Currently, there is limited knowledge about the immunological profiles of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). We used computer-based immunoinformatic analysis and the newly resolved 3-dimensional (3D) structures of the SARS-CoV-2 S trimeric protein, together with analyses of the immunogenic profiles of SARS-CoV, to anticipate potential B-cell and T-cell epitopes of the SARS-CoV-2 S protein for vaccine design, particularly for peptide-driven vaccine design and serological diagnosis. Nine conserved linear B-cell epitopes and multiple discontinuous B-cell epitopes composed of 69 residues on the surface of the SARS-CoV-2 trimeric S protein were predicted to be highly antigenic. We found that the SARS-CoV-2 S protein has a different antigenic profile than that of the SARS-CoV S protein due to the variations in their primary and 3D structures. Importantly, SARS-CoV-2 may exploit an immune evasion mechanism through two point mutations in the critical and conserved linear neutralization epitope (overlap with fusion peptide) around a sparsely glycosylated area. These mutations lead to a significant decrease in the antigenicity of this epitope in the SARS-CoV-2 S protein. In addition, 62 T-cell epitopes in the SARS-CoV-2 S protein were predicted in our study. The structure-based immunoinformatic analysis for the SARS-CoV-2 S protein in this study may improve vaccine design, diagnosis, and immunotherapy against the pandemic of COVID-19.

Using conditional cell reprogramming, we generated a stable cell culture of an extremely rare and aggressive neuroendocrine cervical cancer. The cultured cells contained HPV-16, formed colonies in soft agar and rapidly produced tumors in immunodeficient mice. The HPV-16 genome was integrated adjacent to the Myc gene, both of which were amplified 40-fold. Analysis of RNA transcripts detected fusion of the HPV/Myc genes, arising from apparent microhomologous recombination. Spectral karyotyping (SKY) and fluorescent-in-situ hybridization (FISH) demonstrated coordinate localization and translocation of the amplified Myc and HPV genes on chromosomes 8 and 21. Similar to the primary tumor, tumor cell cultures expressed very high levels of the Myc protein and, in contrast to all other HPV-positive cervical cancer cell lines, they harbored a gain-of-function mutation in p53 (R273C). Unexpectedly, viral oncogene knockdown had no effect on the growth of the cells, but it did inhibit the proliferation of a conventional HPV-16 positive cervical cancer cell line. Knockdown of Myc, but not the mutant p53, significantly inhibited tumor cell proliferation. On the basis of these data, we propose that the primary driver of transformation in this aggressive cervical cancer is not HPV oncogene expression but rather the overexpression of Myc.

Porcine circovirus type 3 (PCV3) has recently been isolated from diseased pigs within the USA. The objective was to detect the presence of PCV3 in dogs. Nested polymerase chain reactions (PCR) with PCV3-specific primers for the capsid gene were used to detect PCV3 genomic DNA in serum samples from dogs (n = 44) in China. There was PCV3 DNA detected in 4 of 44 dogs [all were negative for PCV2 and canine circovirus (CanineCV)]. Based on sequence analysis, positive sequences were grouped into PCV3 genotypes. However, these isolates had close evolutionary relationships with FoxCV (KP941114) and CanineCV (JQ821392). Further investigations of the epidemiology, evolutionary biology, and pathobiology of PCV3 to dogs are warranted.

A newly emerging porcine circovirus, designated PCV3, has been reported in various countries (USA, Poland, South Korea and China) since 2017. Its presence may be associated with porcine dermatitis and nephropathy syndrome (PDNS), reproductive failure, and multisystem inflammation. In this study, we report identification of PCV3 in cases of reproductive failure in various regions in Hunan, China. From January 2015 to December 2016, sera were collected from 190 sows from seven farms with reproductive problems. Specifically, 85 samples were from sows with a history of reproductive failure, whereas the remaining 105 were from healthy sows. The PCV3-positive rate was significantly higher in sows with reproductive failure (45.9%) than in healthy sows (21.9%), based on quantitative PCR (qPCR) assays. Although phylogenetic analysis based on the cap gene suggested that these PCV3 isolates belonged to the clade PCV3a, amino acid sequence variations in the Cap protein still occurred among these isolates, and these might have contributed to antigenic alterations of the Cap protein, based on the Jameson-Wolf antigenic index. Finally, we concluded that PCV3 was circulating in sows in Hunan province, China. However, the association of PCV3 with reproductive failure in sows and its potential for vertical transmission need to be studied further.

Porcine circovirus type 2 (PCV2) causes increased mortality and poor growth or weight loss in apparently healthy swine. Therefore, methods to detect PCV2-specific antibodies in swine serum are important for prevention, diagnosis, and control of PCV2-associated diseases (PCVAD). In this study, PCV2 virus-like particles (VLPs) were used to develop a rapid, simple and economical indirect enzyme-linked immunosorbent assay to detect (with high sensitivity) PCV2-specific antibodies in swine serum. The PCV2 capsid protein (Cap) was overexpressed in E. coli after optimizing the cap gene. Subsequently, the soluble Cap was rapidly purified in one step by automated fast protein liquid chromatography (FPLC). The purified PCV2 Cap was shown by transmission electron microscopy and gel filtration chromatography to be capable of self-assembling into VLPs in vitro. Using the purified VLPs as antigens, optimal operating conditions for the VLP ELISA were determined. The concentration of PCV2 VLPs was 1 µg/ml per well, and the dilution factors for swine serum and horseradish peroxidase (HRP)-labeled goat anti-pig antibody were 1:150 and 1:4000, respectively. Out of 241 serum samples tested with this assay, 83.4 % were found to be positive. Importantly, the VLP ELISA had a total coincidence rate of 97.4 % (74/76) compared to an Ingezim PCV2 ELISA IgG assay. In summary, this rapid, inexpensive VLP ELISA has the potential to greatly facilitate large-scale investigations of PCV2-associated serotypes.

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Background Porcine circovirus type 3 (PCV3), recently widely isolated from pigs with various clinical conditions, is likely globally epidemic. However, development of serological diagnosis for PCV3 in pigs is ongoing. Our objectives were to: 1) establish an indirect ELISA, using PCV3 capsid protein (Cap) prepared by Baculovirus Expression Vector System (BEVS) as a high-quality coating antigen for detection of PCV3-associated antibodies in serum samples; and 2) use this ELISA to conduct a serological survey for PCV3 in various regions of Hunan province, China. Results The PCV3 positive rate to the ELISA assay (total of 190 serum samples) was higher in sows with reproductive failure compared to healthy sows (34/85, 40.0% versus 30/105, 28.6%), with similar results using qPCR assays. Further, in an additional 1038 serum samples collected from January 2016 to May 2018 in various regions of Hunan province and tested with this established ELISA, 20 to 84% were positive for PCV3 (according to region of sera collection), with high PCV3 seroprevalence (> 50%) in herds in Changde, Hengyang and Yueyang. Moreover, among serum samples from herds in Shaoyang and Changde, PCV3 seroprevalence was higher in sows than in other classes of pigs (i.e., suckling piglets, nursery pigs, gilts, growing-finishing pigs and boars). Conclusions We developed a full-length PCV3 Cap-based ELISA using a eukaryotic expression system with excellent potential to elucidate PCV3 epidemiology. Based on this assay, PCV3 has been circulating in Hunan province. PCV3 prevalence was lower in healthy sows than in those with reproductive failure. Further studies are warranted to identify the PCV3 responsible for high seroprevalence in sows and determine pathogenesis of PCV3 in sows with reproductive failure. Electronic supplementary material The online version of this article (10.1186/s12917-019-1810-3) contains supplementary material, which is available to authorized users.