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PubMed2003

Four-gene-combination DNA vaccine protects mice against a lethal vaccinia virus challenge and elicits appropriate antibody responses in nonhuman primates.

Two major infectious forms of vaccinia virus (VACV) have been described: the intracellular mature virion (IMV), and the extracellular enveloped virion (EEV). Due to their stability in the environment, IMVs play a predominant role in host-to-host transmission, whereas EEVs play an important role in dissemination within the host. In a previous report, we demonstrated that mice vaccinated with VACV L1R (IMV immunogen) and A33R (EEV immunogen) were protected from a lethal poxvirus challenge. Vaccination with a combination of both genes conferred greater protection than either gene alone, suggesting that an immune response against both IMV and EEV is advantageous. Here, we report that in mice individually administered DNA vaccines with two different VACV immunogens, A27L (IMV immunogen) or B5R (EEV immunogen), failed to significantly protect; however, vaccination with a combination of both genes conferred a high level of protection. Mice were completely protected when vaccinated with a combination of four VACV genes (A27L + A33R + L1R + B5R). Rhesus macaques vaccinated with this four-gene-combination developed appropriate antibody responses to each protein. Antibody responses elicited by this vaccine cross-reacted with monkeypox virus orthologous proteins. These data indicate that a gene-based vaccine comprised of the VACV A27L + A33R + L1R + B5R genes may be a useful candidate to protect against other orthopoxviruses, including those that cause monkeypox and smallpox.

Amino Acid Sequence↗
PubMed1975

Isolation of poxvirus from an African Rodent.

A poxvirus was isolated from a wild gerbil (Tatera kempii) caught in northern Dahomey, Africa at the time of an epidemic of human smallpox. Electron microscopic appearance and serologic reactions placed it in the vaccinia subgroup of poxviruses. The isolate differed from ectromelia, rabbitpox, vaccinia, monkeypox, and cowpox viruses in pock morphology on chorioallantoic membrane, ceiling temperature, relative innocuity for mice, and cytopathic effect in tissue culture. Like variola minor virus, it had a ceiling temperature of 38 C, produced small hypertrophic foci in tissue culture, and failed to grow in rabbit skin. Inoculated into a rhesus monkey, it caused fever but no skin eruption and produced seroconversion and protection from subsequent challenge with monkeypox virus. The growing list of animal viruses that differ only slightly from smallpox virus suggests the hypothesis that long-term survival of variola virus may be based on inapparent infection in animals as well as virulent spread among humans.

Animals↗
PubMed1979

Structural polypeptides of Orthopoxvirus: their distribution in various members and location within the virion.

The structural polypeptides of accepted species and recently isolated members of the genus Orthopoxvirus have been examined by SDS-polyacrylamide gel electrophoresis. The viruses shared many polypeptides but some differences were found. The viruses could be divided into a vaccinia group (including buffalopox, 'Lenny' and MK-10), an ectromelia group (including elephant virus and Moscow virus), cowpox, camelpox and monkeypox. Minor differences were found in the polypeptides of monkeypox virus strains from human and monkey outbreaks. Controlled degradation of virions showed that the polypeptides which enabled the viruses to be differentiated were located in the surface and sub-surface layers. The cores of the viruses all gave the same complex polypeptide pattern.

Animals↗
PubMed1993

Characterization of the gene encoding the A-type inclusion protein of camelpox virus and sequence comparison with other orthopoxviruses.

A gene was identified in camelpox virus strain CP-1 that is similar to the 160K gene of cowpox virus strain Brighton (BR) that encodes the A-type inclusion body protein (ATIP). The CP-1 gene was mapped, sequenced, and the presence of the ATIP-specific mRNA was demonstrated. The open reading frame [2178 nucleotides (nt)] was found at a similar position in the CP genome as the one reported for the cowpox virus 160K ATI gene. DNA sequence comparison revealed a deletion of two adjacent adenine residues relative to cowpox virus BR, generating a reading frame shift accompanied by the formation of a translational stop codon. An identical deletion has been described for vaccinia virus strain Western Reserve. The DNA sequence of the corresponding region of monkeypox virus strain Copenhagen revealed a deletion leading to a putative stop codon 75 nt upstream of the same stop codons in the camelpox and vaccinia virus genes. These findings are consistent with the expression of truncated ATIPs, of 94K in vaccinia and camelpox viruses and of 92K in monkeypox virus. In addition, a deletion of 789 bp could be localized downstream of the ATI open reading frame in camelpox virus isolates of different origin. This causes the transcription of a shortened ATI-specific mRNA (3.7 kb) relative to vaccinia and cowpox viruses (both 4.5 kb). The similarity observed in ATIP-encoding and flanking sequences might suggest that vaccinia and camelpox viruses are descended from a common ancestor.

Amino Acid Sequence↗
PubMed2001

Vaccinia virus inhibitors as a paradigm for the chemotherapy of poxvirus infections.

Poxviruses continue to pose a major threat to human health. Monkeypox is endemic in central Africa, and the discontinuation of the vaccination (with vaccinia virus) has rendered most humans vulnerable to variola virus, the etiologic agent of smallpox, should this virus be used in biological warfare or terrorism. However, a large variety of compounds have been described that are potent inhibitors of vaccinia virus replication and could be expected to be active against other poxviruses as well. These compounds could be grouped in different classes: (i) IMP dehydrogenase inhibitors (e.g., EICAR); (ii) SAH hydrolase inhibitors (e.g., 5'-noraristeromycin, 3-deazaneplanocin A, and various neplanocin A derivatives); (iii) OMP decarboxylase inhibitors (e.g., pyrazofurin) and CTP synthetase inhibitors (e.g., cyclopentenyl cytosine); (iv) thymidylate synthase inhibitors (e.g., 5-substituted 2'-deoxyuridines); (v) nucleoside analogues that are targeted at viral DNA synthesis (e.g., Ara-A); (vi) acyclic nucleoside phosphonates [e.g., (S)-HPMPA and (S)-HPMPC (cidofovir)]; and (vii) polyanionic substances (e.g., polyacrylic acid). All these compounds could be considered potential candidate drugs for the therapy and prophylaxis of poxvirus infections at large. Some of these compounds, in particular polyacrylic acid and cidofovir, were found to generate, on single-dose administration, a long-lasting protective efficacy against vaccinia virus infection in vivo. Cidofovir, which has been approved for the treatment of cytomegalovirus retinitis in immunocompromised patients, was also found to protect mice, again when given as a single dose, against a lethal aerosolized or intranasal cowpox virus challenge. In a biological warfare scenario, it would be advantageous to be able to use a single treatment for an individual exposed to an aerosolized poxvirus. Cidofovir thus holds great promise for treating human smallpox, monkeypox, and other poxvirus infections. Anecdotal experience points to the efficacy of cidofovir in the treatment of the poxvirus infections molluscum contagiosum and orf (ecthyma contagiosum) in immunosuppressed patients.

Antiviral Agents↗
PubMed1962

Comparative studies of the multiplication of antigenically related poxviruses on the chorioallantoic membrane of the chick embryo.

Hahon, Nicholas (U. S. Army Chemical Corps, Frederick, Md.) and James J. Friel. Comparative studies of the multiplication of antigenically related poxviruses on the chorioallantoic membrane of the chick embryo. J. Bacteriol. 83:837-843. 1962.-A comparison was made of the general growth curves of several poxviruses, variola, alastrim, vaccinia, cowpox, rabbitpox, and monkeypox, on the chorioallantoic membrane of the chick embryo. All curves showed a lag period, followed by a tenfold increase of infectivity each 5 hr for approximately 20 to 25 hr, a peak growth level at 48 to 52 hr (except monkeypox), and a gradual decline after 72 hr. Closer examination of the lag period revealed a rapid decline of virus infectivity within 2 to 4 hr after inoculation and the manifestation of an eclipse phase for 4 and 6 hr with all poxviruses. The lag period was terminated with the detection of intracellular virus at 8 to 12 hr, followed by the appearance of extracellular virus 2 to 6 hr later. The proportion of extracellular to intracellular virus was 0.1 or less throughout the different stages of poxvirus growth. In contrast to findings with the other poxviruses, however, extracellular virus equaled or exceeded the quantity of intracellular virus during the latter stages of the growth curves of variola and alastrim viruses.

Animals↗
PubMed2001

Detection and differentiation of old world orthopoxviruses: restriction fragment length polymorphism of the crmB gene region.

A restriction fragment length polymorphism (RFLP) assay was developed to identify and differentiate Old World, African-Eurasian orthopoxviruses (OPV): variola, vaccinia, cowpox, monkeypox, camelpox, ectromelia, and taterapox viruses. The test uses amplicons produced from virus genome DNA by PCR with a consensus primer pair designed from sequences determined for the cytokine response modifier B (crmB) gene of 43 different OPV strains of known taxonomic origin. The primer pair amplified a single specific product from each of the 115 OPV samples tested. Size-specific amplicons identified and differentiated ectromelia and vaccinia virus strains, which contain a truncated crmB gene, and enabled their differentiation from other OPV species. Restriction digests of amplified products allowed the identification and differentiation of variola, monkeypox, camelpox, vaccinia, and cowpox virus species and strains.

Humans↗
PubMed2004

Real-time PCR system for detection of orthopoxviruses and simultaneous identification of smallpox virus.

A screening assay for real-time LightCycler (Roche Applied Science, Mannheim, Germany) PCR identification of smallpox virus DNA was developed and compiled in a kit system under good manufacturing practice conditions with standardized reagents. In search of a sequence region unique to smallpox virus, the nucleotide sequence of the 14-kDa fusion protein gene of each of 14 variola virus isolates of the Russian World Health Organization smallpox virus repository was determined and compared to published sequences. PCR primers were designed to detect all Eurasian-African species of the genus ORTHOPOXVIRUS: A single nucleotide mismatch resulting in a unique amino acid substitution in smallpox virus was used to design a hybridization probe pair with a specific sensor probe that allows reliable differentiation of smallpox virus from other orthopoxviruses by melting-curve analysis. The applicability was demonstrated by successful amplification of 120 strains belonging to the orthopoxvirus species variola, vaccinia, camelpox, mousepox, cowpox, and monkeypox virus. The melting temperatures (T(m)s) determined for 46 strains of variola virus (T(m)s, 55.9 to 57.8 degrees C) differed significantly (P = 0.005) from those obtained for 11 strains of vaccinia virus (T(m)s, 61.7 to 62.7 degrees C), 15 strains of monkeypox virus (T(m)s, 61.9 to 62.2 degrees C), 40 strains of cowpox virus (T(m)s, 61.3 to 63.7 degrees C), 8 strains of mousepox virus (T(m), 61.9 degrees C), and 8 strains of camelpox virus (T(m)s, 64.0 to 65.0 degrees C). As most of the smallpox virus samples were derived from infected cell cultures and tissues, smallpox virus DNA could be detected in a background of human DNA. By applying probit regression analysis, the analytical sensitivity was determined to be 4 copies of smallpox virus target DNA per sample. The DNAs of several human herpesviruses as well as poxviruses other than orthopoxviruses were not detected by this method. The assay proved to be a reliable technique for the detection of orthopoxviruses, with the advantage that it can simultaneously identify variola virus.

Animals↗
PubMed2005

Countermeasures to the bioterrorist threat of smallpox.

Variola, the agent of smallpox, is a bioterrorist threat, as is monkeypox virus, which also occurs naturally in Africa. Development of countermeasures, in the form of improved vaccines, antiviral drugs, and other therapeutic strategies are a high priority. Recent advances in molecular biology and in animal model development have provided fresh insight into the virulence determinants for smallpox and the pathophysiology of disease. The complex replication cycle for orthopoxviruses, and the pivotal role for viral-specific immunomodulatory proteins which contribute to escape from immunologic surveillance, provide many unique targets for therapeutic intervention. The "toxemia" of smallpox has been elucidated in part by variola-infected primate studies which revealed the central role of apoptosis and the evolution of a cytokine storm leading to hemorrhagic diathesis, resembling fulminent "black" smallpox. This suggests a potential role for therapeutic strategies developed for septic shock, in treatment of smallpox. Drugs licensed for other viruses which share molecular targets with orthopoxviruses (e.g. Cidofovir) or cancer drugs (e.g. Gleevec and other tyrosine kinase inhibitors) have immediate application for treatment of smallpox and monkeypox and provide leads for second generation drugs with higher therapeutic indices. Recent advances in identification of virulence determinants and immune evasion genes facilitate the design of alternative vaccines to replace live vaccinia strains that are unsuitable for a large proportion of individuals in a mass immunization campaign.

Animals↗
PubMed1977

Poxvirus in West African nonhuman primates: serological survey results.

Ten species of nonhuman primates in West African habitat were analysed for variolavaccinia subgroup haemagglutination-inhibition (HI) and neutralization antibodies. The animals were taken in 27 different sampling zones in parts of the Ivory Coast, Mali, and Upper Volta. Of the 195 tested, 15 (8%) had elevated HI antibodies after nonspecific reactions were reduced with potassium periodate pretreatment. Positive neutralization antibodies were found in 21% (44 of 206). Antibodies were detected in serum from monkeys living near two areas where monkeypox cases in humans had occurred. Four samples were tested for monkeypox specific antibodies using an indirect immunofluorescent test; 3 were positive. Despite the prevalence of poxvirus antibodies in monkeys (and other animals) in West Africa, smallpox eradication has been maintained in the area since 1970; thus, animal reservoirs of poxvirus appear to pose no threat to the worldwide smallpox eradication programme.

Africa, Western↗
PubMed1973

Evaluation of virological laboratory methods for smallpox diagnosis.

Between July 1966 and May 1972 the Vesicular Disease Laboratory, Center for Disease Control, Atlanta, Ga., USA, tested specimens from 849 suspected smallpox cases by at least 2 methods, electron microscopy and chick embryo chorioallantoic membrane (CAM) cultures. A smaller number of specimens was tested by each of 4 methods: electron microscopy, CAM culture, agar gel precipitation, and tissue culture. For specimens handled in the field the CAM culture method was less sensitive than electron microscopy because the adverse conditions often inactivated the virus. CAM cultures were valuable for identifying members of the poxvirus subgroups, however, particularly when supplemented by tissue culture. The agar gel precipitation test was the least sensitive but was of value in confirming the results of electron microscopy. The latter was highly effective for the diagnosis of varicella, but dependably identified only about half of the vaccinia infections; for vaccinia, the CAM technique was essential. The occurrence of human monkeypox cases in West Africa emphasized that the usual smallpox diagnostic methods were inadequate. More sophisticated tests, such as the rabbit dermal sensitivity test, are necessary for accurate diagnosis of these cases as monkeypox.

Adult↗
PubMed2026

Genomic Epidemiology and Clinical Characteristics of Mpox Lineage C.1 Outbreak in Thailand, 2023-2024.

Since 2022, human monkeypox virus (hMPXV) has emerged in non-endemic regions, including Thailand. However, the genomic dynamics and clinical correlates of local transmission remain incompletely defined. Whole-genome sequencing was performed on hMPXV from 16 patients in Thailand (2023-2024) using targeted amplicon NGS. Phylogenetic analyses integrated global reference sequences. Mutational profiles, specifically non-synonymous substitutions and APOBEC3-associated signatures, were analyzed in relation to clinical data. Phylogenetic reconstruction identified three temporal phases. Early 2022 cases (clade IIb lineages A and B) were interspersed with global sequences, consistent with multiple introductions. In contrast, 2023-2024 cases were dominated by lineage C.1. All 16 genomes belonged to C.1 (one C.1.1), and formed a distinct mid-2023 cluster, designated C.1/Thai/Cluster, supporting sustained local transmission. APOBEC3-associated mutations were pervasive across the C.1 lineage overall, including within C.1/Thai/Cluster, without evidence of significant enrichment specific to this cluster. The cohort comprised exclusively male patients (81% HIV-positive, MSM), with predominantly genital painful lesions and a median recovery time of 23 days. No significant associations were detected between viral genetic variation and clinical outcomes. Mpox transmission in Thailand evolved from multiple introductions to sustained C.1-dominated local spread, underscoring the importance of continued genomic surveillance.

Humans↗
PubMed2004

Bioterrorism: is it a real threat?

The Geneva Protocol of 1925 commits the signatory nations to refraining from the use of biological weapons. However, the terrorist assaults of September 2001 and, subsequently, the anthrax-containing letters are cause for great concerns: new threats to the security of nations are expected, as terrorist organizations seem to increasingly explore novel ways of spreading terror. In this context, naturally emerging diseases such as SARS, monkeypox or West Nile fever assume new importance because it is difficult to distinguish between natural epidemics and possible bioweapon assaults. Great efforts on the part of governments and public health authorities are necessary to counteract these threats.

Anthrax↗
PubMed2026

Multiomics profiling of plasma reveals lipid-immune dysregulation and exosome remodeling in mpox and mpox-HIV co-infection.

BACKGROUND: Monkeypox virus (MPXV) infects diverse human cell types, and human immunodeficiency virus (HIV) co-infection is common. The immunometabolic consequences of MPXV infection, and how it may be altered by HIV, remain poorly defined. METHODS: We performed quantitative plasma lipidomics and precise metabolomics in a discovery cohort (n = 81) comprising MPXV-monoinfected (MPLWOH), MPXV-HIV-coinfected (MPLWH), and HIV-monoinfected (PLWH) patients and healthy controls, integrating exosome proteomics, cytokine profiling, and transcriptomics of exosome-treated HepG2 and A549 cells for functional interpretation. An independent validation cohort (n = 65) was used to assess cross-cohort reproducibility. FINDINGS: MPXV infection induced broad lipid remodeling, with elevations in phosphatidylserine (PS) and phosphatidylethanolamine (PE) and reductions in phosphatidylcholine (PC), lysophospholipids, cholesteryl ester (CE), and exosomal lecithin-cholesterol acyltransferase (LCAT) and lipoprotein lipase (LPL). These lipid alterations were correlated with tissue injury markers and inflammatory cytokines. The MPLWH group exhibited more severe metabolic disruption, including marked sulfatide (SL) depletion, lower cholesterol and high-density lipoprotein cholesterol (HDL-c), and extensive rewiring of lipid-cytokine associations. SL depletion in MPLWH correlated with abundances of COPI-mediated retrograde trafficking proteins in exosomes. Transcriptomic profiling of exosome-treated cells provided functional validation: MPLWOH exosomes induced lipid metabolism and repair-associated epithelial programs, while MPLWH exosomes drove phospholipid remodeling and acute inflammatory and mucosal barrier-stress responses. CONCLUSIONS: MPXV infection reprograms host lipid metabolism and exosome composition, with HIV co-infection amplifying inflammatory, metabolic, and trafficking disruptions. These convergent multi-omics signatures link systemic lipid dysregulation to exosome-mediated immunomodulation and identify potential targets for host-directed interventions. FUNDING: This study was funded by the Major Project of Guangzhou National Laboratory.

Adult↗
PubMed2025

Mpox: current knowledge and understanding-a scoping review.

Mpox in humans is a rash illness resulting from infection with monkeypox virus (MPXV). In 2022, a public health emergency of international concern (PHEIC) was declared with 115 countries reporting cases of Mpox. Most of these countries had not previously reported cases. This global outbreak was sustained primarily by human-to-human transmission within complex sexual networks. Whilst these cases were similar to previous clade II West African MPXV isolates, they were sufficiently genomically distinct to result in WHO recognizing two subclades within clade II: clade IIa and clade IIb. In 2024, a second PHEIC was declared, resulting from a marked increase in cases of clade I MPXV. In this scoping review, we compare the major clinical, epidemiological, and genomic features of the major mpox lineages and the implications for vaccination, transmission, infection control and treatment..

Humans↗
PubMed1999

Production and characterization of human monoclonal antibody Fab fragments to vaccinia virus from a phage-display combinatorial library.

A combinatorial, phage-display library of human Fab antibody fragments was generated from IgG heavy chain (HC) and light chain (LC) genes cloned from the lymphocytes of a vaccinia virus (VACV)-immune donor. To ascertain the complexity of the library, nucleotide sequences of the variable regions of the HC and LC genes were determined. Fourteen distinct HC and 18 distinct LC (7 kappa and 11 lambda) that formed a combinatorial library of 22 Fabs were identified. Immune-precipitation of radiolabeled VACV revealed that at least six different VACV proteins were recognized by the antibodies. Plaque-reduction neutralization demonstrated that six of the Fabs neutralized VACV in the presence of anti-human antibody. ELISA studies indicated that 15 of the Fabs were cross-reactive with monkeypox virus.

Amino Acid Sequence↗
PubMed1980

Virion polypeptides of poxviruses.

Structural polypeptides from a number of poxviruses were analysed by SDS-polyacrylamide gel electrophoresis. The patterns obtained with orthopoxviruses were generally quite similar to one another, but variola, monkeypox, cowpox and vaccinia viruses could be distinquished by their profiles in the molecular weight (mol. wt.) region around 30,000 to 40,000; some additional variation was found amongst cowpox and vaccinia strains. Whitepox virus was shown to have structural polypeptides indistinguishable from those of variola virus. The structural polypeptides of poxviruses belonging to other genera were different from those of the orthopoxvirus, except those of mol. wt. about 122,000 and 97,000, which were common to all viruses irrespective of genus. A polypeptide of mol. wt. about 25,000 was also observed in all cases, though its position varied slightly with the individual virus.

Chymotrypsin↗
PubMed2005

Multiplex PCR detection and species differentiation of orthopoxviruses pathogenic to humans.

A method for one-stage rapid identification of four orthopoxvirus species pathogenic to humans based on multiplex polymerase chain reaction (MPCR) was developed. Five pairs of oligonucleotide primers--one, genus-specific; and the rest, species-specific for variola, monkeypox, cowpox, and vaccinia viruses, respectively--were used concurrently for MPCR assay of orthopoxvirus DNAs. Specificity and sensitivity of the method developed were evaluated using DNAs of 57 orthopoxvirus strains, including the DNAs isolated from human case clinical materials.

Cowpox virus↗