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Establishment of an In Vitro Model of Persistent Chicken Anemia Virus Infection.

ABSTRACT: Persistent infection of chicken anemia virus (CAV) in chickens has been suspected to result in immunosuppression and exogenous virus contamination within vaccine production. However, no direct evidence for persistent CAV infection has thus far been obtained. In this study, we aimed to establish an in vitro model of persistent CAV infection. CAV-infected MDCC-MSB1 (MSB1) cells, a Marek's disease virus-transformed continuous cell line, were cultured in the presence of both CAV and CAV neutralizing antibody (NA). Cell viability, expression of viral antigens, viral DNA, and recovery of CAV were examined by acridine orange/propidium iodide staining, immunofluorescence measurement, real-time PCR, and viral isolation, respectively. The results indicated that CAV was maintained and possibly replicated in CAV-infected cells cultured in the presence of NA, without affecting host cell viability. It was also shown that persistently infectious CAV induced cell death again after removing NA. The persistent infection of CAV in MSB1 cells was not related to viral gene mutation. In summary, we have herein established a novel model of persistent CAV infection in MSB1 cells cultured in the presence of NA.

SUBMITTER: Van Dong H

PROVIDER: S-EPMC7602425 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Establishment of an In Vitro Model of Persistent Chicken Anemia Virus Infection.

Van Dong Hieu H Tran Giang Thi Huong GTH Trinh Dai Quang DQ Takeda Yohei Y Ogawa Haruko H Imai Kunitoshi K

Pathogens (Basel, Switzerland) 20201015 10

Persistent infection of chicken anemia virus (CAV) in chickens has been suspected to result in immunosuppression and exogenous virus contamination within vaccine production. However, no direct evidence for persistent CAV infection has thus far been obtained. In this study, we aimed to establish an in vitro model of persistent CAV infection. CAV-infected MDCC-MSB1 (MSB1) cells, a Marek's disease virus-transformed continuous cell line, were cultured in the presence of both CAV and CAV neutralizing ...[more]

PMID: 33076262

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Project description:Naturally acquired chicken anemia virus (CAV) infection in chickens frequently occurs from 3 weeks of age onward after maternally derived antibodies have decayed. The oral inoculation of older chickens with CAV was reported to have negative effects on cell-mediated immune function, and pathological changes were identified. To date, there has been no complete illustration of an immunological and persistent infection. To understand the pathogenesis of persistent CAV infection, an immunological study of CAV-infected 3-week-old specific pathogen-free (SPF) chickens was carried out by different routes of inoculation. The weight, packed cell volumes, and organ samples were obtained at 7, 14, 21, and 28 days postinfection (dpi). Here, we compared hematological, immunological, and sequential pathological evaluations and determined the CAV tissue distribution in different organs. Neither a reduction in weight gain nor anemia was detected in either the inoculated or the control group. The immune-pathological changes were investigated by evaluating the body and thymus weight ratio and specific antibody titer. Delayed recovery of the thymus corresponding to a low antibody response was detected in the orally inoculated group. This is different from what was found in chickens intramuscularly infected with the same dose of CAV. The CAV remaining in a wide range of tissues was examined by viral reisolation into cell culture. The absence of the virus in infected tissues was typically found in the intramuscularly inoculated group. These chickens were immediately induced for a protective antibody response. A few viruses replicating in the thymus were found 21 dpi due to the regression in the antibody titer in the orally inoculated group. Our findings support that a natural infection with CAV may lead to the gradual CAV viral replication in the thymus during inadequate antibody production. The results clearly confirmed that virus-specific antibodies were essential for viral clearance. Under CIA-risk circumstances, administration of the CAV vaccine is important for achieving a sufficient protective immune response.

Project description:Equine infectious anemia virus (EIAV) infection of horses is characterized by well-defined waves of viremia associated with the sequential evolution of distinct viral populations displaying extensive envelope gp90 variation; however, a correlation of in vivo envelope evolution with in vitro serum neutralization phenotype remains undefined. Therefore, the goal of the present study was to utilize a previously defined panel of natural variant EIAV envelope isolates from sequential febrile episodes to characterize the effects of envelope variation during persistent infection on viral neutralization phenotypes and to define the determinants of EIAV envelope neutralization specificity. To assess the neutralization phenotypes of the sequential EIAV envelope variants, we determined the sensitivity of five variant envelopes to neutralization by a longitudinal panel of immune serum from the source infected pony. The results indicated that the evolution of the EIAV envelope sequences observed during sequential febrile episodes produced an increasingly neutralization-resistant phenotype. To further define the envelope determinants of EIAV neutralization specificity, we examined the neutralization properties of a panel of chimeric envelope constructs derived from reciprocal envelope domain exchanges between selected neutralization-sensitive and neutralization-resistant envelope variants. These results indicated that the EIAV gp90 V3 and V4 domains individually conferred serum neutralization resistance while other envelope segments in addition to V3 and V4 were evidently required for conferring total serum neutralization sensitivity. These data clearly demonstrate for the first time the influence of sequential gp90 variation during persistent infection in increasing envelope neutralization resistance, identify the gp90 V3 and V4 domains as the principal determinants of antibody neutralization resistance, and indicate distinct complex cooperative envelope domain interactions in defining sensitivity to serum antibody neutralization.

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Establishment of an In Vitro Model of Persistent Chicken Anemia Virus Infection.

Publications

Establishment of an In Vitro Model of Persistent Chicken Anemia Virus Infection.

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