Immune Response Research Articles

Acute Disseminated Encephalomyelitis (ADEM) following viral infections: Investigating triggers, immune response and management

ADEM is diagnosed as an acute disseminated demyelinating disease of the CNS. It is normally initiated by viral infections. The objective of the current research was to elucidate the etiology, immunologic characteristics and management of ADEM in 430 subjects in Hayatabad Medical Complex, Peshawar. Of the viral causes the most commonly identified were influenza, 25%; measles, 20%; and varicella, 18%. Perhaps this may show how distribution of the infective agents common in this region may explain the occurrence of ADEM. They have similarly given out high cytokine concentration particularly IL-6 and TNF-α and similar to past findings they might not only be biomarkers of disease severity but may also be a site of immunomodulation. As for the reaction to the treatment the favorable responses are 80% of the times on corticosteroids and the IVIG and plasmapheresis are great for severe cases or if the prior one failed. Datalocalized at six months follow-up, the majority of the patients had positively improved in seventy percent with the other ten percent patients’ symptoms nearly dropped down. Therefore, the present research suggests that diagnosis of ADEM should be done early and patients ought to be put on working on formulation of tailor made treatment plans that might improve prognosis of ADEM. Findings regarding genetic profile identifying high risk patients who must be prescribed special medicine are still lacking. Therefore, this study will therefore complement information on Immune defenses and therapeutic approaches in ADEM in a region of the world where viral diseases are common.

Lack of WDFY4 leads to impaired immune response and poor cancer prognosis.

WDFY4 plays an essential role in the immune system by regulating B-cell growth and development and participating in antigen processing during cross-presentation. WDFY4 is closely related to asthma and systemic lupus erythematosus; however, its role in cancer remains unclear. The purpose of this study is to use bioinformatics to determine whether abnormal expression of WDFY4 is a risk factor for cancer and to preliminarily analyze the ways in which WDFY4 affects cancer through experiments. R language packages and bioinformatic database were used to mine the potential carcinogenic effect of WDFY4 and analyze the differential WDFY4 expression in cancer, gene mutations, different tumor prognoses, immune cell infiltration, tumor microenvironment, and DNA methylation correlation. H1975 and A549 cell lines were infected with lentiviruses to overexpress WDFY4, and the effect of WDFY4 on the activity, proliferation, apoptosis, and cell cycle of lung cancer cells was analyzed. WDFY4 was differentially expressed in human tumors in unpaired and paired samples. The differential expression of WDFY4 in unpaired and paired or protein samples from the Clinical Proteome Tumor Analysis Consortium of eight cancers was consistent. WDFY4 methylation was downregulated in 17 cancer types and caused prognostic differences in different directions in some cancers. WDFY4 overexpression significantly inhibited the activity and proliferation of lung cancer cell lines, promoted apoptosis, and caused cell cycle arrest. Differential WDFY4 expression in cancers leads to differences in the prognosis of various cancers. WDFY4 can be an independent prognostic factor for glioma, KIRC, and LUSC.

Evaluating the efficacy and safety of the Covishield vaccine: Balancing immune response and side effects in diverse populations

Evaluating the efficacy and safety of the Covishield vaccine: Balancing immune response and side effects in diverse populations

Vaccination in hemophilic children: Challenges, innovations, and future directions

Introduction: Hemophilia, an inherited disorder primarily affecting blood clotting due to deficiencies in clotting factors VIII or IX, presents complex challenges for vaccination. Children with hemophilia require vaccinations to prevent infections that may exacerbate their bleeding disorder. However, traditional intramuscular vaccination poses bleeding risks, necessitating specialized administration methods and careful post-vaccination care. Purpose: This study examines the efficacy and safety of vaccination in pediatric hemophilia patients, emphasizing the importance of individualized protocols to manage bleeding risks effectively. It explores recent advancements, including needle-free technologies, the potential of gene therapy, and mRNA vaccines. Materials and Methods: The study synthesizes current guidelines from global health organizations, hemophilia management protocols, and clinical studies on vaccine safety in hemophilia. It highlights techniques such as subcutaneous administration, clotting factor prophylaxis, and the use of fine-gauge needles to minimize bleeding complications. Results: Findings indicate that with appropriate management, hemophilic children achieve adequate immune responses to vaccines, with reduced bleeding risks. Emerging technologies, such as microneedle patches, jet injectors, and gene therapy, offer promising safer vaccination alternatives. Additionally, mRNA vaccines demonstrate strong immunogenicity without the need for live-virus vectors, reducing complication risks. Conclusions: Vaccination, when adapted to hemophilia-specific needs, remains essential for infection prevention in this vulnerable population. Newer vaccine technologies and individualized care protocols enhance safety and efficacy, contributing to improved quality of life and long-term health outcomes for hemophilic children.

Prediction of prognosis and immune response in lung adenocarcinoma based on mitophagy and lactate-related gene signatures.

Lung adenocarcinoma (LUAD) causes leading death worldwide. Mitophagy and lactate metabolism accumulation are distinctive features of LUAD. We aimed to identify lactate-related genes (LRGs) signatures based on mitophagy for predicting prognosis and immune response in LUAD. The gene expression and clinical data were downloaded from TCGA and GEO database. First, the subtype analysis was analyzed based on 29 mitophagy genes. Survival, immune, and function differences between the different subtypes were analyzed. Then, based on mitophagy genes and 14 LRGs, the best LRGs were screened to construct a risk score model and combined with clinical factors to establish a nomogram for predicting patient survival. Finally, the expression level and molecular function of the key candidate gene OGDH were verified by in vitro experiments. All the LUAD samples were divided into 2 subtypes: sub1 and sub2. The sub2 possessed worse survival. Immune score, immune checkpoint genes, and human leucocyte antigen genes in sub1 were higher than in sub2. Six optimal mitophagy-related LRGs were used to construct a risk score model. A high-risk score indicates poorer survival, higher tumor mutation burden, and higher drug sensitivity. The nomogram was robust in predicting LUAD survival. The experiments in vitro showed that knockdown of OGDH inhibited the proliferation, migration and invasion in LUAD cells. A nomogram based on the construction of the mitophagy-related lactate genes predicts prognosis and immune response in LUAD. These results could help with risk stratification and targeted therapy for LUAD.

Pan-immune-inflammation value predicts immunotherapy response and reflects local antitumor immune response in rectal cancer.

The pan-immune-inflammation value reflects the systemic inflammatory response, and tumor-infiltrating lymphocytes indicate a local immune response in rectal cancer. However, the association between systemic inflammatory response, as indicated by the pan-immune-inflammation value, and local immune responses in rectal cancer remains unclear. This study analyzed 915 treatment-naïve rectal cancer patients from the Peking Union Medical College Hospital and PLA General Hospital (PLAGH) cohorts who underwent radical surgery to investigate the relationship between the pan-immune-inflammation value and immune responses. Lower pan-immune-inflammation value was significantly associated with improved disease-free survival and cancer-specific survival. Multivariate Cox regression models identified the pan-immune-inflammation value as an independent prognostic factor. In the PLAGH cohort, patients with low pan-immune-inflammation values had higher immune cell levels, activated immune pathways, and increased expression of immune checkpoint genes according to RNA sequencing. Hematoxylin and eosin staining and immunohistochemical analysis revealed that lower pan-immune-inflammation value was associated with higher tumor-infiltrating lymphocyte density, more mature tertiary lymphoid structures, increased CD8+ T cells, and elevated human lymphocyte antigen class I expression. Conversely, patients with high pan-immune-inflammation values exhibited pathways linked to tumor progression, such as angiogenesis, epithelial-mesenchymal transition, hypoxia, KRAS signaling, and TGF-ß signaling. Among patients receiving anti-PD-1 therapy, responders had low pre- and post-treatment pan-immune-inflammation values. The pan-immune-inflammation value is a reliable marker associated with distinct immune microenvironment characteristics and can effectively predict disease-free survival, cancer-specific survival, and response to immunotherapy.

Genistein in focus: pharmacological effects and immune pathway modulation in cancer.

Cancer is a significant global health concern, responsible for mortality and morbidity of individuals. It is characterized by uncontrolled cellular growth, tumor formation, and potential metastasis. The immune system is pivotal in recognizing and eliminating cancerous cells, with immune cells such as T cells, B cells, natural killer cells (NK), and dendritic cells playing critical roles. Dysregulation of immune responses can contribute to cancer progression. Phytochemicals, bioactive compounds derived from plants, have gained attention for their potential roles in cancer prevention and therapy due to their antioxidant, anti-inflammatory, and immunomodulatory properties. Genistein, an isoflavone found in soy products, is of particular interest. In this study, genistein's mechanisms of action at the molecular and cellular levels in cancer were demonstrated, highlighting its impact on T and B lymphocytes, NK cells and dendritic cells. Genistein's ability to influence cytokine production, reducing levels of inflammatory cytokines such as TNF-α, IL-6, and IL-1β, is emphasized. Genistein modulates inflammatory response pathways like Toll-like receptors (TLRs), NF-κB, chemokines, and MAPK, inhibiting tumor growth, promoting apoptosis, and reducing metastasis. It shows promise in overcoming chemoresistance, particularly in ovarian and neuroblastoma cancers, by inhibiting autophagy. Genistein also affects T-cell execution markers, including granzyme B, TNF-α, and FAS ligand in cancer by influencing key proteins involved in immune response and apoptosis. Clinical trials have investigated genistein's therapeutic potential, revealing its promise in enhancing the efficacy of traditional cancer treatments while mitigating associated toxicities. Genistein helps overcome chemoresistance in various cancers by inhibiting autophagy and promoting apoptosis. It also enhances immunotherapy by boosting immune responses and modifying antigens, but careful dosing is needed when combined with anti-PD-1 treatments to avoid reducing effectiveness.

Identification and Immunological Characterization of Annexin B8 and Annexin E1 from Spirometra Erinaceieuropaei Spargana.

Sparganosis is a parasitic zoonotic disease that poses a serious threat to public hygiene and human health. Annexin is a phospholipid-binding protein with calcium ion binding activity, serving various important functions, including interaction with the parasite and regulation of the host's immune response. In this study, two annexin (ANX) family genes, Spirometra erinaceieuropaei (S. erinaceieuropaei) Annexin B8 (SeANXB8) and E1 (SeANXE1), isolated from spargana, were cloned and immunologically characterized. Both recombinant S. erinaceieuropaei Annexin B8 (rSeANXB8) and E1 (rSeANXE1) were specifically recognized by serum from rats immunized with the recombinant proteins, displaying strong immunoreactivity. They are also among the major components of sparganum excretion/secretion products (ESPs). SeANXE1 was identified in the parasite's tegument, testis, genital pore, ovary, and eggs, while SeANXB8 was found in the parasite's tegument and eggs. Plasminogen (PLG)-binding assays revealed that the two annexins could bind to human PLG in a concentration-dependent manner, which was blocked by the corresponding antibodies. These findings suggest that SeANXB8 and SeANXE1 may be involved in host-parasite interaction and may influence the host's immune response during sparganosis. They could be potential diagnosis and vaccination targets for sparganosis.

Recombinant Saccharomyces cerevisiae EBY100/pYD1-FaeG: a candidate for an oral subunit vaccine against F4+ ETEC infection.

Diarrheal diseases attributable to multidrug-resistant F4+ enterotoxigenic Escherichia coli (ETEC) are escalating in severity, posing significant risks to the health and safety of both humans and animals. This study used Saccharomyces cerevisiae EBY100 to display the FaeG subunit of F4 colonizing factor as an oral vaccine against F4+ ETEC infection. Mice were orally immunized twice with 108 CFU of EBY100/pYD1-FaeG, followed by a challenge with F4+ ETEC EC6 on day 7 post-immunization. The results showed that the recombinant strain EBY100/pYD1-FaeG orally enhanced the growth of the small intestine villi, significantly boosted the expression of tight junction proteins (ZO-1, Occludin, MUC2, and Claudin) (P < 0.05), and modulated the gut microbiota composition. Additionally, immunization with EBY100/pYD1-FaeG also upregulated the levels of IL-2, IL-4, and IFN-γ in the intestines of mice (P < 0.01), while serum IgG and fecal sIgA titer significantly increased (P < 0.05). These immune responses enhanced the capacity to fight against ETEC, leading to an increased survival rate of mice and relieved damage to tissues and organs of mice infection. In summary, the study suggested that the recombinant Saccharomyces cerevisiae EBY100/pYD1-FaeG could effectively stimulate the immune response and generate specific antibodies against F4+ ETEC, showing its potential to serve as a subunit oral vaccine candidate for preventing F4+ ETEC infection.IMPORTANCEThe multidrug-resistant F4+ enterotoxigenic Escherichia coli (ETEC) strains are the primary clinical pathogens responsible for post-weaning diarrhea in piglets, resulting in substantial economic losses in the pig farming industry. In the study, we developed an oral vaccine candidate, Saccharomyces cerevisiae EBY100/pYD1-FaeG, to prevent diarrhea caused by multidrug-resistant F4+ ETEC. Oral administration of EBY100/pYD1-FaeG significantly enhanced immune responses, improved intestinal health, and provided protection against F4+ ETEC infection in mice. This approach offers a potential application prospect for preventing F4+ ETEC infections that lead to post-weaning diarrhea in clinical settings and provides a promising solution for addressing the growing threat of antibiotic resistance in bacterial pathogens.

LINE-1 hypomethylation characterizes the inflammatory response in coeliac disease associated-intestinal mucosa and small bowel adenocarcinomas.

Long interspersed nuclear elements 1 (LINE-1) are the most abundant and the only autonomous mobile elements in the human genome. When their epigenetic repression is removed, it can lead to disease, such as autoimmune diseases and cancer. Coeliac disease (CeD) is an immune-mediated disease triggered by an abnormal T-cell response to dietary gluten and a predisposing condition of small bowel adenocarcinoma (SBA), frequently characterized by epigenetic alterations. The aim of this work was to assess LINE-1 methylation by bisulphite pyrosequencing and NanoString® gene transcription analysis in 38 CeD-SBAs compared with 25 SBAs associated with Crohn's disease (CrD-SBAs) and 25 sporadic SBAs (S-SBA). Both analyses were also performed in duodenal mucosae from 12 untreated CeD patients (UCD) and 19 treated CeD patients (TCD), and in 11 samples of normal intestinal mucosa to better investigate the role of LINE-1 deregulation in CeD and in CeD-SBA. A significant loss of LINE-1 methylation was observed in CeD-SBAs and in mucosae from UCD patients (with very similar methylation levels) compared with controls. By contrast, a restoration of normal LINE-1 methylation levels was found in TCD mucosae after a strict gluten-free diet. LINE-1 hypomethylation does not lead to expression of ORF1 and ORF2, with the only exception being for one CeD-SBA. The expression analysis of enzymes modulating DNA methylation and inflammatory genes confirmed that CeD-SBA shared a very similar expression profile of UCD mucosae showing a strong upregulation of genes involved in inflammation, immune response, and T-cell activity compared with TCD mucosae. For the first time, this work demonstrates that loss of DNA methylation is an intrinsic epigenetic feature of CeD, accompanying the immune response as a reversible mechanism in patients following a strict gluten-free diet, and suggests the possible role of LINE-1 hypomethylation in promoting cell adaptability during the gliadin-related inflammatory process. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Type 1 Immune Responses Related to Viral Infection Influence Corticosteroid Response in Asthma.

Corticosteroid-responsive type 2 (T2) inflammation underlies the T2-high asthma endotype. However, we hypothesized that type 1 (T1) inflammation, possibly related to viral infection, may also influence corticosteroid response. To determine the frequency and within-patient variability of T1-high, T2-high, and T1/T2-high asthma endotypes and whether virally influenced T1-high disease influences corticosteroid response in asthma. Patients in the Severe Asthma Research Program (SARP)-3 had sputum collected at baseline, after intramuscular (triamcinolone acetonide) corticosteroid treatment, and at 1 and 3-year follow-ups. Sputum cell RNA was used for whole transcriptome gene network and viral metagenomic analyses. We then profiled patients as highly expressing T1 and/or T2 gene networks and established the influence of these endotypes on corticosteroid responsiveness and likelihood of viral transcript detection in the airways. We found that 22% and 35% of asthma patients highly expressed T1 and T2 network genes, respectively, and that 8.5% highly expressed both networks. Asthma severity outcomes were worse in T2-high compared to T1-high asthma and most severe in the T1-high/T2-high subgroup. Corticosteroid treatment strongly suppressed T2 but poorly suppressed T1 gene expression, and corticosteroid-associated improvements in FEV1 occurred only in patients with T1-low/T2-high disease and not in T1-high/T2-high patients. Viral metagenomic analyses uncovered that 24% of asthma sputum samples tested positive for a respiratory virus and high viral carriage was associated with 14-fold increased risk of T1-high disease. Airway type 1 immune responses are relatively common in asthma, are largely corticosteroid-resistant, and are associated with sub-clinical viral infection.

Single-cell analysis reveals expanded CD8+ GZMKhigh T cells in CSF and shared peripheral clones in sporadic amyotrophic lateral sclerosis

Abstract Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease that affects motor neurons in the brain and spinal cord. Despite the crucial role of aberrant immune responses in ALS pathogenesis, studies investigating immunological profiles in the cerebrospinal fluid (CSF) of ALS patients have reported inconsistent findings. Herein, we explored the intrathecal adaptive immune response and features of circulating T cells between CSF and blood of ALS patients, using single-cell RNA and T cell receptor (TCR) sequencing. This study comprised a total of 11 patients with apparently sporadic ALS and three controls with non-inflammatory diseases. We collected CSF from all participants, and for three ALS patients, we additionally obtained paired samples of peripheral blood mononuclear cells (PBMCs). Utilizing droplet-based single-cell RNA and TCR sequencing, we analyzed immunological profiles, gene expression characteristics, and clonality. Furthermore, we examined T cell characteristics in both PBMC and CSF samples, evaluating the shared T cell clones across these compartments. In the CSF, ALS patients exhibited a lower proportion of CD4+ T cells (45.2% vs. 61.2%, p = 0.005), and a higher proportion of CD8+ GZMKhi effector memory T cells (TEMs) than controls (21.7% vs. 16.8%, p = 0.060). Higher clonality was observed in CD8+ TEMs in ALS patients compared to controls. In addition, CSF macrophages of ALS patients exhibited a significant increase in chemokines recruiting CD8+ TEMs. Immunohistochemical analysis showed slightly higher proportions of T cells in the perivascular and parenchymal spaces in ALS patients than in controls, and CD8+ TEMs co-localized with neurons or astrocytes in the motor cortices of ALS patients. Clonally expanded CD8+ GZMKhi TEMs primarily comprised shared T cell clones between CSF and PBMCs. Moreover, the shared CD8+ TEMs of PBMCs exhibited gene expression profiles similar to CSF T cells. ALS patients showed an increase in proportion and clonality of CD8+ GZMKhi TEMs and activated features of macrophages in CSF. The shared T cell clone between CSF and blood was mainly composed of expanded CD8+ GZMKhi TEMs. In conclusion, single-cell immune profiling provided novel insights into the pathogenesis of ALS, characterized by activated macrophages and clonally expanded CD8+ T cells potentially communicate between the central nervous system and peripheral circulation.

Pan-cancer TCGA analysis reveals the potential involvement of B-type lamins in dysregulating chromosome segregation in human cancer.

Lamins play a crucial role in maintaining nuclear structure and function. Our study investigates the expression patterns and clinical implications of B-type lamins with a special focus on lamin B2 across various cancer types using comprehensive RNA sequencing datasets. We found that high expression levels of lamin B1 and lamin B2 are associated with decreased overall and disease-free survival in cancer. This association is further pronounced when both lamins are co-expressed, indicating a compounded negative impact on patient prognosis. Additionally, we highlighted the relationship between B-type lamins and the tumor microenvironment. Lamin B1 mRNA expression shows a strong positive correlation with activated CD4+ T-cells and type 2 T-helper cells (Th2), suggesting its role in immune cell infiltration and response within tumors. Lamin B2 expression also correlates moderately with these immune cells, indicating a potential but lesser role in modulating the immune landscape. Notably, the epigenetic state of lamin B1 significantly affects the tumor microenvironment, suggesting a dual role in structural integrity and immune modulation. We have identified 9 lamin B2 interacting proteins that are co-expressed with B-type lamins in cancerous conditions and modulate cytokinesis and cell division pathways during tumorigenesis. Furthermore, we have identified specific molecular targets of B-type lamins that co-express with them in a range of human cancers and are potentially involved in dysregulating chromosome segregation and mRNA binding. Overexpression of these targets alongside B-type lamins correlates with poor prognosis in multiple cancers. These findings underscore the potential of B-type lamins as biomarkers for poor prognosis and as targets for cancer therapies.

CaMKII-dependent non-canonical RIG-I pathway promotes influenza virus propagation in the acute-phase of infection.

Ca2+/calmodulin-dependent protein kinase II (CaMKII) is one of hundreds of host-cell factors involved in the propagation of type A influenza virus (IAV), although its mechanism of action is unknown. Here, we identified CaMKII inhibitory peptide M3 by targeting its kinase domain using affinity-based screening of a tailored random peptide library. M3 inhibited IAV cytopathicity and propagation in cells by specifically inhibiting the acute-phase activation of retinoic acid-inducible gene I (RIG-I), which is uniquely regulated by CaMKII. Downstream of the RIG-I pathway activated TBK1 and then IRF3, which induced small but sufficient amounts of transcripts of the genes for IFN α/β to provide the capped 5'-ends that were used preferentially as primers to synthesize viral mRNAs by the cap-snatching mechanism. Importantly, knockout of RIG-I in cells almost completely inhibited the expression of IFN mRNAs and subsequent viral NP mRNA early in infection (up to 6 h after infection), which then protected cells from cytopathicity 24 h after infection. Thus, CaMKII-dependent acute-phase activation of RIG-I promoted IAV propagation, whereas the canonical RIG-I pathway stimulated antiviral activity by inducing large amounts of mRNA for IFNs and then for antiviral proteins later in infection. Co-administration of M3 with IAV infection rescued mice from the lethality and greatly reduced proinflammatory cytokine mRNA expression in the lung, indicating that M3 is highly effective against IAV in vivo. Thus, regulation of the CaMKII-dependent non-canonical RIG-I pathway may provide a novel host-factor-directed antiviral therapy.IMPORTANCEThe recent emergence of IAV strains resistant to commonly used therapeutic agents that target viral proteins has exacerbated the need for innovative strategies. Here, we originally identified CaMKII-inhibitory peptide M3, which efficiently inhibits IAV-lethality in vitro and in vivo. M3 specifically inhibited the acute-phase activation of RIG-I, which is a novel pathway to promote IAV propagation. Thus, this pathway acts in an opposite manner compared with the canonical RIG-I pathway, which plays essential roles in antiviral innate immune response later in infection. The CaMKII-dependent non-canonical RIG-I pathway can be a promising and novel drug target for the treatment of infections.

The Association between Systemic Inflammation and the Prostate Cancer: based on the National Health and Nutrition Examination Survey and Mendelian Randomization Analysis.

The purpose of this combination research was to examine the relationship between systemic inflammation and the risk of prostate cancer (PCa) through the National Health and Nutrition Examination Survey (NHANES) cross-sectional study and two-sample Mendelian randomization (MR) analysis. We incorporated NHANES data spanning the years 2001 to 2010, with exposure as systemic inflammation, evaluated using systemic immune-inflammation index (SII) and outcome as PCa, and performed multivariate logistic regression and restricted cubic spline (RCS) to test the correlation between SII and PCa. Further, two-sample MR was used to identify causal associations between specific immune cells and PCa. A total of 7706 participants (age≥40 years) were included in the analysis in the cross-sectional study, including 350 PCa cases, 7356 controls. Higher SII levels were associated with increased odds of PCa (P<.05). The odds ratio (OR) for PCa was 1.51 (95% CI 1.09-2.08) for the highest versus lowest quartile of SII levels in the fully adjusted model. Also, the RCS analysis showed a threshold effect, with SII levels above 8.90 associated with increased odds of PCa. In addition, MR results suggested a causal relationship between CD62L- monocyte, CD62L- HLA DR+ monocyte, CD14+ CD16+ monocyte, CD62L- Dendritic Cell, Monocytic Myeloid-Derived Suppressor Cell, CD28- CD8dim T cell, CD39+ resting CD4 regulatory T cell and PCa (P<.05). This combination analysis provides evidence for a significant causal relationship between systemic inflammation and PCa risk. These findings highlight systemic inflammation and inflammatory immune responses as potential modifiable risk factors for PCa.

Preparation, Characterization and Immune Activity of ViolaPhilippicaPolysaccharide PLGA Nanoparticles.

Recent pharmacological studies have demonstrated that Viola philippicapolysaccharide (VPP) exhibits a modulating effect on immune activity. However, its utilization has been hampered by its large particle size and complex spatial structure. Poly-lactic-co-glycolic acid copolymer (PLGA) is recognized as an effective drug delivery carrier, exhibiting excellent biochemical properties. In this experiment, VPP was encapsulated with PLGA to form Viola philippica polysaccharide PLGA nanoparticles (VPP-PLGA NPs). The morphological structure and immunomodulatory effects of VPP-PLGA NPs were evaluated. The particle size of VPP-PLGA NPs was reduced compared to VPP, and the optimal preparation conditions were as follows: the ratio of the organic phase to the internal aqueous phase was 8:1, the ratio of the external aqueous phase to the primary emulsion was 7:1, and the concentration of PLGA was 20 mg/mL. Additionally, VPP-PLGA NPs significantly increased the NO content, IL-4, and IFN-γ levels in RAW264.7 cells, as well as enhanced their phagocytic activity. Furthermore, VPP-PLGA NPs were found to increase NO content and IFN-γ secretion in bone marrow-derived dendritic cells (BMDCs). These findings suggest that VPP-PLGA NPs could enhance the immune activity and may be utilized as a VPP delivery system for inducing strong immune responses.

Glycoconjugate Vaccines: Platforms and Adjuvants for Directed Immunity.

Central to immune recognition is the glycocalyx, a glycan-rich coat on all cells that plays a crucial role in interactions that enable pathogen detection and activation of immune defenses. Pathogens and cancerous cells often display distinct glycans on their surfaces, making these saccharide antigens prime targets for vaccine development. However, carbohydrates alone generally serve as poor immunogens due to their often weak binding affinities, inability to effectively recruit T cell help, and reliance on adjuvants to induce immune activation. The introduction of glycoconjugate vaccines, initially involving the covalent coupling of carbohydrate antigens to carrier proteins, marked a pivotal advancement by facilitating neutralizing antibody production against these carbohydrate targets. Despite successes in generating glycoconjugate vaccines against certain bacterial diseases, challenges persist in creating effective vaccines against numerous intracellular pathogens and non-communicable diseases such as cancer. New developments in conjugate vaccine platforms aim to overcome these limitations by optimizing the display of glycan and T cell epitope as well as incorporating defined carbohydrate adjuvants to direct tailored immune responses. These advancements promise to improve the effectiveness of carbohydrate-based vaccines and broaden their coverage against a wide range of diseases.

Canine parvovirus NS1 induces host translation shutoff by reducing mTOR phosphorylation.

Autonomous parvoviruses, which possess compact genomes, are obligate intracellular parasites that necessitate host cell division for their replication cycle. Consequently, the modulation of host translation and usurpation of cellular machinery are hypothesized to facilitate immune evasion, enhance viral transmission, and perpetuate long-term infection. Despite the biological significance, the precise mechanisms by which autonomous parvoviruses regulate host translation remain understudied. Our study elucidates that CPV-2 infection induces a shutoff of host translation through the attenuation of mTOR phosphorylation. This mechanism may enable the virus to subvert the host immune response and engender pathogenic effects.

Manganese improves anti-PD-L1 immunotherapy via eliciting type I interferon signaling in melanoma.

The immune checkpoint inhibitor therapy represented by blocking programmed cell death protein 1/ programmed cell death-ligand 1 (PD-1/PD-L1) has made significant progress in melanoma treatment. However, the response rate and therapeutic effect of immunotherapy alone are still not ideal for melanoma. In this study, we aimed to evaluate the defects of treating anti-PD-L1 alone and the therapeutic effect and molecular mechanism of combined therapy with anti-PD-L1 and MnCl2. We detected the changes of immune cell populations after anti-PD-L1 treatment in melanoma xenograft mouse model. Further, we evaluated the regulatory effect of MnCl2 on dendritic cells (DCs) maturation in vitro. Next, we tested the therapeutic effect and regulatory effect on the tumor microenvironment with anti-PD-L1 and MnCl2 via combining treatment with anti-PD-L1 and MnCl2. Anti-PD-L1 therapy has a certain tumor suppressive function, but the effect is not ideal. The results of flow cytometry showed that the number of CD4+ T cells and CD8+ T cells significantly increased after anti-PD-L1 treatment. However, the number of DCs remained basically unchanged after anti-PD-L1 treatment. In vitro, we confirmed that MnCl2 significantly promoted DCs maturation vis activating cGAS-STING signaling pathway. The combination of anti-PD-L1 and MnCl2 displayed the best tumor suppression effect in melanoma xenograft mouse model. In tumor microenvironment, the infiltration of T cells and the maturation of DCs were significantly promoted, demonstrating a strong anti-tumor immune response. In summary, we conclude that combining anti-PD-L1 with MnCl2 is a promising therapeutic strategy for melanoma.

Therapeutic Monoclonal Antibody─Antidrug Antibody Affinity Constant Determination Using Capillary Electrophoresis-Tandem Mass Spectrometry.

Monoclonal antibody (mAbs) therapeutics cannot evade the occurrence of adverse effects. Thus, mAbs are commonly triggering immune responses corresponding to the expression of antidrug antibodies. Antidrug antibodies can neutralize mAbs, leading to their inhibition and hasten clearance, which dramatically hampers their therapeutic effects. Therefore, studies concerning the affinity between a mAbs and the corresponding antidrug antibody are demonstrating a growing interest to further understand the outcome of biotherapeutics after administration. We describe a novel analytical approach for the determination of the dissociation constant (Kd) between a mAb and an antidrug antibody using capillary electrophoresis coupled to mass spectrometry (CE-MS/MS). The CE-MS/MS method employs a competitive assay, followed by the quantification of the residual amount of the active mAbs. The methodology allowed for the measurement of Kd using serum samples without the implementation of immobilization to achieve protein-protein interaction. This characteristic enabled us to generate the interaction in conditions reflecting the physiological environment. A mathematical treatment was developed to calculate Kd from MS/MS data, taking into account the stoichiometry of the mAbs/antidrug antibody complex and the bivalent properties of the two immunoglobulins. Prior to CE-MS/MS analysis, the interaction of the two proteins was studied by using mass photometry (MP) to determine equilibrium conditions and complexation stoichiometry. CE-MS/MS was used to investigate the interaction between infliximab and a neutralizing anti-infliximab antibody. Results allowed to measure a Kd of 14.4 ± 2.9 nM. MS instrumentation sensitivity and specificity showed to be relevant to achieve accurate and robust Kd measurements for strong interactions in the nanomolar range.