ABSTRACT
Objective To examine the relationship between serum vitamin D level and immune imbalance in advanced schistosomiasis patients with liver fibrosis. Methods A total of 120 advanced schistosomiasis patients with liver fibrosis that were admitted to the Department of Schistosomiasis of The First Hospital of Jiaxing City from May 2016 to September 2018 were recruited as the observation group, and 50 healthy volunteers randomly sampled from the hospital during the same period served as the control group. The serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ cell proportion, CD8+ cell proportion, 25-hydroxyvitamin D [25(OH)D] levels were compared between the two groups. Liver fibrosis was classified into grade I, II and III according to the classification criteria of liver fibrosis by ultrasonography, and the serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ proportion, CD8+ proportion, 25(OH)D levels were compared among patients with grade I, II and III liver fibrosis. In addition, all patients were classified into the sufficient group, the insufficient group and the deficient group according to the serum vitamin D level, and the serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ proportion, CD8+ proportion, 25(OH)D levels were compared among these three groups. Moreover, the associations of the serum vitamin D level with these immune indicators were examined. Results The 120 advanced schistosomiasis patients with liver fibrosis included 58 men and 62 women, and had a mean age of (72.00 ± 3.00) years. There were 32 cases with grade I liver fibrosis, 46 cases with grade II liver fibrosis, and 42 cases with grade III liver fibrosis. There were no significant differences between the observation group and the control group in terms of serum D-dimer, total cholesterol (TC), triglyceride (TG), C3 complement or C4 complement levels (t = 2.467, 0.322, 0.790, -2.432 and -2.630, all P values > 0.05); however, there were significant differences seen in alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood calcium, blood phosphorus, IgG antibody, IgA antibody, CD4+ proportion, CD8+ proportion, and 25(OH)D levels (t = 5.130, 6.382, -1.341, 2.361, 8.708, 11.783, -2.995, -6.543 and -3.022, all P values < 0.05). In addition, there were significant differences in AST, ALT, blood phosphorus, IgA antibody, C3 complement, CD8+ cell proportion and 25(OH)D levels among patients with grades I, II and III liver fibrosis (F = 19.704, 16.254, 62.669, 49.347, 5.430, 5.434 and 5.783, all P values < 0.05). There were significant differences in ALT, blood phosphorus, IgA antibody, CD8+ cell proportion and 25(OH)D levels between patients with grades I and III liver fibrosis (all P values < 0.05), and significant differences were seen between patients with grades II and III liver fibrosis in terms of blood phosphorus, IgA antibody and CD8+ cell proportion (all P values < 0.05), while there was a significant difference in the CD8+ cell proportion between patients with grades I and II liver fibrosis (P < 0.05). Moreover, there were significant differences among the sufficient, insufficient and deficient groups in terms of IgG antibody, IgA antibody, C3 complement, CD4+ cell proportion and CD8+ cell proportion (F = 13.303, 59.623, 8.698, 9.969 and 12.805, all P values < 0.05), and there was a significant difference in the CD8+ cell proportion between the insufficient and deficient groups (P < 0.05). Pearson correlation analysis revealed that serum 25(OH)D level were negatively associated with IgG and IgA antibody levels (r = -0.754 and -0.773, both P values < 0.05), and positively associated with C3 complement, CD4+ cell proportion and CD8+ cell proportion in advanced schistosomiasis patients with liver fibrosis (r = 0.827, 0.850 and 0.830, all P values < 0.05). Conclusion Immune imbalance occurs in advanced schistosomiasis patients with liver fibrosis, and serum vitamin D level may correlate with immune imbalance in advanced schistosomiasis patients with liver fibrosis.
ABSTRACT
To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.
Subject(s)
Animals , Humans , Antiviral Agents , Metabolism , Baculoviridae , Genetics , Cell Line , Cloning, Molecular , Gene Expression , Gene Fusion , Genetic Vectors , Immunoglobulin Fc Fragments , Genetics , Immunoglobulin G , Genetics , Insecta , Interferon-alpha , Genetics , Recombinant Fusion Proteins , Genetics , Recombinant Proteins , Genetics , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the distribution of hepatitis delta virus (HDV) marker among hepatitis B virus (HBV) infected patients and to reveal its clinical significance.</p><p><b>METHOD</b>To collect the clinical data and sera samples of HBV infected patients and to detect HDAg, Anti-HDV as well as HBV infection markers by means of enzyme-linked immunosorbnent assay. These data combined with clinical diagnostic results and biochemical index were then analyzed.</p><p><b>RESULT</b>462 samples of HBV infected patients were collected including 210 HBV carriers without symptom, 175 chronic HBV infections, 35 acute HBV infections and 42 liver fibrosis. The HDV infection rate was 4.8% overall. The highest infection rate of 9.5% was found in the group of liver fibrosis whereas the lower rate of 6.9% was found in HBV chronic carriers. HDV infection rate was 7.8% among the population of 40-60 years old, obviously higher than any other age groups.</p><p><b>CONCLUSION</b>HDV infection was significantly higher in the chronic HBV patients and liver fibrosis patients. Because HDV infection was highly associated with the progress of liver disease, we suggest the screen of HDV markers among hepatitis patients and discriminate whether the patient was co-infected with HDV.</p>