Donor hematopoietic cells from transgenic mice that express GFP are immunogenic in immunocompetent recipients.
OBJECTIVE
To study the immunogenicity of hematopoietic cells marked with green fluorescence protein (GFP) while avoiding the potentially confounding effects of viral gene transduction, marked cells from GFP+ transgenic mice were tracked after transplantation into unconditioned immunocompetent recipients.


METHODS
Marrow was harvested from GFP+ transgenic mice that had been crossed onto a BALB/cByJ background. Unconditioned marrow transplantation involved infusion of sex-matched or sex-mismatched cells into female BALB/cByJ hosts. Engraftment and contribution to circulating nucleated blood cells were compared to recipients of donor cells that were not GFP-marked. Donor cells were detected by flow cytometry (GFP) and fluorescence in situ hybridization (FISH) for Y-chromosome sequences.


RESULTS
Donor cells from mice of the same genetic background that did not express GFP were detected for more than four-weeks in unconditioned recipients. In contrast, GFP-marked cells in the blood peaked at one-week, declined to undetectable levels by two-weeks and were not detected in the marrow at sacrifice. In sex-mismatched studies, detection of male GFP+ donor cells by FISH yielded levels similar to those observed by flow cytometry, in contrast to the levels detected for many weeks in mice infused with male cells that did not express GFP. In immunocompetent recipients immunized with irradiated GFP-expressing cells, rechallenge with GFP+ cells resulted in the accelerated loss of donor cells.


CONCLUSIONS
Donor marrow cells from GFP+ transgenic mice were lost after infusion into unconditioned immunocompetent mice and sensitization studies infer an immunologic mechanism. These results are similar to studies of virally transduced cells. Thus, infusion of cells with optimum engraftment potential could not compensate for the loss of donor cells due to immunogenicity.
Bubnic
Simon J
SJ
Osiris Therapeutics Inc., 2001 Aliceanna Street, Baltimore, MD 21231, USA. sbubnic@osiristx.com
Nagy
Andras
A
Keating
Armand
A
D000818:Animals; D001773:Blood Cells / Q000276:immunology*; D016026:Bone Marrow Transplantation; D005260:Female; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology*; D006085:Graft Survival / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000276:immunology*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D019076:Transgenes / Q000235:genetics / Q000276:immunology*; D014998:Y Chromosome / Q000235:genetics / Q000276:immunology
D049452:Green Fluorescent Proteins