scholarly journals 229.Seminal plasma TGFβ activates pro-inflammatory cytokine synthesis in human cervical epithelial cells

2004 ◽  
Vol 16 (9) ◽  
pp. 229 ◽  
Author(s):  
D. J. Sharkey ◽  
S. A. Robertson
Keyword(s):  
Epithelial Cells ◽  
Seminal Plasma ◽  
Model Systems ◽  
Human Seminal Plasma ◽  
Neutralising Antibodies ◽  
Active Constituents ◽  
Gm Csf ◽  
Cytokine Synthesis ◽  
Cervical Epithelial Cells

Exposure to semen at intercourse in women elicits an inflammation-like response characterised by recruitment of inflammatory cells and expression of pro-inflammatory cytokines including GM-CSF, interleukin-6 (IL-6) and IL-8 (1). Studies in animal models have implicated TGFβ as the major active moiety in seminal plasma, and we have shown previously that TGFβ1 and TGFβ3 are present in high concentrations in human seminal plasma (>100 ng/mL), while TGFβ2 is less abundant. To investigate the physiological significance of each of the three TGFβ isoforms as pro-inflammatory agents in human seminal plasma, we have established in vitro model systems to measure human cervical cell cytokine synthesis. Primary cervical epithelial cells prepared from ectocervix of hysterectomy tissues or transformed Ect1 cells were incubated for 12 h with human recombinant TGFβ (isoforms 1, 2 or 3) or with seminal plasma in the presence or absence of isoform-specific TGFβ neutralising antibodies. Epithelial cell supernatants were recovered 24 h later and supernatants were analysed by commercial ELISA to quantify GM-CSF, IL-6 and IL-8 production. Each of the three TGFβ isoforms mimicked seminal plasma and were comparable in their capacity to stimulate >10-fold increases in both GM-CSF and IL-6 expression in a dose-responsive manner. In contrast, unlike seminal plasma none of the TGFβ isoforms induced IL-8 expression. Addition of neutralising antibodies to TFGβ1, TGFβ2 and TGFβ3 each effected >50% reduction in the ability of seminal plasma to induce GM-CSF and IL-6, but did not impair seminal plasma-stimulated IL-8 production. Together these data show that TGFβ1, TGFβ2 and TGFβ3 are major active constituents of seminal plasma, acting to elicit GM-CSF and IL-6 production in cervical epithelial cells. However, TGFβ does not fully account for the pro-inflammatory effects of human seminal plasma, and other active constituents remain to be identified. (1) D. J. Sharkey et al. (2003) Proc. SRB.

Human Immunodeficiency Virus Infection of Early Passage Cervical Epithelial Cultures

1993 ◽  
Vol 4 (6) ◽  
pp. 342-345 ◽  
Author(s):  
S L Patrick ◽  
T C Wright ◽  
H E Fox ◽  
H S Ginsberg
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Female Genital Tract ◽  
Virus Production ◽  
Heterosexual Transmission ◽  
Early Passage ◽  
Epithelial Cultures ◽  
Cervical Epithelial Cells

Women are infected with HIV in increasing numbers; the predominant mode of spread is through heterosexual transmission. Little is known regarding the mechanism of HIV transit through the female genital tract. We investigated whether early passaage cervical epithelial cells could be directly infected with HIV-1LAI*. Virus production was measured using the reverse transcriptase (RT) assay and direct assay for syncytia-forming units. In-situ hybridization was performed on infected cervical cell cultures. Immunostaining was carried out using a monoclonal antibody to leukocyte common antigen (LCA). Virus was recovered in the supernatants of all infected cervical cultures. Localization of HIV infection using in-situ hybridization identified rare cells in the population which gave a strong signal. These infected cells had a lymphoid morphology and were also detected using immunostaining for LAC. Cervical epithelial cells were uninfected in this in vitro model; cells in this population which supported viral replication were most likely of the macrophage/monocyte lineage.

scholarly journals Interleukin 1α and interleukin 6 promote the in vitro growth of both normal and neoplastic human cervical epithelial cells

1997 ◽  
Vol 75 (6) ◽  
pp. 855-859 ◽  
Author(s):  
G Castrilli ◽  
D Tatone ◽  
MG Diodoro ◽  
S Rosini ◽  
M Piantelli ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Interleukin 6 ◽  
In Vitro Growth ◽  
Interleukin 1Α ◽  
Cervical Epithelial Cells

421. Seminal fluid TGFβ regulates follistatin mRNA expression human Ect1 cervical epithelial cells

2008 ◽  
Vol 20 (9) ◽  
pp. 101 ◽  
Author(s):  
D. J. Sharkey ◽  
S. A. Robertson
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Response ◽  
Mrna Expression ◽  
Seminal Plasma ◽  
Seminal Fluid ◽  
Differentially Expressed Gene ◽  
Female Reproductive Tract ◽  
High Concentration ◽  
Human Seminal Plasma ◽  
Local Inflammatory Response

Introduction of seminal fluid into the female reproductive tract following coitus stimulates a local inflammatory response. Inflammatory leukocyte recruitment is regulated by induction of cytokine and chemokine synthesis in female tract epithelial cells by seminal fluid signalling agents. Affymetrix microarray analysis in immortalised ectocervical epithelial (Ect1) cells identified the potent anti-inflammatory cytokine follistatin (FST) as the most strongly differentially expressed gene, with a ~12-fold increase in mRNA expression induced by seminal fluid. Follistatin has recently been implicated as a key cytokine in early pregnancy by studies in female follistatin null mice, which exhibit infertility as a consequence of failure to resolve the uterine post-mating inflammatory response. The aim of this study was to investigate seminal plasma regulation of follistatin in human Ect1 cervical cells, and to examine the role of the major active seminal fluid constituent, TGFβ, in controlling Ect1 cells follistatin mRNA expression. To confirm Affymetrix findings, qRT–PCR experiments were undertaken in Ect1 cells incubated with 10% pooled human seminal plasma (SP). Primers specific for the tissue bound isoform of follistatin (FST288) as well as both FST288 and the circulating 315 isoforms (FSTall) were used. Ect1 cell incubation with 10%SP elicited 3.8-fold and 4-fold increases in FST288 and FSTall respectively. Incubation of Ect1 cells with TGFβ1, TGFβ2 and TGFβ3 showed differential effects of the three isoforms, with rTGFβ2 inducing FST288 and FSTall, while rTGFβ1 and TGFβ3 exerted little effect.. These results suggest that seminal plasma induces follistatin synthesis after coitus and that TGFβ2 is at least partly responsible for this effect. Follistatin induced by seminal fluid may act to limit the course of inflammation after intercourse, and thereby prevent uncontrolled inflammatory damage. Follistatin induced in the female tissues would be augmented by follistatin delivered from the male, since human seminal plasma also contains a high concentration of this cytokine.

scholarly journals Nanomaterials induce different levels of oxidative stress, depending on the used model system: Comparison of in vitro and in vivo effects

2020 ◽  
Author(s):  
Isabel Karkossa ◽  
Anne Bannuscher ◽  
Bryan Hellack ◽  
Wendel Wohlleben ◽  
Julie Laloy ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Alveolar Macrophages ◽  
Model System ◽  
Alveolar Epithelial Cells ◽  
Model Systems ◽  
Alveolar Epithelial ◽  
Different Levels

Abstract Background The immense variety and constant development of nanomaterials (NMs) raise the demand for a facilitated risk assessment, for which knowledge on NMs mode of actions (MoAs) is required. For this purpose, a comprehensive data basis is of paramountcy that can be obtained using omics. Furthermore, the establishment of suitable in vitro test systems is indispensable to follow the 3R concept and to master the high number of NMs. In the present study, we aimed at comparing NM effects in vitro and in vivo using a multi-omics approach. We applied an integrated data evaluation strategy based on proteomics and metabolomics to four silica NMs and one titanium dioxide-based NM. For in vitro investigations, alveolar epithelial cells and alveolar macrophages were treated with different doses of NMs, and the results were compared to effects on rat lungs after short-term inhalations and instillations at varying doses with and without a recovery period.Results Since the production of reactive oxygen species (ROS) is described to be a critical biological effect of NMs, and enrichment analyses confirmed oxidative stress as a significant effect upon NM treatment in vitro in the present study, we focused on different levels of oxidative stress. Thus, we found opposite changes for proteins and metabolites that are related to the production of reduced glutathione in alveolar epithelial cells and alveolar macrophages, illustrating that NMs MoAs depend on the used model system. Interestingly, in vivo, pathways related to inflammation were affected to a greater extent than oxidative stress responses. Hence, the assignment of the observed effects to the levels of oxidative stress was different in vitro and in vivo as well. However, the overall classification of “active” and “passive” NMs was consistent in vitro and in vivo.Conclusions The consistent classification indicates both tested cell lines to be suitable for NM toxicity assessment even though the induced levels of oxidative stress strongly depend on the used model systems. Thus, the here presented results highlight that model systems need to be carefully revised to decipher the extent to which they can replace in vivo testing.

Role of diminished epithelial GM-CSF in the pathogenesis of bleomycin-induced pulmonary fibrosis

2000 ◽  
Vol 279 (3) ◽  
pp. L487-L495 ◽  
Author(s):  
Paul J. Christensen ◽  
Marc B. Bailie ◽  
Richard E. Goodman ◽  
Aidan D. O'Brien ◽  
Galen B. Toews ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Pulmonary Fibrosis ◽  
Animal Studies ◽  
Protective Role ◽  
Alveolar Epithelial Cells ◽  
Type Ii ◽  
Alveolar Epithelial ◽  
Gm Csf ◽  
High Level

Evidence derived from human and animal studies strongly supports the notion that dysfunctional alveolar epithelial cells (AECs) play a central role in determining the progression of inflammatory injury to pulmonary fibrosis. We formed the hypothesis that impaired production of the regulatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) by injured AECs plays a role in the development of pulmonary fibrosis. To test this hypothesis, we used the well-characterized model of bleomycin-induced pulmonary fibrosis in rats. GM-CSF mRNA is expressed at a constant high level in the lungs of untreated or saline-challenged animals. In contrast, there is a consistent reduction in expression of GM-CSF mRNA in the lung during the first week after bleomycin injury. Bleomycin-treated rats given neutralizing rabbit anti-rat GM-CSF IgG develop increased fibrosis. Type II AECs isolated from rats after bleomycin injury demonstrate diminished expression of GM-CSF mRNA immediately after isolation and in response to stimulation in vitro with endotoxin compared with that in normal type II cells. These data demonstrate a defect in the ability of type II epithelial cells from bleomycin-treated rats to express GM-CSF mRNA and a protective role for GM-CSF in the pathogenesis of bleomycin-induced pulmonary fibrosis.

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