AN EPSTEIN-BARR VIRUS-NEGATIVE BURKITT-LYMPHOMA CELL-LINE (SFRAMOS) SECRETES A PROLACTIN-LIKE PROTEIN DURING CONTINUOUS GROWTH IN SERUM-FREE MEDIUM

Pituitary human PRL (hPRL) antiserum inhibits growth of B-lymphoblastoid cells in vitro, but the mechanism of inhibition is unclear. In this study the mechanism of inhibition of human B-cell growth by anti-hPRL was explored with an Epstein-Barr virus nuclear antigen (EBNA)-negative Burkitt lymphoma cell line (sfRamos) that proliferates continuously in serum-free medium with human transferrin as the only protein supplement. The data show that antiserum immunoglobulin fraction G (IgG) to pituitary hPRL, but not nonimmune serum IgG, completely inhibited the growth of sfRamos cells. In addition, anti-hPRL IgG identified a single band (29 kDa) in sfRamos spent medium, but not in fresh serum-free medium or in human transferrin, as demonstrated by sodium dodecyl sulfate-reducing polyacrylamide gel electrophoresis and Western immunoblot analysis. Polyacrylamide gel electrophoresis and Western analysis of a mixture containing sfRamos spent medium and excess pituitary hPRL established that the sfRamos 29-kDa PRL-like protein (PLP29) and pituitary hPRL (23 kDa) were electrophoretically distinct. Finally, sfRamos spent medium, but not fresh serum-free medium, was mitogenic for sfRamos and Nb2, a PRL-sensitive node rat lymphoma cell line. These findings demonstrate that PLP29 is biologically and immunologically like pituitary hPRL, but is electrophoretically different from this hormone. We suggest that PLP29 is secreted as an autocrine growth factor by sfRamos Burkitt lymphoma cells during continuous serum-free growth.