Engraftment of genetically modified bone marrow cells in sensitized hosts.
Expression of retrovirally transduced genes in bone marrow-derived cells can be used to establish stable long-term B- and T-cell tolerance. To determine whether preexisting antibodies may prohibit the use of gene therapy to establish tolerance, we examined the extent to which preexisting antibodies specific for the carbohydrate antigen Gal alpha1-3Gal beta1-4GlcNAc-R (alpha Gal) could affect engraftment and development of bone marrow progenitors expressing the enzyme UDPgalactose:beta-D-galactosyl-1,4-N-acetyl-D-glucosaminide alpha(1-3)galactosyltransferase (E.C. 2.4.1.151), or simply alpha GT, which synthesizes the alpha Gal epitope. Groups of alpha GT knockout mice (GT(0) mice) lacking alpha Gal were presensitized to alpha Gal by immunization and then lethally irradiated and reconstituted with varying numbers of alpha GT-transduced syngeneic bone marrow cells. Whereas unimmunized controls were reconstituted with as few as 2 x 10(6) transduced cells, a significant fraction of immunized mice reconstituted with 2 x 10(6) or 4 x 10(6) alpha GT-transduced cells failed to undergo bone marrow engraftment and died. Immunized mice in which radiation protection was achieved failed to express alpha Gal. However, radiation protection and expression of alpha Gal on bone marrow-derived cells, resulting in tolerance, could be achieved by increasing the number of transduced cells used to reconstitute immunized mice. Thus, although high levels of preexisting antibodies can be a significant barrier to engraftment, this barrier can be overcome by increasing the number of transduced cells used for reconstitution.
Bracy
Jennifer
J
Transplantation Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, 02129, USA.
Iacomini
John
J
D000818:Animals; D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology; D016026:Bone Marrow Transplantation / Q000276:immunology*; D005700:Galactosyltransferases / Q000172:deficiency / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods; D006085:Graft Survival / Q000276:immunology; D007108:Immune Tolerance; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D013552:Swine; D013556:Swine, Miniature; D014183:Transplantation, Heterologous; D014312:Trisaccharides / Q000096:biosynthesis / Q000276:immunology
D014312:Trisaccharides; C076569:alpha-galactosyl epitope; D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase