High Levels Of Cytokines Research Articles

The absolute number of circulating Treg cells is reduced in difficult-to-treat RA patients and is ameliorated by low-dose IL-2

ObjectiveCirculating regulatory T cells (Tregs) are closely related to immune tolerance and maintenance of immune homeostasis. Perhaps, there is a unique immune cell phenotype for difficult-to-treat rheumatoid arthritis (D2T RA). Low-dose interleukin-2 (IL-2) has been considered for the treatment of autoimmune diseases. This study focused on the uniqueness of D2T RA lymphocyte subsets and the feasibility of low-dose IL-2 therapy.MethodsParticipants included 1,042 RA patients who were divided into three groups according to the presence or absence of treatment and their response to treatment in the last 6 months—new group, treated group, and D2T group—and 339 healthy controls (HCs). A total of 381 patients—107, 151, and 123 in each of the three experimental groups—received low-dose IL-2 treatment [0.5 million international units (MIU) per day, subcutaneous injection from day 1 to day 5]. The absolute numbers of peripheral blood lymphocyte subsets were detected by flow cytometry (FCM) and serum cytokine levels were detected by flow cytometry bead array (CBA).ResultsThe absolute number of T, CD4+ T, and Treg cells in the D2T RA group was lower than that in the HC, new, and treated RA groups. Compared with the HC and new RA group, the ratio of Th17/Treg cells in the D2T RA group increased. The new, treated, and D2T RA groups had higher cytokine levels than the HC. The number of Treg cells in RA patients was negatively correlated with the disease activity index. Treg cells in the new, treated, and D2T RA groups could be increased by low-dose IL-2 therapy without any side effects.ConclusionsThe number of lymphocytes and subsets in D2T RA patients was reduced, especially Treg cells, resulting in a shift in the balance of effector T cells/Treg cells toward effector T cells, which is ameliorated by low-dose IL-2 without obvious side effects.

The immunogenicity of PRV ΔgE/TK/UL49.5 three-gene-deleted vaccine in mice

BackgroundPseudorabies (PR) caused by the re-emerging of pseudorabies virus (PRV) variant has outbroken among PRV vaccine immunized swine in many pig farms, which has caused serious social and economic consequences since the end of 2011. The PRV UL49.5 protein can inactivate the transporter associated with antigen processing (TAP), thereby downregulating the cell surface expression of swine leukocyte antigen class I (SLA-I) to evade host immune surveillance.MethodsIn this study, based on the PRV ΔgE/TK strain, PRV ΔgE/TK/UL49.5 triple gene deletion strain was constructed through homologous recombination and deletion of the PRV UL49.5 gene by the Cre-LoxP system. Its growth curve and effect on SLA-I transcription level were determined. Preliminary studies were carried out on serum neutralizing antibody levels, IFN-γ and IL-4 cytokines levels in mice immunized with PRV ΔgE/TK/UL49.5, and the viral load and challenge protection in mice tissues after challenge.ResultsThe growth characteristics of PRV ΔgE/TK/UL49.5 strain were similar to those of PRV ΔgE/TK strain. The level of SLA-I was returned to normal after the deletion of PRV UL49.5 gene. The immunization of PRV ΔgE/TK/UL49.5 did not affect the weight gain of mice. Immunized mice could induce high levels of serum neutralization antibodies and immune cytokines, including IFN-γ and IL-4, which could provide complete protection against virulent PRV challenge. No obvious pathological damage was observed in lung, brain and trigeminal ganglion of mice immunized with PRV ΔgE/TK/UL49.5, and the tissue viral load was the lowest.ConclusionsPRV ΔgE/TK/UL49.5 strain can induce enhanced immunogenicity and had the potential to be used as a candidate strain.

Age-related immune response disparities between adults and children with severe COVID-19: a case–control study in China

BackgroundElucidation of immune response differences is critical for uncovering underlying mechanisms and developing potential intervention measures among adults and children with COVID-19.MethodsIn this retrospective study, we analyzed serum biochemical markers and cytokine profiles among adults and children with COVID-19 in the First People’s Hospital of Chenzhou in Hunan, China from 1 December 2022 to 13 February 2023. A case–control study was conducted using propensity score matching (PSM) to mitigate possible confounding factors.ResultsThe significant differences observed included lymphocyte exhaustion, an increased neutrophil-to-lymphocyte (NEU/LYM) ratio, high levels of C-reactive protein (CRP), and a cytokine storm, characterized by high levels of Th1 proinflammatory cytokines, including interleukin 1β (IL-1β), IL-6, IL-8, interferon type I (IFN-γ), and tumor necrosis factor (TNF-α) in the lung among severe adult COVID-19 patients. Additionally, systemic immune responses were observed in children with COVID-19.ConclusionSignificant differences in immune responses between adults and children with COVID-19 highlight the different mechanisms and potential intervention measures of COVID-19.

Peritoneal fluid microbiota profile of patients with deep endometriosis.

Endometriosis is a chronic gynecological disease that affects 10% of reproductive-aged women and characterized by the presence of endometrial tissue outside the uterus. The disease is linked to a pro-inflammatory environment in the peritoneal fluid of patients, with high levels of cytokines, growth factors, and reactive oxygen species. Changes in the peritoneal fluid, such as altered immune cells and cytokines, can be linked to the immune balance in endometriosis. Immunological changes may be related to the presence of microorganisms in the peritoneal fluid that can activate Toll-like receptor (TLR) signaling and trigger an inflammatory response. A high diversity of TLRs has been found in women with endometriosis, and the presence of specific microorganisms in the fluid is suggested to be responsible for the activation of inflammasomes and inflammatory cytokines involved in the development of endometriosis. The present study was conducted at a hospital in southeastern Brazil to test this hypothesis, using a case-control design. Peritoneal fluid from 50 patients was used in this study. The case group consisted of 27 patients with endometriosis and the control group consisted of 23 patients without endometriosis. The samples were stored in a microbiome transport solution, and DNA was extracted and sent for genetic sequencing to identify the microorganisms present. The obtained sequencing reads were processed using a bioinformatics pipeline involving demultiplexing with the Illumina proprietary software, primer detection and removal, error evaluation, quality filtering, error removal using the Deblur software, amplicon sequence variants grouping, and chimera detection using the VSEARCH software. The sheer abundance of the microbiome made it challenging to discern any notable differences between the two groups. Nevertheless, we highlighted the prevalence of three primary bacteria in the peritoneal fluid from patients with endometriosis: Flavobacterium, Pseudomonas, and Bacillus. The results were established after a rigorous experimental design to eliminate potential contamination from extraction kits and handling. Our findings provide valuable insight into the pathogenesis of this disease and can be useful to understand how microbiota and immune system works in endometriosis.

Peptide-based vaccine design against Hendra virus through immunoinformatics approach.

The Hendra virus (HeV) has resulted in epidemics of respiratory and neurological illnesses in animals. Humans have contracted diseases with high fatality rates as a result of infected domestic animals, but effective vaccinations and therapies are currently not available against HeV. Herein, we analyzed the proteome of HeV and constructed an effective and innovative multi-epitope vaccine using immunoinformatics techniques. The vaccine construct was generated, targeting one matrix protein, with the help of the five selected B and T cell epitopes, linkers, and adjuvants and evaluated for their immunogenic properties. In-silico analysis revealed that the epitopes were able to interact with immune receptors and had high antigenic qualities. The post-translational modifications (PTMs), globular, disordered regions, and the active site of the vaccine were predicted, and the strong interactions between the vaccine and Toll-like receptor 5 were observed in molecular docking, indicating their potential significance in the immune response to the designed vaccine. The structural and dynamic stability of the vaccine were ensured by the molecular dynamic simulations. The results of the immune simulations indicated that the designed vaccine might activate B and T cells, which produce high levels of antibodies and cytokines to fight HeV infection. The developed vaccine is useful due to its non-toxicity, non-sensitization, good immunogenicity, non-allergic, and antigenic properties, accessed by various tools; however, experimental verification is needed to confirm the findings of the current study.

P0067 Exploring potential therapeutic implication of anti inflammatory drugs and Probiotic in Experimental Models of IBD-dependent Sarcopenia

Abstract Background Sarcopenia is a syndrome characterized by progressive and generalized loss of skeletal muscle mass. Primary sarcopenia is associated with aging, while secondary sarcopenia is caused by various factors, including inflammatory conditions such as inflammatory bowel disease (IBD) [1]. The complex interaction between inflammation, intestinal dysbiosis, and malnutrition in IBD patients is referred to as the gut-muscle axis [2]. High levels of cytokines and intestinal dysbiosis in IBD activate pathways that disrupt muscle protein synthesis [3]. However, molecular and microbial mechanisms of sarcopenia in IBD are not extensively studied as well as therapeutic approaches. Methods A murine model of DSS-induced colitis was used. To detect sarcopenia, physical performance was assessed using the rotarod test. Muscle was analyzed via immunohistochemistry, fiber counting, and cross-sectional area evaluation. Additionally, intestinal microbiota and serum LPS levels were assessed. An in vitro model of muscle cells was created using the C2C12 cell line, which was exposed to pro-inflammatory cytokines (TNF-α, IFN-γ, IL-6), lipopolysaccharides (LPS), and drugs (Infliximab, Upadacitinib, Tofacitinib) as well as probiotics (L. plantarum, L. rhamnosus) for 30 min and 6 h. Results The murine model showed decreased body weight and muscle function alongside increased expression of muscle atrophy markers (myostatin and Murf-1) in colitic mouse muscle. Furthermore, there was an upregulation of the STAT3-pSTAT3 pathway associated with muscle wasting and a decrease in pAKT, a pathway associated with muscle growth. Surprisingly, DSS-induced colitis in mice resulted in intestinal dysbiosis characterized by reduced Roseburia genus and increased Sutterella, Turicibacter, and Bacteroides in the DSS group, as well as increased serum LPS levels. In vitro, Upadacitinib and Tofacitinib, in the presence of inflammatory cytokines, reduced the expression of pathways associated with muscle atrophy (STAT3, pSTAT3, and NFκB) and increased pAKT expression. When cells were treated with LPS, NFκB expression did not decrease with the addition of the drugs, suggesting that LPS can activate NFκB independently of the JAK/STAT pathway. Treatment with probiotics reduced STAT3 and pSTAT3 signals, suggesting potential positive modulation by all probiotics. Conclusion These data open new avenues for the gut-muscle axis as a promising research area for managing sarcopenia in IBD. Considering microbiota and LPS along with inflammatory cytokines may provide a more comprehensive understanding, especially as LPS-induced signaling appears to be only partially controlled by drugs and potentially positively influenced by probiotics.

Temporal study of myocardial infarction to optimize the application of cardiosphere-derived cells’ secretome as a therapeutic strategy

Abstract Introduction Myocardial infarction (MI) initiates an acute inflammatory response that is crucial for heart repair. However, an exaggerated inflammatory response can lead to adverse left ventricular remodeling and heart failure. The secretome of cardiosphere-derived cells (CDCs) offers promising immunomodulatory properties. This study seeks to determine the optimal timing for CDC secretome application by evaluating CD163 expression in monocytes and plasma levels post-MI to maximize its immunomodulatory properties. Methods CD163 expression in monocytes was evaluated under basal conditions and after MI at 24, 48, 72 hours, and one week post-MI. Plasma protein levels were also assessed at the same time points. Results The results indicate a decrease in CD163 expression in monocytes immediately following MI, with a progressive increase observed over the subsequent 72 hours. Plasma protein analysis revealed elevated levels of TNF-α and IL-6 within the first 24 hours post-MI, which then gradually declined. These findings suggest that the inflammatory response is most acute within the first 24 hours post-MI, characterized by high pro-inflammatory cytokine levels and reduced CD163 expression. Conclusion The application of CDC-derived secretome treatment could be more beneficial during the first 24 hours post-MI. This study highlights the importance of timing in the application of CDC-based therapies to optimize their immunomodulatory effects and improve outcomes in MI patients.

Anti‐Inflammatory and Antimicrobial Effects of Herbal Formulation Called Apo‐Taat Using Extended‐Spectrum ß‐Lactamase‐Producing Escherichia coli Isolates

Pathogens contaminate drinking water in tropical countries causing diarrheal diseases. The conventional treatment for diarrhea is antibiotics. However, overuse and misuse of antibiotics has enabled pathogens to adapt, causing global antibiotic resistance and proliferation of extended‐spectrum ß‐lactamase‐producing Escherichia coli (ESBL‐E. coli), which causes diarrhea and high levels of inflammatory cytokines. Apo‐taat, consisting of equal proportions of Phyllanthus emblica and Caesalpinia sappan, has been used to treat diarrhea and bloody diarrhea. Its antibacterial activity against E. coli ATCC 25922 has been reported, but its inhibitory effect against ESBL‐E. coli has yet to be documented. This study investigated the antibacterial effect of Apo‐taat extract against ESBL‐E. coli and its anti‐inflammatory activity. Antibacterial activity was determined by the microtiter plate–based method. HPLC was used to determine the brazilin and gallic acid contents in Apo‐taat extract. Effects of herbal extracts on nitric oxide, IL‐6, and TNF‐α were investigated in RAW 264.7 cells. Results were that Apo‐taat extract showed MIC values against ESBL‐E. coli in the range of 0.625 to 2.5 mg/mL. Its 50% inhibitory concentration against nitric oxide and IL‐6 production was 83.96 ± 10.60 and 83.06 ± 2.07 μg/mL, respectively, and it had slight inhibition against TNF‐α. These findings suggest that Apo‐taat may have an antibacterial impact on ESBL‐E. coli and anti‐inflammatory activity. Furthermore, safety and clinical trials should be conducted in the future.

Tocilizumab Increases Serum Lipids but Does Not Increase Arteriosclerosis, As Measured by Intima-Media Thickness, in Patients With Rheumatoid Arthritis.

Objectives Patients with rheumatoid arthritis (RA) have a high incidence of arteriosclerotic disease. These are partly attributed to high levels of C-reactive protein (CRP) and inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6. Tocilizumab (TCZ) is an IL-6 receptor antagonist that lowers CRP levels by directly blocking IL-6 signaling. Tocilizumab has been reported to increase serum lipid levels. However, its effect on arteriosclerosis remains unclear. Therefore, we investigated the effect of TCZ on arteriosclerosis in patients with RA. Methods Eighteen Japanese patients with RA who were administered TCZ were included and assessed at baseline and six and 12 months. The Disease Activity Score-28 with erythrocyte sedimentation rate (DAS28-ESR), CRP, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and carotid intima-media thickness (IMT) were measured and analyzed using high-resolution B-mode ultrasonography. Additionally, the effects of concurrent statin administration on any changes in IMT were evaluated. Results From baseline to six or 12 months, TCZ decreased CRP (P < 0.0005) and DAS28-ESR (P < 0.0005) significantly, whereas TC, LDL-C, and HDL-C levels increased significantly (P < 0.005). However, there was no change in the LDL-C/HDL-C ratio(P= 0.821 at six months and P= 0.168 at 12 months), and carotid IMT (P = 0.6874 at six months and P = 0.6951 at 12 months). Comparison of percentage changes in mean IMT revealed no statistical differences between the patient groups with or without statin administration (P = 0.7208 at six months, P = 0.5928 at 12 months). Conclusions According to the 12-month observation data, no significant association was detected between TCZ use and IMT changes in patients with RA, despite its effects on serum lipids. Further long-term studies are needed to confirm thatIL-6 receptor blockers have cardiovascular effects.

Effects of Herpesviruses on Proinflammatory Cytokines in Chronic Periapical Lesions.

BACKGROUND Apical periodontitis is caused by infected dental pulp and may be associated with dental caries or trauma, which can destroy periradicular tissues. This study included periapical tissue of patients with chronic apical periodontitis and aimed to evaluate the presence of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) using polymerase chain reaction (PCR), and to determine the levels of inflammatory cytokines using enzyme-linked immunoassay (ELISA). MATERIAL AND METHODS A total of 79 patients participated in this study. Periapical lesions were taken from the tooth roots indicated for extraction, and were divided into 2 groups: symptomatic and asymptomatic. PCR was used to identify HCMV and EBV. Interleukin 6 (IL-6), interleukin 8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor beta-1 (TGFß-1) were determined using ELISA. RESULTS The occurrence of HCMV (P<0.05) and dual HCMV/EBV infection (P<0.05) was significantly more frequent in symptomatic lesions compared to asymptomatic. The concentrations of IL-6, IL-8, and TNF-alpha were significantly higher in virus-positive lesions compared to virus-negative ones (P<0.05); especially high cytokine levels were found in lesions with dual HCMV/EBV infection (P<0.05). The level of TGF-ß1 was higher in virus-positive compared to virus-negative lesions, but the difference was significant only in lesions with dual HCMV/EBV infection (P<0.05). CONCLUSIONS The higher prevalence of herpesviruses in symptomatic lesions compared to asymptomatic ones indicates their important role in pathogenesis of periapical lesions. Expression of TNF-alpha, IL-6, and IL-8 may be of importance in the development and clinical features of herpesvirus-infected lesions, while TGF-ß1 appears to be of no significance.

Designing a Multi-Epitope Candidate Vaccine against Zika Virus NS5 Protein Through Immunoinformatics Approach for Producing in Plant Systems

Background: The Zika virus is an infectious virus that belongs to the Flaviviridae family. It is transmitted to humans through mosquito vectors and poses a serious threat to human health. Despite available resources, there is currently no effective and secure vaccine against the Zika virus to prevent such infections Objective: Observation of symptoms including fever, rash, joint pain, and conjunctivitis (red eyes) along with the possibility of severe complications during pregnancy, including birth defects such as microcephaly, is noted in infected individuals. So, it is the need of the hour to develop a potential vaccine that can immunize the human beings against this virus and protect them. For the purpose of vaccine development against the Zika virus, bioinformatics and immunoinformatics approaches are utilized. Materials and methods: Different epitopes from RdRp Protein of Zika virus were predicted including B-Cell and T- Cell epitopes. The selected epitopes were analyzed further in order to check their antigenicity, immunogenicity, solubility and toxicity in correspondence to their ability to initiate immune response in the individuals. The potential epitopes having the ability to trigger a good immune response and having non- toxic nature were utilized for vaccine development. By combining the different epitopes, a potential multi-epitope vaccine construct was developed. Combining of epitopes was done using EAAAK, AAY and GPGPG linkers. A 50S ribosomal protein was added at N terminal to improve the immunogenicity of the vaccine and 6-His tag protein also added at C terminal. Results: The constructed vaccine was found to have good population coverage all across the world. In molecular docking, strong interactions were found between the constructed vaccine and Toll-like receptor 3 (-346.16), suggesting their potential relevance in the immune response to the vaccine and dynamical stabilities were inspected through molecular dynamic (MD) simulation methods. The developed vaccine construct also exhibited good results of immune response in immune simulation and revealed that the constructed vaccine activates B and T lymphocytes which induce high levels of antibodies and cytokines to combat Zika infection. The constructed vaccine is an effective biomarker with non-toxicity, non-allergic properties, good immunogenicity, and antigenicity. However, experimental assays are required to verify the results of the present study. Conclusion: These findings suggest that design vaccine candidate effectively trigger immune response with no side effects against Zika virus. Keywords:- Zika virus vaccine, Epitope prediction, immunoinformatics, Multiepitope vaccine, Molecular docking.

Numerical Modelling of Myocardial Infarction in Multivessel Coronary Lesion. II. Patterns of Formation of Large-Scale Damages and Structures

In this work, the basic mechanisms of myocardial infarction development during its inflammatory phase have been studied using mathematical modelling methods. Complex scenarios associated with multivessel lesions of the coronary bed and variability in baseline indicators of the innate immune system state have been considered. Special attention is paid to methodological issues related to the analysis of the effectiveness of the algorithm for approximate solutions of nonlinear initial-boundary value problems and setting up computational experiments under conditions close to the conditions of laboratory experiments in the field of interest. A numerical analysis was performed of several of the most common variants in laboratory practice of the formation of a large lesion in the left ventricle of the mouse heart, caused by the spatiotemporal heterogeneity of the properties of the environment, the immune reaction and ischemic myocardial damage, including recurrent infarction. The main attention is on the following aspects: – analysis of the mechanism of formation of large-scale infarct damages with an extensive core covering almost the entire infarct focus, or with a relatively small core, and assessment of the role of inflammation in these processes; – analysis of current scenarios of structure formation and the role of nonlinear dynamic structures of demarcation inflammation in the formation of a large but highly structured focus. The obtained data allow us to conclude that there is sufficient conservatism during infarction, evidenced by the basic patterns of the inflammatory phase revealed during modeling. The variability of heart attack scenarios manifests itself as a «memory»-effect about the initial data. We observe the «memory»-effect only at a biologically significant time interval, but we also note a general tendency to switch to the usual scenario of inflammation in large-focal infarction, which eliminates the peculiarities of the initial and individual conditions. The role of inflammation has been investigated in the context of a wave process in which spatial localization and the interaction of density and concentration waves can determine the main features of myocardial damage. In particular, within the framework of the adopted mathematical model, the conditions under which the formation of local zones with a relatively low or, conversely, high level of damage is possible have been described. In our work, we have established a significant interdependence between the formation of quasi-stationary structures and the intensity of the immune response. The high probability of developing severe or even terminal myocardial infarction may be due to the high level of immune system factors, in particular, monocytes-macrophages or cytokines in the reinfarction heart.

Mechanism of Interleukin-17A Regulation of Mesenchymal Stroma/Stem Cell Osteogenic Differentiation.

The immune and musculoskeletal systems closely interplay in bone repair and regeneration. After bone injury, the body produces high levels of cytokines and signaling molecules to balance bone formation and resorption. Interleukin (IL)-17A, a cytokine expressed early in the inflammatory process, profoundly influences osteoprogenitor cell fate, thereby contributing to bone homeostasis. In addition, mesenchymal stromal/stem cells (MSCs) can differentiate into osteoblasts, contributing to bone repair and regeneration. Although IL-17A can influence MSCs to become early osteoprogenitor cells, it also can inhibit bone formation. However, the reasons for these dual roles are not yet fully understood. This review overviews IL-17A signaling and the mechanisms that govern MSCs' osteogenic differentiation and summarizes relevant data from the literature on IL-17A's pro- and anti-osteogenic roles.

Cytokines Levels and Anti-tuberculosis Drug Resistance among HIV Seropositive Patients in Ibadan, Southwest, Nigeria

Susceptibility to TB disease rises on mutations in cytokine receptors which ultimately lead to defective signaling and alter the immune surveillance and bacterial killing. Therefore, this study was undertaken to investigate the association between cytokines and anti-TB drug resistances. A total of 217 people living with human immunodeficiency virus at University College Hospital, Ibadan were recruited. Venous blood and sputum samples were collected. Sputum samples were cultured for Mycobacterium tuberculosis complex and characterized using standard methods. Anti-TB drug resistance was detected using phenotypic and genotypic methods. Cytokines levels (IL-10 and IFN-Y) were estimated using ELISA method. Association between cytokines and anti-TB drug resistance was determined using linear regression analysis. There were 105(48.4%) males and 112(51.6%) females with a mean age of 42.95 ± 8.286 years and a range of 28-65 years. The prevalence of TB among PLHIV was found to be 19.1% with male having higher rate than female (P&lt;0.05), and highest rate among aged ≥40. There was no significant association between cytokines (IL-10 and IFN-y) and sex of the subjects (P&gt;0.05). However, females had higher IFN-y level of 231.06±61.81 IU/ml when compared to 227.53±57.92 IU/ml obtained from males. The prevalence of multi-drug resistance TB was 11.9% with Rifampicin resistance having the highest rate of 5.5%. There was no significant association between cytokines (IL-10 and IFN-y) and sex, but age group &gt;40 years had higher IL-10 and IFN-y level of 226.53±52.39 IU/ml and 232.41±66.63 IU/ml respectively. There was an association between high cytokines levels and anti-TB drug resistance (P&lt;0.05). Highest cytokines levels; IL-10 (401.00±7.07 IU/ml) and IFN-Y (401.00±7.07 IU/ml) were recorded in Levofloxacin resistance. There was an association between high cytokines levels and genotypic anti- TB drug resistance (P&lt;0.05) with highest cytokines levels; IL-10 (400.12±12.37 IU/ml) and IFN-Y (472.87±31.93 IU/ml) recorded in Rifampicin resistance. This entire study revealed that there is high prevalence of MDR-TB among PLHIV, with Rifampicin monoresistance having the highest resistance rate among the drugs tested, and pro and inflammatory cytokines (IL-10 and IFN-y) play significant role in the development of Multi Drug Resistant-Tuberculosis among people living with HIV.

Chronic inflammation degrades CD4 T cell immunity to prior vaccines in treated HIV infection

To date, our understanding of how HIV infection impacts vaccine-induced cellular immunity is limited. Here, we investigate inflammation, immune activation and antigen-specific T cell responses in HIV-uninfected and antiretroviral-treated HIV-infected people. Our findings highlight lower recall responses of antigen-specific CD4 T cells that correlate with high plasma cytokines levels, T cell hyperactivation and an altered composition of the T subsets enriched with more differentiated cells in the HIV-infected group. Transcriptomic analysis reveals that antigen-specific CD4 T cells of the HIV-infected group have a reduced expression of gene sets previously reported to correlate with vaccine-induced pathogen-specific protective immunity and further identifies a consistent impairment of the IFNα and IFNγ response pathways as mechanism for the functional loss of recall CD4 T cell responses in antiretroviral-treated people. Lastly, in vitro treatment with drugs that reduce inflammation results in higher memory CD4 T cell IFNγ responses. Together, our findings suggest that vaccine-induced cellular immunity may benefit from strategies to counteract inflammation in HIV infection.

High-affinity chimeric antigen receptor signaling induces an inflammatory program in human regulatory T cells

Regulatory Tcells (Tregs) are promising cellular therapies to induce immune tolerance in organ transplantation and autoimmune disease. The success of chimeric antigen receptor (CAR) Tcell therapy for cancer has sparked interest in using CARs to generate antigen-specific Tregs. Here, we compared CAR with endogenous Tcell receptor (TCR)/CD28 activation in human Tregs. Strikingly, CAR Tregs displayed increased cytotoxicity and diminished suppression of antigen-presenting cells and effector T (Teff) cells compared with TCR/CD28-activated Tregs. RNA sequencing revealed that CAR Tregs activate Teff cell gene programs. Indeed, CAR Tregs secreted high levels of inflammatory cytokines, with a subset of FOXP3+ CAR Tregs uniquely acquiring CD40L surface expression and producing IFN-γ. Interestingly, decreasing CAR antigen affinity reduced Teff cell gene expression and inflammatory cytokine production by CAR Tregs. Our findings showcase the impact of engineered receptor activation on Treg biology and support tailoring CAR constructs to Tregs for maximal therapeutic efficacy.

Insights into respiratory microbiome composition and systemic inflammatory biomarkers of bronchiectasis patients.

The population of microorganisms on/in the human body resides in distinct local microbiomes, including the respiratory microbiome. It remains unclear what defines a healthy and a diseased respiratory microbiome. We investigated the respiratory microbiome in chronic pulmonary infectious disease, i.e., bronchiectasis, and researched correlations between microbiome composition, systemic inflammatory biomarkers, and disease characteristics. The bronchoalveolar microbiome of 47 patients was sequenced, and their serum inflammatory mediators were quantified. The microbiomes were grouped based on their content and diversity. In addition, patients were also grouped into low- and high-response groups according to their inflammatory biomarkers' levels. Certain microbiome compositions, mainly single-species dominated, were associated with high levels of inflammatory cytokines, whereas others correlated with low inflammatory response and remained diverse. We conclude that respiratory microbiome composition is a valuable resource for the diagnostics and personalized management of bronchiectasis, which may include preserving microbiome diversity and introducing possible probiotics.

Cancer-associated fibroblast subtypes modulate the tumor-immune microenvironment and are associated with skin cancer malignancy

Cancer-associated fibroblasts (CAFs) play a key role in cancer progression and treatment outcome. This study dissects the intra-tumoral diversity of CAFs in basal cell carcinoma, squamous cell carcinoma, and melanoma using molecular and spatial single-cell analysis. We identify three distinct CAF subtypes: myofibroblast-like RGS5+ CAFs, matrix CAFs (mCAFs), and immunomodulatory CAFs (iCAFs). Large-cohort tissue analysis reveals significant shifts in CAF subtype patterns with increasing malignancy. Two CAF subtypes exhibit immunomodulatory properties via different mechanisms. mCAFs sythesize extracellular matrix and may restrict T cell invasion in low-grade tumors via ensheathing tumor nests, while iCAFs are enriched in late-stage tumors, and express high levels of cytokines and chemokines to aid immune cell recruitment and activation. This is supported by the induction of an iCAF-like phenotype with immunomodulatory functions in primary healthy fibroblasts exposed to skin cancer cell secretomes. Thus, targeting CAF variants holds promise to enhance immunotherapy efficacy in skin cancers.

Simultaneous Stimulation of Peripheral Blood Mononuclear Cells with CpG ODN2006 and α-IgM Antibodies Leads to Strong Immune Responses in Monocytes Independent of B Cell Activation.

CpG ODN2006 is widely used both in vitro and in vivo to achieve B cell activation and has been previously applied in clinical trials as an adjuvant and anti-cancer agent. Recent studies have demonstrated the benefit of combining CpG ODN2006 with α-IgM antibodies to obtain optimal B cell activation in vitro. In this study, we expanded the knowledge of how both agents affect other types of peripheral blood mononuclear cells (PBMCs), thereby highlighting beneficial and potentially unfavorable properties of the combination of CpG ODN2006 and α-IgM when applied beyond isolated B cells. We elucidated the effects of both compounds on mixed PBMCs, as well as on B cell- and monocyte-depleted PBMCs, allowing us to distinguish between direct effects and indirect influences mediated by other interacting immune cells. Flow cytometry was used to measure the expression of surface markers and intracellular cytokines, while ELISA and multiplex assays were performed to determine cytokine secretion. Our results revealed that stimulation of mixed PBMCs with CpG ODN2006 and α-IgM strongly increased cytokine secretion, primarily originating from α-IgM-stimulated monocytes. Monocyte activation was confirmed by increased CD86 and HLA-DR expression and occurred independently of B cells. The high level of monocyte-derived cytokines after α-IgM exposure did not affect B cell activation. However, it represents a rather unfavorable property for clinical applications. In conclusion, α-IgM is a potent inducer of cytokine production in monocytes. Based on our findings we hypothesize that significant side effects on monocytes can occur when using α-IgM to enhance CpG ODN2006's efficacy on B cells, particularly in clinical settings.

Features of hyperinflammation link the biology of EBV infection and cytokine storm syndromes

BackgroundOvert immune activation by viral infections can lead to cytokine storm syndromes, such as hemophagocytic lymphohistiocytosis (HLH) and macrophage activation syndrome (MAS). ObjectiveWe aim to compare the immune response to different viral pathogens to understand the connection between infections and cytokine storm syndromes. MethodsWe recruited children who presented to the emergency room with fever for ≥ 3 days. We performed immune profiling using Olink proximity extension assay and flow cytometry. We compared the findings with cases of HLH, MAS, Kawasaki disease (KD), and multisystem inflammatory syndrome in children (MIS-C). ResultsWe enrolled 352 febrile patients and studied 110 cases with confirmed common viral infections. We found that Epstein-Barr virus (EBV) uniquely triggered high levels of multiple cytokines (IL-18, IL-27, tumor necrosis factor, FLT3 ligand, and lymphotoxin alpha) and IFN-γ-induced chemokines (CXCL9/10/11 and CCL19). These patterns are similar to the hyperinflammatory response associated with HLH / MAS, but less consistent with the findings in KD and MIS-C. Flow cytometry analysis revealed that CD38+HLA-DR+ T lymphocytes, which are pathogenic cells responsible for IFN-γ production in HLH / MAS, are vastly expanded in patients with acute EBV infection. Cell sorting identified CD38+HLA-DR+ T cells as atypical lymphocytes that are classically associated with acute EBV infection. ConclusionThis work broadens our understanding of common viral infections in children and provides an immunologic basis for the link between EBV infection and HLH / MAS.