PURIFICATION AND IDENTIFICATION OF CHEMOKINES POTENTIALLY INVOLVED INKIDNEY-SPECIFIC METASTASIS BY A MURINE LYMPHOMA VARIANT - INDUCTION OF MIGRATION AND NF-KAPPA-B ACTIVATION

The ESb-MP cell line is the subclone of a highly malignant variant of murine methylcholanthrene-induced T lymphoma, ESb, When injected in vi vo, ESb-MP cells metastasize to the kidney with high frequency, wherea s a non-adherent variant, ESb tells, rarely form metastatic foci in th e kidney, Our previous results skewed that ESb-MP, but not ESb, cells were able to migrate in response to murine kidney-conditioned media (K CM). In an effort to characterize the tumor cell chemoattractant(s) pr oduced by kidney cells, we found that the murine kidney mesangial cell line MES-13 released more chemotactic activity for ESb-MP cells than present in KCM. A major heparin-binding chemotactic activity was purif ied to homogeneity by sequential fast-performance liquid chromatograph y and reversed phase high-performance liquid chromatography, Amino aci d sequencing of the formic acid-digested active fractions revealed tha t the purified protein was identical to murine MCP-1(JE) and its activ ity was neutralized by an anti-MCP-1(JE) antibody, Another chemokine, RANTES, was also purified from MES-13 cell supernatant. The chemotacti c activity contained in the MES-13 cell supernatant and in murine KCM was neutralized in part by a combination of anti-MCP-1(JE) and anti-RA NTES antibodies. We further examined the differences in the ESb-MP and ESb cells. Binding studies using a variety of radio-iodinated chemoki nes showed that although both ESb-MP and ESb cells expressed substanti al levels of high-affinity binding sites for CC chemokines, only ESb-M P cells migrated in response to CC chemokines and these cells constitu tively expressed higher levels of beta 2 integrin adhesion protein CD1 1b than their parental ESb cells. CC chemokines also activated NF kapp a B in ESb-MP but not its ESb cells. Our results indicate that CC chem okines, selectively chemoattract and activate ESb-MP cells. Thus, loca lly produced chemokines, MCP-1(JE) and RANTES in particular, may contr ibute to the preferential metastasis of ESb-MP cells to the kidneys. ( C) 1998 Wiley-Liss, Inc.