Immunological and molecular characterization of an aggressive murine lymphoma variant: modulation in vitro and in vivo.

The highly metastatic murine ESb-L lymphoma was analyzed with respect to its possible origin and phenotype modulation. By determining the methylation status of the CD8 gene an early thymic origin of the ESb-L lymphoma cells is suggested. It revealed that the precursors of ESb-L cells had at least one CD8 allele expressed during their development. ESb-L tumor cells were found to express ICAM-1, ICAM-2, VLA-4 and Mel14 as adhesion molecules and homing receptors and CD25, CD69 and CD124 (HSA) as T-cell related activation markers. PCR analysis revealed that ESb-L tumor cells express a Th2-like cytokine pattern with mRNAs for IL-4, IL-5, IL-6, IL-10 and IL-13, but not for IL-2 and IFNgamma. In addition mRNA for TNFalpha, LT, IFNalpha and the chemokines MIP1alpha and MIP1beta was found. The expression of the adhesion molecules ICAM-1, ICAM-2, VLA-4 and of the T-cell activation marker CD25 on ESb-L tumor cells could be upregulated by incubating the cells with 10 ng/ml TNFalpha. For CD25 this effect was confirmed also at the mRNA level. Using the lacZ transduced T-cell lymphoma ESb-L-CI we were able to re-isolate live tumor cells from the primary site or from a metastasized liver and to investigate their phenotype ex vivo. MIP1alpha mRNA expression was strongly reduced in ex vivo isolated tumor cells as compared to in vitro grown cells indicating the modulatory role of the tumor microenvironment. The presented data suggest possible roles of TNFalpha and/or other microenvironmental factors modulating the expression of molecules involved in cell migration and adhesion thereby influencing cancer metastasis.