OBJECTIVE: Poxviruses are zoonotic pathogens that infect humans, mammals, vertebrates, and arthropods. However, the specific role of ticks in transmission and evolution of these viruses remains unclear. METHODS: Transcriptomic and metatranscriptomic raw data from 329 sampling pools of seven tick species across five continents were mined to assess the diversity and abundance of poxviruses. Chordopoxviral sequences were assembled and subjected to phylogenetic analysis to trace the origins of the unblasted fragments within these sequences. RESULTS: Fifty-eight poxvirus species, representing two subfamilies and 20 genera, were identified, with 212 poxviral sequences assembled. A substantial proportion of AT-rich fragments were detected in the assembled poxviral genomes. These genomic sequences contained fragments originating from rodents, archaea, and arthropods. CONCLUSION Our findings indicate that ticks play a significant role in the transmission and evolution of poxviruses. These viruses demonstrate the capacity to modulate virulence and adaptability through horizontal gene transfer, gene recombination, and gene mutations, thereby promoting co-existence and co-evolution with their hosts. This study advances understanding of the ecological dynamics of poxvirus transmission and evolution and highlights the potential role of ticks as vectors and vessels in these processes.
Animals ; Poxviridae/physiology* ; Ticks/virology* ; Phylogeny ; Transcriptome ; Evolution, Molecular ; Poxviridae Infections/virology* ; Genome, Viral

Objective: To investigate the clinical, pathological and immunophenotypic features, molecular biology and prognosis of fibrin-associated large B-cell lymphoma (LBCL-FA) in various sites. Methods: Six cases of LBCL-FA diagnosed from April 2016 to November 2021 at the Beijing Friendship Hospital, Capital Medical University, Beijing, China and the First Affiliated Hospital, Wenzhou Medical University, Wenzhou, China were collected. The cases were divided into atrial myxoma and cyst-related groups. Clinical characteristics, pathological morphology, immunophenotype, Epstein Barr virus infection status, B-cell gene rearrangement and fluorescence in situ hybridization of MYC, bcl-2, bcl-6 were summarized. Results: The patients' mean age was 60 years. All of them were male. Three cases occurred in atrial myxoma background, while the others were in cyst-related background, including adrenal gland, abdominal cavity and subdura. All cases showed tumor cells located in pink fibrin clot. However, three cyst-related cases showed the cyst wall with obviously fibrosis and inflammatory cells. All cases tested were non germinal center B cell origin, positive for PD-L1, EBER and EBNA2, and were negative for MYC, bcl-2 and bcl-6 rearrangements, except one case with MYC, bcl-2 and bcl-6 amplification. All of the 5 cases showed monoclonal rearrangement of the Ig gene using PCR based analysis. The patients had detailed follow-ups of 9-120 months, were treated surgically without radiotherapy or chemotherapy, and had long-term disease-free survivals. Conclusions: LBCL-FA is a group of rare diseases occurring in various sites, with predilection in the context of atrial myxoma and cyst-related lesions. Cyst-related lesions with obvious chronic inflammatory background show more scarcity of lymphoid cells and obvious degeneration, which are easy to be missed or misdiagnosed. LBCL-FA overall has a good prognosis with the potential for cure by surgery alone and postoperative chemotherapy may not be necessary.
Humans ; Male ; Middle Aged ; Atrial Fibrillation ; Epstein-Barr Virus Infections ; Fibrin/genetics* ; Herpesvirus 4, Human/genetics* ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse/pathology* ; Myxoma ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Proto-Oncogene Proteins c-bcl-6/genetics*

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β₂m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a Superdex^TM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
Animals ; Capripoxvirus ; Epitopes, T-Lymphocyte/genetics* ; Peptides/genetics* ; Poxviridae Infections ; Sheep ; Sheep Diseases

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

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

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

OBJECTIVETo construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.

METHODSThe flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.

RESULTSThe recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.

CONCLUSIONGreen fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.

Cells, Cultured ; Endothelial Cells ; metabolism ; Gene Deletion ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Orf virus ; Plasmids ; Transfection

Halo nevi (HN), also known as Sutton's nevi or leukoderma acquisitum centrifugum, are nevomelanocytic nevi surrounded by a rim of depigmentation. A 9-year-old female presented with an asymptomatic solitary flesh-colored 0.75-cm-sized papule surrounded by depigmentation on the right cheek 1 month ago. The excisional biopsy demonstrated molluscum contagiosum and decreased basal layer pigmentation on Fontana-Masson stain. The halo phenomenon is often associated with an acquired nevus cell nevus, but has also been associated with a congenital nevus cell nevus, non-neuroectodermal lesions such as angiomas, seborrheic keratoses, lichen planus, sarcoid, psoriasis, involuting flat warts, and histiocytoma. Our patient had an unusual association of molluscum contagiosum with the halo phenomenon. We suppose that the halo phenomenon in this patient might have been provoked by the molluscum contagiosum virus. A halo nevus-like appearance suggests the possibility of disease other than nevus. In such cases, skin biopsy is required for confirmation.
Biopsy ; Cheek ; Child ; Female ; Hemangioma ; Histiocytoma ; Humans ; Keratosis, Seborrheic ; Lichen Planus ; Molluscum contagiosum virus ; Molluscum Contagiosum* ; Nevus ; Nevus, Halo ; Pigmentation ; Psoriasis ; Skin ; Warts

BACKGROUNDGallbladder carcinoma (GBC) has a high mortality rate, requiring synergistic anti-tumor management for effective treatment. The myxoma virus (MYXV) exhibits a modest clinical value through its oncolytic potential and narrow host tropism.

METHODSWe performed viral replication assays, cell viability assays, migration assays, and xenograft tumor models to demonstrate that bone marrow-derived stem cells (BMSCs) may enhance efficiency of intravenous MYXV delivery.

RESULTSWe examined the permissiveness of various GBC cell lines towards MYXV infection and found two supported single and multiple rounds of MYXV replication, leading to an oncolytic effect. Furthermore, we found that BMSCs exhibited tropism for GBC cells within a Matrigel migration system. BMSCs failed to affect the growth of GBC cells, in terms of tumor volume and survival time. Finally, we demonstrated in vivo that intravenous injection of MYXV-infected BMSCs significantly improves the oncolytic effect of MYXV alone, almost to the same extent as intratumoral injection of MYXV.

CONCLUSIONThis study indicates that BMSCs are a promising novel vehicle for MYXV to clinically address gallbladder tumors.

Animals ; Bone Marrow Cells ; cytology ; Cell Movement ; physiology ; Cell Survival ; physiology ; Female ; Gallbladder Neoplasms ; therapy ; virology ; Humans ; Immunohistochemistry ; Mice ; Myxoma virus ; pathogenicity ; Stem Cells ; cytology ; physiology ; Virus Replication ; physiology ; Xenograft Model Antitumor Assays

Traditional approach of inactivated or live-attenuated vaccine immunization has resulted in impressive success in the reduction and control of infectious disease outbreaks. However, many pathogens remain less amenable to deal with the traditional vaccine strategies, and more appropriate vaccine strategy is in need. Recent discoveries that led to increased understanding of viral molecular biology and genetics has rendered the used of viruses as vaccine platforms and as potential anti-cancer agents. Due to their ability to effectively induce both humoral and cell-mediated immune responses, viral vectors are deemed as an attractive alternative to the traditional platforms to deliver vaccine antigens as well as to specifically target and kill tumor cells. With potential targets ranging from cancers to a vast number of infectious diseases, the benefits resulting from successful application of viral vectors to prevent and treat human diseases can be immense.
Adenoviridae ; Alphavirus ; Communicable Diseases ; Disease Outbreaks ; Genetic Vectors ; Humans ; Immunization ; Molecular Biology ; Poxviridae ; Vaccines