title: Immunoresponse against the transgene limits hematopoietic engraftment of mice transplanted in utero with virally transduced fetal liver.
abstract: In utero cell and gene therapies constitute alternative strategies to the postnatal treatment of inherited diseases. Fetal hematopoietic progenitors could be a potential source of donor cells for these strategies. In this study, hematopoietic lineage-negative fetal liver cells from 14.5-day-old fetuses were transduced under different cytokine and culture combinations using a lentiviral vector expressing the enhanced green fluorescent protein (EGFP). When cells were transduced for 6 h in the presence of mSCF, hTPO and FLT3-L in retronectin-coated dishes at a multiplicity of infection of 10 transduction units/cell, up to 70% of granulo-macrophage colony-forming cells expressed the EGFP reporter gene. In utero transplantation experiments revealed that conditions leading to high transduction efficiencies were associated with poor engraftments of syngeneic recipients. Significantly, this effect was associated with the detection of a humoral and cellular immunoresponse against the transgenic protein. Moreover, the humoral response against EGFP was detected not only in in utero transplanted recipients but also in the operated mothers, suggesting the maternal origin of the anti-EGFP immunoresponse. These observations reinforce the necessity of carefully studying the potential immunoresponses in future prenatal gene therapy protocols.
authors.affiliation: Hematopoiesis and Gene Therapy Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas y Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid, Spain.
authors.forename: M E
authors.initials: ME
authors.lastname: Alonso-Ferrero
authors.forename: A
authors.initials: A
authors.lastname: Valeri
authors.forename: R
authors.initials: R
authors.lastname: Yañez
authors.forename: S
authors.initials: S
authors.lastname: Navarro
authors.forename: M I
authors.initials: MI
authors.lastname: Garin
authors.forename: J C
authors.initials: JC
authors.lastname: Ramirez
authors.forename: J A
authors.initials: JA
authors.lastname: Bueren
authors.forename: J C
authors.initials: JC
authors.lastname: Segovia
mesh_terms: D000818:Animals; D000917:Antibody Formation; D005260:Female; D005333:Fetus; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D007111:Immunity, Cellular; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000196:embryology*; D051379:Mice; D016513:Mice, SCID; D011247:Pregnancy; D011270:Pregnancy, Animal; D014161:Transduction, Genetic; D019076:Transgenes / Q000276:immunology*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins