A molecular epidemiological probe for pig microchimerism.
We have cloned and characterized a single-copy DNA sequence from the porcine alpha-1,3-galactosyltransferase gene that corresponds to a 547-base pair intron separating exons 3 and 4 of the protein coding domain. Polymerase chain reaction amplification of this sequence from flanking oligonucleotides generates a species-specific DNA probe (pgt34) capable of recognizing 50 pg chimeric template DNA at a pig to human cellular ratio of 1/10,000. Homologous DNA sequence is not identified in the macaque, baboon, or human genome by Southern hybridization. Analysis of a discordant model of pig to baboon xenotransplantation demonstrates peripheral blood microchimerism in the presence of a functioning pig kidney xenograft and persistence of microchimerism in lymphatic tissue after graft removal. This probe should be useful for tracking the fate of porcine cells in patients undergoing xenotransplantation of whole organs or free tissues such as pancreatic islet cells and should facilitate studies of microchimerism in experimental models of pig to monkey xenotransplantation.
Hoopes
C W
CW
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
Platt
J L
JL
D000818:Animals; D030801:Animals, Genetically Modified; D015342:DNA Probes / Q000032:analysis; D005700:Galactosyltransferases / Q000235:genetics; D006801:Humans; D007438:Introns; D016030:Kidney Transplantation; D010215:Papio; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013552:Swine; D018183:Transplantation Chimera / Q000235:genetics*; D016042:Transplantation, Heterotopic
D015342:DNA Probes; D005700:Galactosyltransferases; C037959:beta-D-galactosyl-N-acetylglucosaminylglycopeptide alpha-1,3-galactosyltransferase