Effects of interleukins on Crouzon fibroblast phenotype in vitro. Release of cytokines and IL-6 mRNA expression

In Crouzon's syndrome, the cranial morphogenic processes are altered due to the early fusion of the facial and basal cranial bones. Evidence that Crouzon fibroblasts have an altered phenotype which is modulated by interleukin 1 (IL-1) and interleukin 6 (IL-6) is provided. 3H]Thymidine incorporation and cell count studies showed that Crouzon fibroblasts have an accelerated proliferation rate. 3H]Glucosamine incorporation studies, followed by chromatography analysis of culture medium samples, revealed that the various classes of glycosaminoglycans (GAG) secreted into the medium were differently distributed in Crouzon and normal fibroblasts. Crouzon fibroblasts secreted less total GAG, particularly hyaluronic acid (HA) and heparan sulfate (HS), but a relatively greater quantity of chondroitin sulfate. Type I and III collagen were raised in Crouzon fibroblast medium whereas the concentration of fibronectin was lower than in normal cells. Interleukin treatment induced changes in cell growth and neosynthesis of extracellular matrix macromolecules. Both IL-1 and IL-6 stimulated proliferation of Crouzon fibroblasts, whereas only IL-6 increased 3H]thymidine incorporation in normal cells. IL-1 provided a drop in HA and a rise in GAG sulfates in normal fibroblasts, but caused an opposite effect in Crouzon fibroblasts. Type I collagen and fibronectin secretions are differently modulated by the cytokines in the two populations. Moreover, level of IL-1 able to stimulate 3H] thymidine incorporation into mouse thymocytes, and level of IL-1 receptor antagonist (IL-1ra), as determined by ELISA, were higher in pathological than in normal fibroblasts. Also evidence that levels of IL-1 alpha and IL-6 proteins measured by ELISA and also IL-6 mRNA expression are enhanced in Crouzon fibroblasts is provided. These novel data suggested that, in Crouzon's syndrome, the modifications in the relative concentrations of the extracellular matrix (ECM) components associated with the abnormal cytokine networks may alter the balance of the microenvironment where the morphogenic events take place.