Short tandem repeat analysis to monitor chimerism in macaca fascicularis.
Chimerism assessment following bone marrow transplantation (BMT) in cynomolgus monkeys (cynos) has been hampered by the lack of good engraftment markers. In human BMT, such markers have been provided by short tandem repeat (STR) loci. We tested the idea that techniques effective for detecting human STR could be readily adapted to cynos. Genomic DNA was extracted from cyno unseparated blood or peripheral cell subsets. With only slight modifications, reagents for detecting human STR alleles were used to amplify and detect cyno STRs and to quantitate allelic mixtures on an automated sequencer. Of the 15 STR loci tested, only CSF1PO, D18S51, and FGA successfully amplified, with seven, seven and two alleles, respectively. CSF1PO and D18S51 heterozygosity (80% and 55%, respectively) allowed use of these two loci for chimerism quantitation after BMT. The successful adaptation of human STR reagents to monitor chimerism in transplanted cynos will facilitate the use of this species in preclinical tolerance studies.
Transplantation Immunology Laboratory, Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, CA, USA.
Macy
M
Lau
Tamara
T
Vayntrub
F Carl
FC
Grumet
Robert
R
Lowsky
Samuel
S
Strober
Richard
R
Hoppe
Michael
M
Larson
Bari
B
Holm
Bruce
B
Reitz
Dominic
D
Borie
D000818:Animals; D016026:Bone Marrow Transplantation / Q000276:immunology*; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016131:Lymphocyte Subsets / Q000276:immunology; D008252:Macaca fascicularis; D020080:Tandem Repeat Sequences; D018183:Transplantation Chimera
D004247:DNA