Cytokine-like effects of prolactin in human mononuclear and polymorphonuclear leukocytes.

Some biochemical events following the binding of prolactin (PRL) to its receptor in normal human leukocytes were investigated. PRL enhanced JAK2 phosphorylation in peripheral blood mononuclear cells (PBMC) but not in granulocytes. PRL also induced phosphorylation of Stat-5 in PBMC and Stat-1 in granulocytes. Subsequent binding of Stat-5- and of Stat-1-like molecules to a GAS responsive element from the beta-casein promoter was detected by EMSA. p38 MAPK (but not p42/p44 MAPK) was activated by PRL in both leukocyte populations. PRL induced iNOS and CIS mRNA expression in granulocytes. Increased expression of IRF-1 and SOCS-2 was observed in granulocytes and of SOCS-3 and iNOS in PBMC. Similar effects were obtained with ovine and human PRL. Antiserum to PRL reduced iNOS and IRF-1 expression induced by PRL in granulocytes and reduced iNOS expression in PBMC. Also, pretreatment of granulocytes with a p38 MAPK inhibitor (SB 203580) prevented in part PRL-induced iNOS and IRF-1 expression. In PBMC, the p38 inhibitor decreased PRL-induced iNOS gene expression. These results indicate that PRL-induced gene regulation in leukocytes requires the activation of at least two different pathways: the Stat and the MAP kinase pathways. Moreover, although PRL activates Stat in both leukocyte types, signal transduction is different in granulocytes and in PBMC. Most importantly, PRL modulates the expression of genes crucial to leukocyte function. The present findings reinforce the concept that PRL has "cytokine-like" activity in human leukocytes.

Effects of prolactin on signal transduction and gene expression: possible relevance for systemic lupus erythematosus.

Receptors for prolactin (PRL-R) are expressed in normal leukocytes from rat and man. PRL signals through PRL-R associated Janus tyrosine kinase (Jak)-2 and signal transducers and activators of transcription (Stat). In addition, in human leukocytes PRL also activates the p38 MAP kinase pathway. PRL, at physiological concentrations, stimulates the expression of the interferon regulatory factor (IRF)-1 gene in rat spleen and bone marrow cells. In man, genes induced by PRL include several members of the 'suppressors of cytokine signaling' (SOCS) family and inducible nitric oxide synthase (iNOS; in mononuclear cells and in granulocytes) and IRF-1 (in granulocytes). Thus, in normal leukocytes, PRL induces the expression of several genes relevant to innate and acquired immune responses. Sex hormones, such as estrogen and PRL, have been implicated in the pathogenesis of murine and human SLE. Also defective signaling in leukocytes is a feature of the disease. What the origin is of aberrant signaling processes in SLE lymphocytes and how they relate to tolerance breakdown and immunopathology is still unknown. It is not unlikely that PRL is a player at some level. The exact contribution of PRL to immune responses in normal subjects and in SLE patients is not known. Further work should also indicate whether PRL might contribute to the onset or progression of the disease and assess the possible benefits of manipulating PRL concentrations in patients.

Prolactin activates interferon regulatory factor-1 expression in normal lympho-hemopoietic cells.

It has been proposed that prolactin (PRL) is a lympho-hemopoietic growth and differentiation factor. We show here by Western blotting that PRL-receptors (PRL-R) are expressed in normal rat bone marrow and spleen cells. We also show that PRL stimulates the phosphorylation of the PRL-R-associated Janus tyrosine kinase (JAK)-2 in rat bone marrow and spleen cells. This leads to the activation and subsequent binding of signal transducer and activator of transcription (Stat) 5b to an interferon regulatory factor-1 (IRF-1) gamma activation sequence (GAS) as visualized by electromobility shift assay. As shown after reverse transcription of mRNA by polymerase chain reaction, PRL, at physiological concentrations (0.01 microg/ml), stimulates the expression of the IRF-1 gene in these normal cells. PRL could thus affect several aspects of the immune response.

Expression of SOCS genes in normal and leukemic human leukocytes stimulated by prolactin, growth hormone and cytokines.

To evaluate the possible role of the recently described family of suppressors of cytokine signaling (SOCS) factors in the human lympho-hemopoietic system, we have monitored SOCS factor expression, both constitutive and induced by either cytokines, prolactin (PRL) or growth hormone (GH), using polymerase chain reaction in normal and leukemic cells. CIS (cytokine-inducible SH2-containing protein), SOCS-2 and SOCS-3 were constitutively expressed in peripheral blood mononuclear leukocytes. SOCS-3 expression was enhanced by PRL or by IFN-gamma. In bone marrow cells and granulocytes, CIS expression was induced and SOCS-2 enhanced by IFN-gamma and by PRL. In tonsillar cells, CIS expression was increased and SOCS-2 was induced by IL-1beta, IL-6, PRL and GH. SOCS-3 expression was enhanced by IL-1beta. The expression of SOCS-7 was increased by IL-6, PRL and GH. In Raji B-lymphoma cells, the expression of SOCS-2 and SOCS-7 was enhanced by IL-1beta. In THP-1 myeloid leukemia cells pretreated with TPA (to induce receptors for IFN-gamma), IFN-gamma induced SOCS-2. Jurkat cells expressed more SOCS-2 when exposed to PRL. Original observations in this work include the first report on SOCS-7 induction by cytokines. Also our data shed new light on the possible involvement of PRL and GH in the cytokine network. These hormones could modulate the transduction of signals originating from receptors for various cytokines.

Prolactin, growth hormone and the immune system in humans.

Prolactin (PRL) and growth hormone (GH) qualify as lymphoid growth and differentiation factors. Yet, long-standing hyper- or hyposecretion of PRL or GH does not induce any significant alteration of the immune system. Subclinical changes in laboratory parameters (such as chemotaxis or phagocytosis by granulocytes or macrophages or natural killer cell activity) have been reported in some of these conditions. The GH-insulin-like growth factor (IGF)-I axis is dysregulated in ageing, in catabolic states and in critical illness. Immune parameters, such as infection rate, are being monitored during clinical trials with GH or IGF-I. Hyperprolactinaemia may play an aggravating role in systemic lupus erythematosus, in autoimmune thyroiditis and in other autoimmune diseases. The patient may benefit from increased alertness about interactions between the endocrine and immune systems.

Plastic phenotype of human oligodendroglial tumour cells in vitro.

Human oligodendroglioma cells cultured in serum-supplemented media lose their oligodendrocytic antigenic markers and acquire astrocytic markers. However, after reimplantation in rodent brain, these cells re-express oligodendrocytic markers. This switch in human oligodendroglioma cell phenotype could result from the interplay of different stimuli in vitro vs in vivo The in vitro differentiation into astrocytes might result from non-physiological culture conditions. It is shown that human oligodendroglioma cells behave in a way similar to that of rodent bipotential 0-2 A progenitor cells which can be driven to differentiate into either oligodendrocytes or type 2-astrocytes depending on the culture medium. Indeed, in serum-supplemented medium, human oligodendroglioma cells proliferated and expressed the GFAP astrocytic marker. In chemically defined medium containing insulin, human oligodendroglioma cells were quiescent and expressed the OI oligodendrocyte-specific marker. In both media, human oligodendroglioma cells expressed the A2B5 membrane marker as well as the SCIP transcription factor specific of 0-2 A cells, further confirming their oligodendrocytic origin. Replacement of insulin by platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF), known to maintain 0-2 A progenitors in a proliferative state, stimulated DNA replication of human oligodendroglioma cells cultured in chemically defined medium.