4.2 Article

Detection of microchimerism by minor histocompatibility antigen HA-1 allele-specific nested polymerase chain reaction

Journal

BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION
Volume 11, Issue 5, Pages 345-348

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/j.bbmt.2005.01.009

Keywords

minor histocompatibility antigen; microchimerism; nested PCR

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A minor histocompatibility antigens (mHags) can induce T-cell reactivities with important consequences for the graft-versus-leukemia effect and the development of graft-versus-host disease in IIIA-matched stem cell transplantation settings. Recently, m-Hag-specific T cells were also demonstrated in multiparous woman and in solid organ transplant recipients. Microchimeric cells have been detected in the latter settings. To study whether microchimerism is instrumental in the induction and/or maintenance of mHag T cells, we developed an HA-1 allele-specific nested polymerase chain reaction. To optimize and validate the reliability of this method at different levels of microchimerism, serial dilutions of HA-1(H) cells titrated into HA-1(R) cells were tested. We demonstrated that tire HA-1(H) allele can be reliably and consistently detected at concentrations as low as 1:10(5) without losing specificity.. The developed HA-1-specific nested polymerase chain reaction is an important tool that facilitates the detection of HA-1 microchimerism in various clinical specimens and that promotes investigation of the effects of microchimerism on induction of mHag-specific T cells in the various settings of immunization. (C) 2005 American Society for Blood and Marrow Transplantation.

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