Metes, D and Logar, A and Rudert, WA and Zeevi, A and Woodward, J and Demetris, AJ and Abu-Elmagd, K and Eghtesad, B and Shapiro, R and Fung, JJ and Trucco, M and Starzl, TE and Murase, N
(2003)
Four-color flow cytometric analysis of peripheral blood donor cell chimerism.
Human Immunology, 64 (8).
787 - 795.
ISSN 0198-8859
Abstract
Passenger leukocytes have been demonstrated to play significant roles in initiating and also regulating immune reactions after organ transplantation. Reliable techniques to detect donor leukocytes in recipients after organ transplantation are essential to analyze the role, function, and behavior of these leukocytes. In this report we describe a simple, reliable method to detect donor cells with low frequencies using peripheral blood samples. Detection of small numbers of major histocompatibility complex (MHC) mismatched cells was first studied using four-color flow cytometry in artificially created cell mixtures. By selecting the CD45+ population and simultaneous staining with several leukocyte lineage markers (CD3, CD4, CD8, CD56, and CD19), MHC-mismatched leukocytes were consistently detected in cell suspensions prepared from directly stained whole blood samples with a threshold sensitivity as low as 0.1%-0.2%. When the fresh peripheral blood mononuclear cells were separated by conventional Ficoll gradient purification, similar, but slightly lower levels of donor cells were detected. Blood samples obtained 1-5 months after liver, kidney, and intestine transplants revealed that the kind of organ allograft influenced levels and lineage pattern of the circulating donor cells. This procedure provided a simple and reliable method in determining early chimerism in transplant recipients. However, the detection of MHC-mismatched leukocytes of all lineages was much lower when frozen peripheral blood mononuclear cells were used. © American Society for Histocompatibility and Immunogenetics, 2003. Published by Elsevier Inc.
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Item Type: |
Article
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Status: |
Published |
Creators/Authors: |
Creators | Email | Pitt Username | ORCID |
---|
Metes, D | dim4@pitt.edu | DIM4 | | Logar, A | | | | Rudert, WA | | | | Zeevi, A | | | | Woodward, J | jew7@pitt.edu | JEW7 | | Demetris, AJ | | | | Abu-Elmagd, K | | | | Eghtesad, B | | | | Shapiro, R | | | | Fung, JJ | | | | Trucco, M | mnt@pitt.edu | MNT | | Starzl, TE | tes11@pitt.edu | TES11 | | Murase, N | | | |
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Centers: |
Other Centers, Institutes, Offices, or Units > Thomas E. Starzl Transplantation Institute |
Date: |
1 August 2003 |
Date Type: |
Publication |
Journal or Publication Title: |
Human Immunology |
Volume: |
64 |
Number: |
8 |
Page Range: |
787 - 795 |
DOI or Unique Handle: |
10.1016/s0198-8859(03)00113-7 |
Institution: |
University of Pittsburgh |
Refereed: |
Yes |
ISSN: |
0198-8859 |
Other ID: |
uls-drl:31735062121011, Starzl CV No. 2195 |
Date Deposited: |
08 Apr 2010 17:37 |
Last Modified: |
22 Jun 2021 13:56 |
URI: |
http://d-scholarship.pitt.edu/id/eprint/5581 |
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