bone homeostasis and combating the development of osteoporosis. Immunoporosis is the term used to describe the recent convergence of research on the immune system's role in osteoporosis. Gut harbors the largest component of the immune system and there is growing evidence that intestinal immunity plays a vital role in regulating bone health. Gut-resident regulatory T cells (GTregs) play an essential role in inhibiting immune responses and preventing various inflammatory manifestations. Our findings show that GTregs have a pivotal role in the pathophysiology of post-menopausal osteoporosis (PMO). We investigated the potential of GTregs in regulating the development of bone cells in vitro. We observed that GTregs significantly enhance osteoblastogenesis with concomitant inhibition of osteoclastogenesis in a cell-ratio-dependent manner. We further report that the deficiency of short-chain fatty acids (SCFAs) in osteoporotic conditions substantially disrupts the composition of GTregs, leading to a loss of peripherally derived Tregs (pTregs) and an expansion of thymus-derived Tregs (tTregs). Moreover, the administration of probiotics Lactobacillus rhamnosus and Bifidobacterium longum modulated the GTregs compartment in an SCFA-dependent manner to mitigate inflammatory bone loss in PMO. Notably, SCFAs-primed GTregs were found to be significantly more effective in inhibiting osteoclastogenesis compared to unprimed GTregs. Altogether our results, for the first time, highlight the crucial role of GTregs in the pathophysiology of PMO, with potential clinical implications in the near future.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"40 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tuberculosis is major challenge to the health care system with TB associated death rates increasing annually. Optimum management of TB (particularly latent or MDR cases) warrants use of immunological approaches like subunit or peptide-based vaccination for tailoring effector immunity in patients. Since MHC class I is a potent enhancer element of host immunity and effective in clearing large variety of intracellular pathogens or tumors. In this context, we explore whether MHC-I restricted peptides from clinical isolates of M. tuberculosis can be used as an adjuvant for augmenting host immune responses. In the present study, we have synthesized various peptides from clinical isolates of M. tuberculosis which were having high affinity for Class I MHC molecules as potential immune enhancer for T cell or iNKT cell populations. We have evaluated the immunogenic potential of various MHC class I restricted epitopes (Rv2588c, Rv1357, Rv0148, Rv2973, Rv2557 and Rv2445) which were derived from clinical isolates of M. tuberculosis on increased proliferation of T or iNKT cells, release of IFN gamma secreted by T cells as well as NO as indicative parameters of immuno-stimulation. As expected, FACS and ELISA data clearly revealed that these peptides were potentially immunogenic for PBMCs from both healthy as well as 10 HC PTB patients. Our data clearly demonstrated a significant immune response in the PBMC from w PTB patients over healthy individuals which mimicked booster response. Our cytokine and nitric oxide data further revealed the influence of these peptides on sensitizing innate immune response as well.
结核病是医疗保健系统面临的一大挑战，与结核病相关的死亡率逐年上升。结核病（尤其是潜伏或 MDR 病例）的最佳治疗需要使用亚单位或多肽疫苗等免疫学方法来调整患者的效应免疫。由于 MHC I 类是宿主免疫的有效增强因子，能有效清除多种细胞内病原体或肿瘤。在此背景下，我们探讨了从结核杆菌临床分离物中提取的 MHC-I 限制肽是否可用作增强宿主免疫反应的佐剂。在本研究中，我们从结核杆菌的临床分离物中合成了多种多肽，这些多肽对 I 类 MHC 分子具有高亲和力，可作为 T 细胞或 iNKT 细胞群的潜在免疫增强剂。我们评估了从结核杆菌临床分离株中提取的各种 MHC I 类受限表位（Rv2588c、Rv1357、Rv0148、Rv2973、Rv2557 和 Rv2445）对 T 细胞或 iNKT 细胞增殖、T 细胞分泌的 IFN γ 释放以及作为免疫刺激指示性参数的 NO 的免疫原性潜力。正如预期的那样，FACS 和 ELISA 数据清楚地表明，这些肽对健康和 10 HC PTB 患者的 PBMC 都有潜在的免疫原性。我们的数据清楚地表明，肺结核患者的 PBMC 比健康人的 PBMC 产生了明显的免疫反应，这与增效反应相似。我们的细胞因子和一氧化氮数据进一步揭示了这些肽对先天性免疫反应敏化的影响。
{"title":"MHC class I / II restricted T cell epitopes from clinical isolate of Mycobacterium tuberculosis: A potential candidate for vaccine development for Tuberculosis","authors":"Niharika Sharma, Bhawna Sharma, Beenu Joshi, Santosh Kumar, Keshar Kunja Mohanty, Hridayesh Prakash","doi":"10.1101/2024.09.13.612852","DOIUrl":"https://doi.org/10.1101/2024.09.13.612852","url":null,"abstract":"Tuberculosis is major challenge to the health care system with TB associated death rates increasing annually. Optimum management of TB (particularly latent or MDR cases) warrants use of immunological approaches like subunit or peptide-based vaccination for tailoring effector immunity in patients. Since MHC class I is a potent enhancer element of host immunity and effective in clearing large variety of intracellular pathogens or tumors. In this context, we explore whether MHC-I restricted peptides from clinical isolates of M. tuberculosis can be used as an adjuvant for augmenting host immune responses. In the present study, we have synthesized various peptides from clinical isolates of M. tuberculosis which were having high affinity for Class I MHC molecules as potential immune enhancer for T cell or iNKT cell populations. We have evaluated the immunogenic potential of various MHC class I restricted epitopes (Rv2588c, Rv1357, Rv0148, Rv2973, Rv2557 and Rv2445) which were derived from clinical isolates of M. tuberculosis on increased proliferation of T or iNKT cells, release of IFN gamma secreted by T cells as well as NO as indicative parameters of immuno-stimulation. As expected, FACS and ELISA data clearly revealed that these peptides were potentially immunogenic for PBMCs from both healthy as well as 10 HC PTB patients. Our data clearly demonstrated a significant immune response in the PBMC from w PTB patients over healthy individuals which mimicked booster response. Our cytokine and nitric oxide data further revealed the influence of these peptides on sensitizing innate immune response as well.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"141 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142268665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-18DOI: 10.1101/2024.09.17.613525
Art Marzok, Jonathan P Mapletoft, Braeden Cowbrough, Daniel B Celeste, Michael R D'Agostino, Jann C Ang, Andrew Chen, Vithushan Surendran, Anna Dvorkin-Gheva, Ali Zhang, Yasmine Kollar, Hannah D Stacey, Sam Afkhami, Mannie Lam, Kevin R Milnes, Matthew S Miller
While several viral infections have been associated with amyotrophic lateral sclerosis (ALS), the mechanism(s) through which they promote disease has remained almost entirely elusive. This study investigated the impact of common, acute viral infections prior to disease onset on ALS progression in the SOD1G93A mouse model. A single sublethal infection prior to onset of ALS clinical signs was associated with markedly accelerated ALS disease progression characterized by rapid loss of hindlimb function. Prior infection resulted in gliosis in the lumbar spine and upregulation of transcriptional pathways involved in inflammatory responses, metabolic dysregulation, and muscular dysfunction. Therapeutic suppression of gliosis with an anti-inflammatory small molecule, or administration of a direct-acting antiviral, was associated with significantly improved ALS clinical signs, akin to what was observed in uninfected animals. This study provides causal and mechanistic evidence that the immune response elicited by acute viral infections may be an important etiological factor that alters ALS disease trajectory, and provides insight into novel therapeutic and preventative strategies for ALS.
虽然有几种病毒感染与肌萎缩性脊髓侧索硬化症（ALS）有关，但它们促进疾病发生的机制却几乎完全不为人知。本研究在 SOD1G93A 小鼠模型中研究了发病前常见急性病毒感染对 ALS 进展的影响。在渐冻症临床症状出现之前的一次亚致死性感染会明显加速渐冻症的进展，其特点是后肢功能迅速丧失。之前的感染会导致腰椎神经胶质增生，并上调涉及炎症反应、代谢失调和肌肉功能障碍的转录通路。用抗炎小分子或直接作用抗病毒药物治疗抑制神经胶质增生与明显改善 ALS 临床症状有关，这与在未感染动物身上观察到的情况类似。这项研究提供了因果关系和机理方面的证据，证明急性病毒感染引起的免疫反应可能是改变 ALS 疾病轨迹的重要病因，并为 ALS 的新型治疗和预防策略提供了启示。
{"title":"Acute Viral Infection Accelerates Neurodegeneration in a Mouse Model of ALS","authors":"Art Marzok, Jonathan P Mapletoft, Braeden Cowbrough, Daniel B Celeste, Michael R D'Agostino, Jann C Ang, Andrew Chen, Vithushan Surendran, Anna Dvorkin-Gheva, Ali Zhang, Yasmine Kollar, Hannah D Stacey, Sam Afkhami, Mannie Lam, Kevin R Milnes, Matthew S Miller","doi":"10.1101/2024.09.17.613525","DOIUrl":"https://doi.org/10.1101/2024.09.17.613525","url":null,"abstract":"While several viral infections have been associated with amyotrophic lateral sclerosis (ALS), the mechanism(s) through which they promote disease has remained almost entirely elusive. This study investigated the impact of common, acute viral infections prior to disease onset on ALS progression in the SOD1G93A mouse model. A single sublethal infection prior to onset of ALS clinical signs was associated with markedly accelerated ALS disease progression characterized by rapid loss of hindlimb function. Prior infection resulted in gliosis in the lumbar spine and upregulation of transcriptional pathways involved in inflammatory responses, metabolic dysregulation, and muscular dysfunction. Therapeutic suppression of gliosis with an anti-inflammatory small molecule, or administration of a direct-acting antiviral, was associated with significantly improved ALS clinical signs, akin to what was observed in uninfected animals. This study provides causal and mechanistic evidence that the immune response elicited by acute viral infections may be an important etiological factor that alters ALS disease trajectory, and provides insight into novel therapeutic and preventative strategies for ALS.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142268661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-18DOI: 10.1101/2024.09.12.612760
Victoria L. Rhodes, Robert M. Waterhouse, Kristin Michel
Innate immunity in mosquitoes has received much attention due to its potential impact on vector competence for vector-borne disease pathogens, including malaria parasites. The nuclear factor (NF)-kB-dependent Toll pathway is a major regulator of innate immunity in insects. In mosquitoes, this pathway controls transcription of the majority of the known canonical humoral immune effectors, mediates anti-bacterial, anti-fungal and anti-viral immune responses, and contributes to malaria parasite killing. However, besides initial gene annotation of putative Toll pathway members and genetic analysis of the contribution of few key components to immunity, the molecular make-up and function of the Toll pathway in mosquitoes is largely unexplored. To facilitate functional analyses of the Toll pathway in mosquitoes, we report here manually annotated and refined gene models of Toll-like receptors and all putative components of the intracellular signal transduction cascade across 19 anopheline genomes, and in two culicine genomes. In addition, based on phylogenetic analyses, we identified differing levels of evolutionary constraint across the intracellular Toll pathway members, and identified a recent radiation of TOLL1/5 within the An. gambiae complex. Together, this study provides insight into the evolution of TLRs and the putative members of the intracellular signal transduction cascade within the genus Anopheles.
{"title":"The molecular Toll pathway repertoire in anopheline mosquitoes","authors":"Victoria L. Rhodes, Robert M. Waterhouse, Kristin Michel","doi":"10.1101/2024.09.12.612760","DOIUrl":"https://doi.org/10.1101/2024.09.12.612760","url":null,"abstract":"Innate immunity in mosquitoes has received much attention due to its potential impact on vector competence for vector-borne disease pathogens, including malaria parasites. The nuclear factor (NF)-kB-dependent Toll pathway is a major regulator of innate immunity in insects. In mosquitoes, this pathway controls transcription of the majority of the known canonical humoral immune effectors, mediates anti-bacterial, anti-fungal and anti-viral immune responses, and contributes to malaria parasite killing. However, besides initial gene annotation of putative Toll pathway members and genetic analysis of the contribution of few key components to immunity, the molecular make-up and function of the Toll pathway in mosquitoes is largely unexplored. To facilitate functional analyses of the Toll pathway in mosquitoes, we report here manually annotated and refined gene models of Toll-like receptors and all putative components of the intracellular signal transduction cascade across 19 anopheline genomes, and in two culicine genomes. In addition, based on phylogenetic analyses, we identified differing levels of evolutionary constraint across the intracellular Toll pathway members, and identified a recent radiation of TOLL1/5 within the An. gambiae complex. Together, this study provides insight into the evolution of TLRs and the putative members of the intracellular signal transduction cascade within the genus Anopheles.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-17DOI: 10.1101/2024.09.14.613008
Xinkang Huan, Hongwei Gao
Since the end of 2019, the coronavirus disease 2019 (COVID-19) has been endemic worldwide for three years, causing more than 6.95 million deaths and having a massive impact on the global political economy. With time, the Severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) is also constantly mutating. Mutations lead to stronger infectivity or virulence of the virus, and some monoclonal antibodies against wild-type SARS-COV-2 are also challenging to play a role. Amubarvimab and Romlusevimab were originally developed against wild-type SARS-COV-2; however, these monoclonal antibodies' neutralizing efficacy and mechanism against these mutants are unknown. In this study, the binding ability of Amubarvimab and Romlusevimab to 7 mutant strains were tested by computer method and the interaction mechanism was explored. Our experimental data show that Amubarvimab can effectively bind most mutations and maintain the stability of the complexes mainly through hydrogen bond interaction; However, the binding efficiency of Romlusevimab was lower than that of Amubarvimab, and the stability of 18 the complexes was maintained mainly through electrostatic interaction. Both Amubarvimab and Romlusevimab show low binding potency against E406W and Q498Y mutations, so there is a certain probability of immune escape in the face of variants carrying E406W and Q498Y mutations when Amubarvimab and Romlusevimab are used in combination.
{"title":"In computer explore The neutralization mechanism of Amubarvimab and Romlusevimab against SARS-COV-2 mutants","authors":"Xinkang Huan, Hongwei Gao","doi":"10.1101/2024.09.14.613008","DOIUrl":"https://doi.org/10.1101/2024.09.14.613008","url":null,"abstract":"Since the end of 2019, the coronavirus disease 2019 (COVID-19) has been endemic worldwide for three years, causing more than 6.95 million deaths and having a massive impact on the global political economy. With time, the Severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) is also constantly mutating. Mutations lead to stronger infectivity or virulence of the virus, and some monoclonal antibodies against wild-type SARS-COV-2 are also challenging to play a role. Amubarvimab and Romlusevimab were originally developed against wild-type SARS-COV-2; however, these monoclonal antibodies' neutralizing efficacy and mechanism against these mutants are unknown. In this study, the binding ability of Amubarvimab and Romlusevimab to 7 mutant strains were tested by computer method and the interaction mechanism was explored. Our experimental data show that Amubarvimab can effectively bind most mutations and maintain the stability of the complexes mainly through hydrogen bond interaction; However, the binding efficiency of Romlusevimab was lower than that of Amubarvimab, and the stability of 18 the complexes was maintained mainly through electrostatic interaction. Both Amubarvimab and Romlusevimab show low binding potency against E406W and Q498Y mutations, so there is a certain probability of immune escape in the face of variants carrying E406W and Q498Y mutations when Amubarvimab and Romlusevimab are used in combination.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-17DOI: 10.1101/2024.09.16.613292
Laure Bosquillon de Jarcy, Dylan Postmus, Jenny Jansen, Julia Melchert, Donata Hoffmann, Victor Max Corman, Christine Goffinet
MPOX virus (MPXV), formerly known as monkeypox virus, led to a rapidly evolving pandemic starting May 2022, with over 90,000 cases reported beyond the African continent. This pandemic outbreak was driven by the MPXV variant Clade IIb. In addition, Clade I viruses circulating in the Democratic Republic of Congo (DRC) are drawing increased attention as cases constantly rise and Clade Ib, first identified in 2023, is now co-circulating with Clade Ia and seems to exhibit enhanced human-to-human transmissibility. While most infected individuals display a self-limiting disease with singular pox-like lesions, some endure systemic viral spread leading to whole-body rash with risks for necrosis, organ loss, and death. Intra-host dissemination and cellular tropism of MPXV are largely unexplored in humans. To establish a potential susceptibility of circulating immune cells to MPXV, we exposed human PBMCs from healthy donors ex vivo to a currently circulating MPXV clade IIb virus isolate in absence and presence of IFN-α2a. qPCR of DNA extracted from cell lysates, but less from supernatants, revealed increasing MPXV DNA quantities that peaked at five to six days post-exposure, suggesting susceptibility of PBMCs to infection. IFN-α2a pretreatment markedly reduced the quantity of MPXV DNA, suggesting that infection is sensitive to type I IFNs. Plaque assays from supernatants showed that infection gave rise to de novo production of infectious MPXV. In virus-inclusive scRNA-sequencing, monocytes, cycling NK cells and regulatory CD4+ T-cells scored positive for viral RNA, suggesting that these are the MPXV-susceptible cell types within the human PBMC population. Analysis of differentially expressed genes displayed a pronounced downregulation of expression pathways driving innate immunity in MPXV-infected cells, a well-established feature of poxviral infection. Pretreatment of PBMCs with current antivirals Cidofovir and Tecovirimat resulted in reduced amounts of viral antigen production and of released infectivity, suggesting suitability of the human PBMC infection model as a platform for evaluation of current and future antivirals and justifying trials to investigate Cidofovir and Tecovirimat as drugs reducing intra-patient viral spread. Together, our data suggest that human PBMCs are productively infected by MPXV which is accompanied by significant modulation of the cellular milieu. Our results have the potential to illuminate aspects of intra-host propagation of MPXV that may involve a lymphohematogenous route for replication and/or intra-host dissemination.
MPOX病毒（MPXV）以前被称为猴痘病毒，从2022年5月开始迅速演变成大流行病，非洲大陆以外报告的病例超过9万例。这次大流行是由 MPXV 变种 IIb 驱动的。此外，随着病例不断增加，在刚果民主共和国（刚果（金））流行的 I 支系病毒越来越受到关注，2023 年首次发现的 Ib 支系病毒目前正与 Ia 支系病毒共同流行，似乎表现出更强的人际传播性。虽然大多数感染者表现为单发痘样病变的自限性疾病，但也有一些人忍受着全身性病毒传播，导致全身皮疹，并有坏死、器官缺失和死亡的风险。MPXV在人体内的宿主内传播和细胞滋养特性在很大程度上尚未得到研究。为了确定循环免疫细胞对 MPXV 的潜在易感性，我们将健康捐献者的人 PBMCs 在体内外暴露于目前循环的 MPXV IIb 支系病毒分离物，在无 IFN-α2a 和有 IFN-α2a 的情况下，对细胞裂解液（但上清液较少）提取的 DNA 进行 qPCR 检测，发现 MPXV DNA 数量不断增加，在暴露后五到六天达到峰值，表明 PBMCs 易受感染。IFN-α2a 预处理显著减少了 MPXV DNA 的数量，表明感染对 IFNs 敏感。从上清液中提取的斑块检测结果表明，感染后会重新产生具有传染性的 MPXV。在包含病毒的 scRNA 测序中，单核细胞、循环 NK 细胞和调节性 CD4+ T 细胞的病毒 RNA 呈阳性，表明这些是人类 PBMC 群体中 MPXV 易感细胞类型。对差异表达基因的分析表明，MPXV 感染细胞中驱动先天性免疫的表达途径明显下调，这是痘病毒感染的一个公认特征。用目前的抗病毒药物西多福韦酯和特考韦酯预处理 PBMC，可减少病毒抗原的产生量和释放的感染性，这表明人类 PBMC 感染模型适合作为评估目前和未来抗病毒药物的平台，并证明了研究西多福韦酯和特考韦酯作为减少患者体内病毒传播药物的试验是合理的。总之，我们的数据表明，人类 PBMC 在受到 MPXV 感染后，细胞环境会发生显著变化。我们的研究结果有可能揭示了 MPXV 在宿主体内传播的各个方面，其中可能涉及到淋巴血源性复制途径和/或宿主体内传播。
{"title":"MPXV Infects Human PBMCs in a Type I Interferon-Sensitive Manner","authors":"Laure Bosquillon de Jarcy, Dylan Postmus, Jenny Jansen, Julia Melchert, Donata Hoffmann, Victor Max Corman, Christine Goffinet","doi":"10.1101/2024.09.16.613292","DOIUrl":"https://doi.org/10.1101/2024.09.16.613292","url":null,"abstract":"MPOX virus (MPXV), formerly known as monkeypox virus, led to a rapidly evolving pandemic starting May 2022, with over 90,000 cases reported beyond the African continent. This pandemic outbreak was driven by the MPXV variant Clade IIb. In addition, Clade I viruses circulating in the Democratic Republic of Congo (DRC) are drawing increased attention as cases constantly rise and Clade Ib, first identified in 2023, is now co-circulating with Clade Ia and seems to exhibit enhanced human-to-human transmissibility. While most infected individuals display a self-limiting disease with singular pox-like lesions, some endure systemic viral spread leading to whole-body rash with risks for necrosis, organ loss, and death. Intra-host dissemination and cellular tropism of MPXV are largely unexplored in humans. To establish a potential susceptibility of circulating immune cells to MPXV, we exposed human PBMCs from healthy donors ex vivo to a currently circulating MPXV clade IIb virus isolate in absence and presence of IFN-α2a. qPCR of DNA extracted from cell lysates, but less from supernatants, revealed increasing MPXV DNA quantities that peaked at five to six days post-exposure, suggesting susceptibility of PBMCs to infection. IFN-α2a pretreatment markedly reduced the quantity of MPXV DNA, suggesting that infection is sensitive to type I IFNs. Plaque assays from supernatants showed that infection gave rise to de novo production of infectious MPXV. In virus-inclusive scRNA-sequencing, monocytes, cycling NK cells and regulatory CD4+ T-cells scored positive for viral RNA, suggesting that these are the MPXV-susceptible cell types within the human PBMC population. Analysis of differentially expressed genes displayed a pronounced downregulation of expression pathways driving innate immunity in MPXV-infected cells, a well-established feature of poxviral infection. Pretreatment of PBMCs with current antivirals Cidofovir and Tecovirimat resulted in reduced amounts of viral antigen production and of released infectivity, suggesting suitability of the human PBMC infection model as a platform for evaluation of current and future antivirals and justifying trials to investigate Cidofovir and Tecovirimat as drugs reducing intra-patient viral spread. Together, our data suggest that human PBMCs are productively infected by MPXV which is accompanied by significant modulation of the cellular milieu. Our results have the potential to illuminate aspects of intra-host propagation of MPXV that may involve a lymphohematogenous route for replication and/or intra-host dissemination.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"54 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252115","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-17DOI: 10.1101/2024.09.12.612619
Simon J Cleary, Longhui Qiu, Yurim Seo, Peter Baluk, Dan Liu, Nina K Serwas, Jason G Cyster, Donald M McDonald, Matthew F Krummel, Mark R Looney
Intravital microscopy has enabled the study of immune dynamics in the pulmonary microvasculature, but many key events remain unseen because they occur in deeper lung regions. We therefore developed a technique for stabilized intravital imaging of bronchovascular cuffs and collecting lymphatics surrounding pulmonary veins in mice. Intravital imaging of pulmonary lymphatics revealed ventilation-dependence of steady-state lung lymph flow and ventilation-independent lymph flow during inflammation. We imaged the rapid exodus of migratory dendritic cells through lung lymphatics following inflammation and measured effects of pharmacologic and genetic interventions targeting chemokine signaling. Intravital imaging also captured lymphatic immune surveillance of lung-metastatic cancers and lymphatic metastasis of cancer cells. To our knowledge, this is the first imaging of lymph flow and leukocyte migration through intact pulmonary lymphatics. This approach will enable studies of protective and maladaptive processes unfolding within the lungs and in other previously inaccessible locations.
{"title":"Intravital imaging of pulmonary lymphatics in inflammation and metastatic cancer","authors":"Simon J Cleary, Longhui Qiu, Yurim Seo, Peter Baluk, Dan Liu, Nina K Serwas, Jason G Cyster, Donald M McDonald, Matthew F Krummel, Mark R Looney","doi":"10.1101/2024.09.12.612619","DOIUrl":"https://doi.org/10.1101/2024.09.12.612619","url":null,"abstract":"Intravital microscopy has enabled the study of immune dynamics in the pulmonary microvasculature, but many key events remain unseen because they occur in deeper lung regions. We therefore developed a technique for stabilized intravital imaging of bronchovascular cuffs and collecting lymphatics surrounding pulmonary veins in mice. Intravital imaging of pulmonary lymphatics revealed ventilation-dependence of steady-state lung lymph flow and ventilation-independent lymph flow during inflammation. We imaged the rapid exodus of migratory dendritic cells through lung lymphatics following inflammation and measured effects of pharmacologic and genetic interventions targeting chemokine signaling. Intravital imaging also captured lymphatic immune surveillance of lung-metastatic cancers and lymphatic metastasis of cancer cells. To our knowledge, this is the first imaging of lymph flow and leukocyte migration through intact pulmonary lymphatics. This approach will enable studies of protective and maladaptive processes unfolding within the lungs and in other previously inaccessible locations.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"40 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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