Use of highly sensitive mitochondrial probes to detect microchimerism in xenotransplantation models.
Chimerism, defined as the co-existence of cells of different origin within the same organism, has received much attention in hematopoietic cell and organ transplantation because of the strict relationship between its establishment and the induction of specific tolerance. Traditional methods applied for chimerism detection, such as immunohistochemistry, cytogenetics, fluorescent-activated cell sorter analysis, and serological and biochemical testing, are limited by their sensitivity. We have established a highly sensitive molecular approach based on the amplification of the mitochondrial cytochrome B gene and tested its specificity and sensitivity level in six different mammalian species, including human, pig, mouse, rat, sheep and rabbit. Increased sensitivity of detection of specific amplification products was obtained by the non-radioactive Southern blot technique. This novel approach allows the detection of one cell against the background of 1 to 4 x 10(6) xenogenec cells and will be helpful for high-sensitivity analysis of donor cell engraftment after xenotransplantation procedures in these animal models.
Centro di Ricerca E. Menni, Fondazione Poliambulanza Istituto Ospedaliero, Brescia, Italy.
Maddalena
M
Soncini
Patrizia Bonassi
PB
Signoroni
Marco
M
Bailo
Daniela
D
Zatti
Alessandra
A
Gregori
Guerino
G
Lombardi
Alberto
A
Albertini
Georg S
GS
Wengler
Ornella
O
Parolini
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D046528:Chimerism; D045303:Cytochromes b / Q000235:genetics / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000032:analysis; D006801:Humans; D008928:Mitochondria / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014183:Transplantation, Heterologous
D015415:Biomarkers; D004272:DNA, Mitochondrial; D045303:Cytochromes b