{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T10:03:00Z","timestamp":1777543380660,"version":"3.51.4"},"update-to":[{"DOI":"10.12688\/f1000research.11432.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2017,6,22]],"date-time":"2017-06-22T00:00:00Z","timestamp":1498089600000}}],"reference-count":62,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2017,6,22]],"date-time":"2017-06-22T00:00:00Z","timestamp":1498089600000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Medical Research Council","award":["G1100356, G060048"],"award-info":[{"award-number":["G1100356, G060048"]}]},{"name":"Medical Research Council","award":["G1100356"],"award-info":[{"award-number":["G1100356"]}]},{"name":"Medical Research Council","award":["G1002033"],"award-info":[{"award-number":["G1002033"]}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns4:p>\n                    <ns4:bold>Background:<\/ns4:bold>\n                    Human mast cells (MCs) are long-lived tissue-resident immune cells characterised by granules containing the proteases chymase and\/or tryptase. Their phenotype is modulated by their tissue microenvironment. The human uterus has an outer muscular layer (the myometrium) surrounding the endometrium, both of which play an important role in supporting a pregnancy. The endometrium is a sex steroid target tissue consisting of epithelial cells (luminal, glandular) surrounded by a multicellular stroma, with the latter containing an extensive vascular compartment as well as fluctuating populations of immune cells that play an important role in regulating tissue function. The role of MCs in the human uterus is poorly understood with little known about their regulation or the impact of steroids on their differentiation status.\n                  <\/ns4:p>\n                  <ns4:p>The current study had two aims: 1) To investigate the spatial and temporal location of uterine MCs and determine their phenotype; 2) To determine whether MCs express receptors for steroids implicated in uterine function, including oestrogen (ER\u03b1, ER\u03b2), progesterone (PR) and glucocorticoids (GR).<\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Methods:<\/ns4:bold>\n                    Tissue samples from women (n=46) were used for RNA extraction (n=26) or fixed (n=20) for immunohistochemistry.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Results:<\/ns4:bold>\n                    Messenger RNAs encoded by\n                    <ns4:italic>TPSAB1<\/ns4:italic>\n                    (tryptase) and\n                    <ns4:italic>CMA1<\/ns4:italic>\n                    (chymase) were detected in endometrial tissue homogenates. Immunohistochemistry revealed the relative abundance of tryptase MCs was myometrium&gt;basal endometrium&gt;functional endometrium. We show for the first time that uterine MCs are predominantly of the classical MC subtypes: (positive, +; negative, -) tryptase+\/chymase- and tryptase+\/chymase+, but a third subtype was also identified (tryptase-\/chymase+). Tryptase+ MCs were of an ER\u03b2+\/ER\u03b1-\/PR-\/GR+ phenotype mirroring other uterine immune cell populations, including natural killer cells.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Conclusions:<\/ns4:bold>\n                    Endometrial tissue resident immune MCs have three protease-specific phenotypes. Expression of both ER\u03b2 and GR in MCs mirrors that of other immune cells in the endometrium and suggests that MC function may be altered by the local steroid microenvironment.\n                  <\/ns4:p>","DOI":"10.12688\/f1000research.11432.2","type":"journal-article","created":{"date-parts":[[2017,6,22]],"date-time":"2017-06-22T09:30:17Z","timestamp":1498123817000},"page":"667","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":26,"title":["Immunoprofiling of human uterine mast cells identifies three phenotypes and expression of ER\u03b2 and glucocorticoid receptor"],"prefix":"10.12688","volume":"6","author":[{"given":"Bianca","family":"De Leo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arantza","family":"Esnal-Zufiaurre","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frances","family":"Collins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hilary O.D.","family":"Critchley","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9051-9380","authenticated-orcid":false,"given":"Philippa T.K.","family":"Saunders","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2017,6,22]]},"reference":[{"key":"ref-1","doi-asserted-by":"publisher","first-page":"5071-4","DOI":"10.1210\/jcem.86.10.8101","article-title":"The glucocorticoid receptor is specifically expressed in the stromal compartment of the 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All the samples in this study were processed according to standard methods approved by our pathologist.","URL":"https:\/\/f1000research.com\/articles\/6-667\/v2#referee-comment-2804","order":1,"name":"referee-comment-2804","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.12343.r23114, Areege M. Kamal, The National Centre for Early Detection of Cancer, Oncology Teaching Hospital, Baghdad Medical City, Baghdad, Iraq, 31 May 2017, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/6-667\/v1#referee-response-23114","order":2,"name":"referee-response-23114","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Philippa Saunders<\/b>; <i>Posted: 17 Jun 2017<\/i>; We thank Dr Kamal for her comments on our paper which we have edited to address some of her concerns.Specific responses:We have now referenced the small study by Mori <i>et al<\/i>&nbsp;(1997) in the Introduction and the study by Engemise <i>et al<\/i> in the Discussion.We have changed the text of the Introduction so that the abbreviations for TPSAB1 and CMA1 are introduced during the first mention of their importance in defining the mast cell phenotype.Methods.The text describing the samples has been changed to highlight the patient information provided in Suppl Table 1 and clarify that although 46 patients were recruited during this study their samples were split between RNA (n=29) and fixation for immunostaining (n=26).Analysis methods for RTPCR data: Although the delta delta CT method is one of the most popular it is based on comparable amplification efficiencies of the endogenous control and gene of interest. As determined by the standard curves the efficiencies of the TPSAB1 and CMA1 were not suitable for analysis by this method and therefore a more appropriate method was the relative standard curve method. The text in M&amp;M has been revised to make this clearer.See: Bustin SA, Benes V, Garson JA etal, the MIQE guidelines: Minimum Information for Publication of Quantitative Real-Time PCR experiments, Clinical Chemistry 55:4 611-622 (2009)These methods are also described in detail in Applied Biosystems manual onlinehttp:\/\/www6.appliedbiosystems.com\/support\/tutorials\/pdf\/performing_rq_gene_exp_rtpcr.pdfThe tonsil sample was chosen as a positive control as this tissue has mast cells which contain tryptase and chymase, the RNA sample was purchased from a commercial supplier and we chose this tissue because we were able to access fixed material from our hospital to use for control experiments to validate the antibodies.For dual immunostaining primary antibodies were applied to sections overnight (16h) and incubated in a fridge - secondary antibodies were applied for 1h at room temperature. This is a standard method used in our laboratory and in our hands it leads to lower rates of non-specific background staining than application of primary antibody at room temperature. We did not quantify the fluorescent staining.Results.The limitations on the amount of individual fixed samples meant that not all of the fixed samples (n=20) were stained with all antibodies and this is reflected in the numbers stated in the figure legends.MCs were considered as ERbeta positive only if the entire nucleus was identified in the section (as highlighted with white arrowheads). In this study the low number of chymase only MCs meant we were not able to conduct a comprehensive staining for ERbeta.In the summary Table 1 we decided to combine the results from the functional and basal layers because the mast cells were very rare in the former and we wanted to provide an overview to complement the more comprehensive data shown in the individual photographs.Our interpretation of the data related to GR positive cells (as illustrated in Figure 3, 6 panels) is that the tryptase positive mast cells that were GR+ appeared to be in resting state i.e. the tryptase appeared to be confined to the cytoplasmic area around the nucleus.Discussion.We used a standard curve method so 'concentration' is more appropriate than 'levels' which is a common term when the delta CT method is used.We have changed the text to make it clearer that this paragraph is discussing mRNAs and hence protein release is not relevant to the argument about constitutive expression.","URL":"https:\/\/f1000research.com\/articles\/6-667\/v2#referee-comment-2803","order":3,"name":"referee-comment-2803","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"PTKS, AZ-E and FC were funded by MRC Programme (grant G1100356) to PTKS, which also paid for the costs of consumables used in this study. BD was in receipt of an MRC PhD studentship funded by the Centre Grant for Reproductive Health (G1002033). Salaries of nurses who recruited patients to the study were paid by MRC grants G1100356 (PI. PTKS) and G0500047 (PI. HODC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":4,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Data associated with the article are available under the terms of the Creative Commons Zero \"No rights reserved\" data waiver (CC0 1.0 Public domain dedication).","order":0,"name":"copyright-info","label":"Copyright"},{"value":"Ana Claudia Zenclussen, Endocrinology DOI: 10.3410\/f.727723558.793539636","URL":"http:\/\/f1000.com\/prime\/727723558#eval793539636","order":0,"name":"recommendation-793539636","label":"Recommendation","group":{"name":"prime-recommendations","label":"F1000Prime article recommendations"}}]}}