Orthopoxvirus vector has a broad prospect in recombinant vaccine research, but the rarely severe side-effect impedes its development. Vaccinia virus and Cowpox virus of Orthopoxvirus have broad host range, and they have typical host range genes as K1L, CP77 and C7L. These three genes affect host range of Vaccinia virus, disturb the cell signaling pathways, suppress immune response and are related to virulence.
Cell Line ; Cowpox virus ; genetics ; immunology ; pathogenicity ; physiology ; Genetic Vectors ; Host Specificity ; genetics ; Orthopoxvirus ; genetics ; immunology ; pathogenicity ; physiology ; Signal Transduction ; Vaccines, Synthetic ; immunology ; Vaccinia virus ; genetics ; immunology ; pathogenicity ; physiology ; Viral Proteins ; genetics ; metabolism ; Viral Vaccines ; immunology ; Virulence

Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used: the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with 1 x 105 cells of P388. Mice were injected at the same site with 5 x 105 PFU of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating 2 x 105 tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, 5 x 106 PFU of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.
Animals ; Breast Neoplasms ; Cell Culture Techniques ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Genes, Viral ; HeLa Cells ; Humans ; Injections, Intraperitoneal ; Interleukin-4* ; Interleukins* ; Mice ; Orthopoxvirus ; T-Lymphocytes ; T-Lymphocytes, Cytotoxic ; Vaccinia virus* ; Vaccinia*

In 2022, the outbreak of human monkeypox (HMPX) occurred in many non-endemic countries. World Health Organization (WHO) assesses that this outbreak is "atypical". The history of monkeypox and HMPX must be reviewed to clearly recognize the "typical" outbreaks to fully understand this comment. Therefore, this paper reviews the epidemiological history of monkeypox, especially HMPX, and discusses and analyzes the atypical manifestations and the possible causes of the present outbreak based on the recent views of WHO, other organizations/institutions, and experts. The text describes the thought-provoking history of the interaction between the monkeypox virus and the human being in the past 64 years, and provides various information and views on the outbreak of HMPX, which is helpful to understand risk assessment and the potential impact of this outbreak on clinical and public health in future.
Animals ; Disease Outbreaks ; Humans ; Monkeypox/epidemiology* ; Monkeypox virus

Recently, monkeypox has become a global concern amid the ongoing COVID-19 pandemic. Monkeypox is an acute rash zoonosis caused by the monkeypox virus, which was previously concentrated in Africa. The re-emergence of this pathogen seems unusual on account of outbreaks in multiple nonendemic countries and the incline to spread from person to person. We need to revisit this virus to prevent the epidemic from getting worse. In this review, we comprehensively summarize studies on monkeypox, including its epidemiology, biological characteristics, pathogenesis, and clinical characteristics, as well as therapeutics and vaccines, highlighting its unusual outbreak attributed to the transformation of transmission. We also analyze the present situation and put forward countermeasures from both clinical and scientific research to address it.
COVID-19 ; Disease Outbreaks/prevention & control* ; Humans ; Monkeypox/epidemiology* ; Monkeypox virus ; Pandemics/prevention & control*

Monkeypox is a zoonosis caused by monkeypox virus. Monkeypox virus belongs to the Orthopoxviruses genus in the Poxviridae family, which is regarded as the most important Orthopoxvirus infection in human beings after the extinction of smallpox. Since the first human monkeypox case was reported in the Democratic Republic of the Congo in 1970, monkeypox has become endemic in Central and West African. From May 6 to July 15, 2022, monkeypox has broken out in many countries. Monkeypox cases have been detected in 62 countries and regions. Moreover, human to human transmission has occurred and attracted high global attention. Monkeypox virus has been discovered for more than 60 years, but the understanding and research of its natural host, epidemiological characteristics and treatment are still relatively limited. Therefore, this study analyzes the epidemic situation, the possible causes of the outbreak and the future key research directions, and puts forward countermeasures to provide scientific basis for the prevention and control of monkeypox.
Animals ; Humans ; Monkeypox virus ; Monkeypox/epidemiology* ; Prevalence ; Poxviridae Infections/epidemiology* ; Zoonoses

Animals ; Disease Outbreaks ; Humans ; Monkeypox ; diagnosis ; epidemiology ; transmission ; Monkeypox virus ; isolation & purification ; United States ; epidemiology

Vaccinia virus complement control protein (VCP) is one of the proteins encoded by vaccinia virus to modulate the host inflammatory response. VCP modulates the inflammatory response and protects viral habitat by inhibiting the classical and the alternative pathways of complement activation. The extended structure of VCP, mobility between its sequential domains, charge distribution and type of residues at the binding regions are factors that have been identified to influence its ability to bind to complement proteins. We report that a Lister strain of vaccinia virus encodes a VCP homolog (Lis VCP) that is functional, glycosylated, has two amino acids less than the well-characterized VCP from vaccinia virus WR strain (WR VCP), and the human smallpox inhibitor of complement enzymes (SPICE) from variola virus. The glycosylated VCP of Lister is immunogenic in contrast to the weak immunogenicity of the nonglycosylated VCP. Lis VCP is the only orthopoxviral VCP homolog found to be glycosylated, and we speculate that glycosylation influences its pattern of complement inhibition. We also correlate dimerization of VCP observed only in mammalian and baculovirus expression systems to higher levels of activity than monomers, observed in the yeast expression system.
Amino Acid Sequence ; Animals ; Cell Line ; Cercopithecus aethiops ; Complement Activation ; drug effects ; immunology ; Complement System Proteins ; metabolism ; Dimerization ; Gene Expression ; Glycosylation ; Humans ; Molecular Sequence Data ; Protein Binding ; Protein Structure, Tertiary ; Recombinant Proteins ; genetics ; metabolism ; Smallpox ; immunology ; metabolism ; Structure-Activity Relationship ; Vaccinia virus ; chemistry ; immunology ; metabolism ; Variola virus ; chemistry ; immunology ; metabolism ; Viral Proteins ; genetics ; metabolism ; pharmacology

OBJECTIVES: The aim of this study was to estimate the medical surge capacity required for mass prophylaxis based on a hypothetical outbreak of smallpox.METHODS: We performed a simulation using the Bioterrorism and Epidemic Outbreak Response Model and varied some important parameters, such as the number of core medical personnel and the number of dispensing clinics.RESULTS: Gaps were identified in the medical surge capacity of the Korean government, especially in the number of medical personnel who could respond to the need for mass prophylaxis against smallpox.CONCLUSIONS: The Korean government will need to train 1,000 or more medical personnel for such an event, and will need to prepare many more dispensing centers than are currently available.
Bioterrorism ; Korea ; Smallpox ; Surge Capacity ; Vaccination ; Variola virus

Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-β-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.
Animals ; Mice ; Monkeypox virus ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Blotting, Western ; Recombinant Proteins ; Escherichia coli/genetics*

We have examined isolation and identification protocols for three virus simulant candidates to biological warfare agents. MS2 phage, a simulant for yellow fever virus and Hantaan virus, was propagated using as a host an E. coli strain with F pilus. MS2 phage genome was examined by reverse transcription and polymerase chain reaction (RT-PCR). Coat protein of the phage preparation was examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric analysis. Cydia pomonella granulosis virus (CpGV) is a virus simulant candidate to smallpox virus. CpGV was isolated from a commercialized CpGV pellet. In this study, we developed new isolation and identification protocols for CpGV. One disadvantage of using CpGV is that it is not easy to determine viability of the virus. Here, we have included T4 phage as an alternative. We established a high titer production protocol and developed an easy genome identification protocol that does not require purified phage DNA. Stability of these virus preparations was also examined under various storage conditions. When the virus preparations were not subjected to freeze drying, MS2 phage was most stable when it was stored in liquid nitrogen but unstable at 4℃. In contrast, T4 phage was most stable when it was stored at 4℃. CpGV was stable at −20℃ but not at 4℃. Stability during or after freeze drying was also investigated. The result showed that 70~80% MS2 survived the freeze drying process. In contrast, only about 15% of T4 phage survived during the freeze drying. CpGV was found to be degraded during freeze drying.
Bacteriophage T4 ; Bacteriophages ; Biological Warfare Agents ; DNA ; Electrophoresis ; Freeze Drying ; Genome ; Granulovirus ; Hantaan virus ; Levivirus ; Nitrogen ; Polymerase Chain Reaction ; Reverse Transcription ; Variola virus ; Yellow fever virus