Immune regulatory cytokines in seminal plasma of healthy men: A scoping review and analysis of variance

OnlinePublObjective: Seminal plasma cytokines are associated with fertility and reproductive health, but progressing their clinical utility is hampered by absence of reference data on concentration ranges of relevant cytokines in healthy men. We employed a systematic approach to assemble current evidence on the concentrations of immune regulatory cytokines present in seminal plasma (SP) of normozoospermic and/or fertile men and evaluated the impact of different platform methodologies for cytokine quantification. Evidence review: A systematic literature search was performed utilising PubMed, Web of Science and Scopus. Databases were searched from inception until 30th June 2022 inclusive, using combinations of keywords pertaining to seminal fluid and cytokines, and was restricted to human participants. Original data with values reported as concentration of specific cytokines in SP of men clearly defined as fertile or normozoospermic were extracted from studies written in English. Results: A total of 3769 publications were initially identified, ofwhich 118 fulfilled the eligibility criteria for inclusion. A total of 51 individual cytokines are detectable in SP of healthy men. The number of studies reporting on each cytokine range from 1 to >20. The reported concentrations for many cytokines linked with fertility status, including IL6, CXCL8/IL8, and TNFA, are highly variable between published studies. This is associated with the different immunoassaymethodologies utilised andmay be exacerbated by a lack of validation of assays to ensure suitability for SP assessment. Due to the large variation between studies, accurate reference ranges for healthy men cannot be determined from the published data. Conclusions: The concentrations of cytokines and chemokines detected in SP is inconsistent and highly variable between studies and cohorts, limiting current capacity to define reference ranges for cytokine concentrations in fertile men. The lack of standardisation in methods used to process and store SP, and variation in platforms used to evaluate cytokine abundance, are factors contributing to the observed heterogeneity. To progress the clinical utility of SP cytokine analysis will require standardisation and validation ofmethodologies so that reference ranges for healthy fertile men can be defined.Hannah E. Lyons, BridgetM. Arman, Sarah A. Robertson, David J. Sharke

MiOXSYS andOxiSperm II assays appear to provide no clinical utility for determining oxidative stress in human sperm—results from repeated semen collections

OnlinePublBackground: Oxidative stress in semen contributes up to 80% of all infertility diagnosis. Diagnostics to measure oxidative stress in semen was recently added to the 6th edition WHO methods manual, although diagnostic predictive values need to be interpreted with caution as there are still several research questions yet to be answered. Objectives: To determine the natural fluctuations in semen redox indicators (MiOXSYS® and OxiSperm® II) within and between men and their association with markers of sperm oxidative stress. Materials and methods: Total, 118 repeat semen samples from 31 generally healthy men aged 18–45 years, over 6 months. Standard semen analysis as per 5th WHO manual. Semen redox levels measured via MiOXSYS® and OxiSperm® II. Additional attributes of sperm quality; HBA® binding assay and sperm hyperactivation and oxidative stress; DNA fragmentation (Halo® Sperm) and lipid peroxidation (BODIPY™ 581/591 C11) were assessed. Results: Samples with high redox-potential (MiOXSYS® ≥1.47 sORP/10⁶ sperm/ml) had lower sperm, motility, morphology and higher DNA fragmentation (P 0.05). Fluctuations in semen redox levels varied greater between men than within men over the study period. Discussion: Neither MiOXSYS® nor OxiSperm® II assays were predictive of sperm function or sperm oxidative stress. This was likely due at least in part to limited understanding of their biochemistry and clinical application. As a result, these assays seem to provide no additional clinical utility beyond that of a standard semen analysis, highlighting the imperative for the development of new robust point-of-care devices for accurately determining sperm oxidative stress. Conclusion: These findings suggest that MiOXSYS® and OxiSperm® II systems for the measurement of sperm oxidative stressmay have limited diagnostic potential.Patience Castleton, Prabin Gyawali, Nicola Mathews, Shadrack Mulinge Mutuku, David James Sharkey, Nicole Olivia McPherso

Sperm modulate uterine immune parameters relevant to embryo implantation and reproductive success in mice

Seminal fluid factors modulate the female immune response at conception to facilitate embryo implantation and reproductive success. Whether sperm affect this response has not been clear. We evaluated global gene expression by microarray in the mouse uterus after mating with intact or vasectomized males. Intact males induced greater changes in gene transcription, prominently affecting pro-inflammatory cytokine and immune regulatory genes, with TLR4 signaling identified as a top-ranked upstream driver. Recruitment of neutrophils and expansion of peripheral regulatory T cells were elevated by seminal fluid of intact males. In vitro, epididymal sperm induced IL6, CXCL2, and CSF3 in uterine epithelial cells of wild-type, but not Tlr4 null females. Collectively these experiments show that sperm assist in promoting female immune tolerance by eliciting uterine cytokine expression through TLR4-dependent signaling. The findings indicate a biological role for sperm beyond oocyte fertilization, in modulating immune mechanisms involved in female control of reproductive investment.John E. Schjenken, David J. Sharkey, Ella S. Green, Hon Yeung Chan, Ricky A. Matias, Lachlan M. Moldenhauer and Sarah A. Robertso

CDKI-73 is a novel pharmacological inhibitor of Rab11 cargo delivery and innate immune secretion

Innate immunity is critical for host defence against pathogen and environmental challenge and this involves the production and secretion of immune mediators, such as antimicrobial peptides and pro-inflammatory cytokines. However, when dysregulated, innate immunity can contribute to multifactorial diseases, including inflammatory rheumatic disorders, type 2 diabetes, cancer, neurodegenerative and cardiovascular diseases and even septic shock. During an innate immune response, antimicrobial peptides and cytokines are trafficked via Rab11 multivesicular endosomes, and then sorted into Rab11 vesicles for traffic to the plasma membrane and secretion. In this study, a cyclin-dependent kinase inhibitor CDKI-73 was used to determine its effect on the innate immune response, based on previously identified targets for this compound. Our results showed that CDKI-73 inhibited the delivery of Rab11 vesicles to the plasma membrane, resulting in the accumulation of large multivesicular Rab11 endosomes near the cell periphery. In addition to the effect on endosome delivery, CDKI-73 down-regulated the amount of innate immune cargo, including the antimicrobial peptide Drosomycin and pro-inflammatory cytokines interleukin-6 (IL-6) and tumour necrosis factor alpha (TNFα). We concluded that CDKI-73 has the potential to regulate the delivery and secretion of certain innate immune cargo, which could be used to control inflammation.Alexandra Sorvina, Tetyana Shandala, Shudong Wang, David J. Sharkey, Emma Parkinson-Lawrence, Stavros Selemidis and Douglas A. Brook

Effect of Intralipid infusion on peripheral blood T cells and plasma cytokines in women undergoing assisted reproduction treatment

Objectives: Intravenous infusion of Intralipid is an adjunct therapy in assisted reproduction treatment (ART) when immune-associated infertility is suspected. Here, we evaluated the effect of Intralipid infusion on regulatory T cells (Treg cells), effector T cells and plasma cytokines in peripheral blood of women undertaking IVF. Methods: This prospective, observational pilot study assessed Intralipid infusion in 14 women exhibiting recurrent implantation failure, a clinical sign of immune-associated infertility. Peripheral blood was collected immediately prior to and 7 days after intravenous administration of Intralipid. Plasma cytokines were measured by Luminex, and T-cell subsets were analysed by flow cytometry. Results: A small increase in conventional CD8+ T cells occurred after Intralipid infusion, but no change was seen in CD4+ Treg cells, or naïve, memory or effector memory T cells. Proliferation marker Ki67, transcription factors Tbet and RORγt, and markers of suppressive capacity CTLA4 and HLA-DR were unchanged. Dimensionality-reduction analysis using the tSNE algorithm confirmed no phenotype shift within Treg cells or other T cells. Intralipid infusion increased plasma CCL2, CCL3, CXCL8, GM-CSF, G-CSF, IL-6, IL-21, TNF and VEGF. Conclusion: Intralipid infusion elicited elevated pro-inflammatory cytokines, and a minor increase in CD8+ T cells, but no change in pro-tolerogenic Treg cells. Notwithstanding the limitation of no placebo control, the results do not support Intralipid as a candidate intervention to attenuate the Treg cell response in women undergoing ART. Future placebo-controlled studies are needed to confirm the potential efficacy and clinical significance of Intralipid in attenuating cytokine induction and circulating CD8+ T cells.Kerrie L Foyle, David J Sharkey, Lachlan M Moldenhauer, Ella S Green, Jasmine J Wilson, Cassandra J Roccisano ... et al

Regulatory T cells are paramount effectors in progesterone regulation of embryo implantation and fetal growth

Published: June 8, 2023Progesterone (P4) is essential for embryo implantation, but the extent to which the pro-gestational effects of P4 depend on the maternal immune compartment is unknown. Here, we investigate whether regulatory T cells (Treg cells) act to mediate luteal phase P4 effects on uterine receptivity in mice. P4 antagonist RU486 administered to mice on days 0.5 and 2.5 post coitum (dpc) to model luteal phase P4 deficiency caused fewer CD4+Foxp3+ Treg cells and impaired Treg functional competence, along with dysfunctional uterine vascular remodeling and perturbed placental development in mid-gestation. These effects were linked with fetal loss and fetal growth restriction, accompanied by a Th1/CD8-skewed T cell profile. Adoptive transfer at implantation of Treg cells - but not T conventional (Tconv) cells - alleviated fetal loss and fetal growth restriction by mitigating adverse effects of reduced P4 signaling on uterine blood vessel remodeling and placental structure, and restoring maternal T cell imbalance. These findings demonstrate an essential role for Treg cells in mediating P4 effects at implantation, and indicate that Treg cells are a sensitive and critical effector mechanism through which P4 drives uterine receptivity to support robust placental development and fetal growth.Ella S. Green, Lachlan M. Moldenhauer, Holly M. Groome, David J. Sharkey, Peck Y. Chin, Alison S. Care, Rebecca L. Robker, Shaun R. McColl, and Sarah A., Robertso

Regulatory T cell proportion and phenotype are altered in women using oral contraception

Advance access publication 04 July 2022Regulatory T (Treg) cells are a specialized CD4+ T cell subpopulation that are essential for immune homeostasis, immune tolerance, and protection against autoimmunity. There is evidence that sex-steroid hormones estrogen and progesterone modulate Treg cell abundance and phenotype in women. Since natural oscillations in these hormones are modified by hormonal contraceptives, we examined whether oral contraception (OC) use impacts Treg cells and related T cell populations. T cells were analyzed by multiparameter flow cytometry in peripheral blood collected across the menstrual cycle from healthy women either using OC or without hormonal contraception and from age-matched men. Compared to naturally cycling women, women using OC had fewer Treg cells and an altered Treg cell phenotype. Notably, Treg cells exhibiting a strongly suppressive phenotype, defined by high FOXP3, CD25, Helios, HLADR, CTLA4, and Ki67, comprised a lower proportion of total Treg cells, particularly in the early- and mid-cycle phases. The changes were moderate compared to more substantial differences in Treg cells between women and men, wherein women had fewer Treg cells—especially of the effector memory Treg cell subset—associated with more T helper type 1 (Th1) cells and CD8+ T cells and lower Treg:Th1 cell and Treg:CD8+ T cell ratios than men. These findings imply that OC can modulate the number and phenotype of peripheral blood Treg cells and raise the possibility that Treg cells contribute to the physiological changes and altered disease susceptibility linked with OC use.Lachlan M. Moldenhauer, Min Jin, Jasmine J. Wilson, Ella S. Green, David J. Sharkey, Mark D. Salkeld, Thomas C. Bristow, M. Louise Hull, Gustaaf A. Dekker, and Sarah A. Robertso

It takes a community to conceive: an analysis of the scope, nature and accuracy of online sources of health information for couples trying to conceive

This study examined the nature and accuracy of information available across online platforms for couples trying to conceive. A consumer simulation-based investigation of English websites and social media (Facebook, Twitter, Instagram) was undertaken using common search terms identified in a pilot study. Claims about fertility and pregnancy health were then extracted from the results and analysed thematically. The accuracy of each claim was assessed independently by six fertility and conception experts, rated on a scale of 1 (not factual) to 4 (highly factual), with scores collated to produce a median rating. Claims with a medianscoreb3 were classified as inaccurate. The use of the terms 'trying to conceive' and '#TTC' were common identifiers on online platforms. Claims were extracted predominantly from websites (n= 89) rather than social media, with Twitter and Instagram comprising commercial elements and Facebook focused on community-based support. Thematic analysis revealed three major themes among the claims across all platforms: conception behaviour and monitoring, lifestyle and exposures, and medical. Fact-checking by the experts revealed that 40% of the information assessed was inaccurate, and that inaccuracies were more likely to be present in the conception behaviour and monitoring advice, the topics most amenable to modification. Since online information is a readily accessible and commonly utilized resource, there is opportunity for improved dissemination of evidence-based material to reach interested couples. Further cross-disciplinary and consumer-based research, such as a user survey, is required to understand how best to provide the 'trying to conceive' community with accurate information.Sophie G.E. Kedzior, Tina Bianco-Miotto, James Breen, Kerrilyn R. Diener, Martin Donnelley, Kylie R. Dunning Megan A.S. Penno, John E. Schjenken, David J. Sharkey, Nicolette A. Hodyl, Tod Fullston, Maria Gardiner, Hannah M. Brown, Alice R. Rumbol

Stomatal responses of Eucalyptus species to elevated CO2 concentration and drought stress

Five species of Eucalyptus (E. grandis, E. urophylla, E. camaldulensis, E. torelliana, and E. phaeotrica), among the ten species most commonly used in large scale plantations, were selected for studies on the effects of elevated CO2 concentration [CO2] and drought stress on stomatal responses of 2.5-month old seedlings. The first three species belong to the subgenus Smphyomyrtus, whereas the fourth species belongs to the subgenus Corymbia and E. phaeotrica is from the subgenus Monocalyptus. Seedlings were grown in four pairs of open-top chambers, arranged to have 2 plants of each species in each chamber, with four replications in each of two CO2 concentrations: 350 ± 30 mumol mol-1 and 700 ± 30 mumol mol-1. After 100 days in the chambers, a series of gas exchange measurements were made. Half the plants in each chamber, one plant per species per chamber, were drought-stressed by withholding irrigation, while the remaining plants continued to be watered daily. Drought stress decreased stomatal conductance, photosynthesis and transpiration rates in all the species. The effect of drought stress on stomatal closure was similar in both [CO2]. The positive effects of elevated [CO2] on photosynthesis and water use efficiency were maintained longer during the stress period than under well-watered conditions. The photosynthetic rate of E. phaeotrica was higher even in the fourth day of the drought stress. Drought stress increased photoinhibition of photosynthesis, as measured by chlorophyll fluorescence, which varied among the species, as well as in relation to [CO2]. The results are in agreement with observed differences in stomatal responses between some eucalyptus species of the subgenera Symphyomyrtus and Monocalyptus

The criminal justice voluntary sector: concepts and an agenda for an emerging field

This is the peer reviewed version of the following article: Tomczak, P. & Buck, G. (2019). The criminal justice voluntary sector: concepts and an agenda for an emerging field. Howard Journal of Crime and Justice, 58(3), which has been published in final form at https://doi.org/10.1111/hojo.12326. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Volunteers and voluntary organisations play significant roles pervading criminal justice. They are key actors, with unrecognised potential to shore up criminal justice and/or collaboratively reshape social justice. Unlike public and for-profit agents, criminal justice volunteers and voluntary organisations (CJVVOs) have been neglected by scholars. We call for analyses of diverse CJVVOs, in national and comparative contexts. We provide three categories to highlight distinctive organising auspices, which hold across criminal justice: statutory volunteers, quasi-statutory volunteers and voluntary organisations. The unknown implications of these different forms of non-state, non-profit justice involvement deserve far greater attention from academics, policymakers and practitioners