A macrophage colony-stimulating factor receptor–green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse

Sasmono, R. Tedjo; Oceandy, Delvac; Pollard, Jeffrey W.; Tong, Wei; Pavli, Paul; Wainwright, Brandon; Ostrowski, Michael C.; Himes, S. Roy; Hume, David

doi:10.1182/blood-2002-02-0569

Cited by 608 publications

(744 citation statements)

References 51 publications

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“…For inducible expression these mice have to be bred with another transgenic mouse, in which rtTA inducible promoter drives expression of "geneofinterest". The c-fms gene promoter is active in macrophages and bone marrow progenitors (Sasmono et al, 2003). Our system is distinct, because the SRA promoter expresses a transgene only in mature macrophages (Horvai et al, 1995) and in bone marrow derived dendritic cells (Pan, unpublished observations).…”

Section: Discussionmentioning

confidence: 99%

Dual-promoter lentiviral system allows inducible expression of noxious proteins in macrophages

Hui

Mostoslavsky

Eruslanov

et al. 2008

Journal of Immunological Methods

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In-depth studies of innate immunity require efficient genetic manipulation of macrophages, which is especially difficult in primary macrophages. We have developed a lentiviral system for inducible gene expression both in macrophage cell lines and in primary macrophages. A transgenic mouse strain C3H.TgN(SRA-rtTA) that expresses reverse tetracycline transactivator (rtTA) under the control of macrophage-specific promoter, a modified human scavenger receptor A (SR-A) promoter was generated. For gene delivery, we constructed a dual-promoter lentiviral vector, in which expression of a "gene-of-interest" is driven by a doxycycline-inducible promoter and the expression of a selectable surface marker is driven by an independent constitutive promoter UBC. This vector is used for transduction of bone marrow-derived macrophage precursors. The transduced cells can be enriched to 95-99% purity using marker-specific monoclonal antibodies, expanded and differentiated into mature macrophages or myeloid dendritic cells. We also successfully used this approach for inducible protein expression in hard to transfect macrophage cell lines.Because many proteins, which are expressed by activated or infected macrophages, possess cytotoxic, anti-proliferative or pro-apoptotic activities, generation of stable macrophage cell lines that constitutively express those proteins is impossible. Our method will be especially useful to study immunity-related macrophage proteins in their physiological context during macrophage activation or infection.

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Section: Discussionmentioning

confidence: 99%

Dual-promoter lentiviral system allows inducible expression of noxious proteins in macrophages

Hui

Mostoslavsky

Eruslanov

et al. 2008

Journal of Immunological Methods

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“…Endometriosis was induced using wild-type and transgenic colony stimulating factor 1 receptoreenhanced green fluorescent protein (Csf1reEGFP) mice (MacGreen; founder stocks were a gift from Bernadette Dutia and Prof. David Hume, The Roslin Institute, The University of Edinburgh, UK) 19 on a C57BL/6 background using a previously validated model of endometriosis, as described. 9 In brief, donor endometrial tissue was recovered during induced menses 20 and injected into the peritoneal cavity of recipient mice.…”

Section: Mouse Model Of Endometriosismentioning

confidence: 99%

Estradiol Is a Critical Mediator of Macrophage-Nerve Cross Talk in Peritoneal Endometriosis

Greaves

Temp

Esnal-Zufiurre

et al. 2015

The American Journal of Pathology

141

121

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Endometriosis occurs in approximately 10% of women and is associated with persistent pelvic pain. It is defined by the presence of endometrial tissue (lesions) outside the uterus, most commonly on the peritoneum. Peripheral neuroinflammation, a process characterized by the infiltration of nerve fibers and macrophages into lesions, plays a pivotal role in endometriosis-associated pain. Our objective was to determine the role of estradiol (E2) in regulating the interaction between macrophages and nerves in peritoneal endometriosis. By using human tissues and a mouse model of endometriosis, we demonstrate that macrophages in lesions recovered from women and mice are immunopositive for estrogen receptor b, with up to 20% being estrogen receptor a positive. In mice, treatment with E2 increased the number of macrophages in lesions as well as concentrations of mRNAs encoded by Csf1, Nt3, and the tyrosine kinase neurotrophin receptor, TrkB. By using in vitro models, we determined that the treatment of rat dorsal root ganglia neurons with E2 increased mRNA concentrations of the chemokine C-C motif ligand 2 that stimulated migration of colony-stimulating factor 1edifferentiated macrophages. Conversely, incubation of colony-stimulating factor 1 macrophages with E2 increased concentrations of brainderived neurotrophic factor and neurotrophin 3, which stimulated neurite outgrowth from ganglia explants. In summary, we demonstrate a key role for E2 in stimulating macrophage-nerve interactions, providing novel evidence that endometriosis is an estrogen-dependent neuroinflammatory disorder. Endometriosis affects 10% of reproductive age women and is associated with persistent pelvic pain. 1 It is defined by the presence of endometrial-like tissue (lesions) found outside the uterus, most commonly on the peritoneum. The mechanisms underlying endometriosis-associated pain are poorly understood, but it has been postulated that estrogen-dependent neuroinflammation may be involved. 2 Notably, the presence of endometrial tissue fragments on the peritoneum elicits an immune response, including recruitment of macrophages, 3 blood vessels, and nerve fibers into the resultant lesions. 4,5 Within the lesions, CD68 þ macrophages have been detected in close association with nerve fibers. 6 Studies investigating macrophage activation and recruitment have revealed that endometriosis-associated macrophages exhibit a phenotype consistent with the alternative end of the macrophage activation spectrum. 7,8 In a mouse model of endometriosis that included cell transfer of polarized macrophages, Bacci et al 7 reported that mice injected with proinflammatory macrophages [macrophage (interferon g)] developed microscopic lesions, but those injected with alternatively activated macrophages [macrophage (IL-4)] developed larger lesions with a well-developed vasculature. Our studies in a mouse model of endometriosis have revealed that macrophages resident in peritoneal lesions can originate from both the peritoneum and the endometrium. 9

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“…EGER-Fms cells were derived from BMC isolated from EGFP23 reporter mice in which EGFP expression is controlled by the proximal promoter of c-fms gene combined with its first intron; this transgene has been demonstrated to be a definitive marker for the mononuclear phagocyte system (Sasmono et al, 2003). Accordingly, in bone marrow from EGFP23 mice, most EGFP-expressing cells corresponded to differentiated myeloid cells, based on high forward scatter (FSC) (size) parameter (Figure 3a, upper panel), and they expressed the myeloid marker MAC1 at their surface ( Figure 3a, middle panel), indicating that the transgene expression is restricted to myeloid cells.…”

Section: Eger-fms Cells Differentiated In Macrophages In Response To mentioning

confidence: 99%

“…The transgene delta6.7fms-EGFP (also called 7.2fms-EGFP), a gift of Dr DA Hume (University of Queensland, Brisbane, Australia), has been described previously (Sasmono et al, 2003). Briefly, the proximal promoter of c-fms combined with the first intron directed consistent expression of the EGFP reporter gene in the same locations as the endogenous gene.…”

Section: Micementioning

confidence: 99%

E2a/Pbx1 oncogene inhibits terminal differentiation but not myeloid potential of pro-T cells

Bourette

Mouchiroud

2006

Oncogene

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A macrophage colony-stimulating factor receptor–green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse

Cited by 608 publications

References 51 publications

Dual-promoter lentiviral system allows inducible expression of noxious proteins in macrophages

Dual-promoter lentiviral system allows inducible expression of noxious proteins in macrophages

Estradiol Is a Critical Mediator of Macrophage-Nerve Cross Talk in Peritoneal Endometriosis

E2a/Pbx1 oncogene inhibits terminal differentiation but not myeloid potential of pro-T cells

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