The advent of CRISPR-based genome editing has revolutionized crop improvement, offering unprecedented precision and efficiency in modifying key agronomic traits. This review comprehensively examines the mechanisms, applications, and future potential of CRISPR technology in enhancing global crop production. CRISPR-Cas systems, originally identified as adaptive immune mechanisms in bacteria and archaea, have been repurposed for targeted genome editing in plants. The CRISPR-Cas9 system, in particular, has emerged as a powerful tool for introducing site-specific double-strand breaks, enabling precise genetic modifications. The three-stage process of adaptation, expression, and interference underlies the CRISPR mechanism, with guide RNAs directing Cas endonucleases to specific genomic loci. Advances in CRISPR technology have expanded its applications beyond gene knockouts, encompassing base editing, prime editing, and epigenome editing. These innovations have facilitated the development of crops with enhanced yield, stress tolerance, disease resistance, nutritional content, and post-harvest quality. However, challenges related to off-target effects, regulatory hurdles, ethical concerns, and public acceptance must be addressed to fully harness the potential of CRISPR in agriculture. Integration of CRISPR with other cutting-edge technologies, such as synthetic biology, artificial intelligence, and high-throughput phenotyping, holds immense promise for accelerating crop improvement efforts. As research continues to refine CRISPR tools and expand their applicability across diverse plant species, this transformative technology is poised to play a pivotal role in shaping a sustainable, resilient, and productive global food system for future generations.

Klebsiella oxytoca is a clinically significant opportunistic bacterium that contributes to global morbidity and mortality. Despite its clinical relevance, the genomic diversity, virulence factors, and immune evasion mechanisms of this pathogen are not yet fully elucidated. This review provides an overview of current knowledge on antibiotic resistance in K. oxytoca, a characteristic that poses a significant challenge to therapeutic options. We discuss available bacterial vaccines and their role in mitigating antimicrobial resistance, reducing infection rates, and lowering antibiotic use. We also examine the challenges of developing a vaccine against K. oxytoca using traditional approaches, including reliance on animal testing, high production costs, and the lengthy timelines associated with vaccine development. We also suggest that emerging tools such as subtractive proteomics, reverse vaccinology, artificial intelligence, and machine learning offer promising effects to accelerate the identification of vaccine candidates against this pathogen even though few studies have applied these tools specifically to K. oxytoca. Overall, despite progress in understanding K. oxytoca, further research is needed to develop innovative therapeutic and preventive strategies, including effective vaccines.

Background Ischemic stroke (IS) induces profound dysregulation of the neuro–molecular innate immune–vascular network, yet the molecular immune states and regulatory mechanisms of key cellular subpopulations remain insufficiently defined. Although traditional Chinese medicine (TCM) exhibits multi-target immunomodulatory potential, its cell-type and cell-state–specific actions within the ischemic brain microenvironment at single-cell resolution remain unclear. Methods Single-cell RNA sequencing was used to construct a cellular atlas of the ischemic mouse brain, followed by integrative bioinformatic analyses to characterize innate immune–related neural cell subpopulations and their regulatory networks. Network pharmacology and molecular docking were applied to identify salidroside (SAL), a major active compound of Rhodiola , and predict its potential molecular targets. In vivo experiments were performed to validate cellular and molecular changes associated with SAL treatment. Results In a mouse model of IS, ischemic injury induced pronounced imbalances across multiple immune and glial cell subpopulations. A transcriptionally defined Notch1 + Hes5 + astrocyte (ASC), enriched for progenitor-like and reparative gene signatures, was markedly reduced after ischemic injury, whereas reactive SerpinA3N + ASC and pro-inflammatory Sell + microglia (MG) were expanded. Additionally, alterations were observed in immune-regulatory cell populations, including Cxcl12 + endothelial cells (ECs) and Gpr34 + Ptgs1 + MG. In vivo validation showed that SAL treatment was associated with modulation of Notch1/Hes5 signaling in ASC, reduced reactive ASC features, and partial normalization of ECs alterations, accompanied by changes consistent with attenuated neuroimmune activation. These effects coincided with altered intercellular communication, particularly involving NOTCH signaling. Conclusions This study provides single-cell–level insights into innate immune microenvironment remodeling following IS and identifies a Notch1 + Hes5 + ASC subpopulation with transcriptional features associated with reparative-related programs and responsiveness to SAL. The findings suggest that SAL-associated neuroprotection was accompanied by modulation of ASC states and immune–glial communication, highlighting the potential of SAL-associated immunoregulatory effects at the single-cell level in IS.

Parkinson's disease (PD) is characterised by α-synuclein aggregation, dopaminergic neuron loss and chronic neuroinflammation. Disruption of the blood-brain barrier enables immune cell infiltration, including dendritic cells (DCs) and CD4+ T-cells, contributing to disease progression. To explore peripheral immune mechanisms in PD, we isolated DCs and CD4+ T-cells from the blood of 17 PD patients and 10 controls using magnetic separation, followed by flow cytometry and single-cell RNA sequencing. Cell-type annotation identified CD4+ T-cell and DC subtypes, including rare DC3 cells. PD patients showed reduced circulating DCs, with no change in CD4+ T-cell levels. Differential gene expression and pathway analysis suggest CD4+ effector memory T-cells (TEMs) and cDC2s as important mediators of immune responses in PD, enriched for immune-related pathways including T-cell activation and antigen presentation. Our findings implicate specific immune subsets in PD-associated neuroinflammation, suggesting cDC2s and CD4+ TEMs as potential targets for immunomodulatory strategies.

Chicken B cell development represents a remarkable evolutionary divergence from mammalian paradigms, featuring unique three-stage ontogeny centered on the bursa of Fabricius, an avian-specific primary B cell lymphoid organ. Unlike mammals where B cells develop continuously in bone marrow, chickens utilize a temporally restricted program spanning pre-bursal (E5-E14), bursal (E8-hatching), and post-bursal phases (hatching-bursal involution), each characterized by distinct molecular mechanisms and anatomical sites. In this review, we documented chicken B cell development in three developmental phases (pre-bursal to post-bursal phases) and compared it with mammalian B cell development mostly in humans as a representative mammalian model. In chicken, while the embryonic bursa of Fabricius serves as the primary B cell receptor (BCR)-dependent B cell developmental organ, it also supports BCR-independent early colonization followed by extensive activation-induced cytidine deaminase (AID)-mediated gene conversion rather than V(D)J recombination for antibody diversification. Recent gene knockout studies reveal paradoxical BCR signaling requirements for post-hatched chicken B cell development, with J H knockout chickens lacking post-hatched B cells, while recombination activating gene 1 ( RAG1 ) knockout chickens maintain post-hatched bursal B cell populations through alternative pathways. Single-cell RNA sequencing has identified previously unrecognized chicken B cell subpopulations and provided molecular signatures for bursal and post-bursal B cells, addressing longstanding phenotypic marker limitations. These findings demonstrate that effective chicken humoral immunity can be achieved through alternative evolutionary strategies, with reduced dependence on RAG1 activity compared to mammalian systems, providing new perspectives on immune system evolution and adaptive immunity mechanisms.

Introduction Ethnobotanically, Andrographis paniculata aerial parts have been used to alleviate the symptoms of viral infections, colds, and sore throats. Although the utility of Andrographis for these indications has had widespread acceptance, multiple knowledge gaps, regarding mechanisms and specific clinical applications, exist for this botanical. Prior rodent studies have limited applicability to inform future clinical applications due to routes of administration used, nonclinical high doses tested, and exclusive evaluation of chemically purified compounds. Methods Experiments used a commercial extract of A. paniculata manufactured from aerial parts that was standardized to contain ≥10% andrographolides. In vitro evaluation measured NF-κB in THP-1 monocytes, iNOS in RAW 264.7 macrophages and Nrf2 in HepG2 cells. In vivo research used a non-lethal influenza A mouse model with two groups ( A. paniculata treatment and vehicle control). Following infection, mice were gavaged daily for 8 days–treatment group received 250 mg extract/kg body weight (dose equivalent to the human label-recommended daily intake). Results The extract inhibited NF-κB and iNOS while activating Nrf2. Although andrographolide contributed to these effects, the findings indicate the presence of additional active constituents that have not yet been characterized. In the mouse model, oral administration of the extract produced a roughly five-fold reduction in lung viral load by day 3 post-infection ( p < 0.021) compared with control animals. Discussion Based on the clinically relevant design of our mouse model, the results lay the groundwork for future preclinical research to determine optimal treatment schedule/dosage and immune mechanisms, thereby informing the design for future resilience efficacy clinical trials.

Metal ions are involved in many biological functions such as enzyme catalysis, signal transduction and gene expression regulation in biological system. They play multiple roles in the pathogenesis and immune escape mechanism of gastric cancer(GC). This review begins by outlining the fundamental biological roles of metal ions, highlighting its significance in tumor development. We focus on elucidating how metal ions modulate the gastric cancer immune landscape by regulating immune cell functions, and participating in specific signaling pathways. Additionally, the potential of metal ion interference as an emerging therapeutic strategy for tumors is discussed, along with the prospects for applying metal-based nanomaterials in the treatment of gastric cancer. Additionally, we discuss the crosstalk between ferroptosis and cuproptosis mediated by metal ions, which provides a novel perspective for understanding metal ion-dependent tumor cell death. The potential of metal ion interference as an emerging therapeutic strategy and the application prospects of metal-based nanomaterials in gastric cancer treatment are summarized. Finally, we point out key future research needs, including clarifying the dynamics of metal ions in the gastric cancer microenvironment, standardizing metal-related biomarkers for clinical stratification, and optimizing the safety and targeting of metal-based therapies. This review comprehensively summarizes the regulatory roles and mechanisms of metal ions in the gastric cancer immune microenvironment, offering theoretical support for the development of precision therapeutic strategies targeting metal ion homeostasis.

Estrogen receptor (ER) positive breast cancer is the most prevalent subtype, commonly responsive to endocrine therapies. Immune checkpoint inhibitors (ICIs) have limited efficacy in ER-positive disease, highlighting the need for the development of combination immunotherapies for these patients. We previously established that nitroso-N-methylurea-induced mammary tumors in outbred Sprague-Dawley rats mimic immune evasive mechanisms and the heterogeneity of ICI response observed in patients. We identified a "luminal growing" gene signature in ER-positive tumors, which correlated with tumor growth and immune-related differences. Here, we evaluated targeting candidates from this signature KMT5B/C and IKBKE using inhibitors A-196 and IKBKEi respectively, alongside anti-estrogen (fulvestrant) and a TGFβ blocking antibody (NIS793), both individually and in combination with αPD-L1, within this rat model. Fulvestrant emerged as the most effective treatment, inducing regression of most existing tumors and reducing on-treatment tumor burden when combined with αPD-L1. A-196, while ineffective as a monotherapy, demonstrated enhanced response when combined with αPD-L1. Comprehensive tumor profiling through polychromatic flow cytometry and single-cell RNA sequencing revealed that A-196 induced a luminal-to-basal shift in tumor epithelial cells, enhancing antigen presentation, whereas epithelial-to-mesenchymal transition was linked to fulvestrant resistance. Our findings underscore the value of the rat mammary tumor model for preclinical studies in ER-positive breast cancer and advocate for the further validation and potential clinical development of KMT5B/C inhibitors to enhance the efficacy and broaden the applicability of ICI therapy in cancer patients.

Introduction The killer-cell immunoglobulin-like receptor ( KIR ) gene cluster exhibits complicated diversity in haplotype content, copy-number variation (CNV), and allelic polymorphism. To date, 2,286 distinct KIR alleles have been released in the IPD-KIR Database. However, little is known about the impact of high-resolution-level KIR allelic polymorphisms on leukemia. Our previous study showed that the KIR AA genotype carrying more inhibitory genes conferred differential protection against leukemia in the Chinese Southern Han population. Herein, we hypothesized the impact of KIR alleles in the KIR A haplotype and cognate human leukocyte antigen (HLA) ligand on leukemia. Methods The study cohort included 318 ALL patients, 336 AML patients, and 306 unrelated healthy controls. All the study samples were subject to HLA-A , - B , and - C sequencing-based genotyping (PCR-SBT) and high-resolution KIR genotyping for all the seven functional KIR genes ( KIR2DL1 , KIR2DL3 , KIR2DL4 , KIR3DL1 , KIR3DL2 , KIR3DL3 , and KIR2DS4 ) on the KIR A haplotype. HLA and KIR genotypes were assigned using Assign 4.7.1 software. Results In the present study, our high-resolution genetic analysis revealed protective KIR – HLA interactions in individuals with the KIR AA genotype. The strong inhibitory KIR2DL1*00201 + C2 interaction reduced ALL risk ( p = 0.01), while KIR2DL1*00302 + C2 ( p = 0.008), KIR2DL3*00201 + C1 ( p = 0.03), and KIR3DL1*00501 + Bw4 80I ( p = 0.008) interactions protected against AML ( p < 0.05). However, the functionally weaker inhibitory KIR2DL1*004 + C2 interaction conferred ALL risk ( p = 0.01) in individuals with the KIR Bx genotype. Notably, we found that the allelic polymorphisms of the structure gene KIR3DL3 were associated with the occurrence of leukemia. KIR3DL3*001 tends to confer protection against AML (8.4% vs. 1.3%, p = 0.004, Pc = 0.06), whereas KIR3DL3*009 conferred susceptibility to AML (29.3% vs. 47.1%, p = 0.001, Pc = 0.016). KIR3DL3*001 differs from KIR3DL3*009 by an amino acid substitution of non-charged asparagine (N) to charged histidine (H) in its transmembrane domain, suggesting that this functional variant site KIR3DL3_N300H may play a critical role in the occurrence of leukemia in the Chinese population. Conclusion These data suggest that KIR AA individuals possess strong inhibitory interactions of KIR alleles and HLA , arming KIR AA + NK cells to meditate stronger alloreactivity and cytotoxicity against leukemia cells with lowered HLA expression. Our findings may provide valuable insights into leukemia pathogenesis and better understanding of the immune mechanisms.

Introduction Insects possess a well-developed innate immune system, which encompasses both cellular and humoral mechanisms. On the basis of the similarities in neuropeptide actions between insects and vertebrates, we assume that neuropeptides such as tachykinin-related peptides ( TRP s) regulate insect immune responses and are themselves modulated following infection. Methods In this study, we examined how immune activation affects the expression of genes encoding TRP precursors and receptors ( TRP and TRPR ) and whether TRPs directly modulate selected immune mechanisms in the pest species Tenebrio molitor . In our experiment, we combined cytokine stimulation, genetic knockdown, pharmacological inhibition, immune profiling, and survival analysis to dissect neuropeptide function. Results Our results revealed two important insights. First, after activation of the immune system, TRP and TRPR genes were significantly downregulated in the nervous system and immune-related cells. These changes are closely correlated with the changes of the expression level of immune genes. We then show, using Spantide II, a potent antagonist of TRPR, and RNAi knock down of TRP and TRPR , the modulation of key processes of the T. molitor humoral response. This includes the over expression of genes encoding antimicrobial peptides and the important arthropod immune effector phenoloxidase activity. Discussion Our findings highlight a compelling association between the TRP and immune regulation in Tenebrio and provide insights into the hormonal regulation of physiological processes in insects. Our research also provides novel insights that can contribute to the development of sustainable pest control strategies amid increasing insecticide resistance.

Spontaneous control of HIV replication has been primarily associated with cellular immune responses. However, it remains multifactorial, and viral determinants, innate and humoral immune responses could be additional relevant contributors. Furthermore, posttreatment control cases reveal new roles for humoral responses. This review describes the direct, indirect and passive roles of humoral responses in HIV control. New evidence supports the role of the humoral responses in the natural control of HIV. Indeed, a strong association has been reported between polyfunctional humoral responses and slow disease progression, highlighting the active role of both neutralizing and nonneutralizing antibodies in natural control. Moreover, broadly neutralizing antibodies (bNAbs) are being considered as therapeutic interventions to directly or indirectly mediate HIV control in cure strategies. Data from the latest clinical trials show that treatment with bNAbs may induce high-quality CD8 T-cell responses, pointing to bNAbs as a major indirect strategy to induce durable HIV control. Finally, humoral responses can serve as biomarkers for monitoring elite controllers and have been useful to identify stable aviremic controllers, providing new clinical monitoring tools. Humoral responses are relevant for understanding immune mechanisms of control, for defining therapeutic interventions and for the clinical follow-up of elite controllers.

Background/Objectives: Vaccination coverage among adolescents remains below the recommended target, highlighting the need for effective educational strategies to improve vaccine knowledge. This study aimed to assess baseline knowledge of vaccines and immune mechanisms among adolescents and to evaluate whether a school-based educational intervention can improve knowledge related to vaccination. Methods: A prospective quasi-experimental pre–post study was conducted between 1 February 2025 and 1 June 2025 among adolescents aged 14–19 years attending high schools in Southern Italy. The intervention was based on the e-Bug educational module and delivered by trained nurses through interactive lessons, gamification, and guided discussions. Vaccine-related knowledge was assessed using a questionnaire administered before and after the intervention. Changes in knowledge scores were analyzed using paired statistical tests, and the effect size was estimated. A stepwise multivariate linear regression model was employed to identify factors associated with post-intervention test scores, with statistical significance set as p ≤ 0.05. Results: Among 386 participants, the majority were female (74.2%), the average age was 15.8, and 15% reported a chronic medical condition. Knowledge gaps were observed at baseline, particularly regarding the items on recommended adolescent vaccinations (37.4%), the definition of innate immunity (25.6%), and the mechanism of vaccines’ action (51%). After the intervention, all the items showed an improvement in correct answers, statistically significant for 5 of the 7 analyzed items (r = 0.364, p < 0.001). The most pronounced improvement was in the awareness of age-specific recommended vaccines (61.2%). The multivariate linear regression analysis showed that those with higher pre-intervention test scores, those who had parents with chronic medical conditions, those whose fathers worked, and those willing to participate in similar future interventions were more likely to achieve higher post-intervention test scores. Conclusions: School-based interventions may represent an effective strategy for enhancing adolescents’ knowledge related to vaccination, but further studies with control groups and long-term follow-up are needed to confirm effectiveness.

Breast cancer remains a leading cause of cancer-related mortality despite advances in targeted therapies and immune checkpoint inhibitors (ICIs). While ICIs have reshaped therapeutic paradigms in multiple solid tumors, their clinical efficacy in breast cancer is predominantly restricted to triple-negative breast cancer (TNBC), underscoring the necessity for innovative immunotherapeutic modalities with broader applicability. Bispecific antibodies (BsAbs) are engineered molecules capable of simultaneously engaging tumor-associated antigens (TAAs) such as EGFR, EpCAM, Trop-2, CEACAM5, MUC1, and PSMA, in addition to B7-H4, HER2, and PD-L1, alongside immune effector cells, primarily CD3+ T lymphocytes. This dual targeting promotes the formation of a cytolytic immunological synapse, leading to T-cell activation, proliferation, and targeted tumor cell lysis via granzyme/perforin pathways. Advances in BsAb design-including optimization of affinity constants, valency, Fc engineering to modulate effector functions, and molecular formats (e.g., BiTEs, DARTs, tandem scFvs)-have improved tumor penetration, pharmacokinetics, and reduced off-target toxicity. Preclinical and early-phase clinical data demonstrate robust antitumor activity of BsAbs, both as monotherapy and in synergistic combinations with chemotherapy, targeted agents, or ICIs, potentially overcoming tumor microenvironment-mediated immunosuppression and immune escape mechanisms. Major challenges include heterogeneity and temporal variability of TAA expression, cytokine release syndrome (CRS) mitigation strategies, pharmacodynamic optimization, and delivery methods to maximize tumor bioavailability while limiting systemic toxicity. This review details the molecular mechanisms, preclinical evidence, clinical trial outcomes, and translational challenges for BsAbs in breast cancer, highlighting their transformative potential in expanding immunotherapeutic efficacy beyond current limitations.

Introduction: Cancer is one of the leading causes of death worldwide, and while chemotherapy and radiotherapy remain primary treatments, they are limited by poor tumour specificity, systemic toxicity and drug resistance. Nanoparticle (NP)-based drug delivery offers a promising alternative, with improved solubility, controlled release, and targeted delivery, to enhance efficacy and minimise adverse effects. However, delivery remains inefficient, often resulting in off-target accumulation, immune clearance, renal filtration, and poor tumour penetration. This review explores how biological and physiological barriers limit NP-based delivery and evaluates strategies such as biomimetic cloaking, synthetic surface modifications, and tumour-responsive targeting to improve therapeutic accumulation and efficacy. Methods: This review looked at strategies to improve NP delivery in cancer by analysing their biodistribution and interaction with biological surfaces and barrier. Peer-reviewed studies were identified through PubMed and Google Scholar using keywords such as “nanoparticle clearance”, “tumour penetration”, “immune evasion”, “biomimetic cloaking”, and “PEGylation”. Articles published between 1990 and 2025 were included if they investigated NP delivery barriers, biodistribution, or design strategies that are specific to cancer. Review articles that provided contextual backgrounds were also considered. Results: Biomimetic cloaking of nanoparticles improves NP circulation and tumour accumulation. Red blood cell membrane-coated NPs evade immune recognition and reduce opsonisation by displaying self-marker proteins that prolong circulation. Platelet membrane cloaking can improve tumour targeting by taking advantage of platelet interactions with damaged vasculature and circulating tumour cells. PEGylation forms a hydrophilic barrier, preventing opsonisation and clearance by macrophages. Zwitterionic coatings neutralise surface charge to reduce renal filtration, while NP size optimisation (50-150 nm) helps balance circulation time and tumour permeability. However, tumour penetration remains a challenge, particularly in solid tumours with high interstitial pressure. Implications: These findings show the importance of integrated NP systems that combine immune evasion, tumour targeting, and local release mechanisms. These innovations align with the shift towards personalised and biomimetic therapies, improving targeted cancer treatments. Continued research is needed to bridge the gap between preclinical findings and clinical applications, improving cancer treatment outcomes. Future work should focus on scalable, low-immunogenic solutions to the systemic and tumour-specific barriers to unlock the full potential of cancer nanomedicine.

The liverwort Marchantia polymorpha has emerged as a model for studying plant immunity in bryophytes, providing unique insights into conserved defense mechanisms across land plants. By contrast, Marchantia-specific immune mechanisms remained largely underexplored. In this study, we investigated the genetic basis of quantitative resistance in M. polymorpha against the fungal pathogen Colletotrichum nymphaeae, a naturally occurring compatible parasite. Through a combination of phenotypic, cytological, and transcriptomic approaches, combined with genome-wide association studies (GWAS), we identified key defense-related genes and pathways. Leveraging the biological and genetic variability present in a collection of natural M. polymorpha accessions, we highlight the role of horizontally transferred microbial-like terpene synthase genes, which may contribute to the exceptional terpene diversity of liverworts and potentially play a role in pathogen resistance. GWAS uncovered candidate loci associated with resistance traits, implicating both core immune components and specialized metabolic pathways. Transcriptomic analyses performed on two accessions with contrasting phenotypes after inoculation with C. nymphaeae revealed the upregulation of accession-specific and horizontally acquired genes. These results provide insights into the specific molecular underpinnings of bryophyte immunity and underscore the evolutionary significance of horizontal gene transfer and specialized metabolites in shaping plant-pathogen interactions.

Condyloma acuminatum (CA), primarily caused by low-risk HPV6/11, is a benign proliferative disease that is difficult to cure and prone to recurrence. However, the molecular and immune mechanisms underlying relapse remain unclear. We combined metabolomic profiling with single-cell RNA sequencing to investigate recurrence-associated changes. Metabolomics revealed dysregulation of ascorbate and aldarate, glycerophospholipid, purine, and arginine/proline metabolism in recurrent CA. Single-cell analysis identified altered expression of metabolism-related genes (AMD1, GSTM3, ALDH3A1, GPX1, GPX4) in keratinocytes, associated with hyperproliferation, impaired differentiation, and ferroptosis resistance. Immune profiling identified transcriptionally distinct myeloid subpopulations in recurrent CA lesions, including M2 macrophages and dendritic cells. KEGG analysis indicated enrichment of antigen processing, phagosome, and endocytosis pathways in M2 macrophages, and antigen processing and viral carcinogenesis in dendritic cells, suggesting altered immune regulatory states. Notably, the key polyamine biosynthesis regulator AMD1 was downregulated in both M2 macrophages and dendritic cells in recurrent lesions, paralleling metabolic evidence of altered arginine-polyamine pathways. These findings suggest that recurrent CA involves coordinated metabolic dysregulation across keratinocytes and immune cells, highlighting potential targets for immunometabolic intervention.

Filament-driven activation of the Kongming antiviral system by deoxyinosine triphosphate.

A Group A Streptococcus (GAS, Streptococcus pyogenes) is an exclusively human pathogen whose virulence is driven by a diverse array of surface structures, secreted toxins, and immune evasion mechanisms. Central to its pathogenicity is the M protein, a surface-anchored molecule that inhibits phagocytosis by interfering with complement deposition and binding host factors such as fibrinogen. GAS also secretes a wide range of toxins and enzymes that damage tissues and disrupt host defences. Streptolysin O and streptolysin S are potent cytolysins that lyse immune cells and contribute to tissue necrosis. Pyrogenic exotoxins (such as SpeA and SpeC) act as superantigens, triggering massive, dysregulated T cell activation and cytokine release, an underlying mechanism in streptococcal toxic shock syndrome. Additional factors like DNases and streptokinase facilitate bacterial spread by breaking down host tissue and counteracting neutrophil extracellular traps (NETs). Immune evasion is further supported by the production of enzymes that interfere with complement functions, like the cleavage of chemokines and the targeting of antibodies. Together, these virulence determinants allow GAS to cause a wide spectrum of diseases, ranging from uncomplicated pharyngitis and impetigo to invasive conditions like necrotising fasciitis and sepsis. This review provides a timely overview of the important GAS virulence factors and an update on the current vaccine landscape.

This research paper addresses the hypothesis that the in vivo criterion of bovine somatic cell count (SCC)<200,000 cells/ml milk as a diagnostic marker for healthy mammary tissue is not suitableto be adopted to milk samples taken post slaughter. To study immune mechanisms associated with intramammary infections, we developed a mammary explant model. As SCC is routinely applied to differentiate between healthy and inflamed mammary tissue, donor cows were selected based on their milk SCC obtained in vivo. Furthermore, milk cell differentiation for early mastitis detection via flow cytometry allows identification of leucocyte subpopulations and complements SCC. To replace in vivo examination and allow for post mortem selection of donor cows, this explorative study aimed to investigate how slaughter influences the reliability of SCC and differential milk cell count (DMCC) and to assess their validity as diagnostic markers for udder health in bovine milk samples obtained post slaughter. Therefore, quarter milk samples from cows were obtained in vivo and post mortem and analysed to determine SCC and DMCC and identify major mastitis pathogens. The logarithmized numbers of SCC, non-viable cells, viable cells, lymphoid cells, polymorphonuclear (PMN) and large cells per ml milk were compared using linear mixed-effects models in milk samples obtained from cows in vivo and post mortem. The number of lymphoid cells, PMN and large cells was significantly higher in milk samples obtained post mortem than in vivo, with PMN being the most prominent cell population. Higher milk SCC values measured post mortem might be explained by migration of leucocytes into the periphery during slaughter. This should be considered when modelling intramammary infection in vitro using udder tissue. Reflecting these findings, it is not feasible to endorse SCC as a reliable marker for post mortem selection of donor cows with healthy mammary tissue for in vitro models.

Chronic rhinosinusitis (CRS) has aprevalence of up to 11% in Europe and the USA, making it one of the most common chronic diseases. Classification based on immunological endotypes is increasingly being integrated into the disease definition, particularly for chronic rhinosinusitis with nasal polyps (CRSwNP). Depending on the specific mechanisms underlying chronic tissue inflammation, different endotypes are characterized. Genetic and epigenetic changes in the mucosal immune system play asignificant role in this context. Identifying endotypes can help to better understand disease heterogeneity and develop personalized treatment approaches. In part1 of this publication, we discussed the immunological classifications of typeIV hypersensitivity reactions (T1-, T2-, and T3-endotypes), while part2 focused on typeV hypersensitivity reactions (epithelial barrier defects). The aim of part3 is to describe typeVI hypersensitivity immune reactions and highlight their implications for extended diagnostics and treatment. The European Academy of Allergy and Clinical Immunology (EAACI) recently published aposition paper presenting an updated nomenclature for immunological hypersensitivity reactions, now encompassing nine distinct immunological reaction types. The antibody-mediated reactions originally classified by Coombs and Gell as typesI, II, andIII have been expanded and described in greater detail. Direct cellular and inflammatory responses to chemical substances are defined as typeVI hypersensitivity reactions and are the focus of this third part of the publication series. In CRSwNP patients with typeVI hypersensitivity immune reactions, an imbalance between cyclooxygenase (COX) isoforms1 and2 can be observed. This imbalance is exacerbated by COX-1-inhibiting drugs, leading to reduced prostaglandin E2 (PGE2) synthesis and overproduction of leukotrieneC4 (LTC4) and PGD2. As aresult of the chronic mucosal inflammation, alarmins such as interleukin-33 (IL-33) and thymic stromal lymphopoietin (TSLP) are released. These cytokines activate Th2 lymphocytes and type2 innate lymphoid cells (ILC2s), prompting the release of cytokines such as IL‑4, IL‑5, and IL-13. Expansion of the immunological classification to include typeVI hypersensitivity reactions represents an important step toward abetter understanding of the pathophysiology of CRSwNP. Identifying these specific reaction patterns-particularly those triggered by chemical substances-highlights the complexity of the underlying immune mechanisms and emphasizes the need for endotype-based diagnostics. Incorporating these insights into clinical practice allows for more targeted individualized therapies and marks another step toward personalized medicine in CRS.