A method for improved yield and purity in extracting lymphocytes from lung tumors.
Tumor-infiltrating lymphocytes (TIL) from human pulmonary tumors have been studied as a model for local depression of cell-mediated immunity at the tumor site. To better characterize these lymphoid infiltrates, a new method of lymphocyte extraction of high yield and purity was developed. The TIL from 11 human lung tumors of varying histologic types were prepared. After mechanical disaggregation to a single cell suspension, the TIL, contaminated with tumor cells and macrophages, were incubated with carbonyl iron to deplete macrophages. Further purification was performed in a unit gravity sedimentation apparatus utilizing a continuous 2-4% Ficoll gradient. Usable TIL recovery was 41.2 +/- 7.7% of input cells and was minimally contaminated by tumor cells and macrophage/monocytes. The high yield and purity of TIL preparations have resulted in a higher proportion of tumors suitable for immunological analysis. An example of the type of experiments possible with such preparations is shown.
D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D004950:Esterases / Q000032:analysis; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology*; D008214:Lymphocytes; D008264:Macrophages / Q000201:enzymology
D004950:Esterases