Microchimerism and tolerance afterin UteroBone marrow transplantation in mice
Journal of Surgical Research, 1998•Elsevier
Background. Donor-specific tolerance has been induced after both fetal and neonatal
hematopoietic stem cell (HSC) transplantation in mice. However, the relationship between
hematopoietic microchimerism and tolerance in these models has not been defined due to
the insensitivity of donor cell detection methodology. To address this problem we developed
a semiquantitative polymerase chain reaction (PCR)-based assay for detection of
microchimerism after major histocompatibility (MHC) class I disparate HSC transplantation …
hematopoietic stem cell (HSC) transplantation in mice. However, the relationship between
hematopoietic microchimerism and tolerance in these models has not been defined due to
the insensitivity of donor cell detection methodology. To address this problem we developed
a semiquantitative polymerase chain reaction (PCR)-based assay for detection of
microchimerism after major histocompatibility (MHC) class I disparate HSC transplantation …
Background
Donor-specific tolerance has been induced after both fetal and neonatal hematopoietic stem cell (HSC) transplantation in mice. However, the relationship between hematopoietic microchimerism and tolerance in these models has not been defined due to the insensitivity of donor cell detection methodology. To address this problem we developed a semiquantitative polymerase chain reaction (PCR)-based assay for detection of microchimerism after major histocompatibility (MHC) class I disparate HSC transplantation. This assay was used to examine the relationship between microchimerism and tolerance after fetal and neonatal transplantation of fully allogeneic bone marrow cells.
Materials and methods
C57BL/6 mice (H2-Kb) were used as adult bone marrow donors and Balb/c mice (H2-Kd) were used as fetal or newborn recipients. A dose of 1010BM cells/kg was injected intraperitoneally into recipient animals. Peripheral blood of animals which survived beyond 3 weeks of age was analyzed by PCR for the presence of donor MHC class I DNA. Tolerance was tested by placement of donor-specific skin grafts after determination of chimerism status.
Results
Our assay was found to be specific for H2-Kb donor cells in an H2-Kd background with a sensitivity of <0.0001%. Of 49 animals injectedin utero19 (38%) had donor DNA present in peripheral blood at low levels (<0.1%) whereas only 1 of 18 neonatally injected animals had detectable donor cells (P< 0.01). Tolerance to donor-specific skin grafts was found in 6 of 9 animals which were chimeric afterin uteroHSC transplantation whereas none of the 18 neonatally injected animals including the chimeric animal were tolerant.
Conclusions
Our results indicate the following. (1) Hematopoietic microchimerism can be detected by PCR in peripheral blood afterin uteroinjection of fully allogeneic HSCs. (2) Fetal injections yield a higher incidence of microchimerism than newborn injections. (3) Tolerance can be induced across a fully allogeneic barrier byin uteroHSC transplantation and this is associated with the presence of peripheral blood microchimerism.
Elsevier