Escherichia coli A0 34/86 (EcO83) is a probiotic strain used in newborns to prevent nosocomial infections and diarrhoea. This bacterium stimulates both pro- and anti-inflammatory cytokine production and its intranasal administration reduces allergic airway inflammation in mice. Despite its benefits, there are concerns about the use of live probiotic bacteria due to potential systemic infections and gene transfer. Extracellular vesicles (EVs) derived from EcO83 (EcO83-EVs) might offer a safer alternative to live bacteria. This study characterizes EcO83-EVs and investigates their interaction with host cells, highlighting their potential as postbiotic therapeutics. EcO83-EVs were isolated, purified, and characterised following the Minimal Information of Studies of Extracellular Vesicles (MISEV) guidelines. Ex vivo studies conducted in human nasal epithelial cells showed that EcO83-EVs increased the expression of proteins linked to oxidative stress and inflammation, indicating an effective interaction between EVs and the host cells. Further in vivo studies in mice demonstrated that EcO83-EVs interact with nasal-associated lymphoid tissue, are internalised by airway macrophages, and stimulate neutrophil recruitment in the lung. Mechanistically, EcO83-EVs activate the NF-κΒ signalling pathway, resulting in the nitric oxide production. EcO83-EVs demonstrate significant potential as a postbiotic alternative to live bacteria, offering a safer option for therapeutic applications. Further research is required to explore their clinical use, particularly in mucosal vaccination and targeted immunotherapy strategies.

• Klíčová slova
• EVs, Ec083, NF‐κΒ signalling, bacterial extracellular vesicles, macrophage, nitric oxide, postbiotics, probiotic,
• MeSH
• aplikace intranazální * MeSH
• epitelové buňky metabolismus   MeSH
• Escherichia coli * metabolismus   MeSH
• extracelulární vezikuly * metabolismus   MeSH
• lidé MeSH
• lymfoidní tkáň metabolismus   MeSH
• makrofágy metabolismus   MeSH
• myši MeSH
• NF-kappa B metabolismus   MeSH
• oxidační stres MeSH
• plíce mikrobiologie   metabolismus   MeSH
• probiotika * aplikace a dávkování   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• NF-kappa B MeSH

BACKGROUND: The domestic cat (Felis catus) is an important companion animal and is used as a large animal model for human disease. However, the comprehensive study of adaptive immunity in this species is hampered by the lack of data on lymphocyte antigen receptor genes and usage. The objectives of this study were to annotate the feline T cell receptor (TR) loci and to characterize the expressed repertoire in lymphoid organs of normal cats using high-throughput sequencing. RESULTS: The Felis catus TRG locus contains 30 genes: 12 TRGV, 12 TRGJ and 6 TRGC, the TRB locus contains 48 genes: 33 TRBV, 2 TRBD, 11 TRBJ, 2 TRBC, the TRD locus contains 19 genes: 11 TRDV, 2 TRDD, 5 TRDJ, 1 TRDC, and the TRA locus contains 127 genes: 62 TRAV, 64 TRAJ, 1 TRAC. Functional feline V genes form monophyletic clades with their orthologs, and clustering of multimember subgroups frequently occurs in V genes located at the 5' end of TR loci. Recombination signal (RS) sequences of the heptamer and nonamer of functional V and J genes are highly conserved. Analysis of the TRG expressed repertoire showed preferential intra-cassette over inter-cassette rearrangements and dominant usage of the TRGV2-1 and TRGJ1-2 genes. The usage of TRBV genes showed minor bias but TRBJ genes of the second J-C-cluster were more commonly rearranged than TRBJ genes of the first cluster. The TRA/TRD V genes almost exclusively rearranged to J genes within their locus. The TRAV/TRAJ gene usage was relatively balanced while the TRD repertoire was dominated by TRDJ3. CONCLUSIONS: This is the first description of all TR loci in the cat. The genomic organization of feline TR loci was similar to that of previously described jawed vertebrates (gnathostomata) and is compatible with the birth-and-death model of evolution. The large-scale characterization of feline TR genes provides comprehensive baseline data on immune repertoires in healthy cats and will facilitate the development of improved reagents for the diagnosis of lymphoproliferative diseases in cats. In addition, these data might benefit studies using cats as a large animal model for human disease.

• Klíčová slova
• Expressed repertoire, Feline, T cell receptor, TRA/TRD, TRB, TRG, V/J usage,
• MeSH
• adaptivní imunita genetika   MeSH
• fylogeneze MeSH
• genetické lokusy genetika   MeSH
• genomika metody   MeSH
• kočky genetika   imunologie   MeSH
• lidé MeSH
• lymfoidní tkáň metabolismus   MeSH
• receptory antigenů T-buněk klasifikace   genetika   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• zvířata MeSH
• Check Tag
• kočky genetika   imunologie   MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• receptory antigenů T-buněk MeSH

The proliferation, differentiation and function of immune cells in vertebrates, as well as in the invertebrates, is regulated by distinct signalling pathways and crosstalk with systemic and cellular metabolism. We have identified the Lime gene (Linking Immunity and Metabolism, CG18446) as one such connecting factor, linking hemocyte development with systemic metabolism in Drosophila. Lime is expressed in larval plasmatocytes and the fat body and regulates immune cell type and number by influencing the size of hemocyte progenitor populations in the lymph gland and in circulation. Lime mutant larvae exhibit low levels of glycogen and trehalose energy reserves and they develop low number of hemocytes. The low number of hemocytes in Lime mutants can be rescued by Lime overexpression in the fat body. It is well known that immune cell metabolism is tightly regulated with the progress of infection and it must be supported by systemic metabolic changes. Here we demonstrate that Lime mutants fails to induce such systemic metabolic changes essential for the larval immune response. Indeed, Lime mutants are not able to sustain high numbers of circulating hemocytes and are compromised in the number of lamellocytes produced during immune system challenge, using a parasitic wasp infection model. We therefore propose the Lime gene as a novel functional link between systemic metabolism and Drosophila immunity.

• Klíčová slova
• CG18446, Drosophila, Immunity, Leptopilina boulardii, Metabolism,
• MeSH
• buněčná diferenciace MeSH
• Drosophila melanogaster imunologie   metabolismus   MeSH
• energetický metabolismus MeSH
• hemocyty cytologie   metabolismus   MeSH
• imunita * MeSH
• jaderné proteiny metabolismus   MeSH
• larva metabolismus   MeSH
• lymfoidní tkáň metabolismus   MeSH
• mutace genetika   MeSH
• proteiny Drosophily metabolismus   MeSH
• tukové těleso metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• jaderné proteiny MeSH
• Lime protein, Drosophila MeSH Prohlížeč
• proteiny Drosophily MeSH

Monocytes play an essential role in the defense against bacterial pathogens. Bone marrow (BM) and peripheral blood (PB) monocytes in pigs consist of the main "steady-state" subpopulations: CD14 hi/CD163-/SLA-DR- and CD14 low/CD163+/SLA-DR+. During inflammation, the subpopulation of "inflammatory" monocytes expressing very high levels of CD163, but lacking the SLA-DR molecule (being CD14 low/CD163+/SLA-DR-) appears in the BM and PB and replaces the CD14 low/CD163+/SLA-DR+ subpopulation. However, current knowledge of monocyte migration into inflamed tissues in pigs is limited. The aim of the present study was to evaluate the distribution of "inflammatory" CD14 low/CD163+/SLA-DR- monocytes during experimental inflammation induced by Actinobacillus pleuropneumoniae (APP) and a possible role for chemokines in attracting "inflammatory" CD14 low/CD163+/SLA-DR- monocytes into the tissues. Monocyte subpopulations were detected by flow cytometry. Chemokines and chemokine receptors were detected by RT-qPCR. The "steady-state" monocytes were found in the BM, PB, spleen and lungs of control pigs. After APP-infection, "inflammatory" monocytes replaced the "steady-state" subpopulation in BM, PB, spleen and moreover, they appeared in an unaffected area, demarcation zone and necrotic area of the lungs and in tracheobronchial lymph nodes. They did not appear in mesenteric lymph nodes. Levels of mRNA for various chemokines with their appropriate receptors were found to be elevated in BM (CCL3-CCR1/CCR5, CCL8-CCR2/CCR5, CCL19-CCR7), necrotic area of the lungs (CCL3-CCR1, CCL5-CCR1/CCR3, CCL11-CCR3, CCL22/CCR4) and tracheobronchial lymph nodes (CCL3-CCR1) and therefore they could play a role in attracting monocytes into inflamed tissues. In conclusion, "inflammatory" monocytes appear in different lymphoid tissues and the lungs after APP infection in pigs. Various chemokines could drive this process.

• MeSH
• Actinobacillus pleuropneumoniae fyziologie   MeSH
• antigen CD163 MeSH
• antigeny diferenciační myelomonocytární metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• chemokiny genetika   metabolismus   MeSH
• infekce bakteriemi rodu Actinobacillus imunologie   mikrobiologie   veterinární   MeSH
• lymfoidní tkáň metabolismus   MeSH
• messenger RNA genetika   MeSH
• monocyty cytologie   metabolismus   MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• plíce metabolismus   MeSH
• prasata MeSH
• průtoková cytometrie veterinární   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• receptory chemokinů genetika   metabolismus   MeSH
• zánět mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigen CD163 MeSH
• antigeny diferenciační myelomonocytární MeSH
• CD antigeny MeSH
• chemokiny MeSH
• messenger RNA MeSH
• receptory buněčného povrchu MeSH
• receptory chemokinů MeSH

OBJECTIVE: Experimental and clinical studies have shown that autoimmunity-causing diabetes may be abrogated by immune intervention. Several anti-T-lymphocyte antibodies focus on distinct T-cell targets. We tested the effect of murine anti-thymocyte globulin (ATG; Genzyme, Framingham, MA) in peripheral lymphoid organs of non-obese diabetic (NOD) mice after the onset of hyperglycemia. METHODS: Diabetic NOD mice were treated with two doses of ATG (1 mg totally) or maintained without treatment as controls. Blood glucose levels were monitored twice a week. The mice were terminated at day 0, 7, 14, or 28 after the initiation of the study. Subpopulations of T-lymphocytes and FoxP3+ (forkhead box P3 positive) regulatory T-cells were analyzed among elements isolated from the spleen and pancreatic lymph nodes. RESULTS: Mice with blood glucose levels greater than 13 mmol/L were included in the study. Diabetes remission occurred in 16% (3/19) of mice treated with ATG. Only one case of remission was observed in the control group (6%; 1/16). ATG therapy a significantly decreased the CD8+/CD4+ T-lymphocyte ratio. Among splenocytes, a significant difference was detected only on day 7 (0.069 versus 0.198 T-lymphocyte ratio); in lymph nodes, a decrease was observed on day 28 (0.21 versus 0.51 T-lymphocytes ratio). The regulatory T-cells population increased after ATG administration compared with the control group at day 7 (16.2% versus 10.8% in CD4+ splenocytes; 20.7% versus 10.3% in CD4+ lymph node cells). However, the increased FoxP3+ cell population was not durable. CONCLUSIONS: ATG treatment of diabetic NOD mice showed an immunoregulatory effect in peripheral lymphoid tissue with a significantly deceased CD8+/CD4+ ratio, which, however, did not normalize the metabolic parameters in a short period after the onset of overt diabetes.

• MeSH
• antilymfocytární sérum terapeutické užití   MeSH
• autoimunita MeSH
• časové faktory MeSH
• diabetes mellitus 1. typu imunologie   terapie   MeSH
• experimentální diabetes mellitus imunologie   terapie   MeSH
• glukózový toleranční test MeSH
• hyperglykemie imunologie   MeSH
• imunitní systém MeSH
• krevní glukóza metabolismus   MeSH
• lymfoidní tkáň metabolismus   MeSH
• myši inbrední NOD MeSH
• myši MeSH
• průtoková cytometrie metody   MeSH
• výsledek terapie MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antilymfocytární sérum MeSH
• krevní glukóza MeSH

Due to the persisting threat of development of new highly pathogenic influenza A subtypes, a mucosal vaccination which would induce a potent and cross-protective reaction is desirable. We succeeded in mucosal immunization of mice with an inactivated influenza A virus by using delipidated Bacillus firmus (DBF) as adjuvant. The mechanism of adjuvant effect was followed in NALT by comparing the response after intranasal immunization by inactivated influenza virus type A (H1N1) alone, adjuvant alone (DBF), or by a mixture of virus+DBF. Expression of selected gene groups was tested via qPCR at 7 different time-points: cytokines (IL-2, IFN-γ, IL-4, IL-6, and IL-10), type I interferons (IFN-α4, IFN-α11, IFN-α12, and IFN-β), toll-like receptors (TLR2, TLR3, TLR7, and TLR9), iNOS and CCR7. Intranasally administered DBF and the mixture of virus+DBF induced an elevated expression of IFN-γ, IL-6 and IL-10 cytokines, type I interferons, iNOS, and pDC markers in NALT. Multimarker qPCR data was analyzed by relative quantification and by principal component analysis. DBF has been shown to be a very efficient adjuvant for the stimulation of innate immunity after IN immunization. DBF accelerated, increased, and prolonged the antiviral response.

• MeSH
• adjuvancia imunologická aplikace a dávkování   MeSH
• analýza hlavních komponent MeSH
• aplikace intranazální MeSH
• Bacillus imunologie   MeSH
• časové faktory MeSH
• cytokiny genetika   MeSH
• exprese genu účinky léků   MeSH
• imunizace metody   MeSH
• interferon typ I genetika   MeSH
• interleukin-10 genetika   MeSH
• interleukin-2 genetika   MeSH
• lymfoidní tkáň metabolismus   MeSH
• membránové glykoproteiny genetika   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nazofarynx metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• synergismus léků MeSH
• toll-like receptor 2 genetika   MeSH
• toll-like receptor 3 genetika   MeSH
• toll-like receptor 7 genetika   MeSH
• toll-like receptor 9 genetika   MeSH
• toll-like receptory genetika   MeSH
• vakcíny proti chřipce aplikace a dávkování   imunologie   MeSH
• virus chřipky A, podtyp H1N1 imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adjuvancia imunologická MeSH
• cytokiny MeSH
• interferon typ I MeSH
• interleukin-10 MeSH
• interleukin-2 MeSH
• membránové glykoproteiny MeSH
• Tlr2 protein, mouse MeSH Prohlížeč
• TLR3 protein, mouse MeSH Prohlížeč
• Tlr7 protein, mouse MeSH Prohlížeč
• toll-like receptor 2 MeSH
• toll-like receptor 3 MeSH
• toll-like receptor 7 MeSH
• toll-like receptor 9 MeSH
• toll-like receptory MeSH
• vakcíny proti chřipce MeSH

A key molecule necessary for activation of T lymphocytes through their antigen-specific T cell receptor (TCR) is the transmembrane adaptor protein LAT (linker for activation of T cells). Upon TCR engagement, LAT becomes rapidly tyrosine phosphorylated and then serves as a scaffold organizing a multicomponent complex that is indispensable for induction of further downstream steps of the signaling cascade. Here we describe the identification and preliminary characterization of a novel transmembrane adaptor protein that is structurally and evolutionarily related to LAT and is expressed in B lymphocytes, natural killer (NK) cells, monocytes, and mast cells but not in resting T lymphocytes. This novel transmembrane adaptor protein, termed NTAL (non-T cell activation linker) is the product of a previously identified WBSCR5 gene of so far unknown function. NTAL becomes rapidly tyrosine-phosphorylated upon cross-linking of the B cell receptor (BCR) or of high-affinity Fcgamma- and Fc epsilon -receptors of myeloid cells and then associates with the cytoplasmic signaling molecules Grb2, Sos1, Gab1, and c-Cbl. NTAL expressed in the LAT-deficient T cell line J.CaM2.5 becomes tyrosine phosphorylated and rescues activation of Erk1/2 and minimal transient elevation of cytoplasmic calcium level upon TCR/CD3 cross-linking. Thus, NTAL appears to be a structural and possibly also functional homologue of LAT in non-T cells.

• MeSH
• adaptorové proteiny signální transdukční * MeSH
• aktivace lymfocytů MeSH
• B-lymfocyty imunologie   metabolismus   MeSH
• buněčné linie MeSH
• buňky NK imunologie   metabolismus   MeSH
• fosfoproteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• fosforylace MeSH
• lidé MeSH
• lymfoidní tkáň cytologie   metabolismus   MeSH
• membránové mikrodomény chemie   metabolismus   MeSH
• membránové proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• monocyty imunologie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• proteiny * MeSH
• receptory antigenů B-buněk metabolismus   MeSH
• receptory Fc metabolismus   MeSH
• receptory IgE metabolismus   MeSH
• receptory IgG metabolismus   MeSH
• sekvence aminokyselin MeSH
• signální transdukce * MeSH
• T-lymfocyty imunologie   metabolismus   MeSH
• transportní proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adaptorové proteiny signální transdukční * MeSH
• fosfoproteiny MeSH
• LAT protein, human MeSH Prohlížeč
• Lat protein, mouse MeSH Prohlížeč
• LAT2 protein, human MeSH Prohlížeč
• LAT2 protein, mouse MeSH Prohlížeč
• membránové proteiny MeSH
• proteiny * MeSH
• receptory antigenů B-buněk MeSH
• receptory Fc MeSH
• receptory IgE MeSH
• receptory IgG MeSH
• transportní proteiny MeSH

In this study, the immunohistochemical expression of bcl-2 protein in benign and malignant lymphoid aggregates in bone marrow biopsies was investigated in order to estimate its significance in distinguishing the biologic nature of the aggregates. Paraffin-embedded tissues of 46 bone marrow biopsies were stained with a monoclonal antibody to bcl-2 protein using the supersensitive streptavidin biotin immunoperoxidase method after a microwave heating of the sections. Bcl-2 protein immunoreactivity was observed in various proportions of lymphoid cells in both reactive and malignant lymphoid bone marrow aggregates. The percentage of bcl-2 positive cells in malignant aggregates was substantially higher (mean value 78%) than that observed in reactive nodules (mean value 60%). The presence of bcl-2 protein has been confirmed both in malignant and benign bone marrow lymphoid aggregates. Thus, the bcl-2 protein expression should not be used as a discriminating criterion for the malignant nature of lymphoid aggregates.

• MeSH
• diferenciální diagnóza MeSH
• hyperplazie MeSH
• kostní dřeň chemie   metabolismus   patologie   MeSH
• lidé MeSH
• lymfoidní tkáň chemie   metabolismus   patologie   MeSH
• nádorové biomarkery chemie   MeSH
• nehodgkinský lymfom chemie   diagnóza   MeSH
• protoonkogenní proteiny c-bcl-2 biosyntéza   chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nádorové biomarkery MeSH
• protoonkogenní proteiny c-bcl-2 MeSH

The effect of indomethacin--a nonsteroid antiinflammatory drug with potential antitumor activity--on the development of radiation-induced changes was followed in blood, bone marrow, spleen, thymus and testes of rats. Indomethacin administered in drinking water (0.7-1.0 mg/kg per day) during a continuous 7-day irradiation with gamma rays (dose rate of 2.055 Gy/day, total accumulated dose of 14.385 Gy) caused a higher and more rapid incorporation of 3H-thymidine into blood DNA, and an increase in blood RNA concentration. The results suggest some stimulation of hemopoiesis recovery by indomethacin treatment in continuously irradiated rats.

• MeSH
• časové faktory MeSH
• dávka záření MeSH
• DNA analýza   krev   MeSH
• inbrední kmeny potkanů MeSH
• indomethacin farmakologie   MeSH
• kostní dřeň účinky léků   metabolismus   účinky záření   MeSH
• krysa rodu Rattus MeSH
• lymfoidní tkáň účinky léků   metabolismus   účinky záření   MeSH
• radioprotektivní látky * MeSH
• RNA analýza   krev   MeSH
• spektrofotometrie MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA MeSH
• indomethacin MeSH
• radioprotektivní látky * MeSH
• RNA MeSH

Further study of the response to chronic stress stimulation in the early postnatal phase showed that the i.p. injection of physiological saline (stress stimulation) induced lymphopenia, a 50% decrease in the incorporation of 3H-leucine into isolated lymphocytes and a decrease in the weight of the thymus in 7-day-old male rats. No such changes were observed in adult animals. If repeated doses of phenobarbital were administered to stressed young rats, however, lymphopenia did not occur and the rate of the incorporation of 3H-leucine into isolated lymphocytes was not different from the control value; the protein content of the lymphocytes was significantly raised, however. In adult animals, phenobarbital increased the rate of incorporation of 3H-leucine into the lymphocytes. The repeated administration of phenobarbital reduced the weight of the thymus in both young and adult animals, but a decrease in spleen weight was recorded only in the young animals. A single i.p. injection of ACTH or dexamethasone caused lymphopenia and slowed down the incorporation of 3H-leucine into the lymphocytes of both young and adult animals. The results show that the striking decrease observed in the rate of the liver metabolism of corticosterone in suckling young rats not injured by repeated stress stimulation is accompanied by significant changes in the lymphoid tissue.

• MeSH
• fenobarbital farmakologie   MeSH
• fyziologický stres metabolismus   patologie   MeSH
• inbrední kmeny potkanů MeSH
• kortikosteron metabolismus   MeSH
• krysa rodu Rattus MeSH
• lymfocyty účinky léků   metabolismus   MeSH
• lymfoidní tkáň účinky léků   metabolismus   patologie   MeSH
• proteiny metabolismus   MeSH
• věkové faktory MeSH
• velikost orgánu účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fenobarbital MeSH
• kortikosteron MeSH
• proteiny MeSH