Lack of LTR and ENV genetic variation during bovine leukemia virus-induced leukemogenesis.
Genetic variation of the Bovine Leukemia Virus (BLV) appears to be limited in vitro and during the latent phase of the disease. However, cells in tumors often harbor deleted proviruses that are defective for expression. In order to gain insight into the involvement of viral genetic variation during pathogenesis, the BLV LTR and the env proviral sequences were analyzed in tumor tissues. A sheep (M230) was injected with the cloned BLV provirus 344 and became persistently infected with circulating lymphocytes reaching 345,000/mm3. After 11 months, this infected sheep developed leukemia-lymphoma. DNA was extracted from peripheral blood leukocytes at the time of tumor development and the LTR and the env gene were amplified, using the polymerase chain reaction procedure, cloned, and sequenced. Twenty independent LTR and twenty env clones were analyzed. It appeared that the in vivo mutation rate in the env gene was 0.043% (eight mutations including seven transitions out of 18,300 bp). Five point mutations (all transitions) were identified in the LTR, corresponding to 0.041% modifications (four mutations out of 9740 bp). These mutation rate values (0.043 and 0.041) were close to those due to the Taq DNA polymerase errors (0.030%). Altogether, these data demonstrate the lack of genetic variation in the LTR and the env gene during this case of BLV-induced pathogenesis in vivo. They confirm that the defectiveness of some BLV proviruses in vivo, thus, is not a mandatory step in the leukemogenic process.