Insertion of reticuloendotheliosis virus long terminal repeat into a bacterial artificial chromosome clone of a very virulent Marek's disease virus alters its pathogenicity.
Co-cultivation of the JM/102W strain of Marek's disease virus (MDV) with reticuloendotheliosis virus (REV) resulted in the generation of a recombinant MDV containing the REV long terminal repeat (LTR) named the RM1 strain of MDV, a strain that was highly attenuated for oncogenicity but induced severe bursal and thymic atrophy. We hypothesize that the phenotypic changes were solely due to the LTR insertion. Furthermore, we hypothesize that insertion of REV LTR into an analogous location in a different MDV would result in a similar phenotypic change. To test these hypotheses, we inserted the REV LTR into a bacterial artificial chromosome (BAC) clone of a very virulent strain of MDV, Md5, and designated the virus rMd5-RM1-LTR. The rMd5-RM1-LTR virus and the rMd5 virus were passaged in duck embryo fibroblast cells for up to 40 passages before pathogenicity studies. Susceptible chickens were inoculated intra-abdominally at hatch with the viruses rMd5-RM1-LTR, rMd5 BAC parental virus, wild-type strain Md5, or strain RM1 of MDV. The rMd5-RM1-LTR virus was attenuated at cell culture passage 40, whereas the rMd5 BAC without RM1 LTR retained its pathogenicity at cell culture passage 40. Using polymerase chain analysis, the RM1 LTR insert was detected in MDV isolated from buffy coat cells collected from chickens inoculated with rMd5-RM1-LTR, but only at 1 week post inoculation. The data suggest that the presence of the RM1 LTR insert within MDV genome for 1 week post inoculation with virus at hatch is sufficient to cause a reduction in pathogenicity of strain Md5 of MDV.
Avian Disease and Oncology Laboratory, US Department of Agriculture, Agricultural Research Service, East Lansing, MI 48823, USA.
Jody K
JK
Mays
Robert F
RF
Silva
Taejoong
T
Kim
Aly
A
Fadly
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002478:Cells, Cultured; D002645:Chickens; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D005260:Female; D008297:Male; D025922:Mardivirus / Q000235:genetics* / Q000472:pathogenicity*; D008380:Marek Disease / Q000821:virology*; D016254:Mutagenesis, Insertional / Q000379:methods; D016133:Polymerase Chain Reaction; D017986:Reticuloendotheliosis Viruses, Avian / Q000235:genetics*; D020079:Terminal Repeat Sequences / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
D000914:Antibodies, Viral