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Color-coded tracking method helps scientists analyze outcomes of newly transplanted tissue
Date:5/27/2010

uld express natural Treg cells (nTreg) in a fluorescent green protein and another in which Teff cells express a fluorescent red protein. (A third color, yellow, also came into play when the red effector T cells commit to the induced Treg phenotype, expressing both red and green fluorescent proteins and thereby appearing as yellow cells. This approach enables ready distinction between natural and induced Treg cells.)

The BIDMC investigators then partnered with a group of physicists at Massachusetts General Hospital led by Charles Lin, PhD. Lin and his team had developed a novel imaging technique that coupled in vivo flow cytometry with endoscopic confocal microscopy.

"Genetically mismatched insulin-producing islet cells were transplanted beneath the thin capsule surrounding the animals' kidneys," explains Koulmanda. "Infiltration of T cells into the transplant was then visualized in untreated recipient mice in which vigorous rejection occurred. These results were then compared with mice that had received a short course of treatment enabling them to permanently accept the transplant [i.e. immune tolerance]."

Using the new imaging process, the authors were able to clearly see that the ratio of protective T cells to destructive T cells differed markedly in the transplant tolerant mice compared with the mice in which transplant tissue was rejected. Among the "rejection mice," red cells rushed into the transplant far in advance of infiltration into the transplant by the tolerance hosts.

"As the events of rejection proceeded, the number of transplanted infiltrating T cells vastly exceeded those present in the tolerant transplants, even though the numbers of 'protective' yellow and green cells were equal in these groups at a later point in time," explains Koulmanda. "This is the first time that we have been able to monitor transplanted allograft tissue in a live lab animal. While static images of cells have been captured in the past, ou
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Contact: Bonnie Prescott
bprescot@bidmc.harvard.edu
617-667-7306
Beth Israel Deaconess Medical Center
Source:Eurekalert

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