Centrosomes play a crucial role during immune cell interactions and initiation of the immune response. In proliferating cells, centrosome numbers are tightly controlled and generally limited to one in G1 and two prior to mitosis. Defects in regulating centrosome numbers have been associated with cell transformation and tumorigenesis. Here, we report the emergence of extra centrosomes in leukocytes during immune activation. Upon antigen encounter, dendritic cells pass through incomplete mitosis and arrest in the subsequent G1 phase leading to tetraploid cells with accumulated centrosomes. In addition, cell stimulation increases expression of polo-like kinase 2, resulting in diploid cells with two centrosomes in G1-arrested cells. During cell migration, centrosomes tightly cluster and act as functional microtubule-organizing centers allowing for increased persistent locomotion along gradients of chemotactic cues. Moreover, dendritic cells with extra centrosomes display enhanced secretion of inflammatory cytokines and optimized T cell responses. Together, these results demonstrate a previously unappreciated role of extra centrosomes for regular cell and tissue homeostasis.

• MeSH
• centrozom * metabolismus   MeSH
• chemotaxe MeSH
• cytokiny metabolismus   MeSH
• dendritické buňky * metabolismus   MeSH
• kontrolní body buněčného cyklu MeSH
• lidé MeSH
• mitóza MeSH
• organizační centrum mikrotubulů MeSH
• pohyb buněk MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• T-lymfocyty metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• PLK2 protein, human MeSH Prohlížeč
• protein-serin-threoninkinasy MeSH

The fertilization of freshwater fish occurs in an environment that may negatively affect the gametes; therefore, the specific mechanisms triggering the encounters of gametes would be highly expedient. The egg and ovarian fluid are likely the major sources of these triggers, which we confirmed here for rainbow trout (Oncorhynchus mykiss). The ovarian fluid affected significantly spermatozoa performance: it supported high velocity for a longer period and changed the motility pattern from tumbling in water to straightforward moving in the ovarian fluid. Rainbow trout ovarian fluid induced a trapping chemotaxis-like effect on activated male gametes, and this effect depended on the properties of the activating medium. The interaction of the spermatozoa with the attracting agents was accompanied by the "turn-and-run" behavior involving asymmetric flagellar beating and Ca2+ concentration bursts in the bent flagellum segment, which are characteristic of the chemotactic response. Ovarian fluid created the optimal environment for rainbow trout spermatozoa performance, and the individual peculiarities of the egg (ovarian fluid)-sperm interaction reflect the specific features of the spawning process in this species.

• Klíčová slova
• Oncorhynchus mykiss, chemotaxis, fertilization, ovarian fluid, sperm motility,
• MeSH
• chemotaxe fyziologie   MeSH
• fertilizace fyziologie   MeSH
• Oncorhynchus mykiss metabolismus   MeSH
• ovarium cytologie   metabolismus   MeSH
• spermie cytologie   metabolismus   MeSH
• vápníková signalizace fyziologie   MeSH
• zvířata MeSH
• zygota cytologie   metabolismus   MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

To improve the treatment of psoriasiform inflammation, we developed actively targeted nanocarriers loaded with the phosphodiesterase 4 inhibitor AN2728. Methods: Phospholipid-poly(lactic-co-glycolic acid) nanohybrids were prepared and conjugated with monovalent anti-desmoglein 3 antibody to bind keratinocytes. Results: The actively targeted nanohybrids were 229 nm in mean size with a nearly neutral surface charge. Flow cytometry and confocal microscopy showed a 9-fold increase in keratinocyte uptake of targeted nanohybrids relative to non-targeted nanoparticles. The nanoparticles localized mainly in lysosomes after internalization. AN2728-loaded antibody-conjugated nanocarriers inhibited cytokine/chemokine overexpression in activated keratinocytes without affecting cell viability. The targeted nanohybrids also suppressed neutrophil migration by reducing CXCL1 and CXCL2 release from keratinocytes. Following subcutaneous administration in mice, the nanohybrids distributed to the epidermis and hair follicles. In a psoriasis-like skin mouse model, the actively targeted nanoparticles were superior to free drug and non-targeted nanoparticles in mitigating skin inflammation. Intervention with the targeted nanosystem reduced the epidermal thickness of the psoriasiform lesion from 191 to 42 µm, decreased the Psoriasis Area Severity Index by 74%, restored barrier function, and returned chemokine levels to baseline. Conclusions: Our developed nanosystem was safe and demonstrated efficient targeting properties for the treatment of cutaneous inflammation.

• Klíčová slova
• active targeting, desmoglein 3, keratinocyte, lipid-polymer nanohybrid, monovalent antibody, psoriasis,
• MeSH
• bicyklické sloučeniny heterocyklické aplikace a dávkování   farmakologie   MeSH
• buněčné linie keratinocytů HaCaT MeSH
• chemokin CXCL1 účinky léků   imunologie   MeSH
• chemokin CXCL2 účinky léků   imunologie   MeSH
• chemotaxe účinky léků   MeSH
• desmoglein 3 imunologie   MeSH
• epidermis MeSH
• fosfolipidy * MeSH
• imunokonjugáty farmakologie   MeSH
• inhibitory fosfodiesterasy 4 aplikace a dávkování   farmakologie   MeSH
• keratinocyty účinky léků   imunologie   MeSH
• kopolymer kyseliny glykolové a mléčné * MeSH
• lidé MeSH
• lyzozomy metabolismus   ultrastruktura   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• nanočástice * MeSH
• neutrofily účinky léků   MeSH
• nosiče léků MeSH
• protilátky imunologie   MeSH
• psoriáza imunologie   patologie   MeSH
• sloučeniny boru aplikace a dávkování   farmakologie   MeSH
• systémy cílené aplikace léků MeSH
• vlasový folikul MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bicyklické sloučeniny heterocyklické MeSH
• chemokin CXCL1 MeSH
• chemokin CXCL2 MeSH
• crisaborole MeSH Prohlížeč
• desmoglein 3 MeSH
• fosfolipidy * MeSH
• imunokonjugáty MeSH
• inhibitory fosfodiesterasy 4 MeSH
• kopolymer kyseliny glykolové a mléčné * MeSH
• nosiče léků MeSH
• protilátky MeSH
• sloučeniny boru MeSH

Cancer development is a highly complicated process in which tumour growth depends on the development of its vascularization system. To support their own growth, tumour cells significantly modify their microenvironment. One of such modifications inflicted by tumours is stimulation of endothelial cell migration and proliferation. There is accumulating evidence that extracellular vesicles (EVs) secreted by tumour cells (tumour-derived EVs, TEVs) may be regarded as "messengers" with the potential for affecting the biological activities of target cells. Interaction of TEVs with different cell types occurs in an auto- and paracrine manner and may lead to changes in the function of the latter, e.g., promoting motility, proliferation, etc. This study analysed the proangiogenic activity of EVs derived from human pancreatic adenocarcinoma cell line (HPC-4, TEVHPC) in vitro and their effect in vivo on Matrigel matrix vascularization in severe combined immunodeficient (SCID) mice. TEVHPC enhanced proliferation of HPC-4 cells and induced their motility. Moreover, TEVHPC stimulated human umbilical vein endothelial cell (HUVEC) proliferation and migration in vitro. Additionally, TEVHPC influenced secretion of proangiogenic factors (IL-8, VEGF) by HUVEC cells and supported Matrigel matrix haemoglobinization in vivo. These data show that TEVs may support tumour propagation in an autocrine manner and may support vascularization of the tumour. The presented data are in line with the theory that tumour cells themselves are able to modulate the microenvironment via TEVs to maximize their growth potential.

• MeSH
• autokrinní signalizace MeSH
• buněčné dělení MeSH
• chemotaxe MeSH
• duktální karcinom pankreatu patologie   MeSH
• endoteliální buňky pupečníkové žíly (lidské) MeSH
• extracelulární vezikuly patologie   MeSH
• fixní kombinace léků MeSH
• interleukin-8 genetika   metabolismus   MeSH
• kolagen MeSH
• laminin MeSH
• lidé MeSH
• messenger RNA biosyntéza   MeSH
• myši SCID MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• nádory slinivky břišní patologie   MeSH
• patologická angiogeneze etiologie   MeSH
• proteoglykany MeSH
• transplantace nádorů MeSH
• vaskulární endoteliální růstový faktor A genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fixní kombinace léků MeSH
• interleukin-8 MeSH
• kolagen MeSH
• laminin MeSH
• matrigel MeSH Prohlížeč
• messenger RNA MeSH
• proteoglykany MeSH
• vaskulární endoteliální růstový faktor A MeSH
• VEGFA protein, human MeSH Prohlížeč

Despite the shared pattern of surface antigens, neoplastic cells in chronic lymphocytic leukemia (CLL) are highly heterogeneous in CD5 expression, a marker linked to a proliferative pool of neoplastic cells. To further characterize CD5high and CD5low neoplastic cells, we assessed the chemokine receptors (CCR5, CCR7, CCR10, CXCR3, CXCR4, CXCR5) and adhesion molecules (CD54, CD62L, CD49d) on the CD5high and CD5low subpopulations, defined by CD5/CD19 coexpression, in peripheral blood of CLL patients (n = 60) subgrouped according to the IgHV mutational status (IgHV mut, n = 24; IgHV unmut, n = 36). CD5high subpopulation showed a high percentage of CXCR3 (P < 0.001), CCR10 (P = 0.001), and CD62L (P = 0.031) and high levels of CXCR5 (P = 0.005), CCR7 (P = 0.013) compared to CD5low cells expressing high CXCR4 (P < 0.001). Comparing IgHV mut and IgHV unmut patients, high levels of CXCR3 on CD5high and CD5low subpopulations were detected in the IgHV mut patients, with better discrimination in CD5low subpopulation. Levels of CXCR3 on CD5low subpopulation were associated with time to the next treatment, thus further confirming its prognostic value. Taken together, our analysis revealed higher CXCR3 expression on both CD5high and CD5low neoplastic cells in IgHV mut with a better prognosis compared to IgHV unmut patients. Contribution of CXCR3 to CLL pathophysiology and its suitability for prognostication and therapeutic exploitation deserves future investigations.

• MeSH
• antigeny CD5 metabolismus   MeSH
• biologické markery MeSH
• chemotaxe imunologie   MeSH
• chronická lymfatická leukemie diagnóza   genetika   metabolismus   terapie   MeSH
• imunofenotypizace MeSH
• Kaplanův-Meierův odhad MeSH
• lidé středního věku MeSH
• lidé MeSH
• molekuly buněčné adheze metabolismus   MeSH
• mutace * MeSH
• pohyb buněk MeSH
• prognóza MeSH
• receptory chemokinů metabolismus   MeSH
• receptory CXCR3 genetika   metabolismus   MeSH
• ROC křivka MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• těžké řetězce imunoglobulinů genetika   MeSH
• variabilní oblast imunoglobulinu genetika   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny CD5 MeSH
• biologické markery MeSH
• CXCR3 protein, human MeSH Prohlížeč
• molekuly buněčné adheze MeSH
• receptory chemokinů MeSH
• receptory CXCR3 MeSH
• těžké řetězce imunoglobulinů MeSH
• variabilní oblast imunoglobulinu MeSH

Selectins belong to a group of adhesion molecules that fulfill an essential role in immune and inflammatory responses and tissue healing. Selectins are glycoproteins that decode the information carried by glycan structures, and non-covalent interactions of selectins with these glycan structures mediate biological processes. The sialylated and fucosylated tetrasaccharide sLex is an essential glycan recognized by selectins. Several glycosyltransferases are responsible for the biosynthesis of the sLex tetrasaccharide. Selectins are involved in a sequence of interactions of circulated leukocytes with endothelial cells in the blood called the adhesion cascade. Recently, it has become evident that cancer cells utilize a similar adhesion cascade to promote metastases. However, like Dr. Jekyll and Mr. Hyde's two faces, selectins also contribute to tissue destruction during some infections and inflammatory diseases. The most prominent function of selectins is associated with the initial stage of the leukocyte adhesion cascade, in which selectin binding enables tethering and rolling. The first adhesive event occurs through specific non-covalent interactions between selectins and their ligands, with glycans functioning as an interface between leukocytes or cancer cells and the endothelium. Targeting these interactions remains a principal strategy aimed at developing new therapies for the treatment of immune and inflammatory disorders and cancer. In this review, we will survey the significant contributions to and the current status of the understanding of the structure of selectins and the role of selectins in various biological processes. The potential of selectins and their ligands as therapeutic targets in chronic and acute inflammatory diseases and cancer will also be discussed. We will emphasize the structural characteristic of selectins and the catalytic mechanisms of glycosyltransferases involved in the biosynthesis of glycan recognition determinants. Furthermore, recent achievements in the synthesis of selectin inhibitors will be reviewed with a focus on the various strategies used for the development of glycosyltransferase inhibitors, including substrate analog inhibitors and transition state analog inhibitors, which are based on knowledge of the catalytic mechanism.

• Klíčová slova
• cancer, carbohydrate determinants, glycosyltransferases, inflammation, inhibitors, selectin ligands, selectin-ligand interactions, selectins, sialyl Lewis x, transition state analog inhibitors,
• MeSH
• buněčná adheze * MeSH
• leukocyty metabolismus   patologie   MeSH
• lidé MeSH
• nádorové proteiny metabolismus   MeSH
• nádory metabolismus   patologie   MeSH
• rolling leukocytů * MeSH
• selektiny metabolismus   MeSH
• zánět metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• nádorové proteiny MeSH
• selektiny MeSH

SCOPE: The docosahexaenoic acid ester of hydroxy linoleic acid (13-DHAHLA) is a bioactive lipid with anti-inflammatory properties from the family of fatty acid esters of hydroxy fatty acids (FAHFA). METHODS AND RESULTS: To explore the biosynthesis of 13-DHAHLA from dietary oils, C57BL/6N mice are gavaged for 8 days with various corn oil/marine oil mixtures containing the same amount of DHA. Plasma levels of omega-3 FAHFAs are influenced by the lipid composition of the mixtures but do not reflect the changes in bioavailability of polyunsaturated fatty acids in plasma. Triacylglycerol-bound DHA and linoleic acid serve as more effective precursors for 13-DHAHLA synthesis than DHA bound in phospholipids or wax esters. Both 13(S)- and 13(R)-DHAHLA inhibit antigen and PGE2 -induced chemotaxis and degranulation of mast cells to a comparable extent and 13(S)-DHAHLA is identified as the predominant isomer in mouse adipose tissue. CONCLUSION: Here, the optimal nutritional source of DHA is identified, which supports production of anti-inflammatory FAHFAs, as triacylglycerol-based marine oil and also reveals a possible role of triacylglycerols in the synthesis of FAHFA lipokines.

• Klíčová slova
• calanus, fatty acids, krill, mast cells, stereoisomers,
• MeSH
• antiflogistika nesteroidní krev   farmakokinetika   MeSH
• biologická dostupnost MeSH
• chemotaxe účinky léků   MeSH
• kyseliny dokosahexaenové farmakokinetika   MeSH
• kyseliny linolové chemie   MeSH
• kyseliny mastné omega-3 farmakokinetika   farmakologie   MeSH
• mastocyty účinky léků   MeSH
• myši inbrední C57BL MeSH
• oleje chemie   farmakokinetika   MeSH
• stereoizomerie MeSH
• triglyceridy chemie   MeSH
• vodní organismy MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 13-hydroxylinoleic acid MeSH Prohlížeč
• antiflogistika nesteroidní MeSH
• kyseliny dokosahexaenové MeSH
• kyseliny linolové MeSH
• kyseliny mastné omega-3 MeSH
• oleje MeSH
• triglyceridy MeSH

A better understanding of the molecular mechanisms leading to mast cell migration and chemotaxis is the long-term goal in mast cell research and is essential for comprehension of mast cell function in health and disease. Various techniques have been developed in recent decades for in vitro and in vivo assessment of mast cell motility and chemotaxis. In this chapter, three microscopy assays facilitating real-time quantification of mast cell chemotaxis and migration are described, focusing on individual cell tracking and data analysis.

• Klíčová slova
• Cell migration, Cell tracking, Chemoattractant, Chemokine, Chemotaxis, Mast cells,
• MeSH
• analýza buněčné migrace metody   MeSH
• biotest metody   MeSH
• buněčný tracking metody   MeSH
• chemotaxe fyziologie   MeSH
• fibronektiny metabolismus   MeSH
• lidé MeSH
• mastocyty cytologie   fyziologie   MeSH
• mikroskopie metody   MeSH
• myši MeSH
• počítačové systémy MeSH
• pohyb buněk fyziologie   MeSH
• prostředí kontrolované MeSH
• sefarosa MeSH
• software MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fibronektiny MeSH
• sefarosa MeSH

Fertilization is a multiple step process leading to the fusion of female and male gametes and the formation of a zygote. Besides direct gamete membrane interaction via binding receptors localized on both oocyte and sperm surface, fertilization also involves gamete communication via chemical molecules triggering various signaling pathways. This work focuses on a mouse taste receptor, mTAS1R3, encoded by the Tas1r3 gene, as a potential receptor mediating chemical communication between gametes using the C57BL/6J lab mouse strain. In order to specify the role of mTAS1R3, we aimed to characterize its precise localization in testis and sperm using super resolution microscopy. The testis cryo-section, acrosome-intact sperm released from cauda epididymis and sperm which underwent the acrosome reaction (AR) were evaluated. The mTAS1R3 receptor was detected in late spermatids where the acrosome was being formed and in the acrosomal cap of acrosome intact sperm. AR is triggered in mice during sperm maturation in the female reproductive tract and by passing through the egg surroundings such as cumulus oophorus cells. This AR onset is independent of the extracellular matrix of the oocyte called zona pellucida. After AR, the relocation of mTAS1R3 to the equatorial segment was observed and the receptor remained exposed to the outer surroundings of the female reproductive tract, where its physiological ligand, the amino acid L-glutamate, naturally occurs. Therefore, we targeted the possible interaction in vitro between the mTAS1R3 and L-glutamate as a part of chemical communication between sperm and egg and used an anti-mTAS1R3-specific antibody to block it. We detected that the acrosome reacted spermatozoa showed a chemotactic response in the presence of L-glutamate during and after the AR, and it is likely that mTAS1R3 acted as its mediator.

• Klíčová slova
• L-glutamate, TAS1R family, acrosome reaction, chemoattractant, chemotaxis, gamete, mTAS1R3 receptor, mouse, sperm,
• MeSH
• buněčná diferenciace MeSH
• chemotaxe MeSH
• exprese genu MeSH
• glutamáty metabolismus   MeSH
• interakce spermie a vajíčka * MeSH
• messenger RNA genetika   MeSH
• mezibuněčná komunikace * MeSH
• myši MeSH
• receptory spřažené s G-proteiny genetika   metabolismus   MeSH
• spermie cytologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glutamáty MeSH
• messenger RNA MeSH
• receptory spřažené s G-proteiny MeSH
• taste receptors, type 1 MeSH Prohlížeč

The systemic anaphylactic reaction is a life-threatening allergic response initiated by activated mast cells. Sphingolipids are an essential player in the development and attenuation of this response. De novo synthesis of sphingolipids in mammalian cells is inhibited by the family of three ORMDL proteins (ORMDL1, 2, and 3). However, the cell and tissue-specific functions of ORMDL proteins in mast cell signaling are poorly understood. This study aimed to determine cross-talk of ORMDL2 and ORMDL3 proteins in IgE-mediated responses. To this end, we prepared mice with whole-body knockout (KO) of Ormdl2 and/or Ormdl3 genes and studied their role in mast cell-dependent activation events in vitro and in vivo. We found that the absence of ORMDL3 in bone marrow-derived mast cells (BMMCs) increased the levels of cellular sphingolipids. Such an increase was further raised by simultaneous ORMDL2 deficiency, which alone had no effect on sphingolipid levels. Cells with double ORMDL2 and ORMDL3 KO exhibited increased intracellular levels of sphingosine-1-phosphate (S1P). Furthermore, we found that concurrent ORMDL2 and ORMDL3 deficiency increased IκB-α phosphorylation, degranulation, and production of IL-4, IL-6, and TNF-α cytokines in antigen-activated mast cells. Interestingly, the chemotaxis towards antigen was increased in all mutant cell types analyzed. Experiments in vivo showed that passive cutaneous anaphylaxis (PCA), which is initiated by mast cell activation, was increased only in ORMDL2,3 double KO mice, supporting our in vitro observations with mast cells. On the other hand, ORMDL3 KO and ORMDL2,3 double KO mice showed faster recovery from passive systemic anaphylaxis, which could be mediated by increased levels of blood S1P presented in such mice. Our findings demonstrate that Ormdl2 deficiency potentiates the ORMDL3-dependent changes in mast cell signaling.

• Klíčová slova
• FcϵRI, ORMDL family, mast cells, passive cutaneous anaphylactic reaction, passive systemic anaphylaxis, sphingolipids, sphingosine-1-phosphate,
• MeSH
• anafylaxe etiologie   metabolismus   MeSH
• biologické markery MeSH
• chemotaxe imunologie   MeSH
• cytokiny metabolismus   MeSH
• exprese genu MeSH
• lysofosfolipidy krev   metabolismus   MeSH
• mastocyty imunologie   metabolismus   MeSH
• membránové proteiny chemie   nedostatek   genetika   metabolismus   MeSH
• multigenová rodina MeSH
• myši knockoutované MeSH
• myši MeSH
• náchylnost k nemoci MeSH
• pasivní kožní anafylaxe genetika   imunologie   MeSH
• sekvence aminokyselin MeSH
• sfingolipidy krev   metabolismus   MeSH
• sfingosin analogy a deriváty   krev   metabolismus   MeSH
• signální transdukce * MeSH
• vápník metabolismus   MeSH
• vápníková signalizace MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• cytokiny MeSH
• lysofosfolipidy MeSH
• membránové proteiny MeSH
• ORMDL3 protein, mouse MeSH Prohlížeč
• sfingolipidy MeSH
• sfingosin MeSH
• sphingosine 1-phosphate MeSH Prohlížeč
• vápník MeSH