Purification and identification of chemokines potentially involved in kidney-specific metastasis by a murine lymphoma variant: induction of migration and NFκB activation
✍ Scribed by Ji Ming Wang; Oleg Chertov; Paul Proost; Jian-Jian Li; Patricia Menton; Luoling Xu; Silvano Sozzani; Alberto Mantovani; Wanghua Gong; Volker Schirrmacher; Jo Van Damme; Joost J. Oppenheim
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- French
- Weight
- 158 KB
- Volume
- 75
- Category
- Article
- ISSN
- 0020-7136
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✦ Synopsis
The ESb-MP cell line is the subclone of a highly malignant variant of murine methylcholanthrene-induced T lymphoma, ESb. When injected in vivo, ESb-MP cells metastasize to the kidney with high frequency, whereas a non-adherent variant, ESb cells, rarely form metastatic foci in the kidney. Our previous results showed that ESb-MP, but not ESb, cells were able to migrate in response to murine kidney-conditioned media (KCM). In an effort to characterize the tumor cell chemoattractant(s) produced 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 purified to homogeneity by sequential fast-performance liquid chromatography and reversed phase high-performance liquid chromatography. Amino acid sequencing of the formic acid-digested active fractions revealed that the purified protein was identical to murine MCP-1(JE) and its activity was neutralized by an anti-MCP-1(JE) antibody. Another chemokine, RANTES, was also purified from MES-13 cell supernatant. The chemotactic 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-RANTES antibodies. We further examined the differences in the ESb-MP and ESb cells. Binding studies using a variety of radio-iodinated chemokines showed that although both ESb-MP and ESb cells expressed substantial levels of high-affinity binding sites for CC chemokines, only ESb-MP cells migrated in response to CC chemokines and these cells constitutively expressed higher levels of 2 integrin adhesion protein CD11b than their parental ESb cells. CC chemokines also activated NFB in ESb-MP but not in ESb cells. Our results indicate that CC chemokines selectively chemoattract and activate ESb-MP cells. Thus, locally produced chemokines, MCP-1(JE) and RANTES in particular, may contribute to the preferential metastasis of ESb-MP cells to the kidneys.