Number Of Immune Cells Research Articles

SURG-48. LASER INTERSTITIAL THERMAL THERAPY REMODELS THE TUMOR IMMUNE LANDSCAPE IN A MOUSE GLIOMA MODEL

Abstract BACKGROUND Laser interstitial thermal therapy (LITT), a minimally invasive surgical technique for tumor ablation, is increasingly being used to treat primary and recurrent glioma. While the primary goal of using LITT has been to destroy tumor tissue, little is known about the impact of LITT on the overall immune response. To investigate changes in the tumor immune landscape, we utilized a mouse model of LITT and applied it to a poorly immunogenic mouse model of glioma. METHODS SB28 glioma cells were implanted in the right frontal lobe of C57BL/6 mice, and on day 7, an optical fiber is inserted into the tumor, paired with a thermocouple for real-time temperature monitoring. A 30W laser was activated in pulses to maintain a peri-lesional temperature of 44C for 150-180 seconds. The tumor and LITT-associated immune response was analyzed by multiparameter flow cytometry, single cell RNA-seq, and multiplex immunofluorescence. RESULTS LITT (versus sham control) triggered an increase in neutrophils in the tumor during the acute phase at day 3 following treatment. Total immune cell numbers increased over time in both sham- and LITT-treated mice, but there was a significant increase at post-LITT day 7 in select adaptive immune cell populations including type 1 conventional dendritic cells, NK cells, and gamma delta T cells. Monocytes/macrophages comprised the majority of the immune infiltrate, and multiplex immunofluorescence revealed these populations localized to the perimeter of the LITT lesion. scRNA-seq also revealed enrichment of a Fabp5- and Gpnmb-expressing macrophage subset following LITT. CONCLUSIONS LITT modifies the tumor immune infiltrate with increases in neutrophils as well as select adaptive cell and macrophage subsets. Future studies will explore adjunct therapies to further augment the immune response in the setting of LITT.

Immune cell changes in Helicobacter pylori infection-induced glandular epithelial cell damage of the gastric mucosa

Objective The purpose of this study was to investigate the relationship between Helicobacter pylori (H. pylori) infection-induced changes in gastric mucosal immune cells and glandular epithelial cell damage and the histopathological characteristics of these changes. Methods We performed a detailed histomorphometry and immunohistochemical analysis of a total of 1635 H. pylori-infected gastric mucosal specimens. Results Stage-wise features were as follows: Early stage of infection: H. pylori was colonized in the mucous layer, and very few neutrophils were visible in the layer. Gastric surface epithelial cell infection stage: H. pylori specifically and selectively adhered to the cytoplasm of the surface mucous cells, with the presence of a small number of neutrophils, eosinophils, and lymphocytes infiltration, which is termed early immune response. Compensatory mucous neck cell hyperplasia stage: proliferation of stem cells in the deep gastric pit and infiltration of a large number of lymphocytes in the superficial layer of lamina propria were observed, which is termed immune cell mobilization counterinsurgency. Lamina propria lesion stage: excessive upward migration of the proliferative area. Infiltration of a large number of lymphocytes, plasma cells, monocytes, macrophages, and other immune cells was observed in the whole layer of the gastric mucosa, which is termed automatic replication of immune cells. Abnormal proliferative transformation stage: presence of intranuclear milky globular body cells or signet-ring cell-like heterotypic cells at the junction of the gastric pit and the gastric gland, with proliferation of large numbers of immune cells and vacuolar degeneration of immune cells, which is termed the proliferation and degeneration of immune cells. Intraepithelial neoplasia stage: clonal hyperplasia of epithelial cells and immunosuppression. Conclusion For controlling the occurrence and development of gastric cancer and effective immune intervention, it is essential to grasp the relationship between H. pylori infection-induced changes in gastric mucosal immune cells and glandular epithelial cell damage and its histopathological characteristics.

Influence of saponin tauroside Sx1 on lifespan and expression of vitamin D receptors in the liver tissue of mice with influenza infection

Introduction. Respiratory infections, including influenza, are often accompanied by hepatitis in humans which pathogenesis is not fully understood. According to the available datan D deficiency is presumably a risk factor in the occurrence of acute respiratory viral infections due to its modulation of immune response. Recent studies indicate that several plant compounds can interact with vitamin D receptors (VDRs) and modulate the activity of VDRs. The biologically active components saponins have found widespread use in clinical practice due to their wide range of biological and pharmacological effects which mechanisms are still largely unclear.Aim. To study the effect of oral administration of saponin tauroside Sx1, obtained from Crimean ivy, on life expectancy and activation of vitamin D receptors in the liver of mice during experimental viral infection of varying severity.Materials and methods. The 11 subgroups were formed from male BALB/c mice, were used in the experiment depending on the infectious dose of influenza virus, 5 LD50 virus or 10 LD50 virus, respectively, including control. Saponin tauroside Sx1 was used as a corrector. Immunohistochemical studies were carried out automatically in a BOND-MAX immunohistainer (Leica, Germany). Primary rabbit polyclonal antibodies to the vitamin D receptor were used.Results and discussion. Due to the administration of tauroside Sx1 at a dose of 200 μg/mouse/day increases the average life expectancy of animals receiving saponin by 4.6 days. Reducing the infectious dose of IV from 10 LD50 to 5 LD50 also changed the onset of death of animals by 2 days in both groups. With different infectious doses of the virus, on the 10th day of the experiment in the subgroups, the expression of vitamin D receptors changes without correction. In subgroup 2V, the number of total positive cells was lower than the control group. Moreover, in the 2Vir subgroup, VDR expression was significantly higher than the control group.Conclusion. The saponin tauroside Sx1 at a dose of 200 µg/mouse/day has a fairly pronounced antiviral effect during experimental infection of mice with the influenza A/H1N1 virus, which is manifested by an increase in the average life expectancy of animals (for 4.6 days) and a decrease in the mortality rate during severe influenza infection, compared with the control group, where 100 % mortality was observed by the 14th day of the experiment. The introduction of saponin on the 4th day of the experiment in all subgroups reduces the total number of immune cells that intensively express VDR.

Severe herpes simplex virus − 1 Kaposi varicelliform eruption and SARS-CoV-2 infection in atopic dermatitis treated with dupilumab

Herpes Simplex Virus-1 (HSV-1) Kaposi varicelliform eruption (KVE) is a rare and severe cutaneous manifestation, clinically characterized by the presence of widespread vesicles and pustules. A case report details a patient with a history of Atopic Dermatitis (AD) and recent SARS-CoV-2 infection who developed a severe KVE subsequent to the viral illness. The patient, a 35-year-old male, presented with severe atopic dermatitis (AD) subsequent to a SARS-CoV-2 infection. In a period of four months, the dermatological eruption underwent a rapid progression to a severe state, characterised by the presence of extensive vesicles and pustules, in addition to the emergence of symptoms. In conjunction with a chest CT scan, plasma and antigen testing, the patient was confirmed to be positive for HSV-1 positive. The patient exhibited elevated levels of IgE and a notable reduction in the absolute number of immune cells. The patient was treated with valaciclovir, piperacillin-tazobactam, IVIG at the same time. Within seven days of treatment, the blisters had dried up and the scabs had fallen off without any pain, pruritus, or fever. This case highlights the potential for severe viral eruptions, such as KVE in individuals with underlying dermatological conditions following viral infections.

The pivotal role of osteopontin in UV-induced skin inflammation in a mouse model.

Osteopontin (OPN) is a pro-inflammatory protein that influences bone remodelling, wound healing, angiogenesis, allergic inflammation, and skin diseases such as psoriasis, contact dermatitis and skin cancer. However, the role of OPN in the skin remains unclear. Therefore, this study aimed to investigate the role of OPN in the skin, particularly in the context of ultraviolet (UV) irradiation-induced inflammation. OPN expression and its effects on inflammatory modulators were assessed in human skin, in a mouse model and in vitro, using a UV source emitting both UVB and UVA radiation, which collectively contribute to UV-induced skin inflammation. OPN expression increased in human and mouse skin after UV irradiation. Compared with wild-type mice, UV irradiation-induced skin phenotypes, such as erythema and skin thickening, were alleviated in OPN-/- mice. In addition, the number of immune cells recruited to the skin after UV irradiation and the expression of inflammatory cytokines and matrix metalloproteinases (MMPs) were observed to be decreased in the skin of OPN-/- mice compared with that of wild-type mice. By contrast, the degree of skin inflammation was higher in the hOPN KI mice than in wild-type mice. Treatment with recombinant OPN increased the expression of MMP-1 and inflammatory cytokines in human dermal fibroblasts and epidermal keratinocytes in vitro. Our results suggest that OPN may play a regulatory role in UV-induced skin inflammation.

Wire injury induced moderate aortic valve stenosis in mice is accompanied by a chronic systemic inflammatory reaction

Abstract Background Patients with aortic valve stenosis (AS) have elevated serum levels of inflammation markers, and blood cytokine levels correlate with ventricular recovery after transcatheter aortic valve replacement. While presence of inflammatory processes in stenotic aortic valves is acknowledged, no systematic characterization of the systemic immune reaction upon AS has been performed, yet. Purpose Hypothesis of this study was that AS induces a systemic inflammatory reaction linked with local processes in the heart. Methods AS was induced by wire injury (WI) or sham surgery was performed in 3-4 months old male C57Bl/6J mice. AS severity, left ventricular (LV) function and hypertrophy indices were assessed with echocardiography (echo). After four weeks, explanted organs were weighted, and flow cytometry identified total leukocyte numbers in blood, heart, and peripheral immune organs (spleen, liver, lymph nodes) and uptake of Cy5-labelled perfluorocarbon nanoemulsions by whole blood leukocytes. Costimulatory molecules (flow cytometry, RT-qPCR) and cytokines (RT-qPCR, Multiplex Immunoassay) were analysed in heart, peripheral immune organs and blood. Results R-values were positive for spleen weight correlation with AS severity, cardiac function and mass suggesting cardiac hypertrophy to preserve heart function. Myocardial myeloid and T cell r-values were positive or negative, respectively, for correlation with spleen and liver weight, hinting at T cell recruitment from peripheral stores. Immune cell numbers in peripheral lymph nodes and spleen showed negative r-values for correlation with heart hypertrophy indices; number of liver myeloid cells correlated negatively with LV wall thickness (monocytes=-0.8571, p=0.0238) suggesting immune cell recruitment to hypertrophic hearts. Cytokine mRNA levels trended mainly towards increase in heart and regional lymph nodes and reduction in spleen and liver after WI. Correlation with echo was more homogeneous after WI, with r-values mainly positive, except of T cell activation markers, for aortic valves, but negative for myocardium, hinting at mechanisms preserving heart function. Correlating unchanged cytokine protein levels in myocardium and plasma with echo, r-values were mostly positive for myocardium, and were more positive than in sham for plasma. Echo and costimulatory molecule correlation showed a more homogeneous pattern in WI, with mostly positive r-values in myocardium and periphery, while most r-values were negative in sham. Together with reduced PFC-uptake by lymphatic cells, this hints at systemic immune cell activation in AS. Conclusion The results suggest that number and activation state of leukocytes in peripheral organs are directly linked to cardiac processes in AS. Considering the pathological value of inflammation, it is crucial to in future studies find out if modulation of the systemic inflammatory reaction relieves severity of AS and prevents development of heart failure.

Distant Metastases of Breast Cancer Resemble Primary Tumors in Cancer Cell Composition but Differ in Immune Cell Phenotypes.

Breast cancer is the most commonly diagnosed cancer in women, with distant metastasis being the main cause of breast cancer-related deaths. Elucidating the changes in the tumor and immune ecosystems that are associated with metastatic disease is essential to improve understanding and ultimately treatment of metastasis. Here, we developed an in-depth, spatially resolved single-cell atlas of the phenotypic diversity of tumor and immune cells in primary human breast tumors and matched distant metastases, using imaging mass cytometry to analyze a total of 75 unique antibody targets. While the same tumor cell phenotypes were typically present in primary tumors and metastatic sites, suggesting a strong founder effect of the primary tumor, their proportions varied between matched samples. Notably, the metastatic site did not influence tumor phenotype composition, except for the brain. Metastatic sites exhibited a lower number of immune cells overall, but had a higher proportion of myeloid cells as well as exhausted and cytotoxic T cells. Myeloid cells showed distinct tissue-specific compositional signatures and increased presence of potentially matrix remodeling phenotypes in metastatic sites. This analysis of tumor and immune cell phenotypic composition of metastatic breast cancer highlights the heterogeneity of the disease within patients and across distant metastatic sites, indicating myeloid cells as the predominant immune modulators that could potentially be targeted at these sites.

Landscape of the immune infiltration and identification of molecular diagnostic markers associated with immune cells in patients with kidney transplantation

Rejection seriously affects the success of kidney transplantations. However, the molecular mechanisms underlying this rejection remain unclear. The GSE21374 and GSE36059 datasets were downloaded from the Gene Expression Omnibus (GEO) database. Next, the Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT) algorithm was used to infer the proportions of 22 immune cells. Moreover, infiltrating immune cell-related genes were identified using weighted gene co-expression network analysis (WGCNA), and enrichment analysis was conducted to observe their biological functions. Extreme Gradient Boosting (XGBoost) and Least Absolute Shrinkage and Selection Operator (LASSO) logistic regression algorithms were used to screen hub genes. Quantitative real-time PCR was conducted to verify the number of immune cells and hub gene expression levels. The rejection and non-rejection groups showed significantly different distributions (P < 0.05) of eight immune cells (B cell memory, Plasma cells, mast cells, follicular helper T cells, T CD8 cells, Macrophages M1, T Cells CD4 memory activated, and gamma delta T cells). Subsequently, CD8A, CRTAM, GBP2, WARS, and VAMP5 were screened as hub genes using the XGBoost and LASSO algorithms and could be used as diagnostic biomarkers. Finally, differential analysis and quantitative real-time PCR suggested that CD8A, CRTAM, GBP2, WARS, and VAMP5 were upregulated in rejection samples compared to non-rejection samples. The present study identified five key infiltrating immune cell-related genes (CD8A, CRTAM, GBP2,WARS, and VAMP5) involved in kidney transplant rejection, which may explain the molecular mechanism of rejection in kidney transplantation development.

The role of lipids in regulating macrophage antifungal immunity.

Macrophages are critical components of the antifungal immune response. Disturbance in the number or function of these innate immune cells can significantly increase susceptibility to invasive fungal infections. Pathogenic fungi cause billions of infections every year and have an unmet clinical need, with many infections associated with unacceptably high mortality rates that primarily affect vulnerable patients with underlying immune defects. Lipid metabolism has been increasingly appreciated to significantly influence macrophage function, particularly of macrophages residing in lipid-rich organs, such as the brain, or macrophages specialized at clearing dead cells including alveolar macrophages in the lungs. In this review, we provide an overview of macrophage lipid metabolism, and discuss how lipid recycling and dysregulation affect key macrophage functions relevant for antifungal immunity including phagocytosis, functional polarization, and inflammasome activation. We focus on the fungal pathogen Cryptococcus neoformans, as this is the most common cause of death from fungal infection in humans and because several lines of evidence have already linked lipid metabolism in the regulation of C. neoformans and macrophage interactions.

Single-cell transcriptomics and tissue metabolomics uncover mechanisms underlying wooden breast disease in broilers

Accompanied by the accelerated growth rate of chickens, the quality of chicken meat has deteriorated in recent years. Wooden breast (WB) is a severe myopathy affecting meat quality, and its pathophysiology depends on gene expression and intercellular interactions of various cell types, which are not yet fully understood. We have performed a comprehensive transcriptomic and metabolomic atlas of chicken WB muscle. Our data showed a significant increase in the number of immune cells, WB muscle displayed a unique cluster of macrophages (cluster 11), distinct from the M1 and M2 macrophages. Regarding the myocytes, the most significant differences were the decrease in cell number and the intensification of fatty deposits. Satellite cells were involved in muscle repair and regeneration producing more collagen. Interestingly, the interaction network in the WB group was weaker compared to that in normal breast muscle. Additionally, we found six key differential metabolites across 22 pathways. When WB occurs, myocytes and endothelial cells undergo apoptosis, macrophages are activated and exert immune functions, satellite cells participate in muscle rebuilding and repair, and the content of metabolites undergoes significant changes. This cell transcriptome profile provides an essential reference for future studies on the development and remodeling of WB.

Concurrent occurrence of ulcerative duodenitis and ulcerative colitis displaying unique responses to golimumab and ustekinumab.

Recent studies have reported the occurrence of upper gastrointestinal (UGI) inflammation in patients with ulcerative colitis (UC). However, whether UC-associated UGI and colorectal lesions share pathogenic cytokine profiles and responses to biologics remains unknown. Herein, we report a case of concurrent UC and ulcerative duodenitis (UD) that displayed unique responses to biologic treatment. Although treatment with prednisolone (PSL) failed to induce remission in both disorders, golimumab (GLM) and ustekinumab (UST) were effective against UD and UC, respectively, and remission of both disorders was achieved using UST. Immunofluorescence analyses revealed that numbers of immune cells expressing TNF-α were comparable in both duodenal and rectal mucosa before the treatment. GLM or UST treatment markedly decreased numbers of TNF-α-expressing duodenal immune cells, suggesting the presence of correlation between TNF-α expression and disease activity of UD. In contrast, TNF-α expression was not parallel to disease activity of UC because GLM or PSL failed to induce remission despite a marked reduction in TNF-α expression. Responsiveness to GLM or UST together with immunofluorescence studies suggests that TNF-α and IL-12/23p40 are pathogenic cytokines causing UD and UC, respectively, in the present case.

The impact of supervised and home exercise activity intervention on circulating immune cell numbers in cancer patients

IntroductionMalignant diseases challenge clinicians to balance treatment intensity and patient quality of life. Regular physical activity positively impacts mental and physical health, benefiting sleep patterns, heart rate, and overall health. Moreover, telehealth physical exercise training represents a viable option for maintaining intrinsic capacity. The American Cancer Society highlights exercise's role in helping patients cope with anti-cancer treatment side effects. In the Czech Republic, there is no fitness-promoting protocol for cancer patients, despite recognized benefits. Exercise may also enhance immune function, with moderate-intensity exercise potentially positively affecting immune cell counts. ObjectiveThis study aimed to analyze the long-term effects of exercise on circulating immune cells in patients undergoing treatment for solid malignancies. Patients and methods49 participants were recruited at the Masaryk Memorial Cancer Institute, Czech Republic, starting September 2021. Participants were randomized into an experimental (SAPA = undergoing monitored exercise program) group (N = 16; madian age: 46,6; median BMI: 25,1) and a control (CO) group (N = 33; madian age: 52,0; median BMI: 25,3). Flow cytometry was used to examine cellular immunological profiles. The exercise program involved thrice-weekly sessions conducted online. ResultsSAPA group showed stable lymphocyte counts post-exercise (percentage chance: +0,5 %; p = 0,256; effect size r = −0,284), while the CO group exhibited a significant drop (percentage chance: −23,0 %; p = 0,015; effect size r = −0,423). B lymphocyte numbers were significantly higher in the SAPA group post-exercise compared to the CO group (p = 0,003; effect size r = −0,422). The number of Th-lymphocytes, T-c lymphocytes, T-gamma/delta lymphocytes, and NK cells remained stable in SAPA but dropped in CO group. ConclusionExercise's impact on the immune system in cancer patients shows promise, with differences noted between acute and chronic exercise effects. Previous studies on acute exercise indicate a rise in immune cell counts, supporting our findings of stable or increased immune cells with controlled exercise in cancer patients. Controlled physical activity stabilizes or increases certain immune cell populations in patients undergoing chemotherapy for solid malignancies, highlighting the potential benefits of incorporating exercise into cancer treatment protocols.

Characterization of pathological features and immune microenvironment in hepatic tuberculosis and pulmonary tuberculosis.

Hepatic tuberculosis (HTB) is rare extrapulmonary tuberculosis that is clinically similar to liver malignancy, making it difficult for correct diagnoses. Pathology is the gold standard for tuberculosis diagnosis. However, there are few reports on the pathological features of HTB. A total of 32 HTB cases were considered and the differences in pathological features and drug resistance were analyzed and compared with those for pulmonary tuberculosis (PTB). Enhanced CT scans showed ring-shaped delayed enhancement during the arterial, venous, and delayed phases. Most HTB cases were single lesions, with the highest incidence in the right lobe, and the average lesion volume was smaller than that of PTB. The frequency of granuloma in pathological changes, the overall share of the lesion area in the HTB group, and the number of foxp3+ cells were significantly higher than in the PTB group. However, no statistically significant differences were observed between the two groups' other pathological features and immune cell numbers. The immune microenvironment of the normal tissues surrounding the lesion was further analyzed. The findings showed that the number of macrophages and foxp3+ cells in the HTB group was significantly higher than in the PTB group. No significant difference in drug resistance was detected between the HTB and PTB groups. In conclusion, there are substantial differences in the characterization of pathological feature and immune microenvironment between HTB and PTB. The frequency of granuloma and subsequent overall share of the lesion area was significantly higher in HTB compared to PTB.

Analysis of key indicators of chronic stress in cats and dogs

Stress is an integral part of the life of every organism. This issue has become especially important now, during wartime, when stress affects both humans and animals. Military events have led to unprecedented changes in the lives of both humans and animals, affecting their daily routines, social interactions, and stress levels. The study was conducted on 12 dogs and 14 cats. The effects of stressors on cortisol, glucose, total leukocytes, and eosinophils levels were shown. The study’s results show that during chronic stress, the studied animals showed an increase in cortisol and glucose levels, indicating increased stress in these animals in response to changes in their daily lives. Total leukocyte counts in dogs and cats were also higher than reference levels, indicating an immediate activation of the immune system in response to stressors. The differential response of eosinophils in animals underscores the complexity of the immune system’s response to stress. Dogs, as social animals, may experience more pronounced immune modulation in response to stressors, potentially making them more sensitive to fluctuations in immune cell numbers. The study revealed important behavioral changes in dogs and cats. Behavioral manifestations are the most visible indicators of an animal’s emotional well-being. Changes in behavior, including anxiety, hiding, vocalization, and altered social interactions, may reflect the emotional and psychological effects of stress. Our findings underscore the importance of considering individualized strategies for managing animal welfare in emergencies

Changes in conjunctival mononuclear phagocytes and suppressive activity of regulatory macrophages in desiccation induced dry eye

PurposeTo evaluate the effects of dry eye on conjunctival immune cell number and transcriptional profiles with attention to mononuclear phagocytes. MethodsExpression profiling was performed by single-cell RNA sequencing on sorted conjunctival immune cells from non-stressed and C57BL/6 mice subjected to desiccating stress (DS). Monocle 3 modeled cell trajectory, scATAC-seq assessed chromatin accessibility and IPA identified canonical pathways. Inflammation and goblet cells were measured after depletion of MRC1+ MΦs with mannosylated clodronate liposomes. ResultsMononuclear phagocytes (monocytes, MΦs, DCs) comprised 72 % of immune cells and showed the greatest changes with DS. Distinct DS induced gene expression patterns were seen in phagocytes classified by expression of Ccr2 and [Timd4, Lyve1, Folr2 (TLR)]. Expression of phagocytosis/efferocytosis genes increased in TLF+CCR2- MΦs. Monocytes showed the highest expression of Ace, Cx3cr1, Vegfa, Ifngr1,2, and Stat1 and TLF−CCR2+ cells expressed higher levels of inflammatory mediators (Il1a, Il1b, Il1rn, Nfkb1, Ccl5, MHCII, Cd80, Cxcl10, Icam1). A trajectory from monocyte precursors branched to terminate in regulatory MΦs or in mDCs via transitional MΦ and cDC clusters. Activated pathways in TLF+ cells include phagocytosis, PPAR/RXRα activation, IL-10 signaling, alternate MΦ activation, while inflammatory pathways were suppressed. Depletion of MRC1+ MΦs increased IL-17 and IFN-γ expression and cytokine-expressing T cells, reduced IL-10 and worsened goblet loss. ConclusionsDryness stimulates distinct gene expression patterns in conjunctival phagocytes, increasing expression of regulatory genes in TLF+ cells regulated in part by RXRα, and inflammatory genes in CCR2+ cells. Regulatory MΦs depletion worsens DS induced inflammation and goblet cell loss.

Immune regulatory and anti-resorptive activities of tanshinone IIA sulfonate attenuates rheumatoid arthritis in mice.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation and painful joint destruction. Current treatments are helpful in RA remission, but strong immunosuppressive activity and patient resistance are clinical issues. This study explores a dual-action inhibitor, possessing both anti-inflammatory and anti-resorptive properties, as a novel treatment for RA. Therapeutic efficacy and mechanisms of ectosteric (tanshinone IIA sulfonate [T06]) and active site-directed (odanacatib [ODN]) inhibitors of cathepsin K (CatK) were evaluated in RA mouse models. Pathology was assessed through biochemical analyses and histopathological examination. Flow cytometry analysis was performed to characterize immune cells. Anti-inflammatory effects of T06 on nuclear factor kappa beta (NF-κB) pathway were studied in macrophages. T06 effectively lowered the number of joint-resident immune cells, accompanied by significantly reduced production of inflammatory cytokines and collagenolytic proteases. This also included the suppression of Th17 cells and IL-17, resulting in the reduction of osteoclasts in arthritic joints and amplification of the overall anti-resorptive effect of T06, which has been attributed to its selective inhibition of the collagenolytic activity of CatK by preventing its oligomerization. The anti-inflammatory mechanism of T06 was based on blocking the phosphorylation of IκBα in the NF-κB pathway, resulting in reduced activation and expression of inflammatory cytokines. In contrast, ODN had no effect on inflammation and disease progression and was limited to the inhibition of CatK. The combined anti-resorptive and anti-inflammatory activities characterize T06 as a novel therapeutic agent for RA.

Age-associated changes in innate and adaptive immunity: role of the gut microbiota.

Aging is generally regarded as an irreversible process, and its intricate relationship with the immune system has garnered significant attention due to its profound implications for the health and well-being of the aging population. As people age, a multitude of alterations occur within the immune system, affecting both innate and adaptive immunity. In the realm of innate immunity, aging brings about changes in the number and function of various immune cells, including neutrophils, monocytes, and macrophages. Additionally, certain immune pathways, like the cGAS-STING, become activated. These alterations can potentially result in telomere damage, the disruption of cytokine signaling, and impaired recognition of pathogens. The adaptive immune system, too, undergoes a myriad of changes as age advances. These include shifts in the number, frequency, subtype, and function of T cells and B cells. Furthermore, the human gut microbiota undergoes dynamic changes as a part of the aging process. Notably, the interplay between immune changes and gut microbiota highlights the gut's role in modulating immune responses and maintaining immune homeostasis. The gut microbiota of centenarians exhibits characteristics akin to those found in young individuals, setting it apart from the microbiota observed in typical elderly individuals. This review delves into the current understanding of how aging impacts the immune system and suggests potential strategies for reversing aging through interventions in immune factors.

Abstract A062: Neurotropic fibroblast population increases following exposure to chemotherapy in pancreatic adenocarcinoma

Abstract Introduction Pancreatic adenocarcinoma (PDA) will be the second leading cause of cancer related death by 2030 and patient response to chemotherapy is poor with a strong likelihood of cancer recurrence. While past studies have investigated pre- and post-chemotherapy patient samples, there is a paucity of studies looking at fresh, patient matched samples which allow us to observe interpatient heterogeneity in chemotherapy response. Methods Here, we study the effects of chemotherapy on five different patients using sc-RNA sequencing of matched treatment naïve and post treatment tissue biopsies to compare the effects of chemotherapy, not only on the tumor cells, but also on the tumor microenvironment – a key contributor to tumor survival and proliferation. We utilized the Seurat v4 pipeline and CellChat v2 to observe transcriptomic differences in epithelial cells, myeloid cells, t-Cells, and fibroblasts and their corresponding putative interactions in pre- and post- treatment states. Results Unsurprisingly, we found insufficient tumor epithelial cells post-treatment to perform further analysis on due to the intended cytotoxic effects of chemotherapy. Therefore, we turned our focus to the tumor microenvironment, where we had sufficient numbers of stromal and immune cells, to investigate the effects of chemotherapy. We identified that fibroblasts changed the most consistently across all patients in response to chemotherapy. The fibroblasts clustered into three distinct groups: an immunomodulatory group, a myofibroblastic and neurotropic group, as well as a mixed group expressing genes from all three categories. Interestingly, the neurotropic fibroblast signature was universally enriched after chemotherapy in all five patients, despite having a small and clinically heterogeneous cohort. Conclusions Through an analysis of each fibroblast group in the treatment naïve and post treatment categories, we observed that a specific set of fibroblasts become less myofibroblastic and increasingly neurotropic following treatment, leading us to posit that fibroblasts altered by exposure to chemotherapy may potentially contribute to a ‘pro-tumor’ environment that supports recurrence. To test this in vitro, we treated patient derived fibroblasts with Gemcitabine and used RTPCR to analyze their relative changes in gene expression. We specifically probed for genes that were definitive of myofibroblasts and neurotropic fibroblasts and saw that post-chemotherapy exposure, neurotropic genes such as NEGR1 and NFIA were upregulated. This validated our scRNA conclusions that fibroblasts become increasingly neurotropic following treatment and then differentially interact with their surrounding environment. In future studies, we will further analyze the role these neurotropic fibroblasts play in chemoresponse and tumor recurrence. Matched samples are critical to studies of chemotherapy effects on PDA due to the fact that they allow us to deconvolute one aspect of PDA heterogenity - interpatient heterogeneity - which is a large limitation in studies of this disease. Citation Format: Aylin Z Henstridge, Padma Kadiyala, Ahmed M Elhossiny, Jianhua Liu, Vaibhav Sahai, Nicole Peterson, Jorge Machicado, Richard Kwon, Allison Schulman, Erik Wamsteker, George Philips, Martin Fernandez-Zapico, Mark Truty, Costas Lyssiotis, Timothy Frankel, Filip Bednar, Marina Pasca di Magliano, Eileen S Carpenter. Neurotropic fibroblast population increases following exposure to chemotherapy in pancreatic adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A062.

Research Progress on the Effects of PI3K/Akt Pathway on Immune Escape

The PI3K/Akt pathway can promote immune escape in tumors, making cancer cells more difficult for the immune system to attack. It is usually possible to regulate the number and function of immune cells by inhibiting apoptosis and T cell activation, and generate immune escape mutations to make cancer cells more resistant to immune monitoring. In-depth understanding of the role of PI3K/Akt pathway in immune escape will help to reveal the mechanism of tumor development, improve the effectiveness of cancer treatment. development, improve the effectiveness of cancer treatment, and restore the immune system's monitoring and attack on cancer cells by inhibiting the activity of PI3K/Akt. In this review, we review the research progress on the effects of G protein-coupled receptor-activated PI3K/Akt pathway on immune escape. This review aims to summarize domestic and international studies that reveal how the PI3K/Akt pathway plays a key role in immune escape, and through in-depth understanding of the role of the PI3K/Akt pathway in immune escape. This review aims to summarize domestic and international studies that reveal how the PI3K/Akt pathway plays a key role in immune escape, and through in-depth understanding of the molecular mechanism, new therapeutic targets can be identified. Scientists from abroad have explored various cancer forms, such as lung, ovarian, and endometrial cancers, etc., so as to provide targeted treatment strategies and increase the applicability of immunotherapy for different cancers; By studying the role of PI3K/Akt pathway, we can explore the potential application of PI3K/Akt pathway for different cancers. By studying the role of PI3K/Akt pathway, we can explore the potential application of existing drugs or new drugs in immunotherapy, so as to improve the diversity of treatment. techniques are combined with clinical data to discover associations between the PI3K/Akt pathway and treatment response in patients to support personalized medicine. Current studies have found that the PI3K/Akt pathway is a key mechanism for immune monitoring of tumor cell escape, and in-depth study of its role is expected to improve the quality of treatment. Current studies have found that the PI3K/Akt pathway is a key mechanism for immune monitoring of tumor cell escape, and in-depth study of its role is expected to improve current immunotherapy strategies and improve patient survival. In the future, whether there are new therapeutic opportunities for PI3K/Akt pathway in immune escape studies, so as to help optimize cancer treatment strategies, improve the efficacy of immunotherapy in future, whether there are new therapeutic opportunities for PI3K/Akt pathway in immune escape studies, so as to help optimize cancer treatment strategies, improve the efficacy of immunotherapy, and improve patient treatment outcomes.

Research on curcumin mediating immunotherapy of colorectal cancer by regulating cancer associated fibroblasts.

The objective was to investigate curcumin's (Cur) function and associated molecular mechanisms in regulating tumor immunity in colon cancer. Primary cancer-associated fibroblasts (CAFs) from mouse CT26 colon cancer tumors were isolated. Validation of primary CAFs using immunofluorescence assay was done. Cell Counting Kit-8 experiments, real-time quantitative PCR (qPCR), and enzyme linked immunosorbent assay experiments were conducted to investigate how curcumin affected the growth and cytokine secretion functions of CAFs. The effect of curcumin on regulating PD-L1 expression on CT26 cells through CAFs in vitro was explored through coculture of CAFs and tumor cells, qPCR, and western blot experiments. A mouse colon cancer cell model was established in Balb/c nude mice to explore the effect of curcumin on colon tumor cells. Changes in the tumor microenvironment were detected by flow cytometry to explore the synergistic effect of curcumin combined with anti-PD-1 monoclonal antibody in the treatment of mouse colon cancer. In vitro, curcumin prevented the growth and TGF-β secretion of CT26 cells. At the same time, curcumin inhibited the secretion of TGF-β by CAFs, thereby downregulating the PD-L1 expression of CT26 cells. In vivo, curcumin combined with anti-PD-1 antibodies can further enhance the inhibitory effect of PD-1 antibodies on tumors and increase the number of tumor-suppressing immune cells in the tumor microenvironment, such as M1 macrophages and CD8 T cells, thus inhibiting tumors. Immune M2 macrophages, regulatory T cells, and other cells were reduced. In conclusion, curcumin reduces the expression of PD-L1 in colon cancer cells and improves the tumor immune microenvironment by inhibiting the proliferation of CAFs and the secretion of TGF-β. Curcumin and anti-PD-1 treatment have synergistic inhibitory effects on colon cancer.