Analysis of testes and semen from rabbits treated by intravenous injection with a retroviral vector encoding the human factor VIII gene: no evidence of germ line transduction
| Autor: | Dale E. Johnson, William G. Glass, Judith Greengard, Martin Giedlin, Edgar Kamantigue, Martha E. I. Leibbrandt, Kim Boekelheide, Douglas J. Jolly, Nancy Sajjadi, Holger Roehl |
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| Rok vydání: | 2000 |
| Předmět: |
Male
Pathology medicine.medical_specialty Time Factors Genetic enhancement Genetic Vectors Oligonucleotides Semen Testicle Biology Models Biological Polymerase Chain Reaction Sensitivity and Specificity Virus Viral vector Andrology Transduction Genetic Gene expression Testis Genetics medicine Animals Tissue Distribution Spermatogenesis Molecular Biology Factor VIII Models Statistical Reproducibility of Results medicine.anatomical_structure Retroviridae Cell culture Molecular Medicine Rabbits |
| Zdroj: | Human gene therapy. 11(18) |
| ISSN: | 1043-0342 |
| Popis: | In a phase 1 clinical trial, we are evaluating a murine leukemia virus (MuLV)-based retroviral vector encoding the human factor VIII gene [hFVIII(V)], administered intravenously, as a therapy for hemophilia A. Preclinical biolocalization studies in adult rabbits revealed vector-specific PCR signals in testis tissue at low levels. In follow-up animal studies we used PCR to (1) estimate the frequency with which a given cell in testis tissue is transduced, and (2) determine whether a positive PCR signal could be detected in semen samples from animals treated with hFVIII(V). Using the 99% confidence bound, results indicate that the probability that a given cell within the testis was transduced is less than 1/709,000 (97 days after treatment). This probability decreased with time after hFVIII(V) administration. Moreover, the rate of provector sequence detection in semen samples collected weekly throughout two cycles of spermatogenesis was 3/4281 reactions (0.07%), which is lower than the rate of false positives (1/800, 0.125%) observed for control animals. Using PCR assays with single-copy sensitivity, we have shown that the small number of transduced cells present in testis tissue does not give rise to detectable transduced cells in semen. |
| Databáze: | OpenAIRE |
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