Triggering Receptor Expressed on Myeloid Cells 2 Mediates the Involvement of M2-Type Macrophages in Pulmonary Tuberculosis Infection.

Background: Macrophage play a significant work in the development of tuberculosis. This study aims to investigate the relationship between TREM2 and macrophage polarization, as well as the related cytokines. Methods: This study involved 43 pulmonary tuberculosis patients and 37 healthy controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of M1/M2 macrophage-related cytokines IL-10 and IL-12 in the peripheral blood of pulmonary tuberculosis patients. The relative mRNA expression levels of TREM2, IL-10 and IL-12 were detected using quantitative real-time PCR (qRT-PCR). Additionally, Spearman rank correlation analysis was used to preliminarily assess the correlation between TREM2 and M1 / M2 macrophages. Hematoxylin-eosin (HE) staining was performed to observe the pathological manifestations of pulmonary tuberculosis lesions. Immunohistochemical (IHC) staining was used to observe the localization of the macrophage-specific molecule CD68, the M1 specific molecule iNOS, the M2 specific molecule CD163, and TREM2. Results: The lesions of pulmonary tuberculosis patients showed Langhans multinucleated macrophages and tuberculous granulomas. The ELISA results indicated that the expression levels of IL-10 and IL-12 were significantly increased in peripheral blood of pulmonary tuberculosis patients. Additionally, the relative mRNA expression levels of TREM2, IL-10 and IL-12 were also significantly higher in the pulmonary tuberculosis group. Furthermore, a positive correlation was observed between TREM2 and IL-10, which are secreted by M2 macrophages. IHC revealed significant positivity of TREM2 and macrophage-related markers in tuberculous granuloma. Specifically, TREM2 and M2 macrophage marker CD163 were significantly expressed in the cytoplasm and membrane of Langhans multinucleated macrophages. Conclusion: The role of macrophage polarization in pulmonary tuberculosis is significant, and further investigation is needed to understand relationship between TREM2 and M2 macrophages.

C18ORF54 promotes immune infiltration and poor prognosis as a potential biomarker for hepatocellular carcinoma.

OBJECTIVE: The morbidity of hepatocellular carcinoma (HCC) is increasing annually. The aim of this study is to investigate the molecular mechanisms of upregulated genes in HCC using bioinformatic methods, so as to identify new potential biological markers. METHODS: The Gene Expression Omnibus database (GEO database) was mined for HCC datasets, which were screened for hub genes and subjected to (Gene Ontology) GO and (Kyoto Encyclopedia of Genes and Genomes) KEGG enrichment analysis. The hub genes were analyzed in terms of Receiver Operating Characteristic (ROC) and methylation levels. Validation of hub genes was completed through basic pathological alterations based on the protein and gene expression level of hub genes. The correlation of genes with immune infiltration in HCC was analyzed based on the database Timer 2.0, and the prognosis as well as survival of hub genes in HCC was analyzed using R studio software. Finally, we performed a gene combination drug analysis on the potential therapeutic targets in HCC. RESULTS: Expression-up-regulated genes were screened via differential analysis, which were mainly enriched in cell cycles and DNA replication pathways. Five hub genes, BRCA1 associated RING domain 1 (BARD1), Mismatch Repair Protein (MSH2), Recombinant H2A Histone Family, Member X (H2AFX), Recombinant H2A Histone Family, Member z (H2AFZ) and Chromosome 18 Open Reading Frame 54 (C18orf54) were identified using a Protein-Protein Interaction Networks (PPI). After a comprehensive analysis of ROC curves and methylation gene mutation sites, C18orf54 was localized followed by basic experiments, so as to verify the C18orf54 upregulated in HCC. Based on the online database Timer 2.0, the immune infiltration of C18orf54 gene in HCC was analyzed, which was found to be negatively correlated with CD4+ T cells and macrophages in HCC, meanwhile a further refinement of the immune checkpoint correlation analysis revealed that C18orf54 was mainly correlated with Hepatitis A virus cellular receptor 2 (HAVCR2), T cell immunoreceptor with Ig and ITIM domains (TIGIT) and Cytotoxic T lymphocyte associate protein-4 (CTLA4). The prognosis and survival of patients with HCC expressing C18orf54 were also analyzed, and it was found that such patients had a higher incidence of adjacent liver tissue inflammation, a higher child-Pugh grade score and a higher rate of residual tumor recurrence. Similarly, the prognosis was worse in the subset of patients with C18orf54. Finally, we performed a combined genetic analysis, which suggested that cyclosporine, quercetin, testosterone and calcitriol might be effective in reducing C18orf54 mRNA expression. CONCLUSION: C18orf54 is involved in the immune infiltration and promotes the poor prognosis of HCC, which could be a candidate biomarker for HCC.

Study on the Correlation between Interleukin-27 and CXCL10 in Pulmonary Tuberculosis.

Objective: To investigate the correlation between interleukin-27 and CXCL10 and other cytokines in pulmonary tuberculosis and to further explore the related miRNAs through bioinformatics. Methods: Collect the lesion tissue and peripheral blood of pulmonary tuberculosis patients and the peripheral blood of healthy controls. Immunohistochemical staining and qRT-PCR were used to observe the expression of interleukin-27, CXCL9, CXCL10, and CXCL11. Then, predict the key miRNA, qRT-PCR was used to verify the expression of miRNA in the peripheral blood and evaluated the correlation between them. Results: Both immunohistochemical staining and qRT-PCR indicated that the expressions of IL-27, CXCL9, CXCL10, and CXCL11 were significantly increased in tuberculosis patients, and IL-27 was significantly correlated with CXCL10 (r = 0.68). Key molecules such as has-let-7b-5p, has-miR-30a-3p, and has-miR-320b were screened out. Among them, has-let-7b-5p was significantly downregulated, and has-miR-30a-3p was significantly upregulated; they were related to interleukin-27 and CXCL10. Conclusion: Our data shows that interleukin-27 and CXCL10 are significantly related in pulmonary tuberculosis, and has-let-7b-5p and has-miR-30a-3p are also related to interleukin-27 and CXCL10. It laid the foundation for subsequently exploiting the potential biomarkers in tuberculosis disease.

Low Tumor Infiltrating Mast Cell Density Reveals Prognostic Benefit in Cervical Carcinoma.

Objectives: Research on the role of mast cells (MCs) in cervical tumor immunity is more limited. Therefore, our study aimed to evaluate the prognostic value of MCs and their correlation with the immune microenvironment of cervical carcinoma (CC). Methods: The Cancer Genome Atlas (TCGA) data was utilized to obtain the degree of immune infiltration of MCs in CC. Meanwhile, this study retrospectively collected patient clinical characteristic data and tissue specimens to further verify the relevant conclusions. Mast cell density (MCD) was measured by the CIBERSORT algorithm in TCGA data and immunohistochemical staining of tryptase in CC tissues. Finally, differentially expressed genes (DEGs) of TCGA data were performed using "limma" packages and key gene modules were identified using the MCODE application in Cytoscape. Results: The results showed MCs were diffusely distributed in CC tissues. Moreover, we found that low tumor-infiltrating MCD was beneficial for overall survival (OS) in the TCGA cohort. Consistent conclusions were also obtained in a clinical cohort. In addition, a total of 305 DEGs were analyzed between the high tumor-infiltrating MCD and low tumor-infiltrating MCD group. Seven key modules, a total of 34 genes, were screened through the MCODE plug-in, which was mainly related to inflammatory response and immune response and closely correlated with cytokines including CSF2, CCL20, IL1A, IL1B, and CXCL8. Conclusion: In short, high tumor-infiltration MCs in CC tissue was associated with worse OS in patients. Furthermore, MCs were closely related to cytokines in the tumor microenvironment, suggesting that they collectively played a role in the immune response of the tumor. Therefore, MCD may be a potential prognostic indicator and immunotherapy target of CC.

Bioinformation Analysis Reveals IFIT1 as Potential Biomarkers in Central Nervous System Tuberculosis.

OBJECTIVE: Central nervous system tuberculosis is the most serious form of extrapulmonary tuberculosis. We aim to discover potential biomarkers involved in the development of the disease. METHODS: Through gene difference analysis, construction of a protein interaction network and tissue specific analysis and other bioinformatics analysis methods, we found out the relatively high expression of important substances in the central nervous system, interferon induced protein with tetratricopeptide repeats 1. Subsequently, the lesion tissue and the resection margin tissue away from the lesion were collected from the 6 cases of central nervous system tuberculosis patients diagnosed from 2019 to 2020, and the pathological manifestations were observed by Hematoxylin and Eosin (H&E) staining, and the expression of IFIT1 was verified by immunohistochemistry. RESULTS: A total of 101 differential genes were analyzed between extrapulmonary tuberculosis patients and normal people, and they were mainly enriched in the interferon pathway. The protein interaction network unearthed 34 key genes. Through tissue specific analysis, it was found that IFIT1 is relatively high in the central nervous system. H&E staining showed the expression of multinucleated macrophages, and immunohistochemistry showed that IFIT1 was significantly positively expressed in the lesion tissue. CONCLUSION: IFIT1 is an important substance involved in central nervous system tuberculosis.

Clinical characteristics and chest computed tomography findings related to the infectivity of pulmonary tuberculosis.

AIM: This study mainly evaluates the clinical characteristics and chest chest computed tomography (CT) findings of AFB-positive and AFB-negative pulmonary tuberculosis (PTB) patients to explore the relationship between AFB-positive and clinico-radiological findings. METHODS: A retrospective analysis of 224 hospitalized tuberculosis patients from 2018 to 2020 was undertaken. According to the AFB smear results, they were divided into AFB-positive pulmonary tuberculosis (positive by Ziehl-Neelsen staining) and AFB-negative pulmonary tuberculosis and patients' CT results and laboratory test results were analyzed. RESULTS: A total of 224 PTB patients were enrolled. AFB-positive (n = 94, 42%) and AFB-negative (n = 130, 58%). AFB-positive patients had more consolidation (77.7% vs. 53.8%, p < 0.01), cavity (55.3% vs. 34.6%, p < 0.01), calcification (38.3% vs. 20%, p < 0.01), bronchiectasis (7.5% vs. 1.5%, p < 0.05), bronchiarctia (6.4% vs. 0.8%, p < 0.05), and right upper lobe involvement (57.5% vs. 33.1%, p < 0.01), left upper lobe involvement (46.8% vs. 33.1%, p < 0.05) and lymphadenopathy (58.5% vs. 37.7%, p < 0.01). CONCLUSION: The study found that when pulmonary tuberculosis patients have consolidation, cavity, upper lobe involvement and lymphadenopathy on chest CT images, they may have a higher risk of AFB-positive tuberculosis.

Study on Virulence Genes of Type III Secretion System of Pseudomonas aeruginosa in Xinjiang Province.

BACKGROUND: To investigate the distribution of virulence genes exoS and exoU of Pseudomonas aeruginosa type III secretion system and their antimicrobial resistance characteristics in Xinjiang Province. METHODS: A total of 228 isolates of Pseudomonas aeruginosa were collected from January 2017 to April 2017 in our hospital. The VITEK2-compac system was used for strain identification and antimicrobial susceptibility test. The disk diffusion method was used for antimicrobial susceptibility supplementation. PCR method was used for detection of exoS and exoU virulence gene. RESULTS: Among 228 isolates of Pseudomonas aeruginosa, 178 (78.07%) were positive for exoS gene, 91 (39.91%) were positive for exoU gene, and 21.49% of the isolates carried both genes (exoU+/exoS+). A total of 30 MDR strains were detected, accounting for 13.16%. The antimicrobial resistance of the exoU+ group was 76.67%, which was significantly higher than that of the exoU-group (23.33%). The difference was statistically significant (p < 0.001). The detection rate of fluoroquinolone-insensitive strains in exoU+ group was as high as 57.45%, which was significantly higher than 42.55% in exoU-group and the difference was statistically significant (p < 0.05). The 30-day mortality rate of the exoU+ group was 8.79%, which was higher than that of the exoU-group (4.38%), and the difference was statistically significant (p < 0.05). CONCLUSIONS: The expression of exoU gene is associated with multidrug resistance, fluoroquinolone resistance, and prognosis. We should enhance the detection of drug resistance and study the pathogenesis and regulation mecha-nism of T3SS, in order to provide new ideas for the design of reasonable treatment strategies and the development of new therapeutic drugs.

Analysis of therapeutic targets and prognostic biomarkers of CXC chemokines in cervical cancer microenvironment.

BACKGROUND: The tumor microenvironment (TME) has received an increasing amount of attention. CXC chemokines can regulate immune cell transport and tumor cell activity to exert anti-tumor immunity. However, studies on the expression and prognosis of CXC chemokines in cervical cancer (CC) are more limited. METHODS: The study investigated the role of CXC chemokines in TME of CC by using public databases. Moreover, quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC) of CXC chemokines were performed to further verify. RESULTS: The transcriptional levels of CXCL1/3/5/6/8/9/10/11/13/16/17 in CC tissues were significantly elevated while the transcriptional levels of CXCL12/14 were significantly reduced. We reached a consistent conclusion that the expression of CXCL9/10/11/13 was verified by quantitative real-time PCR and immunohistochemistry. Moreover, CC patients with low transcriptional levels of CXCL1/2/3/4/5/8 were significantly associated with longer overall survival (OS). The CCL family was related to CXC chemokines neighboring alteration. RELA, NFKB1, LCK and PAK2 were the key transcription factors and kinase targets of CXC chemokines, respectively. We also found there were significant correlations between the expression of CXCL9/10/11 and the infiltration of immune cells (CD8+ T cell, CD4+ T cell, neutrophils and dendritic cells). CONCLUSIONS: In brief, we conducted a comprehensive analysis of CXC chemokines via clinical data and some online public databases. Our results may provide a new idea for the selection of immunotherapeutic targets and prognostic biomarkers for cervical cancer.

Clinical Significance of Procalcitonin, C-Reactive Protein, and Interleukin-6 in Helping Guide the Antibiotic Use for Patients with Acute Exacerbations of Chronic Obstructive Pulmonary Disease.

BACKGROUND: Currently, standards of antibiotic use in acute exacerbations of chronic obstructive pulmonary disease (AECOPD) patients are controversial. OBJECTIVE: The aim of the present study was to analyze the value of procalcitonin (PCT), C-reactive protein (CRP), and interleukin-6 (IL-6) levels to guide the antibiotic treatment of AECOPD patients. METHODS: A total of 371 patients with COPD or AECOPD were included in the study. Clinical and laboratory data were obtained at admission, 325 AECOPD patients and 46 sCOPD patients treated with antibiotics. The receiver operating curve (ROC) was used to evaluate the relationship between CRP, PCT, and IL-6. RESULTS: This study included medical record/case control 1, the COPD group (n = 46) and the AECOPD group (n = 325), and medical record control 2, the nonchanged antibiotic group (n = 203) and the changed antibiotic group (n = 61). In case 1, CRP, PCT, and IL-6 levels in the AECOPD group were higher than that in the control group (P < 0.05), while the result of ROC showed that IL-6 had higher AUC values (0.773) and higher sensitivity (71.7%) than other indicators. The specificity of PCT (93.5%) is higher than other indicators. In case 2, ROC curve results showed that the AUC value of IL-6 (0.771) was slightly higher than PCT and CRP. The sensitivity (85.2%) and specificity (65.5%) of CRP were higher than other indicators. CONCLUSIONS: IL-6 and PCT were elevated in AECOPD patients, resulting in a higher diagnostic value for AECOPD. CRP had a higher diagnostic value for antibiotic use in AECOPD patients.

Study on the relationship between CXCR3 and its ligands and tubal tuberculosis.

OBJECTIVE: Chemokines play an important role in Mycobacterium tuberculosis infection. We aimed to investigate CXCR3, CXCL9, CXCL10 and CXCL11 to explore the correlation between the severity of tubal tuberculosis and chemokines. METHODS: 26 patients with tubal tuberculosis diagnosed in our hospital from 2016 to 2019 were selected as the experimental group, and 18 female patients who underwent high-risk pregnancy supervision in our hospital from 2016 to 2018 were selected as the control group. The pathological manifestations of tubal tuberculosis were observed by HE staining, the expressions of CXCR3 and its ligands in fallopian tubes were detected by immunohistochemistry. RESULTS: Typical granulomatous structure of tubal tuberculosis was observed by HE staining and most of them were accompanied by massive necrosis in the experimental group, while no granulomatous lesions were found in the control group. The results of immunohistochemical staining showed that CXCR3 and its ligands were expressed in the cytoplasm and nucleus of oviduct epithelial cells and inflammatory cells, in the granuloma area. CXCL9, CXCL10 and CXCL11 were related to the severity of the disease. KEY FINDINGS: CXCR3 and its ligands were positively expressed in tubal tuberculosis, especially CXCL9, CXCL10 and CXCL11 were positively correlated with the severity of fallopian tube disease. SIGNIFICANCE: It is helpful for clinical diagnosis and treatment detection, and provides a new therapeutic target for the study of female genital tuberculosis in the future.

Diagnostic value of CXCR3 and its ligands in spinal tuberculosis.

The present study aimed to investigate whether C-X-C motif chemokine receptor 3 (CXCR3) and its ligands may aid in diagnosing spinal tuberculosis (ST). A total of 36 patients with ST and 20 healthy controls were enrolled in the present study. The morphology of tuberculous granuloma in spinal tissue was observed by hematoxylin and eosin staining. The presence and distribution of acid-fast bacilli (AFB) were observed by Ziehl-Neelsen (ZN) staining. The protein expression of Ag85B, IFN-γ, and CXCR3 and its ligands (CXCL9 and CXCL10) were detected by immunohistochemistry. The levels of IFN-γ, CXCR3, CXCL9 and CXCL10 in peripheral blood of patients with ST and healthy controls were detected by reverse transcription-quantitative polymerase chain reaction and ELISA. Typical tuberculous granuloma was observed in the ST close tissue. AFB was observed by ZN staining. Positive expression of Ag85B was found in the surrounding caseous necrotic tissue of the tuberculous granuloma. IFN-γ, CXCR3, CXCL9 and CXCL10 were expressed in the tissue surrounding the tuberculous granuloma and their expression levels were markedly higher than those in the distant tissues. The levels of IFN-γ, CXCR3, CXCL9 and CXCL10 in peripheral blood of patients with ST were significantly higher than those in the healthy controls. Receiver operating characteristic curve analysis demonstrated that IFN-γ, CXCR3 and CXCL10 were more reliable diagnostic markers in terms of sensitivity and specificity. IFN-γ, CXCR3, CXCL9 and CXCL10 were highly expressed in the lesion tissue and peripheral blood samples of patients with ST, and IFN-γ, CXCR3 and its ligands aided in diagnosing ST.

The role of CXCR3 and its ligands expression in Brucellar spondylitis.

AIM: Brucellar spondylitis (BS) is one of the most serious complications of brucellosis. CXCR3 is closely related to the severity of disease infection. This research aimed to study the degree of BS inflammatory damage through analyzing the expression levels of CXCR3 and its ligands (CXCL9 and CXCL10) in patients with BS. METHODS: A total of 29 BS patients and 15 healthy controls were enrolled. Real-Time PCR was used to detect the mRNA expression levels of IFN-γ, CXCR3, CXCL9 and CXCL10 in peripheral blood mononuclear cells (PBMCs) of BS patients and healthy controls. Hematoxylin-Eosin staining was used to show the pathological changes in BS lesion tissues. Immunohistochemistry staining was used to show the protein expression levels of Brucella-Ab, IFN-γ, CXCR3, CXCL9 and CXCL10 in BS lesion tissues. At the same time, ELISA was used to detect the serum levels of IFN-γ, CXCL9 CXCL10 and autoantibodies against CXCR3 in patients with BS. RESULTS: In lesion tissue of BS patients, it showed necrosis of cartilage, acute or chronic inflammatory infiltration. Brucella-Ab protein was abundantly expressed in close lesion tissue. And the protein expression levels of IFN-γ, CXCR3 and CXCL10 were highly expressed in close lesion tissue and serum of BS patients. At the same time, the mRNA expression levels of IFN-γ, CXCR3 and CXCL10 in PBMCs of BS patients were significantly higher than those in controls. CONCLUSION: In our research, the expression levels of IFN-γ, CXCR3 and its ligands were significantly higher than those in controls. It suggested that high expression levels of IFN-γ, CXCR3 and its ligands indicated a serious inflammatory damage in patients with BS.

Clinical Significance of M1/M2 Macrophages and Related Cytokines in Patients with Spinal Tuberculosis.

BACKGROUND: Macrophages are important immune cells involved in Mycobacterium tuberculosis (M.tb) infection. To further investigate the degree of disease development in patients with spinal tuberculosis (TB), we conducted research on macrophage polarization. METHODS: Thirty-six patients with spinal TB and twenty-five healthy controls were enrolled in this study. The specific morphology of tuberculous granuloma in spinal tissue was observed by hematoxylin-eosin (H&E) staining. The presence and distribution of bacilli were observed by Ziehl-Neelsen (ZN) staining. Macrophage-specific molecule CD68 was detected by immunohistochemistry (IHC). M1 macrophages play a proinflammatory role, including the specific molecule nitric oxide synthase (iNOS) and the related cytokine tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). M2 macrophages exert anti-inflammatory effects, including the specific molecule CD163 and related cytokine interleukin-10 (IL-10). The above markers were all detected by quantitative real-time PCR (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and IHC. RESULTS: Typical tuberculous granuloma was observed in the HE staining of patients with spinal TB. ZN staining showed positive expression of Ag85B around the caseous necrosis tissue and Langerhans multinucleated giant cells. At the same time, IHC results indicated that CD68, iNOS, CD163, IL-10, TNF-α, and IFN-γ were expressed around the tuberculous granuloma, and their levels were obviously higher in close tissue than in the distant tissue. RT-PCR and ELISA results indicated that IL-10, TNF-α, and IFN-γ levels of TB patients were also higher than those of the healthy controls. CONCLUSION: The report here highlights that two types of macrophage polarization (M1 and M2) are present in the tissues and peripheral blood of patients with spinal TB. Macrophages also play proinflammatory and anti-inflammatory roles. Macrophage polarization is involved in spinal TB infection.

Study on the association between TGF-ß1 and liver fibrosis in patients with hepatic cystic echinococcosis.

The present study aimed to determine the role of the cytokine transforming growth factor-ß1 (TGF-ß1) in liver fibrosis among patients with hepatic cystic echinococcosis (hepatic CE). Hepatic tissue specimens and serum samples from 30 patients with hepatic CE were collected and TGF-ß1 levels were compared between the two groups. The degree of liver fibrosis was assessed by Masson staining. The expression levels of cytokine TGF-ß1 in liver tissue and serum were detected by immunohistochemistry and ELISA, respectively. Masson staining of liver lesion tissue in patients with hepatic CE indicated different degrees of fibrosis in the liver and the World Health Organization classification was positively correlated with the severity of liver fibrosis (P<0.05). In addition, the expression of cytokine TGF-ß1 was higher in liver lesion tissue specimens compared with that in the adjacent control samples (P<0.05). At the same time, cytokine TGF-ß1 in serum specimens of patients was higher than that in the healthy control group (P<0.05). In conclusion, the expression of TGF-ß1 is upregulated in patients with hepatic CE, which was closely associated to liver fibrosis.

The expression and clinical significance of serum IL-17 in patients with primary biliary cirrhosis.

BACKGROUND: We aimed to investigate the expression and clinical significance of interleukin 17 (IL-17) in patients with primary biliary cirrhosis (PBC). METHODS: PBC patients (n=127), patients without PBC (n=100) were selected from January 2015 to December 2015.The measure of IL-17 level was performed by cytometric beads array (CBA), immunohistochemistry and real-time PCR (QRT-PCR). RESULTS: The expression levels of serum IL-17, IL-6, IFN-γ, TNF-α and IL-10 in PBC groups were significantly higher than control group, a positively correlation between IL-17 and ALT, ALP, GGT, CIV was observed in PBC patients (r=0.350, P=0.013; r=0.373, P=0.008; r=0.337, P=0.017; r=0.349, P=0.021). In addition, IL-17 mRNA expression level in PBC group was higher than control group. Immunohistochemical results suggest that positive cells did not appear in normal tissues, while they appeared in the PBC liver tissue, mainly in the bile duct. CONCLUSIONS: This study shows that IL-17 over expressed in PBC patients, it played a pro-inflammatory effect in the pathogenesis of PBC, most probably as a targeting drug research.

Effect of different types of macrophages on hepatic fibrosis in Echinococcus Granulosus mice.

OBJECTIVE: The purpose of this study is to illustrate the therapeutic effect of which kind of polarized macrophages-based cell therapy in hepatic fibrosis caused by cystic echinococcosis. METHODS: The isolation culture and polarization induction of mouse bone marrow-derived macrophages (BMDM) are established in an in vitro environment. A model of Echinococcus granulosus infection is established by direct injection of the Echinococcus granulosus suspension into the left hepatic lobe. The macrophages are labeled in vitro and the localization of the returned macrophages in the liver of the mice is determined by in vivo tracing. Macrophages of different polarization types are injected into the successfully modeled mice through the tail vein, and the results of HE, Masson, Sirius Red, Desmin immunohistochemistry and Hyp content are inspected to evaluate by liver tissue. Liver pathology and changes in the degree of fibrosis. RESULTS: Bone marrow-derived macrophages have been successfully obtained and induced into M1 and M2 macrophages by different conditions; a model of Echinococcus granulosus infection was successfully established. Macrophages labeled in vitro were returned to the model through the tail vein and they can be located in the liver; a variety of experimental results show that compared with the PBS group, the degree of fibrosis in the M0 group and the M1 group have been reduced, with statistical difference, and the M1 is better than M0 in terms of the therapeutic effect. There is no significant change in the degree of fibrosis in the M2 group. CONCLUSION: Both M1 and M0 macrophages can alleviate liver fibrosis caused by persistent infection of Echinococcus granulosus, but the treatment effect of M1 macrophages is more significant. Cell therapy based on M1 macrophages may be a new idea for treating liver fibrosis caused by persistent infection of Echinococcus granulosus.

CD19+CD24hiCD38hi Regulatory B cells Involved in Hepatic Alveolar Hydatid Infection in Humans.

In larval Echinococcus multilocularis infections causing alveolar echinococcosis (AE) in humans, immune tolerance and/or down-regulation of protective immunity is a marked characteristic of this chronic disease. In order to probe whether CD19+CD24hiCD38hi regulatory B cells (Bregs) took part in the immune suppression, the frequencies of these cells in peripheral blood were analyzed by flow cytometry, and the level of IL-10 and TGF-ß were detected in serum through ELISA of AE patients and healthy individuals. The distribution of IL-10 and TGF-ß in hepatic tissue close to or distant from the lesion of AE patients was detected by immunohistochemistry separately. The results showed that the frequencies of Bregs and the level of IL-10 and TGF-ß increased significantly in AE patients compared with healthy individuals and the distribution of IL-10 and TGF-ß in hepatic tissue close to the lesion was more abundant than in the hepatic tissue distant from lesion. Taken together, the Bregs frequency and the interrelated cytokines (IL-10 and TGF-ß) have a positive correlation to the hepatic alveolar hydatid infection in humans.

Identification of combined T-cell and B-cell reactive Echinococcus granulosus 95 antigens for the potential development of a multi-epitope vaccine.

BACKGROUND: Identification of combined T-cell and B-cell reactive Eg95 antigens for the potential development of a multi-epitope vaccine against Echinococcus granulosus (EG), the causative agent of cystic echinococcosis (CE). METHODS: This study involved the recombinant expression of Eg95 along with associated immune rabbit antiserum preparation. Bioinformatics technology was used to facilitate the analysis of Eg95 molecules. PCR was subsequently used to amplify genetic sequences of the epitopes encoding the T-cell and B-cell reactive peptide fragments. SDS-PAGE was used to assess the expression levels of three proteins. Eg95 serum and patient antiserum, which were assessed using Western blot in order to identify suitable antigenic epitope peptides. ELISA detection assay facilitated comparison of the immune reactivity of the short peptide epitopes. The assay results could be used to determine an EG epitope-based vaccine candidate list from suitably reactive Eg95 epitopes. RESULTS: Eg95 molecules have 3 T-B table. The phage display systems were successfully built using the M13KE carrier. Expression of the three fusion protein peptides were detected. Western blot showed Eg95 antiserum against EG facilitated identification of the three T-cell and B-cell reactive epitopes. After the reaction intensities analyzed by the ELISA, both of the short peptide epitopes Eg95-2 and Eg95-3 showed strong signal strength and associated antigenicity when combined with patient serum and rabbit anti-rEg95 serum. CONCLUSIONS: This study used bioinformatics methods to construct successfully a T-cell and B-cell epitope phage display system for the Eg95 antigen from EG. The two epitopes of Eg95-2 and Eg95-3 demonstrated strong antigenicity with potential applications for peptide vaccine development.