Individual and combined effect of granulocyte–macrophage colony‐stimulating factor and prolactin on maturation of dendritic cells from blood monocytes under serum‐free conditions

• Published in: Immunology Vol. 100; no. 1; pp. 29 - 36
• Main Authors: Matera, L., Galetto, A., Geuna, M., Vekemans, K., Ricotti, E., Contarini, M., Moro, F., Basso, G.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-05-2000; Wiley Subscription Services, Inc; Blackwell Science Inc
• Subjects: Antigen Presentation; Antigens, Surface - metabolism; Cell Culture Techniques; Cell Differentiation - drug effects; Culture Media, Serum-Free; Cytokines - immunology; Dendritic Cells - cytology; Dendritic Cells - drug effects; Dendritic Cells - immunology; Dose-Response Relationship, Immunologic; Down-Regulation - drug effects; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Humans; Isoantigens - metabolism; Lymphocyte Culture Test, Mixed; Monocytes - cytology; Monocytes - drug effects; Original; prolactin; Prolactin - pharmacology; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor - metabolism
• ISSN: 0019-2805; 1365-2567
• DOI: 10.1046/j.1365-2567.2000.00996.x

Summary
Prolactin (PRL) shares structural and functional features with haemopoietic factors and cytokine peptides. Dendritic cells (DC) are involved in both initiating the primary and boosting the secondary host immune response and can be differentiated in vitro from precursors under the effect of granulocyte–macrophage colony‐stimulating factor (GM‐CSF) plus other factors. Because PRL has been shown to functionally interact with GM‐CSF, we have addressed its role on GM‐CSF‐driven differentiation of DC. Monocytic DC precursors from peripheral blood mononuclear cells (PBMC) were enriched either by adhesion to a plastic surface or CD14‐positive selection and cultured for 7 days in serum‐free medium containing GM‐CSF, interleukin (IL)‐4 and PRL, alone or in combination. Cells with large, veiled cytoplasm, expressing major histocompatibility complex (MHC) class II and the costimulatory molecules CD80, CD86 and CD40 and lacking the monocyte marker CD14, were considered as having the phenotype of cytokine‐generated DC. Functional maturation was assessed by proliferation and interferon‐γ (IFN‐γ) release of allogeneic T lymphocytes. Physiological (10–20 ng/ml) concentrations of PRL interacted synergistically with GM‐CSF and the effect was similar to that induced by IL‐4 on GM‐CSF‐driven DC maturation. When used alone, the physiological concentrations of PRL were inhibitory, whereas higher concentrations (80 ng/ml) were stimulatory. The synergistic effect of PRL may in part be caused by its ability to counteract the down‐modulation of the GM‐CSF receptor observed in serum‐free conditions. These data provide further evidence of the significance of PRL in the process of T lymphocyte activation.