The immune microenvironment in inflammatory breast cancer (IBC) is poorly characterised, and molecular and cellular pathways that control accumulation of various immune cells in IBC tissues remain largely unknown. Here, we discovered a novel pathway linking the expression of the tetraspanin protein CD151 in tumour cells with increased accumulation of macrophages in cancerous tissues. It is notable that elevated expression of CD151 and a higher number of tumour-infiltrating macrophages correlated with better patient responses to chemotherapy. Accordingly, CD151-expressing IBC xenografts were characterised by the increased infiltration of macrophages. In vitro migration experiments demonstrated that CD151 stimulates the chemoattractive potential of IBC cells for monocytes via mechanisms involving midkine (a heparin-binding growth factor), integrin α6β1, and production of extracellular vesicles (EVs). Profiling of chemokines secreted by IBC cells demonstrated that CD151 increases production of midkine. Purified midkine specifically stimulated migration of monocytes, but not other immune cells. Further experiments demonstrated that the chemoattractive potential of IBC-derived EVs is blocked by anti-midkine antibodies. These results demonstrate for the first time that changes in the expression of a tetraspanin protein by tumour cells can affect the formation of the immune microenvironment by modulating recruitment of effector cells to cancerous tissues. Therefore, a CD151-midkine pathway can be considered as a novel target for controlled changes of the immune landscape in IBC. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

• MeSH
• antigeny CD151 imunologie   metabolismus   MeSH
• chemokiny metabolismus   MeSH
• lidé MeSH
• makrofágy metabolismus   patologie   MeSH
• midkin metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí fyziologie   MeSH
• zánětlivé nádory prsu metabolismus   patologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The physiological importance of CD151 tetraspanin is known from somatic cells and its outside-in signalling through integrins was described. In male germ cells, two tetraspanins, CD9 and CD81, are involved in sperm-egg membrane fusion, and similarly to integrins, they occupy characteristic regions. We report here on a newly discovered presence of CD151 in sperm, and present its expression and distribution during spermatogenesis and sperm transition during the acrosome reaction. We traced CD151 gene and protein expression in testicular cell subpopulations, with strong enrichment in spermatogonia and spermatids. The testicular and epididymal localization pattern is designated to the sperm head primary fusion site called the equatorial segment and when compared to the acrosome vesicle status, CD151 was located into the inner acrosomal membrane overlying the nucleus. Moreover, we show CD151 interaction with α6 integrin subunit, which forms a dimer with β4 as a part of cis-protein interactions within sperm prior to gamete fusion. We used mammalian species with distinct sperm morphology and sperm maturation such as mouse and bull and compared the results with human. In conclusion, the delivered findings characterise CD151 as a novel sperm tetraspanin network member and provide knowledge on its physiology in male germ cells.

• MeSH
• antigeny CD151 chemie   genetika   metabolismus   MeSH
• exprese genu * MeSH
• fluorescenční protilátková technika MeSH
• integrin alfa6 chemie   metabolismus   MeSH
• konformace proteinů MeSH
• lidé MeSH
• molekulární modely MeSH
• myši MeSH
• spermie metabolismus   MeSH
• testis metabolismus   MeSH
• transport proteinů MeSH
• vazba proteinů MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zárodečné buňky metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Cardiotrophin-1 (CT-1), a member of the IL-6 superfamily, is elevated in the serum of patients with ischemic and valvular heart disease. In this study, we hypothesized that CT-1 induces endothelial cell angiogenesis and that the ADMA/DDAH pathway plays an important role in the process. METHODS: pEGFP-N1-CTF1-GFP and pEGFP-N1 were constructed and used to transiently transfect to HUVECs, mediated by LipofectamineTM 2000. After transfection, the expression of CT-1 was examined by qRT-PCR and western blotting. Endothelial cell proliferation assay was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) method. Migration assay was performed using transwell, tube formation test was examined on Matrigel, eNOSmRNA expression was assayed by qRT-PCR, DDAH I, DDAHII and VEGF expression were detected by western blotting, the level of ADMA and the activity of DDAH were measured by High Performance Liquid Chromatography, NOS activity and the concentration of NO were assayed by L-[3H] citrulline production from L-[3H]arginine. RESULTS: Overexpression of CT-1, increased endothelial cell proliferation, migration and formation of blood vessels, upregulated the expression of eNOSmRNA, DDAHI, DDAHII and VEGF, elevated the activity of DDAH and NOS, decreased the level of ADMA and promoted NO synthesis. In contrast, ADMA partially inhibited the effects of CT-1 induction. CONCLUSIONS: Overexpression of CT-1 increases cell proliferation, migration and formation of blood vessels. This result also suggests that CT-1 may regulate angiogenesis through the ADMA/DDAH pathway.

• MeSH
• amidohydrolasy metabolismus   MeSH
• analýza buněčné migrace MeSH
• antigeny CD151 metabolismus   MeSH
• arginin analogy a deriváty   metabolismus   MeSH
• cévní endotel cytologie   MeSH
• cytokiny metabolismus   fyziologie   MeSH
• endoteliální buňky pupečníkové žíly (lidské) cytologie   MeSH
• fyziologická neovaskularizace fyziologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• oxid dusnatý metabolismus   MeSH
• pohyb buněk fyziologie   MeSH
• proliferace buněk MeSH
• signální transdukce fyziologie   MeSH
• synthasa oxidu dusnatého, typ III metabolismus   MeSH
• transfekce MeSH
• vaskulární endoteliální růstový faktor A metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

It is known that tetraspanin proteins are involved in many physiological somatic cell mechanisms. Additionally, research has indicated they also have a role in various infectious diseases and cancers. This review focuses on the molecular interactions underlying the tetraspanin web formation in gametes. Primarily, tetraspanins act in the reproductive tract as organizers of membrane complexes, which include the proteins involved in the contact and association of sperm and oocyte membranes. In addition, recent data shows that tetraspanins are likely to be involved in these processes in a complex way. In mammalian fertilization, an important role is attributed to CD molecules belonging to the tetraspanin superfamily, particularly CD9, CD81, CD151, and also CD63; mostly as part of extracellular vesicles, the significance of which and their potential in reproduction is being intensively investigated. In this article, we reviewed the existing knowledge regarding the expression of tetraspanins CD9, CD81, CD151, and CD63 in mammalian spermatozoa, oocytes, and embryos and their involvement in reproductive processes, including pathological events.

• MeSH
• embryonální vývoj MeSH
• lidé MeSH
• oocyty fyziologie   MeSH
• rozmnožování * MeSH
• savci fyziologie   MeSH
• spermie fyziologie   MeSH
• tetraspaniny fyziologie   MeSH
• zvířata MeSH
• zygota fyziologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The plasma membrane contains diverse and specialized membrane domains, which include tetraspanin-enriched domains (TEMs) and transmembrane adaptor protein (TRAP)-enriched domains. Recent biophysical, microscopic, and functional studies indicated that TEMs and TRAP-enriched domains are involved in compartmentalization of physicochemical events of such important processes as immunoreceptor signal transduction and chemotaxis. Moreover, there is evidence of a cross-talk between TEMs and TRAP-enriched domains. In this review we discuss the presence and function of such domains and their crosstalk using mast cells as a model. The combined data based on analysis of selected mast cell-expressed tetraspanins [cluster of differentiation (CD)9, CD53, CD63, CD81, CD151)] or TRAPs [linker for activation of T cells (LAT), non-T cell activation linker (NTAL), and phosphoprotein associated with glycosphingolipid-enriched membrane microdomains (PAG)] using knockout mice or specific antibodies point to a diversity within these two families and bring evidence of the important roles of these molecules in signaling events. An example of this diversity is physical separation of two TRAPs, LAT and NTAL, which are in many aspects similar but show plasma membrane location in different microdomains in both non-activated and activated cells. Although our understanding of TEMs and TRAP-enriched domains is far from complete, pharmaceutical applications of the knowledge about these domains are under way.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Cíl: Primárním cílem bylo zjistit frekvenci alel a genotypů nových polymorfizmů spojovaných podle GWAS studií s výskytem žilní trombózy u zdravých osob kavkazské rasy v regionu středních Čech. Metody: Genotypizace trombofilních mutací SERPINC1 IVS +141G>A, GP6 13254T>C a CYP4V2 (Lys259Gln) byla provedena u 1 527 zdravých osob pomocí robotické izolace DNA a následnou amplifikací PCR s analýzou křivky tání (LightCycler 480 System, Roche). Výsledky: Frekvence rizikové alely A polymorfizmu SERPINC1 (IVS +141G>A) byla u sledovaného souboru 11,3 % a frekvence genotypů byla GG 78,36 %, GA 20,66 % a AA 0,98 %. Frekvence rizikové alely T polymorfizmu GP6 13254T>C byla 87,7 % a frekvence genotypů TT 77,14 %, TC 21,15 % a CC 1,70 %. Frekvence rizikové alely A polymorfizmu CYP4V2 (Lys259Gln) byla 65,2 % a frekvence genotypů CC 12,25 %, CA 45,12 % a AA 42,63 %. U obou pohlaví se tyto frekvence nelišily. Závěr: Zjištěné frekvence genotypů a alel potvrzují relativně vysokou prevalenci dalších hereditárních trombofilií v regionu střední Čechy. Jejich klinický význam však zatím není plně znám.

Objective: The aim of study was to determine frequencies of alleles and genotypes of new thrombophilia polymorhisms associated according to GWAS studies with venous thrombosis in Caucasian healthy people in Central Bohemia. Methods: Genotyping of thrombophilic mutations SERPINC1 IVS +141G>A, GP6 13254T>C and CYP4V2 (Lys259Gln) was performed in 1,527 healthy subjects using a robotic DNA isolation and subsequent PCR amplification with melting curve analysis (LightCycler480 System, Roche). Results: For the reference group was the frequency of risk allele A of SERPINC1 (IVS +141G>A) polymorphism 11.3% and frequencies of genotypes were GG 78.36%, GA 20.66% and AA 0.98%. Frequency of risk allele T of GP6 13254T>C polymorphism was 87.7% and frequencies of genotypes were TT 77.14%, TC 21.15% and CC 1.70%. Frequency of risk allele A of CYP4V2 polymorphism (Lys259Gln) was 65.2% and frequencies of genotypes were CC 12.25%, CA 45.12% and AA 42.63%. Those frequencies were not differ for both genders. Conclusion: The results confirm relatively high prevalence of other hereditary thrombophilia polymorphisms in Central Bohemia. But their clinical significance is not fully known yet.

• Klíčová slova
• SERPINC1 (IVS +141G>A), GP6 (13254T>C), CYP4V2 (Lys259Gln),
• MeSH
• dárci krve MeSH
• dospělí MeSH
• genotypizační techniky * metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• nedostatek antitrombinu III * genetika   MeSH
• polymorfismus genetický genetika   MeSH
• prevalence MeSH
• rizikové faktory * MeSH
• systém (enzymů) cytochromů P-450 genetika   MeSH
• trombofilie * epidemiologie   genetika   MeSH
• žilní trombóza * epidemiologie   genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH