Hi Norbert- As far as I know, no tracking dye is approved for use in humans. BrdU was used in the '80's to track in vivo proliferation, particularly in leukemia. I do not know if you could get an investigator-sponsored IND in the States now, but at least there is precedent. I have even less of an idea of how this would fly in Germany. The down side is that you are administering the BrdU in vivo and labeling all cells that are in cycle, not prelabeling the cells as you propose with CFSE or the PKH dyes. The other approach is through the administration of deuterated glucose. I don't know the regulatory status, but this appears to be pretty innocuous stuff. The down side: it requires cell purification and gas chromatography- mass spec. A less quantitative, but less invasive, way to approach this problem is to simultaneously measure tetramer+ events, and a marker of proliferation like Ki67 (intracellular) or CD71. Since the background of both of these markers is low in peripheral T cells, you may see a signal in your tetramer+ events if they are cycling in vivo. Best of luck. This sounds like great stuff. Warmest regards, Albert Albert D. Donnenberg, Ph.D. Professor of Medicine -----Original Message----- From: Norbert Meidenbauer [mailto:norbert.meidenbauer@klinik.uni-regensburg.de] Sent: Thursday, August 22, 2002 3:34 AM To: cyto-inbox Subject: Proliferation analysis of T cells in vivo Dear flowers, we are doing adoptive T cell therapy with antigen-specific T cells for malignant melanoma, tracking down the transferred T cells with tetramer-assays. In average, we are seeing frequencies of antigen- specific T cells between 0,5-2% of CD8+ T cells in the peripheral blood after transfer. We now would like to know, if the transferred T cells still proliferate in the patients. Does anybody now about a dye to label the T cells before transfer, which is approved for use in humans. CFSE, PKH and the Mitotrackers are not approved for human use according to the MSDS (as far as I know). For any hints and suggestions I would be very thankful. Norbert Meidenbauer Dept. Hematology/Oncology University of Regensburg 93042 Germany e-mail: Norbert.meidenbauer@klinik.uni-regensburg.de
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