Bone marrow-derived cells as carriers of recombinant immunomodulatory cytokine genes to lymphoid organs.
The purpose of this study was to determine the feasibility of cytokine gene delivery to lymphatic tissue using transduced bone marrow-derived cells. MBAE and pBABE retroviral vectors carrying the genes for murine interleukin-4 and the selection marker neomycin phosphotransferase (neo) were used to transduce bone marrow-derived dendritic cells (DC) and hematopoietic stem cells (HSC). A transduction efficiency of 11-33% for HSC and 2-10% for DC was achieved. Transduced HSC and DC released 55-170 pg of recombinant interleukin-4 per 1 x 10(6) cells/mL in vitro. To study the migration of the cells in vivo, we introduced the transduced cells into syngenic mice. DC were injected subcutaneously into the front limbs of unconditioned mice and HSC were intravenously administered to irradiated mice. The distribution of the transduced cells was studied by quantitative polymerase chain reaction for the neo gene as a marker. After 3 days, DC migrated to the axillary lymph nodes in the drainage area of the injection site and were detectable up to 5 days. After intravenous administration of transduced HSC, the neo gene could be found in up to 100 copies/5 x 10(3) cells in mesenterial lymph node, spleen, bone marrow, thymus, and liver. The distribution of the transduced cells was heterogenous: in different mice, different organs showed high copies of the neo gene after 10 and 13 days. After 39 days, two of three mice were negative for neo in all organs analyzed. In conclusion, bone marrow-derived cells can be genetically engineered ex vivo to deliver recombinant cytokine genes to lymphoid organs in vivo. In particular, DC might be candidate cells for use in immunomodulatory gene therapy for autoimmune diseases and cancer.
Fiehn
C
C
Department of Medicine V, University of Heidelberg, Germany. Christoph_Fiehn@med.uni-heidelberg.de
Wettschureck
N
N
Krauthoff
A
A
Haas
R
R
Ho
A D
AD
D000818:Animals; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000378:metabolism*; D017931:DNA Primers; D005260:Female; D005822:Genetic Vectors; D015847:Interleukin-4 / Q000235:genetics*; D019868:Kanamycin Kinase / Q000235:genetics; D008198:Lymph Nodes / Q000378:metabolism*; D008297:Male; D051379:Mice; D023421:Models, Animal; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics; D014161:Transduction, Genetic
D017931:DNA Primers; D011994:Recombinant Proteins; D015847:Interleukin-4; D019868:Kanamycin Kinase