Immunology: Investigations on the cell type responsible for the endometrial secretion of complement component 3 (C3)

It has been shown that and human endometria have the capacity to produce complement component 3 (C3). In rats, endometrial C3 is an oestrogen-dependent protein produced and secreted by glandular cells. The cell responsible for the synthesis and secretion of human endometrial C3 has not been clearly defined. Our study was aimed at answering this question. Samples of endometrium obtained from hysterectomies were either immunostained for C3 or digested with collagenase; then the stromal and glandular cells were separated and immunopurified (or not) with an antibody to CD45 coupled to magnetic beads to eliminate the endometrial lymphomyeloid cells. Cells were cultured for 2 weeks and C3 measured in the medium by an in-house radioimmunoassay. Glandular as well as stromal cells stained positively for C3 and released C3 in vitro. The release of C3 from both cell types could be inhibited by cycloheximide. Epithelial cells produced significantly more C3 than stromal cells, and endometrial C3 production was higher for both cell types when these were obtained from secretory as compared to proliferative endometria. Lymphomyeloid cells were possibly a source of C3 since after immunoadsorption of these cells, the remaining stromal or glandular cells produced significantly less C3. We conclude that endometrial stromal, glandular and lymphomyeloid cells all produce C

Inhibition of EGFR/MAPK signaling reduces microglial inflammatory response and the associated secondary damage in rats after spinal cord injury

<p>Abstract</p> <p>Background</p> <p>Emerging evidence indicates that reactive microglia-initiated inflammatory responses are responsible for secondary damage after primary traumatic spinal cord injury (SCI); epidermal growth factor receptor (EGFR) signaling may be involved in cell activation. In this report, we investigate the influence of EGFR signaling inhibition on microglia activation, proinflammatory cytokine production, and the neuronal microenvironment after SCI.</p> <p>Methods</p> <p>Lipopolysaccharide-treated primary microglia/BV2 line cells and SCI rats were used as model systems. Both C225 and AG1478 were used to inhibit EGFR signaling activation. Cell activation and EGFR phosphorylation were observed after fluorescent staining and western blot. Production of interleukin-1beta (IL-1β) and tumor necrosis factor alpha (TNFα) was tested by reverse transcription PCR and ELISA. Western blot was performed to semi-quantify the expression of EGFR/phospho-EGFR, and phosphorylation of Erk, JNK and p38 mitogen-activated protein kinases (MAPK). Wet-dry weight was compared to show tissue edema. Finally, axonal tracing and functional scoring were performed to show recovery of rats.</p> <p>Results</p> <p>EGFR phosphorylation was found to parallel microglia activation, while EGFR blockade inhibited activation-associated cell morphological changes and production of IL-1β and TNFα. EGFR blockade significantly downregulated the elevated MAPK activation after cell activation; selective MAPK inhibitors depressed production of cytokines to a certain degree, suggesting that MAPK mediates the depression of microglia activation brought about by EGFR inhibitors. Subsequently, seven-day continual infusion of C225 or AG1478 in rats: reduced the expression of phospho-EGFR, phosphorylation of Erk and p38 MAPK, and production of IL-1β and TNFα; lessened neuroinflammation-associated secondary damage, like microglia/astrocyte activation, tissue edema and glial scar/cavity formation; and enhanced axonal outgrowth and functional recovery.</p> <p>Conclusions</p> <p>These findings indicate that inhibition of EGFR/MAPK suppresses microglia activation and associated cytokine production; reduces neuroinflammation-associated secondary damage, thus provides neuroprotection to SCI rats, suggesting that EGFR may be a therapeutic target, and C225 and AG1478 have potential for use in SCI treatment.</p