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.
The present study aimed to determine the role of the cytokine transforming growth factor-β1 (TGF-β1) 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-β1 levels were compared between the two groups. The degree of liver fibrosis was assessed by Masson staining. The expression levels of cytokine TGF-β1 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-β1 was higher in liver lesion tissue specimens compared with that in the adjacent control samples (P<0.05). At the same time, cytokine TGF-β1 in serum specimens of patients was higher than that in the healthy control group (P<0.05). In conclusion, the expression of TGF-β1 is upregulated in patients with hepatic CE, which was closely associated to liver fibrosis. Materials and methods Case source and grouping. A total of 30 patients with hepatic CE admitted to the First Affiliated Hospital of Xinjiang Medical University (Urumqi, China) between July 1, 2013 and June 1, 2017 were enrolled in the present study. The age of the patients ranged from 9 to 74 years (median age, 28 years) and the male-to-female ratio was 12/18. Paired liver lesion tissue and normal tissue were obtained from each patient.
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.
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.
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