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 Expression of IL-23 in Gilt Endometrium and Oviduct After Insemination With Seminal Plasma, Spermatozoa or Semen Extender

2020 ◽  
Author(s):  
Anna Svensson ◽  
Jatesada Jiwakanon ◽  
Caroline Fossum ◽  
Anne-Marie Dalin
Keyword(s):  
Inflammatory Response ◽  
Mrna Expression ◽  
Seminal Plasma ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Female Reproductive Tract ◽  
Control Group ◽  
Treatment Groups ◽  
Semen Extender ◽  
Interleukin 23

Abstract Background: Signaling between seminal fluid and the female reproductive tract is required for a successful pregnancy to occur. Insemination with spermatozoa, seminal plasma and extender, is known to cause a rapid inflammatory response in the pig endometrium, which is characterized by an influx of neutrophils into the uterus. The temporary inflammatory response to semen involves induction of cytokines. In this study, potential functions for Interleukin-23 (IL-23) in the inflammatory response to different insemination treatments were examined by studying mRNA expression and immunostaining in samples of gilt oviduct (isthmus and infundibulum) and endometrium collected 35-40 h after insemination. Insemination was performed with either seminal plasma (SP, n = 4), spermatozoa in the extender Beltsville thawing solution (BTS) (SPZ, n = 4), or BTS alone (n = 4). In control gilts (n = 4) an insemination catheter was inserted without anything being inseminated. Any relation between expression of IL-23 and the presence of polymorphonuclear neutrophilic granulocytes (PMNs) in the endometrium was examined. Results: Results showed that IL-23 mRNA was expressed in the oviduct and in the endometrium. There was a significantly lower IL-23 mRNA expression in samples from gilts in the SPZ, SP and BTS treatment groups, compared with the controls. The control group also displayed significantly more neutrophils in samples collected 35-40 h after insemination compared with samples from the SP group. IL-23 immunolabelling was detected in a small number of separate cells as well as in the sub-epithelial connective tissue of the endometrium, the endosalpinx of the isthmus and infundibulum.Conclusions: All fluids used for insemination decreased the expression of IL-23 mRNA in the endometrium compared to catheter-insertion alone, indicating a possible role for IL-23 in the inflammatory response after insemination in gilts.

scholarly journals The High Content of Fructose in Human Semen Competitively Inhibits Broad and Potent Antivirals That Target High-Mannose Glycans

2020 ◽  
Vol 94 (9) ◽  
Author(s):  
Jacklyn Johnson ◽  
Manuel G. Flores ◽  
John Rosa ◽  
Changze Han ◽  
Alicia M. Salvi ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Seminal Fluid ◽  
Cell Attachment ◽  
Sexual Transmission ◽  
Antiviral Agents ◽  
High Concentration ◽  
Human Seminal Plasma ◽  
Sexual Transmission Of Hiv ◽  
Hiv 1

ABSTRACT Semen is the primary transmission vehicle for various pathogenic viruses. Initial steps of transmission, including cell attachment and entry, likely occur in the presence of semen. However, the unstable nature of human seminal plasma and its toxic effects on cells in culture limit the ability to study in vitro virus infection and inhibition in this medium. We found that whole semen significantly reduces the potency of antibodies and microbicides that target glycans on the envelope glycoproteins (Envs) of HIV-1. The extraordinarily high concentration of the monosaccharide fructose in semen contributes significantly to the effect by competitively inhibiting the binding of ligands to α1,2-linked mannose residues on Env. Infection and inhibition in whole human seminal plasma are accurately mimicked by a stable synthetic simulant of seminal fluid that we formulated. Our findings indicate that, in addition to the protein content of biological secretions, their small-solute composition impacts the potency of antiviral microbicides and mucosal antibodies. IMPORTANCE Biological secretions allow viruses to spread between individuals. Each type of secretion has a unique composition of proteins, salts, and sugars, which can affect the infectivity potential of the virus and inhibition of this process. Here, we describe HIV-1 infection and inhibition in whole human seminal plasma and a synthetic simulant that we formulated. We discovered that the sugar fructose in semen decreases the activity of a broad and potent class of antiviral agents that target mannose sugars on the envelope protein of HIV-1. This effect of semen fructose likely reduces the efficacy of such inhibitors to prevent the sexual transmission of HIV-1. Our findings suggest that the preclinical evaluation of microbicides and vaccine-elicited antibodies will be improved by their in vitro assessment in synthetic formulations that simulate the effects of semen on HIV-1 infection and inhibition.

scholarly journals Effect of Lipopolysaccharides (LPS) and Lipoteichoic acid (LTA) on the Inflammatory Response in Rumen Epithelial Cells (REC) and the Impact of LPS on Claw Explants

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2058
Author(s):  
Nicole Reisinger ◽  
Dominik Wendner ◽  
Nora Schauerhuber ◽  
Elisabeth Mayer
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Response ◽  
Structural Integrity ◽  
Animal Health ◽  
Lipoteichoic Acid ◽  
Local Inflammatory Response ◽  
Tissue Integrity ◽  
Separation Force ◽  
The Impact ◽  
Rumen Epithelial Cells

Endotoxins play a crucial role in ruminant health due to their deleterious effects on animal health. The study aimed to evaluate whether LPS and LTA can induce an inflammatory response in rumen epithelial cells. For this purpose, epithelial cells isolated from rumen tissue (RECs) were stimulated with LPS and LTA for 1, 2, 4, and 24 h. Thereafter, the expression of selected genes of the LPS and LTA pathway and inflammatory response were evaluated. Furthermore, it was assessed whether LPS affects inflammatory response and structural integrity of claw explants. Therefore, claw explants were incubated with LPS for 4 h to assess the expression of selected genes and for 24 h to evaluate tissue integrity via separation force. LPS strongly affected the expression of genes related to inflammation (NFkB, TNF-α, IL1B, IL6, CXCL8, MMP9) in RECs. LTA induced a delayed and weaker inflammatory response than LPS. In claw explants, LPS affected tissue integrity, as there was a concentration-dependent decrease of separation force. Incubation time had a strong effect on inflammatory genes in claw explants. Our data suggest that endotoxins can induce a local inflammatory response in the rumen epithelium. Furthermore, translocation of LPS might negatively impact claw health.

scholarly journals Inflammatory Response Elicited by Ureaplasma parvum colonization in human cervical epithelial, stromal, and immune cells

Reproduction ◽  
2021 ◽  
Author(s):  
Ourlad Alzeus Gaddi Tantengco ◽  
Talar Kechichian ◽  
Kathleen L Vincent ◽  
Richard B Pyles ◽  
Paul Mark B Medina ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Epithelial Cells ◽  
Inflammatory Response ◽  
Stromal Cells ◽  
Female Reproductive Tract ◽  
Ureaplasma Parvum ◽  
Cervical Epithelial Cells ◽  
Anti Inflammatory ◽  
The Impact

Ureaplasma parvum is a commensal bacterium in the female reproductive tract but has been associated with pregnancy complications such as preterm prelabor rupture of membranes and preterm birth (PTB). However, the pathologic effects of U. parvum in the cervix, that prevents ascending infections during pregnancy, are still poorly understood. To determine the impact of U. parvum on the cervix, ectocervical (ecto) and endocervical (endo) epithelial and stromal cells were incubated with U. parvum. Macrophages were also tested as a proxy for cervical macrophages to determine the antigenicity of U. parvum. The effects of U. parvum, including influence on cell cycle and cell death, antimicrobial peptide production, epithelial-to-mesenchymal transition (EMT), and inflammatory cytokine levels, were assessed. U. parvum colonized cervical epithelial and stromal cells 4 hours post-infection. Like uninfected control, U. parvum neither inhibited cell cycle progression and nor caused cell death in cervical epithelial and stromal cells. U. parvum increased the production of the antimicrobial peptides (AMPs) cathelicidin and human β-defensin 3 and exhibited weak signs of EMT evidenced by decreased cytokeratin 18 and increased vimentin expression in cervical epithelial cells. U. parvum induced a pro-inflammatory environment (cytokines) and increased MMP-9 in cervical epithelial cells but promoted pro- and anti-inflammatory responses in cervical stromal cells and macrophages. U. parvum may colonize the cervical epithelial layer, but induction of AMPs and anti-inflammatory response may protect the cervix and may prevent ascending infections that can cause PTB. These findings suggest that U. parvum is a weak inducer of inflammation in the cervix.

scholarly journals Anovel Immunological Technique for Identification of Human Seminal Fluid

2010 ◽  
Vol 7 (2) ◽  
pp. 1023-1027
Author(s):  
Baghdad Science Journal
Keyword(s):  
Seminal Plasma ◽  
Human Body ◽  
Seminal Fluid ◽  
Cross Reactivity ◽  
Body Fluids ◽  
Vaginal Fluid ◽  
Human Semen ◽  
Human Seminal Plasma ◽  
Ear Wax ◽  
Immunological Technique

An immunological technique was investigated for the detection of human semen in forensic analysis.This technique included a preparation of anti-human seminal plasma antibodies, by immunizing rabbits with treated human semen. The human semen was treated with an acid to prevent cross reactivity with other human body fluids. The antibody produced was tested against different animal,s seminal fluid samples (dog, goat ,sheep, cow) and human body fluids( saliva, blood , vaginal fluid, ear wax and human semen). It was found that using this developed technique was only selectively responsed with human semen . The prepered kit was evaluated and tested in Forensic laboratory- Ministry of Health. Finally, results were obtained in a comparison with the recommended techniques.

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.

scholarly journals Expression of IL-23 in gilt endometrium and oviduct after insemination with seminal plasma, spermatozoa or semen extender

2021 ◽  
Author(s):  
Anna Svensson ◽  
Jatesada Jiwakanon ◽  
Caroline Fossum ◽  
Anne-Marie Dalin
Keyword(s):  
Connective Tissue ◽  
Inflammatory Response ◽  
Mrna Expression ◽  
Seminal Plasma ◽  
Fold Increase ◽  
Potential Functions ◽  
Treatment Groups ◽  
Cytokine Induction ◽  
Semen Extender ◽  
Interleukin 23

Abstract Objective: Insemination with spermatozoa, seminal plasma and extender, cause a rapid inflammatory response in the pig endometrium, which is characterized by an influx of neutrophils into the uterus. The transient inflammatory response to semen involves cytokine induction. Potential functions for Interleukin-23 (IL-23) in the inflammatory response to different insemination treatments were examined by studying mRNA expression and immunostaining in gilt oviduct and endometrium 35-40 h after insemination. Insemination was performed with seminal plasma (SP), spermatozoa (SPZ) without SP in the extender Beltsville thawing solution (BTS), or BTS alone. In control gilts an insemination catheter was inserted without anything being inseminated. Results: Results showed that IL-23 mRNA was expressed in oviduct and endometrium. There was an approximate 2-4-fold increase in expression of IL-23 mRNA in catheter-only control compared with SPZ, SP and BTS treatment groups. IL-23 immunolabelling was detected in a small number of separate cells as well as in the sub-epithelial connective tissue of the endometrium, the endosalpinx of the isthmus and infundibulum. In conclusion, all fluids used for insemination decreased the expression of IL-23 mRNA in the endometrium compared to catheter-insertion alone, indicating a possible role for IL-23 in the inflammatory response after insemination in gilts.

Transforming growth factor-? (TGF?) in porcine seminal plasma

2011 ◽  
Vol 23 (6) ◽  
pp. 748 ◽  
Author(s):  
Sean O'Leary ◽  
David T. Armstrong ◽  
Sarah A. Robertson
Keyword(s):  
Growth Factor ◽  
Seminal Plasma ◽  
Reproductive Tract ◽  
Transforming Growth Factor ◽  
Seminal Fluid ◽  
Transforming Growth Factor Β ◽  
Interferon Γ ◽  
Female Reproductive Tract ◽  
Active Constituent ◽  
Sperm Density

Bioactive factors in seminal plasma induce cellular and molecular changes in the female reproductive tract after coitus. An active constituent of seminal plasma in mice and humans is the potent immune-modulating cytokine transforming growth factor-β (TGFβ). To investigate whether TGFβ is present in boar seminal plasma, TGFβ1 and TGFβ2 were measured by immunoassay. High levels of TGFβ1 and TGFβ2 were detected in 100% of seminal fluid samples from 73 boars. Both were predominantly in the active, not latent form. Interferon-γ (IFNγ) and lipopolysaccharide (LPS), agents that interact with TGFβ signalling, were detectable in 5% and 100% of samples, respectively. TGFβ1 and TGFβ2 concentrations varied widely between boars, but correlated with each other and with sperm density, and remained relatively constant within individual boars over a 6-month period. Frequent semen collection substantially diminished the concentration of both TGFβ isoforms. Using retrospective breeding data for 44 boars, no correlation between TGFβ content and boar reproductive performance by artificial insemination (AI) with diluted semen was found. It is concluded that TGFβ is abundant in boar seminal plasma, leading to the speculation that, in pigs, TGFβ may be a male–female signalling agent involved in immune changes in the female reproductive tract elicited by seminal fluid.

scholarly journals Bovine endometrial cells do not mount an inflammatory response to Leptospira

2021 ◽  
Author(s):  
Paula C. C. Molinari ◽  
Jarlath E Nally ◽  
John J Bromfield
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Response ◽  
Outer Membrane ◽  
Reproductive Tract ◽  
The United States ◽  
Reproductive Failure ◽  
Female Reproductive Tract ◽  
Human Monocytes ◽  
Endometrial Cells ◽  
Endometrial Epithelial Cells

Leptospirosis causes abortion, premature birth, and stillbirth in cattle, but the mechanisms remain unclear. Infected cattle shed Leptospira intermittently and present a range of clinical symptoms, making diagnosis difficult. The primary route of Leptospira transmission in any animal is colonization of the renal tubule and excretion by urine, however Leptospira can also colonize the female reproductive tract of cows and can be transmitted by semen. Vaccination against Leptospira in the United States is routine in cattle, but immunity is not guaranteed. The cell wall of Leptospira contains Toll-like receptor agonists including peptidoglycan and lipopolysaccharide. The capacity of Leptospira to initiate an innate inflammatory response from uterine endometrial cells is unknown but may be a cause of reproductive failure. Using cell culture, we tested the capacity of bovine endometrial epithelial cells or human monocytes to elicit an inflammatory response to Leptospira borgpetersenii serovar Hardjo strain TC273. Cells were exposed to either heat-killed Leptospira, Leptospira outer membrane, Escherichia coli lipopolysaccharide, Pam3CSK4 or medium alone for 2 to 24 hours. Exposure of bovine endometrial epithelial cells or human monocytes to heat-killed Leptospira or Leptospira outer membrane did not induce the expression of IL1A, IL1B, IL6 or CXCL8, while exposure to E. coli lipopolysaccharide or Pam3CSK4 increased expression of IL1A, IL1B, IL6 and CXCL8 compared to control cells. This data suggests that Leptospira does not trigger a classical inflammatory response in endometrial cells. Understanding the interaction between Leptospira and the female reproductive tract is important in determining the mechanisms of Leptospirosis associated reproductive failure.

scholarly journals Chinese herbal Jin-Ying-Tang attenuates the inflammatory response by inhibiting the activation of TLR4/MyD88/TRAF-6/NIK pathway at the mRNA level in LPS-stimulated mouse mammary epithelial cells

2016 ◽  
Vol 60 (1) ◽  
pp. 71-80
Author(s):  
Qiong Yi ◽  
Xin Li ◽  
Yuan-Fang Li ◽  
Hang Yang ◽  
Xiao-Yi Zhang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Response ◽  
Mrna Expression ◽  
Tumour Necrosis Factor ◽  
Tumour Necrosis ◽  
Mammary Epithelial Cells ◽  
Mrna Level ◽  
Mammary Epithelial ◽  
Tnf Α ◽  
Necrosis Factor

Abstract Introduction: The effects of Jin-Ying-Tang (JYT) on Toll-like Receptor 4 (TLR4) signalling transduction of lipopolysaccharide (LPS)-stimulated mouse mammary epithelial cells (MECs) in vitro were examined. Material and Methods: The cytotoxicity of JYT (0.06-62.50 mg/mL) on mouse MECs was determined by MTT assay. The MECs were co-cultured with LPS in the presence or absence of JYT (39.10 μg/mL, 391 μg/mL, 3910 μg/mL). The concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in the culture supernatants were detected by ELISA. The mRNA expression of TLR4 and downstream TLR4 signalling molecules such as myeloid differentiation factor 88 (MyD88), tumour necrosis factor receptor associated factor 6 (TRAF-6), inhibitor κB (IκB), and nuclear factor κB inducing kinase (NIK) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The results showed that the IC50 of JYT on MECs was 12.25 mg/mL and JYT could significantly decrease the concentrations of IL-6 and TNF-α in LPS-stimulated MECs (P < 0.05). The mRNA expression of TLR4, MyD88, TRAF-6, IκB, and NIK was also significantly decreased when the LPS-stimulated MECs were cocultured at appropriate concentrations of JYT (P < 0.05, P < 0.01). Conclusion: These observations indicate a potential mechanism through which JYT attenuates the systemic inflammatory response to LPS-stimulated mouse mammary epithelial cells by inhibiting the activation of TLR4/MyD88/ TRAF-6/NIK pathway at the mRNA level.

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