Session Time: 6:00pm-7:00pm
Presentation Time: 6:00pm-7:00pm
Location: Hall 4EF
Background: Chimerism, defined as coexistence of donor and recipient cells, has been associated with induction and maintenance of tolerance to allotransplants. Although most studies have measured chimerism using flow cytometry, we proposed precision in readings can be improved with complimentary polymerase chain reaction (PCR) on specific short tandem repeat.
Method: A search in the Rat Genome Database (http://rgd.mcw.edu/) identified the short tandem repeat (STR), D10RAT25, which showed size difference between the two rat strains widely applied in allotransplantation studies, namely Lewis and BN. Chimerism was measured by PCR amplification of D10RAT25 and followed by quantitation with capillary electrophoresis.
Result: A standard curve was constructed with purified Lewis and BN DNA. Chimerism can be measured accurately from 0.4%~100%. Complementation with RT1Bβ can detect as low as 0.1% BN DNA. Good agreement was shown between the current D10Rat25-PCR method and flow cytometry. When applied to rat vascularized composite allotransplantation (VCA), the relationship of systematic (in peripheral blood) or local (at specific organ/tissues) chimerism to allograft outcome was delineated. Furthermore, we demonstrated that the magnitude of changes of VCA skin chimerism during early rejection (set as the parameter DChimerismPOM1-ER) had outstanding discriminative power on VCA outcome. The ROC analysis identified the optimum cut-off value to be 17.7%.
Conclusion: A reliable method to quantify the percentage of BN cells/DNA in BN-LEW chimeras was developed. We have characterized the detection limit, and ensured the matched accuracy with flow cytometry. This method can be widely applied to rat VCA and organ transplantation studies with precision.
CITATION INFORMATION: Cheng H., Huang X., Lin C-.F., AL Deek N., Shih L., Lin C-.H., Wei F-.C. Predicting Outcomes of Rat Vascularized Composite Allotransplants through Quantitative Measurement of Chimerism with PCR-Amplified Short Tandem Repeat Am J Transplant. 2017;17 (suppl 3).
To cite this abstract in AMA style:Cheng H, Huang X, Lin C-F, Deek NAL, Shih L, Lin C-H, Wei F-C. Predicting Outcomes of Rat Vascularized Composite Allotransplants through Quantitative Measurement of Chimerism with PCR-Amplified Short Tandem Repeat [abstract]. https://atcmeetingabstracts.com/abstract/predicting-outcomes-of-rat-vascularized-composite-allotransplants-through-quantitative-measurement-of-chimerism-with-pcr-amplified-short-tandem-repeat/. Accessed July 8, 2020.
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