Replication kinetics of Marek's disease vaccine virus in feathers and lymphoid tissues using PCR and virus isolation

Baigent, Susan J.; Smith, Lorraine P.; Currie, Richard J. W.; Nair, Venugopal

doi:10.1099/vir.0.81299-0

Cited by 64 publications

(61 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The genomes of these strains are highly similar to the oncogenic strains, but crucial differences exist . The MD vaccine viruses replicate in host cells and transmit to other birds [Baigent et al, 2005;Islam et al, 2007;Tan et al, 2007;Gimeno et al, 2011;Islam et al, 2013;], but do not induce disease or lymphomas [Calnek and Witter, 1997]. Despite improved understanding of vaccine-related immunity [Witter, 1984;Osterrieder et al, 2006], an explanation for the anti-tumor effect of vaccination has not yet been established.…”

mentioning

confidence: 99%

Vaccination and Host Marek's Disease-Resistance Genotype Significantly Reduce Oncogenic Gallid alphaherpesvirus 2 Telomere Integration in Host Birds

McPherson

Cheng²,

Smith

et al. 2018

Cytogenet Genome Res

View full text Add to dashboard Cite

Marek's disease (MD) is an infectious disease characterized by lymphomas and high mortality in susceptible chickens. The causative and ubiquitous alpha-herpesvirus known as MD virus (MDV) integrates into host telomeres during early infection through latency, known to be an important phase for oncogenic transformation. Herein, we sought to determine the influence of vaccination and host genetics on the temporal dynamics of MDV-host genome interactions. We studied integration profiles using 2 MD vaccines that vary in protective efficacy in 2 genetic lines that differ in MD resistance/susceptibility. Virus integration of both oncogenic MDV and vaccine strains was observed in both MD susceptible and resistant birds, however, the lines differed in their dynamic telomere-integration profiles. Notably, the resistant host genotype exhibited a smaller percentage of replicating cells with the virus telomere-integrated only phenotype as compared to the susceptible genotype. Vaccination with Rispens, the most protective MD vaccine, also reduced the establishment of the virus telomere-integrated only phenotype, suggesting a significant role of the phenotype in MD lymphoma development. The effect of Rispens vaccination was most dramatic in the susceptible genotype. These results suggest important connections between vaccinal immunity, MDV telomere integration, virus-induced oncogenesis, and virus-host genome interactions in the context of host genetics and disease susceptibility.

show abstract

mentioning

confidence: 99%

Vaccination and Host Marek's Disease-Resistance Genotype Significantly Reduce Oncogenic Gallid alphaherpesvirus 2 Telomere Integration in Host Birds

McPherson

Cheng²,

Smith

et al. 2018

Cytogenet Genome Res

View full text Add to dashboard Cite

show abstract

“…This enabled specific detection of Md5 without detection of the vaccine viruses because the U S 2 gene is absent from HVT (serotype 3), SB-1 (serotype 2), and the BAC-cloned viruses pCVI988-10 and pRB-1B-2 (in which the U S 2 gene is replaced by the BAC F plasmid sequence). Duplex real-time q-PCR assays were performed to quantify both the MDV-1 U S 2 gene and the chicken reference gene ovotransferrin (ovo) in the same reaction using 40 cycles on an ABI 7500 instrument (Applied Biosystems, Foster City, CA), essentially as previously described (4,5). All reactions contained 10 mg bovine serum albumin to overcome the inhibitory effect of melanin pigment (4).…”

Section: Methodsmentioning

confidence: 99%

“…Here, MDV undergoes fully productive replication in the feather follicle epithelium. Characteristic lesions are observed in the feather pulp (13) and MDV DNA can be detected at this site (5). Cell-free MDV is shed into the environment with skin and feather debris from about 7 days postinfection (dpi) onward (20), and shedding continues throughout the life of an infected bird (10).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Relationship Between Levels of Very Virulent MDV in Poultry Dust and in Feather Tips from Vaccinated Chickens

et al. 2013

Self Cite

View full text Add to dashboard Cite

SUMMARY. To assess the effect of various vaccine strains on replication and shedding of virulent Marek's disease virus from experimentally infected chickens, quantitative PCR (q-PCR) methods were developed to accurately quantify viral DNA in infected chickens and in the environment in which they were housed. Four groups of 10 chickens, kept in poultry isolators, were vaccinated at 1 day old with one of four vaccines covering each of the three vaccine serotypes, then challenged with very virulent MDV strain Md5 at 8 days of age. At regular time-points, feather tips were collected from each chicken and poultry dust was collected from the air-extract prefilter of each isolator. DNA was extracted from feather and dust samples and subjected to real-time q-PCR, targeting the U S 2 gene of MDV-1, in order to measure Md5 level per 10 4 feather tip cells or per microgram of dust. Accuracy of DNA extraction from dust and real-time q-PCR were validated by comparing either q-PCR cycle threshold values or the calculated MDV genome level; for use in q-PCR, DNA was extracted from serial dilutions of MDV-infected dust diluted with noninfected dust, or DNA from MDV-infected dust was diluted with DNA from noninfected dust. The results confirmed the accuracy and sensitivity of dust DNA extraction and subsequent q-PCR and showed that differences in virus levels between dust samples truly reflect differences in shedding. Vaccination delayed both replication of Md5 in feather tips and shedding of Md5. First detection of Md5 in feather tips always preceded or coincided with first detection in dust in each group. pCVI988 and HVT+SB-1 were the most efficient vaccines in reducing both replication and shedding of Md5. There was close correlation between mean virus level in feathers of each group and mean virus level in the dust shed by that group. This relationship was similar in each of the vaccinated groups, demonstrating that measurement of the virus in dust can be used to monitor accurately both the infection status of the chickens and environmental contamination by MDV. RESUMEN. Relación entre los niveles del virus de Marek muy virulento en el polvo de instalaciones avícolas y en cañones de las plumas en pollos vacunados.Para evaluar el efecto de diferentes cepas de vacunas sobre la replicación y diseminación del virus muy virulento de la enfermedad de Marek en pollos infectados experimentalmente, se desarrollaron métodos cuantitativos de PCR (q-PCR) para determinar con precisión el ADN viral en los pollos infectados y en el entorno en el que se alojaron. Se alojaron cuatro grupos de 10 pollos por unidad de aislamiento, estas aves fueron vacunadas al primer día de edad con una de cuatro vacunas que incluyen los tres serotipos vacunales. Posteriormente fueron desafiados a los ocho días de edad con la cepa muy virulenta del virus de la enfermedad de Marek, Md5. En intervalos de tiempo regulares, se recolectaron cañones de plumas de cada pollo y también se recolectó el polvo del prefiltro del extractor de cada unidad de aislamiento. Se ...

show abstract

“…MDV expression culminates 14-28 dpi and then gradually decreases. The virus genome load was greater in feathers than in other tissues, such as the spleen and lymphoid tissues (Abdul-Careem et al, 2006;Baigent et al, 2005). Due to their important roles in the release and transmission of viruses, host responses induced in the feather follicle epithelium and the skin have recently attracted significant attention.…”

Section: Introductionmentioning

confidence: 99%

Differential protein analysis of chicken skin infected with Marek΄s disease virus

Chen¹,

Hu²,

Ch³

et al. 2014

View full text Add to dashboard Cite

Summary. -The skin and feather follicle epithelia of birds infected with Marek's disease virus (MDV) are the sites of infectious virus particle formation and shedding. However, the host responses and protein networks involved in the production of virus particles in the skin of MDV-infected chickens are poorly understood. This current study aimed to analyze the differential protein expression patterns in skin between MDV-infected and uninfected specific pathogen-free (SPF) chickens 28 days post infection (dpi) by combining two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) analyses. Through 2-DE analysis, our results revealed 23 proteins whose expression changed significantly following infection, of which 16 proteins were confirmed by MS. The identified proteins were functionally classified into 5 groups: immune-related, cell regulatory, cytoskeletal, metabolism-related and transport proteins. A single protein, beta 2-microglobulin, was further confirmed by real-time quantitative PCR. Beta 2-microglobulin expression was significantly increased in the infected group 28 dpi. This indicates that beta 2-microglobulin might play very important roles in the viral evasion from host immune response.Keywords: Marek΄s disease virus; skin; two-dimensional electrophoresis; beta 2-microglobulin; real time quantitative PCR * Corresponding author. E-mail: aijian@yzu.edu.cn; phone: +86-514-87979217. Abbreviations: MD = Marek΄s disease; MDV = MD virus; FFE = feather follicle epithelium; MHC I = major histocompatibility complex class I; SPF = specific pathogen-free; dpi = days post infection; 2-DE = two-dimensional electrophoresis; CEFs = chicken embryo fibroblasts; β2M = beta 2-microglobulin

show abstract

Replication kinetics of Marek's disease vaccine virus in feathers and lymphoid tissues using PCR and virus isolation

Cited by 64 publications

References 36 publications

Vaccination and Host Marek's Disease-Resistance Genotype Significantly Reduce Oncogenic <b><i>Gallid alphaherpesvirus 2</i></b> Telomere Integration in Host Birds

Vaccination and Host Marek's Disease-Resistance Genotype Significantly Reduce Oncogenic <b><i>Gallid alphaherpesvirus 2</i></b> Telomere Integration in Host Birds

Relationship Between Levels of Very Virulent MDV in Poultry Dust and in Feather Tips from Vaccinated Chickens

Differential protein analysis of chicken skin infected with Marek΄s disease virus

Contact Info

Product

Resources

About