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Frontiers in Immunology 2022Orf virus (ORFV)-based vectors are attractive for vaccine development as they enable the induction of potent immune responses against specific transgenes. Nevertheless,...
BACKGROUND
Orf virus (ORFV)-based vectors are attractive for vaccine development as they enable the induction of potent immune responses against specific transgenes. Nevertheless, the precise mechanisms of immune activation remain unknown. This study therefore aimed to characterize underlying mechanisms in human immune cells.
METHODS
Peripheral blood mononuclear cells were infected with attenuated ORFV strain D1701-VrV and analyzed for ORFV infection and activation markers. ORFV entry in susceptible cells was examined using established pharmacological inhibitors. Using the THP1-Dual™ reporter cell line, activation of nuclear factor-κB and interferon regulatory factor pathways were simultaneously evaluated. Infection with an ORFV recombinant encoding immunogenic peptides (PepTrio-ORFV) was used to assess the induction of antigen-specific CD8+ T cells.
RESULTS
ORFV was found to preferentially target professional antigen-presenting cells (APCs) , with ORFV uptake mediated primarily by macropinocytosis. ORFV-infected APCs exhibited an activated phenotype, required for subsequent lymphocyte activation. Reporter cells revealed that the stimulator of interferon genes pathway is a prerequisite for ORFV-mediated cellular activation. PepTrio-ORFV efficiently induced antigen-specific CD8+ T cell recall responses in a dose-dependent manner. Further, activation and expansion of naïve antigen-specific CD8+ T cells was observed in response.
DISCUSSION
Our findings confirm that ORFV induces a strong antigen-specific immune response dependent on APC uptake and activation. These data support the notion that ORFV D1701-VrV is a promising vector for vaccine development and the design of innovative immunotherapeutic applications.
Topics: Antigen-Presenting Cells; Humans; Leukocytes, Mononuclear; Membrane Proteins; Orf virus; T-Lymphocytes; Transgenes
PubMed: 35615366
DOI: 10.3389/fimmu.2022.873351 -
Open Veterinary Journal 2022Orf is a highly contagious viral skin disease in sheep and goats caused by Orf virus (ORFV) in the genus Parapoxvirus. Although sheep and goats are considered an...
BACKGROUND
Orf is a highly contagious viral skin disease in sheep and goats caused by Orf virus (ORFV) in the genus Parapoxvirus. Although sheep and goats are considered an essential food resource, particularly in Africa, ORFV infection represents an increasing challenge to animal productivity causing high economic losses.
AIM
This study aimed to detect and characterize the ORFV in suspected clinically diseased goats in two neighboring Egyptian governorates, Al-Sharkia and Ismailia, flocks during April 2020 and July 2021by using PCR and phylogenetic analysis of partial B2L sequence.he present study indicate the necessity for establishing normal heart values in conscious and anaesthetized individuals.
METHODS
Kids from two Egyptian governorates showed the clinical picture of ORFV infection. Samples were collected (n = 15) from two different flocks during April 2020 and July 2021. PCR was carried out to detect the ORFV by targeting a highly conserved sequence within ORFV (B2L) gene. To determine the phylogenetic relationship with other ORFV strains, sequencing and phylogenetic analysis were performed.
RESULTS
ORFV infection was confirmed in 12 samples of oral scabs (80%) by PCR targeting a highly conserved sequence within B2L gene. Sequencing of DNA products was performed and obtained sequences revealed 100% identity at the nucleotide level. Two ORFVs, one from each outbreak showed 98.2% nucleotide identity with a previous Egyptian ORFV (KP984529) whereas our isolates showed higher nucleotide identities, 99.1% and 98.7% with ORFV strains from neighboring countries, Sudan and Ethiopia, respectively. The phylogenetic tree grouped isolates into two main clusters, cluster I included isolates of this study and foreign ones mainly from China, India, and Sudan. Interestingly, the vaccine strains of ORF used in different countries were grouped in cluster II with previous Egyptian isolate (KP984529), Ethiopian and Israeli ORFV isolates.
CONCLUSION
Molecular characterization of B2L gene of ORFV isolates revealed higher sequence identities and more close genetic relationships with other ORFV strains circulating in neighboring countries than with the Egyptian isolates. These findings provide an insight into the genetic diversity of field ORFV isolates circulating in goats in the Egyptian governorates.
Topics: Animals; Ecthyma, Contagious; Egypt; Goat Diseases; Goats; Nucleotides; Orf virus; Phylogeny; Sheep; Sheep Diseases
PubMed: 35603066
DOI: 10.5455/OVJ.2022.v12.i2.16 -
Journal of Veterinary Research Mar 2022Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked...
INTRODUCTION
Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection.
MATERIAL AND METHODS
The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (), six beluga whales (), three Pacific white-sided dolphins (), and ten bottlenose dolphins () from an aquarium in Japan were examined using the ELISA.
RESULTS
Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal.
CONCLUSION
The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.
PubMed: 35582482
DOI: 10.2478/jvetres-2022-0005 -
Archives of Virology Jul 2022Contagious pustular dermatitis is a disease that primarily infects small ruminants and possesses zoonotic potential. It is caused by orf virus (ORFV), a member of the...
Recombination drives the emergence of orf virus diversity: evidence from the first complete genome sequence of an Indian orf virus isolate and comparative genomic analysis.
Contagious pustular dermatitis is a disease that primarily infects small ruminants and possesses zoonotic potential. It is caused by orf virus (ORFV), a member of the genus Parapoxvirus. In this study, we evaluated an ORFV outbreak in goats in Madhya Pradesh, a state in central India, during 2017. The transboundary potential of this virus was evaluated by constructing phylogenetic trees. The complete genome sequence of an ORFV isolate named Ind/MP/17 was found to be 139,807 bp in length with 63.7% GC content and 132 open reading frames (ORFs) flanked by 3,910-bp inverted terminal repeats (ITRs). An investigation into evolutionary parameters such as selection pressure (θ = dN/dS) and nucleotide diversity (π) demonstrated that ORFV has undergone purifying selection. A total of 40 recombination events were identified, 21 of which were evident in the Ind/MP/17 genome, indicating its ability to generate new variants.
Topics: Animals; Ecthyma, Contagious; Genomics; Goats; Orf virus; Phylogeny; Recombination, Genetic; Sheep
PubMed: 35546377
DOI: 10.1007/s00705-022-05443-5 -
Veterinary Research Forum : An... Dec 2021Mastitis is the most important disease in dairy cattle industry because of its high economic losses both in herd management, milk and milk products. The aim of this...
Mastitis is the most important disease in dairy cattle industry because of its high economic losses both in herd management, milk and milk products. The aim of this study was to determine the efficacy of inactivated (IPPVO) Para immune activator as a prophylaxis against mastitis and as the therapy for subclinical mastitis in dairy cattle. The prophylactic effects of IPPVO were investigated in California mastitis test (CMT). Healthy Holstein cows were divided into A1 (n = 30) and A2 (n = 30) subgroups. In addition, 90 subclinical mastitis Holstein cows were divided into subgroups of B1, B2, and B3 to investigate the efficacy of IPPVO treatment. A significant difference in CMT levels was observed ( ˂ 0.01) 30 day after treatment in A groups. The difference in somatic cell count (SCC) levels between the A groups 15, 30 day after treatment was significant ( ˂ 0.01). The results of the CMT among the B groups showed no statistically significant difference ( > 0.05). The results of the SCC tests showed no statistically significant difference ( > 0.05) among the B groups on days 0, 9 and 15 after treatment. Coagulase-negative (n = 53) and (n = 30) were the most prevalent bacteria isolated in this study. In conclusion, IPPVO, although had no additional effect when used in combination with antibiotics could possibly be used instead of antibiotics and to protect cattle from subclinical mastitis, however, it is not known how long this prophylaxis effect could last.
PubMed: 35529817
DOI: 10.30466/vrf.2019.110810.2628 -
Frontiers in Microbiology 2022The parapoxvirus Orf virus (ORFV) has long been recognized as one of the valuable vectors in researches of oncolytic virus. In order to develop a potential therapeutic...
The parapoxvirus Orf virus (ORFV) has long been recognized as one of the valuable vectors in researches of oncolytic virus. In order to develop a potential therapeutic strategy for breast cancer based on the oncolytic virotherapy ORFV, firstly we explore the oncolytic effects of ORFV. Our research showed that ORFV exerts anti-tumor effects by inducing breast cancer cell G2/M phase arrest and cell apoptosis. experiments were carried out, in which we treated 4T1 tumor-bearing BALB/C mice intratumoral injection of ORFV. ORFV can exert anti-tumor activity by regulating tumor microenvironment (TME) and inducing a host immune response plus directly oncolytic effect. The CRISPR-Cas9 knockout library targeting 507 kinases was used to screen out PAK4, which is beneficial to the anti-tumor effect of ORFV on breast cancer cells. PF-3758309 is a potent PAK4-targeted inhibitor. Co-using of ORFV and PF-3758309 as a combination treatment produces its anti-tumor effects through inhibition of cell viability, induction of apoptosis and suppression of cell migration and invasion . The results of experiments showed that the tumor growth of mice in the combination treatment group was significantly inhibited, which proved that the combination treatment exerts an effective anti-tumor effect . In summary, we have clarified the oncolytic effect of ORFV on breast cancer, and found that the combination of ORFV and PAK4 inhibitor can effectively improve the oncolytic effect of ORFV. We hope our research could provide a new idea for the development of new treatment strategies for breast cancer.
PubMed: 35401439
DOI: 10.3389/fmicb.2022.845259 -
Journal For Immunotherapy of Cancer Mar 2022Novel therapies are needed to improve outcomes for women diagnosed with ovarian cancer. Oncolytic viruses are multifunctional immunotherapeutic biologics that...
BACKGROUND
Novel therapies are needed to improve outcomes for women diagnosed with ovarian cancer. Oncolytic viruses are multifunctional immunotherapeutic biologics that preferentially infect cancer cells and stimulate inflammation with the potential to generate antitumor immunity. Herein we describe (Orf virus (OrfV)), an oncolytic poxvirus, as a viral immunotherapy for ovarian cancer.
METHODS
The immunotherapeutic potential of OrfV was tested in the ID8 orthotopic mouse model of end-stage epithelial ovarian carcinoma. Immune cell profiling, impact on secondary lesion development and survival were evaluated in OrfV-treated mice as well as in Batf3 knockout, mice depleted of specific immune cell subsets and in mice where the primary tumor was removed. Finally, we interrogated gene expression datasets from primary human ovarian tumors from the International Cancer Genome Consortium database to determine whether the interplay we observed between natural killer (NK) cells, classical type 1 dendritic cells (cDC1s) and T cells exists and influences outcomes in human ovarian cancer.
RESULTS
OrfV was an effective monotherapy in a murine model of advanced-stage epithelial ovarian cancer. OrfV intervention relied on NK cells, which when depleted abrogated antitumor CD8 T-cell responses. OrfV therapy was shown to require cDC1s in experiments with BATF3 knockout mice, which do not have mature cDC1s. Furthermore, cDC1s governed antitumor NK and T-cell responses to mediate antitumor efficacy following OrfV. Primary tumor removal, a common treatment option in human patients, was effectively combined with OrfV for optimal therapeutic outcome. Analysis of human RNA sequencing datasets revealed that cDC1s correlate with NK cells in human ovarian cancer and that intratumoral NK cells correlate positively with survival.
CONCLUSIONS
The data herein support the translational potential of OrfV as an NK stimulating immunotherapeutic for the treatment of advanced-stage ovarian cancer.
Topics: Animals; Carcinoma, Ovarian Epithelial; Cell Line, Tumor; Disease Models, Animal; Female; Humans; Killer Cells, Natural; Licensure; Mice; Oncolytic Virotherapy; Oncolytic Viruses; Orf virus; Ovarian Neoplasms; Sheep
PubMed: 35296558
DOI: 10.1136/jitc-2021-004335 -
Northern Clinics of Istanbul 2021Orf, ecthyma contagiosum, is a zoonotic viral infectious disease caused by parapoxvirus that affects particularly sheep and goats. Human may be infected with direct...
Orf, ecthyma contagiosum, is a zoonotic viral infectious disease caused by parapoxvirus that affects particularly sheep and goats. Human may be infected with direct contact with contagious animals or by handling contaminated animal products. Lesions are localized mostly on the hands and fingers, but atypical localizations such as head or face have been rarely reported. Herein, we report a case of orf disease on the eyebrow with clinical follow-up images. Physicians should be aware of the possibility of this entity based on contact anamnesis with infected animals and clinical appearance.
PubMed: 35284790
DOI: 10.14744/nci.2020.59254 -
Brazilian Journal of Microbiology :... Jun 2022The pseudocowpox virus (PCPV) is recognized for causing exanthematic lesions in cattle and humans. The diagnosis is important because it is a zoonosis and its clinical...
The pseudocowpox virus (PCPV) is recognized for causing exanthematic lesions in cattle and humans. The diagnosis is important because it is a zoonosis and its clinical signs can be confused with foot-and-mouth disease, a high-impact bovine disease in livestock. The objective of this work is to validate a SYBR Green qPCR and a conventional PCR for virus detection in bovine samples. Detection limit tests, repeatability, reproducibility, sensitivity, and specificity were compared. When two analysts were compared, results demonstrated that training and pipetting influence the repeatability. The qPCR was more sensitive than conventional PCR but showed nonspecific reactions distinguishable by the melting curve. Both showed high repeatability and reproducibility.
Topics: Animals; Cattle; Cattle Diseases; Pathology, Molecular; Poxviridae Infections; Pseudocowpox Virus; Real-Time Polymerase Chain Reaction; Reproducibility of Results; Sensitivity and Specificity
PubMed: 35220553
DOI: 10.1007/s42770-021-00664-3 -
Veterinary Medicine and Science Mar 2022Several viruses belonging to the family Poxviridae can cause infections in humans and animals. In Corsica, livestock farming (sheep, goats, pigs, and cattle) is mainly...
BACKGROUND
Several viruses belonging to the family Poxviridae can cause infections in humans and animals. In Corsica, livestock farming (sheep, goats, pigs, and cattle) is mainly mixed, leading to important interactions between livestock, wildlife, and human populations. This could facilitate the circulation of zoonotic diseases, and makes Corsica a good example for studies of tick-borne diseases.
OBJECTIVES
To gain understanding on the circulation of poxviruses in Corsica, we investigated their presence in tick species collected from cattle, sheep, horses, and wild boar, and characterized them through molecular techniques.
METHODS
Ticks were tested using specific primers targeting conserved regions of sequences corresponding to two genera: parapoxvirus and orthopoxvirus.
RESULTS
A total of 3555 ticks were collected from 1549 different animals (687 cattle, 538 horses, 106 sheep, and 218 wild boars). They were tested for the presence of parapoxvirus DNA on one hand and orthopoxvirus DNA on the other hand using Pangeneric real-time TaqMan assays. Orthopoxvirus DNA was detected in none of the 3555 ticks. Parapoxvirus DNA was detected in 6.6% (36/544) of ticks collected from 23 cows from 20 farms. The remaining 3011 ticks collected from horses, wild boars, and sheep were negative. The infection rate in cow ticks was 8.0% (12/148) in 2018 and 6.0% (24/396) in 2019 (p = 0.57). Parapoxvirus DNA was detected in 8.5% (5/59) of Hyalomma scupense pools, 8.2% (15/183) of Hyalomma marginatum pools, and 6.7% (16/240) of Rhipicephalus bursa pools (p = 0.73). We successfully amplified and sequenced 19.4% (7/36) of the positive samples which all corresponded to pseudocowpox virus.
CONCLUSIONS
Obviously, further studies are needed to investigate the zoonotic potential of pseudocowpox virus and its importance for animals and public health.
Topics: Animals; Cattle; Female; Goat Diseases; Horse Diseases; Horses; Ixodidae; Parapoxvirus; Sheep; Sheep Diseases; Swine; Swine Diseases; Tick Infestations; Tick-Borne Diseases; Ticks
PubMed: 35092189
DOI: 10.1002/vms3.700