OR49 Detecting chimerism following lung transplant with graft vs. host disease

Autor: Amit D. Parulekar, Lauren Clark, Peter T. Jindra, Ronald H. Kerman, Angela Hoover, Goutham Dronavalli, Nick Woolley
Rok vydání: 2016
Předmět:
Zdroj: Human Immunology. 77:28
ISSN: 0198-8859
DOI: 10.1016/j.humimm.2016.07.047
Popis: Graft-versus-host disease (GVHD) is a major post-transplant complication, but is rarely found in solid organ transplant. A 50 year old male presented with skin rash and GI symptoms consistent with GVHD two months after a lung transplant, and skin and gastrointestinal biopsies were performed that confirmed the diagnosis. Subsequently, we evaluated a peripheral blood sample (PBL) from the patient for the percentage T cells and B cells expressing donor HLA antigens. A previously described method was used to determine the donor’s typing by SSP (Hahn, 2000). In order to determine the relative amount of donor cells that were present in the recipient, crossmatches were carried out using cells acquired from PBL and biopsy samples of the recipient’s colon. Cells from the colon biopsy were then isolated using a tissue grinder often utilized for spleen lymphocyte isolation. After washing, the cells were re-suspended in McCoy’s 5A Medium and used untreated for a flow crossmatch. During flow crossmatch the cells were incubated with serum (negative control, positive control, the patient’s own serum, and donor specific anti-HLA A1 serum), washed, and later stained with CD3 and CD19 for T-cells and B-cells respectively. The HLA-A1 antigen was solely donor specific, therefore, if there were any HLA-A1 positive cells present in the PBL or biopsy, the donor T-cells/B-cells would deviate from the negative control. This method allows for measurement of live cells that are donor derived, which can be assessed before and after treatment to determine efficacy. The crossmatch performed before treatment displayed a deviation in the T-cells when incubated with anti-HLA-A1 serum that was then measured as 5% of the total T-cells gated. Therefore, 5% of the CD3+ T cell population in the colon was of donor origin. After treatment with solumedrol, another colon biopsy was performed and the crossmatch showed 0% of the CD3+ T-cells were donor derived. As a result, the crossmatch showed the treatment was working and the patient’s symptoms had resolved. A follow-up PBL crossmatch 355 days post-transplant was performed and found 22% donor HLA-A1 expressing CD3+ T cells suggesting the transplanted lung can act as a reservoir for donor T cells or a semi-indirect recognition pathway may present donor HLA on recipient T cells.
Databáze: OpenAIRE
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