Axolotl (Ambystoma mexicanum) lymphocytes produce and are growth-inhibited by transforming growth factor-beta

Recombinant (r)TGF-beta 5, an isoform of TGF-beta thus far identified only in the frog, Xenopus' laevis, inhibited phytohemagglutinin (PHA)-induced mitogenesis of salamander (axolotl) splenocytes and thymocytes, and T cell growth factor-induced proliferation of splenic lymphoblasts. This inhibition could be reversed by incubating (r)TGF-beta 5 with an anti-TGF-beta 5 antibody, but not with an antibody directed against TGF-beta 2, another Xenopus-produced TGF-beta isoform. Acid-treated supernatants from cultures of PHA-stimulated axolotl splenocytes (PHA-SNs) inhibited proliferation of mink lung fibroblasts (MLF), cells whose growth is sensitive to all known isoforms of TGF-beta. This inhibition was reversed by incubating the supernatants with the anti-pan-specific TGF-beta antibody which neutralizes TGF-beta 1, TGF-beta 2, TGF-beta 3 and TGF-beta 5. This inhibition was unaffected by antibodies that specifically neutralize either TGF-beta 1, TGF-beta 2, TGF-beta 3 or TGF-beta 5. Acid-treated axolotl PHA-SNs also inhibited proliferation of PHA-stimulated freshly harvested axolotl spleen cells. Once again, this inhibition could be reversed by treating supernatants with an anti-pan-specific TGF-beta antibody but not with anti-TGF-beta 2 or anti-TGF-beta 5 antibodies. All these data are consistent with the hypothesis that axolotl lymphocytes secrete a previously unknown TGF-beta isoform.