Accumulation of MFG-E8/lactadherin on exosomes from immature dendritic cells

Véron, Philippe; Ségura, Elodie; Sugano, Gaël; Amigorena, Sébastian; Théry, Clotilde

doi:10.1016/j.bcmd.2005.05.001

Cited by 116 publications

(91 citation statements)

References 28 publications

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“…Carcinomas were classified according to the 2004 WHO classification of carcinomas of the urinary bladder (pTNM) (Eble et al, 2004): carcinoma cells confined to the epithelial layer were qualified as in situ carcinoma ¼ pTis or pTa, infiltrating the lamina propria as pT1 and infiltrating the muscularis propria as XpT2. Expression of gene-trapped Mfge8 was evaluated on frozen bladder sections by b-galactosidase reaction as previously described (Silvestre et al, 2005;Veron et al, 2005).…”

Section: Mice and Carcinogenesismentioning

confidence: 99%

“…MFGE8/lactadherin binds to phosphatidylserine at the surface of membrane vesicles (Oshima et al, 2002;Veron et al, 2005) and apoptotic cells (Hanayama et al, 2002) through its FactorVIII-like domain (Shi et al, 2004), and exposes its EGF-like domain for recognition by avb3/b5 integrins (Andersen et al, 2000). MFGE8 displays three types of functions.…”

Section: Introductionmentioning

confidence: 99%

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Milk fat globule—epidermal growth factor—factor VIII (MFGE8)/lactadherin promotes bladder tumor development

Sugano

Bernard‐Pierrot

et al. 2010

Oncogene

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Milk fat globule-epidermal growth factor-factor VIII (MFGE8), also called lactadherin or SED1, is a secreted integrin-binding protein that promotes elimination of apoptotic cells by phagocytes leading to tolerogenic immune responses, and vascular endothelial growth factor (VEGF)-induced angiogenesis: two important processes for cancer development. Here, by transcriptomic analysis of 228 biopsies of bladder carcinomas, we observed overexpression of MFGE8 during tumor development, correlated with expression of genes involved in cell adhesion or migration and in immune responses, but not in VEGF-mediated angiogenesis. To test whether MFGE8 expression was instrumental in bladder tumor development, or a simple consequence of this development, we used genetic ablation in a mouse model of carcinogen-induced bladder carcinoma. We showed that Mfge8 was also upregulated in mouse carcinoma, and that in its absence, Mfge8-deficient animals developed less advanced tumors. Angiogenesis was similar in carcinogen-treated Mfge8-expressing or -deficient bladders, thus ruling out a major role of the proangiogenic function of Mfge8 for its protumoral role. By contrast, the tumorpromoting role of Mfge8 was not observed anymore in mice devoid of adaptive immune system, and human tumors overexpressing MFGE8 where invaded with macrophages and regulatory T cells, thus suggesting that MFGE8/ lactadherin favors development of bladder tumors at least partly by an immune system-dependent mechanism. Our observations suggest future use of MFGE8-inhibiting molecules as therapies of bladder carcinomas, and of a limited number of other human cancers, in which our analysis of public databases also revealed overexpression of MFGE8.

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Section: Mice and Carcinogenesismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Milk fat globule—epidermal growth factor—factor VIII (MFGE8)/lactadherin promotes bladder tumor development

Sugano

Bernard‐Pierrot

et al. 2010

Oncogene

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“…In this way, the EGF like domain serves as a scaffold for the RGD sequence, which is proposed to promote cell adhesion by binding cell surface integrin receptors, such as αvβ3 or αvβ5 (39)(40)(41). The coagulation factor V/VIII like domains bind to phosphatidylserine (PS) exposed on the surface of apoptotic cells (42). Binding of MFG-E8 to PS on apoptotic cells opsonizes them for a complete engulfment by macrophages via αvβ3-or αvβ5-integrins.…”

Section: Discussionmentioning

confidence: 99%

Expression and characterization of recombinant human milk fat globule-EGF factor VIII

Qiang

et al. 2011

Int J Mol Med

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Abstract. Apoptosis plays an important role in the pathobiology of sepsis. The opsonizing protein milk fat globule-EGF factor VIII (MFG-E8) is involved in apoptotic cell clearance. Our previous studies have shown that administration of rat MFG-E8-containing exosomes or recombinant murine MFG-E8 (rmMFG-E8) is protective in a rat model of sepsis induced by cecal ligation of puncture (CLP). However, one obstacle hampering the development of MFG-E8 as a therapeutic agent for septic patients is the potential immunogenicity of animal proteins in humans. The purpose of this study, therefore, was to express recombinant human MFG-E8 (rhMFG-E8) and characterize its biological activity. Using an E. coli system, we successfully expressed and purified the mature molecule of human MFG-E8 (Leu24-Cys387). The purity of rhMFG-E8 was over 99% and it was immunoreactive for specific antihuman MFG-E8 antibodies. Amino acid sequence analysis by LC-MS/MS identified the purified protein as human MFG-E8. Using primary rat peritoneal macrophages, we showed that rhMFG-E8 markedly increased peritoneal macrophage phagocytosis of apoptotic thymocytes, which was as effective as commercial rmMFG-E8. To determine the biological activity of rhMFG-E8 in vivo, male adult rats were subjected to sepsis by CLP. rhMFG-E8 or rmMFG-E8 were administered intravenously at the time of CLP. Our results demonstrated that both rhMFG-E8 and rmMFG-E8 reduced thymocyte apoptosis and plasma levels of lactate and IL-6 at 20 h after CLP, and improved the 10-day survival rate. Thus, we have successfully expressed and purified biologically active rhMFG-E8. Our newly-expressed rhMFG-E8 is highly effective in the rat model of sepsis.

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“…For the secreted MFGE8 protein quantification, the 8 ml of media previously collected was initially subjected to ultracentrifugation at 100,000 x g (50TI rotor, Beckman Coulter, Brea, CA) for 2 hours followed by PBS wash and repeat ultracentrifugation for another 2 hours following a similar protocol to other previously published studies [42][43]. The entire pellet was sonicated and then loaded onto the gel followed by the protocol mentioned above.…”

Section: Protein Extraction Sds-page and Immunoblottingmentioning

confidence: 99%

“…We looked for EREs of 17 bases length [42] (2 flanking nt -1 st half site-3nt spacer-2 nd half site-2 flanking nt) on 11 ortholog promoters of MFGE8 gene. Table 1A shows the summary of ERE predictions in these promoters at the tool sensitivity of 87%.…”

Section: Prediction Of Eres and Pres In The Mfge8 Genementioning

confidence: 99%

Human endometrial milk fat globule-epidermal growth factor 8 (MFGE8) is up regulated by estradiol at the transcriptional level, and its secretion via microvesicles is stimulated by human chorionic gonadotropin (hCG)

Sarhan¹,

Bocca²,

Yu³

et al. 2013

Cell Signal Traffic

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Objective: We have recently showed that MFGE8, a novel epithelial cell protein in the human endometrium, upregulated during the window of implantation. We hypothesized that MFGE8 may act as a key modulator of endometrial remodeling and trophoblast invasion. The aims of this study were (i) to investigate the in vitro regulation of human endometrial epithelial cells MFGE8 transcription, translation, and secretion by sex steroids and hCG; and (ii) to examine the possibility of MFGE8 secretion via microvesicles. Design: Experimental in vitro study using Ishikawa cells. Setting: University center. Interventions: Treatment with estradiol (E2), progesterone (P4), and human chorionic gonatropin (hCG). Main outcome measures: MFGE8 mRNA and protein expression, and identification of secreted microvesicles by mass spectrometry (MS) and immunoblotting. Results: E 2 , but not P4 or hCG, significantly upregulated MFGE8 mRNA expression. hCG significantly increased MFGE8 secretion. Microvesicels obtained after ultracentrifugation were visualized with atomic force microscopy ranging from ~100 to 200 nm. In addition to the expected 46 kD protein, the microvesicles contained a second form of secreted MFGE8 measuring ~30 kD which was confirmed by MS. Conclusions: We demonstrated (i) dual effects of E 2 and hCG on the regulation of MFGE8, and (ii) MFGE8 protein secretion in association with microvesicles. MFGE8 has the potential to modulate endometrial function and implantation via exocrine and/ or paracrine-autocrine effects. To the best of our knowledge, this is the first demonstration of microvesicular secretion of any regulatory protein by endometrial epithelial cells, providing initial evidence suggestive of microvesicular participation in cellular trafficking information in the non-pregnant and pregnant endometrium.

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Accumulation of MFG-E8/lactadherin on exosomes from immature dendritic cells

Cited by 116 publications

References 28 publications

Milk fat globule—epidermal growth factor—factor VIII (MFGE8)/lactadherin promotes bladder tumor development

Milk fat globule—epidermal growth factor—factor VIII (MFGE8)/lactadherin promotes bladder tumor development

Expression and characterization of recombinant human milk fat globule-EGF factor VIII

Human endometrial milk fat globule-epidermal growth factor 8 (MFGE8) is up regulated by estradiol at the transcriptional level, and its secretion via microvesicles is stimulated by human chorionic gonadotropin (hCG)

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