mSphere最新文献

{"title":"Type 2 diabetes mellitus exacerbates vaginal group B <i>Streptococcus</i> colonization via impaired mucosal cytokine response.","authors":"Clare M Robertson, Vicki Mercado-Evans, Addison B Larson, Holly Branthoover, Samantha Ottinger, Marlyd E Mejia, Zainab A Hameed, Lindsey A Gonzalez, Camille Serchejian, Libbie Ogilvie, Jacob J Zulk, Kathryn A Patras","doi":"10.1128/msphere.00027-26","DOIUrl":"10.1128/msphere.00027-26","url":null,"abstract":"<p><p>Type 2 diabetes mellitus (T2D) is a metabolic disorder that confers increased risk of microbial infections, including those caused by the opportunistic pathogen group B <i>Streptococcus</i> (GBS). Asymptomatic GBS vaginal carriage is a notable reservoir for infection, but the impact of T2D on the vaginal mucosa and GBS colonization is not fully understood. We employed a diet-induced mouse model of T2D to investigate the impact of diabetes on glucose availability, vaginal microbiome composition, and vaginal cytokines at baseline and in response to GBS. We observed enhanced susceptibility of diabetic mice to GBS vaginal colonization and reproductive tract dissemination. Despite experiencing hyperglycemia, diabetic mice did not exhibit elevated glucose in the reproductive tract. Regarding the vaginal microbiota, diabetic mice had minimal compositional differences, with decreased <i>Mammaliicoccus</i> being the only significant taxonomic variance. Vaginal cytokine profiling revealed consistently depressed cytokines in diabetic mice, beginning with KC at baseline and expanding to eight pro-inflammatory cytokines post-GBS infection. Diabetic mice exhibited decreased proportions of uterine neutrophils and, following GBS exposure, also displayed an expanded vaginal γδ T cell compartment compared with controls. Pairing cytokine observations with GBS colonization revealed a correlation between delayed vaginal IL-1α induction and persistent vaginal GBS, suggesting that vaginal cytokine deficiency may contribute to diabetic GBS phenotypes. Intravaginal supplementation with rIL-1α resolved GBS burden differences between diabetic mice and controls, confirming that deficient vaginal cytokines contribute to diabetic GBS vaginal persistence. These findings advance our understanding of diabetic vaginal mucosal susceptibility to pathogens and support the potential for immunological intervention.IMPORTANCEPeople with T2D are more susceptible to microbial infections, but there is limited understanding of the mechanisms that drive this vulnerability. One possibility is that T2D enhances the colonization of opportunistic pathogens, like GBS, in mucosal reservoirs as a precursor to infection. In this study, we used a diabetic mouse model to test whether diabetes alters the vaginal mucosa to promote GBS colonization. We found that increased vaginal GBS colonization in diabetic mice was not linked to tissue glucose availability or changes in the vaginal microbiome but instead was associated with impaired vaginal immune responses. These findings provide a foundation for translational approaches to reduce GBS persistence and dissemination in at-risk individuals.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0002726"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410995/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148422862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: Damage in RNA virus infection-a feature or a bug?","authors":"Katherine C Barnett","doi":"10.1128/msphere.00255-26","DOIUrl":"10.1128/msphere.00255-26","url":null,"abstract":"<p><p>Katherine C. Barnett studies cell death and damage in viral infections, focusing on how these processes activate innate immunity and shape viral pathogenesis. In this mSphere of Influence article, she highlights how the work of Aguirre et al. (Nat Microbiol 2:17037, 2017, https://doi.org/10.1038/nmicrobiol.2017.37) changed her perception of how RNA viruses interact with pattern recognition receptors and how later the work of Wang et al. (Nature 616:152-158, 2023, https://doi.org/10.1038/s41586-023-05851-w) shaped her concept of cell death in viral infections. Framing these findings in different models of pathogen detection by the innate immune system, she reflects on how her understanding of the role of damage in RNA virus infection has evolved throughout her career.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0025526"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410963/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148422874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunization with N2 neuraminidase can protect mice against a heterologous influenza A virus challenge even in the absence of cross-NA inhibiting antibodies.","authors":"Laura Amelinck, Anouk Smet, Tine Ysenbaert, Koen Sedeyn, William Warren, Raul Gomila, Thorsten U Vogel, Xavier Saelens, João Paulo Portela Catani","doi":"10.1128/msphere.00300-26","DOIUrl":"10.1128/msphere.00300-26","url":null,"abstract":"<p><p>Neuraminidase (NA) inhibition (NAI) titers have been identified as an independent correlate of protection against influenza. Few studies, however, have investigated the breadth of NA-based immune protection. Previously, we have reported that N2 NAs derived from human H3N2 viruses that circulated between 2009 and 2017 can be subdivided into four antigenic groups. Here, we (i) immunized mice with recombinant soluble tetrameric NA from H3N2 strains representing those four antigenic groups or (ii) passively transferred N2 NA immune serum into naïve mice to evaluate the breadth of protection against a heterologous HxN2 influenza virus challenge. We show that the breadth of protection goes beyond the breadth of NAI and is associated with the presence of cross-reactive antibodies. Interestingly, in the absence of cross-reactive antibodies, the immunization of DBA/2J mice with heterologous recombinant N2 NA from A/Perth/16/2019 was associated with an early onset of disease upon challenge with HxN2 virus with NA from A/Indiana/08/2011 (H3N2) or with H2N2 A/Singapore/1/1957. This early onset of disease was unidirectional and was not observed when mice that had been immunized with recombinant NA from A/Indiana/08/2011 were challenged with HxN2 virus with NA from A/Perth/16/2009 (H3N2).IMPORTANCEAlthough seasonal influenza vaccines mainly target hemagglutinin (HA), the viral surface protein neuraminidase (NA) can also induce protective immunity. In an earlier work, we profiled neuraminidase inhibition across a broad panel of H3N2 viruses and defined four major NA antigenic groups. Here, we tested whether immunizing mice with NA elicits cross protection against challenge viruses carrying NAs from different antigenic groups. Our data separate the predictive value of NA-binding and NA-inhibiting antibody responses and show that protection can extend beyond measurable NA inhibition when cross-reactive antibodies are present. These findings support the inclusion of standardized NA antigens as vaccine components to enhance the breadth of protection offered by seasonal influenza vaccine.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0030026"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410959/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148251884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: Understanding atopic disease through the dynamics of skin-microbe interactions.","authors":"Flavia G Costa","doi":"10.1128/msphere.00797-25","DOIUrl":"10.1128/msphere.00797-25","url":null,"abstract":"<p><p>In this mSphere of Influence article, Flavia G. Costa explores dynamics of host-microbe interactions on the skin through three papers that have shaped her perspectives in this field. She first discusses a publication by Radhouani et al., which showed how <i>S. aureus</i> skin colonization promotes lung inflammation through eosinophil reprogramming. She then highlights the potential protective benefits of understudied skin commensals as described by Myles et al., finally concluding with a recent study demonstrating how byproducts from industrialization can disrupt these beneficial interactions as described by Zeldin et al. This body of work has shaped her approach toward understanding the microbial dynamics that contribute to the rise in incidence of atopic diseases.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0079725"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410751/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148422886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: The bacterial growth law-revisiting the old to discover the new.","authors":"Katsuya Fuchino","doi":"10.1128/msphere.00046-26","DOIUrl":"10.1128/msphere.00046-26","url":null,"abstract":"<p><p>Katsuya Fuchino works in the field of bacterial cell biology and applied microbiology. In this mSphere of Influence article, he reflects on how the studies \"Dependency on medium and temperature of cell size and chemical composition during balanced growth of <i>Salmonella typhimurium</i>\" by Schaechter et al. and \"A metabolic sensor governing cell size in bacteria\" by Weart et al. inspired the field of bacterial cell size homeostasis and his current research. Additionally, he highlights the value of reading classic, original papers that influenced the field, as a means to reflect on paradigm shifts and generate new ideas.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0004626"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410753/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148339409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Applying PCR cycle autonormalization to PacBio full-length 16S rRNA library preparations: impacts on error rates and sequence distributions.","authors":"Charles J Mason, Mikinley Weaver, Karma R Kissinger, Melissa A Johnson, Duan C Copeland, Kirk E Anderson, Scott M Geib","doi":"10.1128/msphere.00295-26","DOIUrl":"10.1128/msphere.00295-26","url":null,"abstract":"&lt;p&gt;&lt;p&gt;The bacterial 16S rRNA gene is widely used to characterize host-associated and environmental microbiomes, most commonly through sequencing short hypervariable regions. Recent improvements in PacBio sequencing chemistry and concatenation approaches can now enable high-throughput, full-length 16S rRNA gene sequencing with high accuracy and depth. However, errors introduced during library preparation remain a major limitation, particularly during PCR amplification of full-length amplicons, where error accumulation may be elevated due to longer sequence lengths. These challenges are amplified when samples vary widely in microbial biomass, making it difficult to select a single optimal number of PCR cycles. Here, we evaluated PCR cycle autonormalization for PacBio Kinnex full-length 16S rRNA gene sequencing across seven agriculturally relevant specimen types. We compared conventional fixed-cycle PCR protocols (20, 24, and 30 cycles) with an autonormalization approach in which individual reactions were terminated during exponential amplification based on real-time fluorescence thresholds. Under the workflow tested here, autonormalized libraries generally retained a high proportion of sequences following denoising and chimera removal, exhibited low residual error rates (&lt;0.005%), and yielded relatively even read distributions across heterogeneous sample inputs. Overamplified reactions (30 cycles) showed elevated residual error rates and greater sequence loss, particularly in samples with higher microbial biodiversity, whereas low-cycle libraries produced more variable read output among specimens. Importantly, the PCR protocol had relatively minor effects on overall community composition compared with specimen type. These results support PCR cycle autonormalization as a useful workflow strategy for heterogeneous full-length 16S library preparation, while also highlighting the importance of library design, pooling strategy, and downstream processing in shaping technical outcomes.IMPORTANCEAmplicon-based sequencing of the 16S rRNA gene is a foundational tool in microbiome research, yet PCR amplification remains a major source of library-preparation error. This challenge is magnified for full-length 16S rRNA sequencing and for workflows that process specimen types with widely varying microbial biomass. Selecting a single PCR cycle number can underamplify low-biomass samples or overamplify high-titer samples, increasing artifacts and sequence loss during downstream processing. Here, we show that PCR cycle autonormalization can be integrated into a PacBio full-length 16S rRNA workflow and, under the conditions tested, provides low residual error rates and relatively even sample representation across heterogeneous inputs. Autonormalization also enables blind pooling of amplicons without post-PCR quantification or equimolar normalization, reducing hands-on time and sample loss. These benefits make cycle autonormalization particularly valuable for high-thr","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0029526"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410752/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148302458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative analysis of mortality risk in SFTS using machine learning and genetic approaches.","authors":"Helin Zha, Lianzi Wang, Haoyang Sun, Jie Li, Yajing Wang","doi":"10.1128/msphere.00831-25","DOIUrl":"10.1128/msphere.00831-25","url":null,"abstract":"<p><p>This study presents a novel approach to predicting mortality risk in patients with severe fever with thrombocytopenia syndrome (SFTS) by integrating machine learning models and genetic analysis techniques. Utilizing clinical data from 107 confirmed cases, we developed several predictive models and identified the extreme gradient boosting (XGBoost) algorithm as the most effective, achieving an area under the curve (AUC) of 0.950. Further analysis was performed based on the 15 robust features selected and performance-optimized through the Boruta algorithm, integrating <u>Sh</u>apley <u>a</u>dditive <u>e</u>x<u>p</u>lanations (SHAP) plots to identify key variables such as IL-6, TNF-α, PLR, and AST. While Mendelian randomization did not reveal definitive causal pathways across all examined factors, notable associations were observed in specific metabolic-inflammatory pathways. These findings offer a foundation for enhanced predictive accuracy and mechanistic understanding of SFTS mortality.IMPORTANCESevere fever with thrombocytopenia syndrome (SFTS) is a tick-borne viral disease with high mortality, posing a major threat to public health in East Asia. This study develops a machine learning (ML) model to quickly predict SFTS patients' mortality risk using routine clinical tests, helping doctors stratify risks early. Moreover, by exploring causal pathways between bodily indicators and disease outcomes, it provides new clues for understanding why SFTS becomes severe, which could guide better treatments in the future.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0083125"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410981/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148258793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of gut viromes in four penguin species reveals diverse novel viruses and host-associated differences.","authors":"Kang Qi, Shuang Zhang, Xiaodong Su, Jiaheng Chen, Shiyin Huang, Yue Chen, Wang Li, Guanying Ni, Jieji Duo, Shixing Yang, Quan Shen, Xiaochun Wang, Yuwei Liu, Ping Wu, Hongfeng Yang, Likai Ji, Xiaolong Wang, Wen Zhang","doi":"10.1128/msphere.00848-25","DOIUrl":"10.1128/msphere.00848-25","url":null,"abstract":"<p><p>Penguins, as distinctive marine birds, play important roles in polar and sub-Antarctic ecosystems, yet the diversity and species-specific distribution of their gut viromes remain insufficiently understood. Here, we used viral metagenomics to characterize the cloacal viromes of four penguin species-<i>Spheniscus humboldti</i> (<i>S. humboldti</i>), <i>Pygoscelis papua</i> (<i>P. papua</i>), <i>Pygoscelis adeliae</i> (<i>P. adeliae</i>), and <i>Aptenodytes forsteri</i> (<i>A. forsteri</i>)-collected at Chimelong Ocean Kingdom. A total of 219 viral sequences representing potentially novel lineages were identified, with more than 94% sharing <80% amino acid similarity with previously known viruses. These sequences were assigned to several viral families, including Parvoviridae, Caliciviridae, Anelloviridae, Circoviridae, and Microviridae, among others. Marked interspecies differences in virome composition were observed: Parvoviridae dominated in <i>S. humboldti</i>, Microviridae were enriched in <i>P. papua</i>, Caliciviridae accounted for a substantial proportion in <i>A. forsteri</i>, and <i>P. adeliae</i> displayed the greatest overall virome diversity. Multiple-virus co-detections, particularly involving Parvoviridae, were frequent in <i>S. humboldti</i>. Phylogenetic analyses showed that many penguin-associated viruses clustered with viruses infecting other avian and fish hosts, suggesting possible dietary or environmental origins of some detected viral sequences. These findings expand current knowledge of penguin gut virome diversity and host-associated differences and provide a valuable foundation for evaluating the ecological roles, health implications, and transmission risks of penguin-associated viruses.IMPORTANCEThis study uncovers significant diversity in the gut viromes of four penguin species, revealing over 219 viral sequences representing potentially novel lineages, many of which showed host-associated distribution patterns. Using viral metagenomics, we identified notable interspecies differences, with Parvoviridae predominating in Spheniscus humboldti and Microviridae being enriched in Pygoscelis papua. These findings highlight the complexity of viral community structures in penguins, including frequent viral co-detections, which could impact host health and ecological adaptation. Additionally, novel bacteriophage communities were identified, emphasizing their potential role in shaping the gut microbiome and influencing viral dynamics. This work provides new insights into viral diversity in wildlife and lays the groundwork for future studies on viral transmission risks and ecological conservation.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0084825"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410756/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148387675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deciphering the <i>in vitro</i> mucin-driven interaction dynamics of a synthetic gut bacterial community.","authors":"Chong Yu, Jiayi Ao, Mengran Long, Ziqiong Xu, Shaohui You, Yanlei Jiao, Songling Zhu, Shu-Lin Liu, Shuang Wang, Hongxia Bao","doi":"10.1128/msphere.00289-26","DOIUrl":"10.1128/msphere.00289-26","url":null,"abstract":"<p><p>The gut microbiota is a complex microbial community that plays a crucial role in host health. Environmental and biological factors within the ecosystem influence the dynamic interactions among its members. Although dietary and host-derived nutrient availability play a key role in shaping microbial ecology and interaction patterns, the dynamics of these interactions within the mucus layer remain poorly understood. In this study, we analyzed a synthetic community comprised of six species with variable abilities to utilize mucin. We performed <i>in vitro</i> growth analyses and monitored the interactions among community members in monoculture, co-culture, and community batch culture under different nutrient conditions. Our results showed that positive interactions were prevalent among bacteria when mucin served as the sole carbon source. In contrast, the addition of glucose or high nutrient availability significantly increased inter-bacterial competition. These findings suggest that mucin mitigates competitive antagonism and potentially promotes community diversity. Further <i>in vivo</i> studies supported the role of mucin in increasing community diversity and modulating bacterial metabolic patterns. Deciphering these intricate relationships is essential for understanding how gut microbiota stability is maintained, and what factors might disrupt this delicate balance.IMPORTANCEThe gut microbiota is essential for host health, yet microbial interactions within the intestinal mucus layer remain poorly understood. Current understanding of gut microbial ecology is largely based on nutrient-rich media that do not accurately reflect the mucosal environment. Here, we demonstrate that when bacteria rely solely on mucin as a carbon source, cooperative interactions predominate. In contrast, the introduction of simple sugars shifts the balance toward intensified interbacterial competition. Mucin mitigates competitive antagonism, promotes resource utilization, and enhances community diversity. By demonstrating that mucus actively shapes microbial interaction patterns, this study provides a mechanistic framework for understanding gut ecosystem resilience. Furthermore, these findings support the development of more physiologically relevant <i>in vitro</i> models for predicting gut microbial dynamics and may guide microbiome-based therapies.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0028926"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410983/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148422731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Plasmodium falciparum</i> leucine-rich repeat 5 disruption alters the transcription progression during asexual and sexual stage development.","authors":"Camilla V Pires, Chengqi C Q Wang, Min Zhang, Chiara Micchelli, Prem Prakash, Jenna Oberstabler, Julian C Rayner, Thomas D Otto, John H Adams","doi":"10.1128/msphere.00186-26","DOIUrl":"10.1128/msphere.00186-26","url":null,"abstract":"<p><p>How <i>Plasmodium falciparum</i> responds to stresses such as antimalarial drug pressure and uses them to trigger developmental decisions such as the production of sexual gametocytes remains poorly understood. Here, we investigated the function of the leucine-rich repeat protein LRR5 (PF3D7_1432400). Previous genome-wide <i>piggyBac</i> mutagenesis established that disruptions of LRR5 increase sensitivity to artemisinin and heat-shock and enhance gametocyte production. Using an optimized single-cell RNA sequencing approach applied to unsynchronized parasite cultures, we profiled transcriptional heterogeneity across the intraerythrocytic developmental cycle in wild-type NF54 parasites and an isogenic LRR5 <i>piggyBac</i> mutant (LRR5pB). Disruption of LRR5 reshaped the parasite transcriptome across multiple stages, with marked downregulation of pathways involved in protein folding, protein export, and host-cell remodeling, including Maurer's cleft components, the PTEX export machinery, and the exported chaperone HSP70-x. In contrast, cytoplasmic heat-shock proteins and translation-associated genes were upregulated, suggesting a compensatory stress response. Notably, LRR5pB parasites displayed premature activation of gametocyte-associated transcriptional programs, including elevated expression of the early gametocyte marker Pfs16, which was confirmed at the protein level. Pseudotime analysis further revealed accelerated transcriptional progression with mistimed expression of trophozoite-, merozoite-, and gametocyte-associated gene clusters. Together, these findings suggest LRR5 as a link between proteostasis, host-cell remodeling, and developmental timing in <i>P. falciparum</i>.IMPORTANCEMalaria parasites must survive environmental stress while deciding whether to continue asexual replication in the blood stream or instead commit to sexual gametocytes for transmission. Understanding how these processes are coordinated is critical for developing new strategies to block both disease and transmission. In this study, we used single-cell transcriptomics to examine how disruption of the <i>P. falciparum</i> protein LRR5 alters parasite development. By analyzing individual parasites from unsynchronized cultures, we uncovered hidden changes in stress-response pathways, host-cell remodeling, and early activation of genes linked to sexual development. Our findings show that loss of LRR5 weakens the parasite's ability to manage stress while prematurely priming parasites for gametocyte formation. These results suggest LRR5 as a link to stress adaptation to transmission potential.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0018626"},"PeriodicalIF":4.0,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13410765/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148252534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}