Combined immunophenotyping and FISH with sex chromosome-specific DNA probes for the detection of chimerism in epidermal Langerhans cells after sex-mismatched bone marrow transplantation |
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Authors: | Harald Hessel Johann Mittermüller Horst Zitzelsberger Heinz-Ulrich Weier Manfred Bauchinger |
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Institution: | 1. GSF-National Research Center for Environment and Health, Institute of Radiobiology, Postfach 1129, D-85758, Oberschlei?heim, Germany 2. GSF-National Research Center for Environment and Health, Institute of Clinical Haematology, Germany 3. Strahlenbiologisches Institut, Ludwig Maximilians Universit?t München, Germany 4. Life Science Division, Lawrence Berkeley Laboratory, Center for Molecular Cytogenetics, Berkeley, California, USA
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Abstract: | Langerhans cells (LC) of the skin represent bone marrow-derived dendritic antigen-presenting cells and are therefore important
in pathophysiological processes such as rejection, graft-versus-host disease, and graft-versus-leukemia-reaction after bone
marrow transplantation (BMT). For understanding of these diseases, the evaluation of the chimeric status of LC following BMT
is of great interest. To analyze the sex chromosome constitution of LC in the skin, we established a modified and refined
technique of combined immunophenotyping and fluorescence in situ hybridization (FISH) and investigated frozen sections of
skin biopsies from nine patients after allogeneic sex-mismatched BMT and of two healthy donors for control. LC were specifically
labeled using a fluorescent CD1a antibody and hybridized simultaneously with X and Y chromosome-specific DNA probes. The results
of this practical application on nine leukemia patients show the appearance of donor-type LC and the persistence of host-type
LC at various times (36 up to 1395 days) after sex-mismatched BMT. Complete chimerism of LC could not be detected in any case.
The frequency of recipient-specific LC ranged from 7% to 92% and showed no correlation with time postgrafting. We conclude
from our results of 1461 analyzed LC that combined immunophenotyping and interphase cytogenetic analysis by FISH is the method
of choice for the assessment of chimerism in a particular cell type after sex-mismatched BMT. Its practical application on
other tissues affected by BMT-related pathophysiological processes reveals further knowledge of the time-dependent course
of chimeric patterns after BMT.
Accepted: 18 July 1996 |
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