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Tissue distribution of adoptively transferred adherent lymphokine-activated killer cells assessed by different cell labels
Authors:P. Basse  R. B. Herberman  M. Hokland  R. H. Goldfarb
Affiliation:(1) Pittsburgh Cancer Institute and Departments of Pathology and Medicine, University of Pittsburgh, 15 213 Pittsburgh, PA, USA;(2) Present address: Institute of Medical Microbiology, University of Aarhus, DK-8000 Aarhus C, Denmark
Abstract:Summary Assessment of the tissue distribution of adoptively transferred adherent lymphokine-activated killer A-LAK) cells by use of51Cr indicated that these effector cells, after an initial phase in the lungs, distributed in high numbers to liver and spleen (30% and 10% of injected dose, respectively). However, when this experiment was repeated with125IdUrd as cell label, fewer than 2% and 0.5% of the injected cells distributed into liver and spleen respectively. To analyse this discrepancy, we compared the tissue distribution of51Cr- and125IdUrd-labelled A-LAK cells with that indicated by alternative direct visual methods for identification of the injected cells, such as fluorescent dyes (rhodamine and H33342) or immunohistochemical staining of asialo-GM1-positive cells. The number of i. v. injected A-LAK cells found in the liver by all visual methods ranged from 1% to 5% of the injected dose, supporting the data obtained with125IdUrd, whereas 25%–30% of the51Cr label was consistently found in this organ. Autoradiography of the liver 24 h after i. v. injection of51Cr-labelled cells revealed a background activity that was four- to fivefold higher than the control level, indicating substantial non-specific accumulation in the liver of51Cr released from A-LAK cells. We conclude that51Cr cannot be reliably used in investigations of cell traffic to the liver because of non-specific accumulation of the51Cr label, particularly in this organ. In contrast, labelling with125IdUrd or rhodamine and immunohistochemical staining of asialo-GM1-positive cells appear to be reliable and essentially equivalent methods for investigations of the fate of adoptively transferred A-LAK cells. Using these methods, we found that only few A-LAK cells redistribute to the liver upon i. v., i. e. systemic, injection, whereas 40%–50% of locally (intraportally) injected A-LAK cells remain in the liver for at least 24 h.
Keywords:LAK migration  Asialo-GM1 antibody  Fluorescence  Rhodamine  Hoechst 33342
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