Objective To predict and analyze the physicochemical properties, structural characteristics, and antigenic epitopes of viral protein (VP) VP1 of Coxsackievirus A10 (CV-A10) by bioinformatics methods. Methods The physicochemical properties and structural characteristics of CV-A10 VP1 were predicted by ProtParam, SOPMA, SWISS-MODEL, PDBsum, and ProSA-web. The antigenic epitopes of CV-A10 VP1 were predicted and analyzed by DNAstar, ABCpred, Bepipred 2.0, ElliPro, DiscoTope-2.0, NetMHCpan-4.1, NetMHCIIpan-4.0, Consurf, VaxiJen v.2.0, AllerTOP v.2.0, ToxinPred2, and IEDB immunogenicity. Results Bioinformatics analysis showed that CV-A10 VP1 was a basic, unstable, and hydrophilic protein, of which the secondary structure mainly consisted of random coil. The analysis revealed that CV-A10 VP1 had multiple potential B and T cell antigenic epitopes as well as a dominant antigenic epitope based on the potential epitope. Conclusion CV-A10 VP1 has multiple potential sites that induce specific humoral and cellular immunity, providing important support for its experimental identification, molecular epidemiological studies, and vaccine development.

Objective To investigate the influence of interleukin-6(IL-6) on the expression and function of programmed death-1(PD-1) in patients with hepatocellular carcinoma (HCC) by measuring the plasma level of IL-6 and the expression of PD-1 in peripheral blood CD8 + T cells from HCC patients. Methods A total of 44 HCC patients who attended Shaanxi Provincial People's Hospital or The Second Affiliated Hospital of Air Force Medical University & Tangdu Hospital of Fourth Military Medical University from January to September 2019 were enrolled as HCC group, and 19 healthy controls, matched for age and sex, were enrolled as HC group. Peripheral blood was collected, and plasma and peripheral blood mononuclear cells were isolated to separate CD8 + T cells. ELISA was used to measure the plasma level of IL-6, and flow cytometry was used to measure the expression level of PD-1 in CD8 + T cells. The separated CD8 + T cells were stimulated with anti-IL-6 neutralizing antibody for 24 hours; CCK-8 assay was used to measure cell proliferation, ELISA was used to measure the levels of interferon-γ (IFNγ) and tumor necrosis factor-α (TNFα) in supernatant, real-time PCR was used to measure the relative mRNA expression levels of perforin, granzyme B, and granulysin, and Western blot was used to measure the phosphorylation levels of STAT3 and Src. The t -test or the paired t -test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between groups. Results Compared with the HC group, the HCC group had a significant increase in the plasma level of IL-6 (99.67±20.92 pg/mL vs 81.05±16.76 pg/mL, t =3.427, P =0.001 1). There was no significant difference in the percentage of CD3 + CD8 + T cells between the HCC group and the HC group, while there was a significant increase in the percentage of PD-1 + CD8 + cells in HCC patients (3.79%±1.36% vs 2.20%±0.47%, t =5.335, P < 0.000 1). In the patients with HCC, although anti-IL-6 neutralizing antibody for inhibiting IL-6 in CD8 + T cells did not affect cell proliferation, it downregulated the expression of PD-1 (2.67%±0.91% vs 3.33%±1.12%, t =2.177, P =0.035) and increased the secretion of IFNγ (13.50±3.82 pg/mL vs 10.82±1.37 pg/mL, t =3.170, P =0.002 8), and there were also significant increases in the relative mRNA expression levels of perforin and granzyme B ( t =6.161 and 14.140, both P < 0.000 1) and a significant reduction in the level of phosphorylated STAT3 ( P < 0.000 1). Conclusion Anti-IL-6 neutralizing antibody can enhance the function of CD8 + T cells in HCC patients possibly by increasing the levels of perforin and granzyme B, improving the secretion of cytokines, and inhibiting the expression of PD-1.