Anti-IL11 monoclonal antibody, clone 33727

Fibroblast growth factor 2 (FGF2) and cyclic AMP (cAMP) play critical roles in controlling the differentiation of osteoblasts and mineralization of bone. We have previously reported that each of FGF2 and forskolin (FSK) alone increase transcription of the bone sialoprotein (BSP) gene, and that together (FGF/FSK) they upregulate BSP gene expression synergistically in rat osteoblast-like ROS 17/2.8 cells. However, other genes that are upregulated after stimulation by FGF2, FSK or FGF/FSK remain unclear. In the present study, we investigated candidate genes associated with mineralization after stimulation by FGF2, FSK and FGF/FSK in two kinds of osteoblast-like cells using microarray and real-time PCR. In ROS17/2.8 cells, FGF2 and FSK each increased the gene expression of c-FOS (7.2-fold and 10.7-fold, respectively). However, FGF/FSK did not induce c-FOS gene expression. FGF2 increased the expression of the dentin matrix protein 1 (DMP1, 129.8-fold) gene. In contrast, FGF/FSK increased the expression of the amphiregulin (AREG, 73-fold) gene maximally. In human osteoblast-like Saos2 cells, FGF2 increased the expression of the osteopontin (SPP1, 16.7-fold), interleukin-8 (IL8, 6.4-fold) and IL11 (4.8-fold) genes. FSK induced the expression of the IL6 (2.6-fold), IL11 (4.0-fold), chemokine ligand 13 (CXCL13, 2.8-fold) and bone morphogenetic protein 2 (BMP2, 2.5-fold) genes. These results suggest that FGF2 and FSK might be crucial regulators of mineralization and bone formation.

BACKGROUND: Human trophoblast invasion and differentiation are essential for a successful pregnancy outcome. Dysregulation of these processes can lead to placental pathologies such as pre-eclampsia. The molecular mechanisms; however, are poorly understood. Interleukin (IL)11-a cytokine that regulates endometrial epithelial cell adhesion, trophoblast motility and invasion during implantation-may be involved in some of these processes. METHODS AND RESULTS: The effect of IL11 on protein expression was investigated in trophoblastic HTR8/SVneo cells and primary extravillous trophoblasts (EVTs) purified from first-trimester placentas. Two-dimension (2D)-differential in-gel electrophoresis analyses revealed that 731 spots were significantly differentially regulated by IL11 in HTR8/SVneo cells: seven spots were analyzed by liquid chromatography-tandem mass spectrometry and 14 unique proteins identified. Protein disulfide isomerase family A, member 3 (PDIA3; endoplasmic reticulum p57) and glucose-regulated protein 78 (GRP78) were further validated to be regulated by IL11 in HTR8/SVneo and primary EVT. One dimension western blot analysis confirmed that PDIA3 was down-regulated in EVT. 2D western blot analysis revealed that GRP78 was post-translationally modified following IL11 treatment. Moreover, IL11 stimulated the secretion of GRP78 in EVT. CONCLUSIONS: Data suggest that IL11, possibly via signal transducers and activators of transcription 3 signaling pathway, regulates PDIA3 protein expression and modification/secretion of GRP78. This is the first study to identify PDIA3 and GRP78 as IL11 targets in invasive trophoblasts and identifies a possible mechanism by which IL11 regulates trophoblast function.