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Following organ transplantation, it is essential that immune tolerance is induced in the graft recipient to reduce the risk of rejection and avoid complications associated with the long-term use of immunosuppressive drugs. Immature dendritic cells (DCs) are considered to promote transplant tolerance and may minimize the risk of graft rejection. The aim of the study was to evaluate the effects of immunosuppressive agents: rapamycin (Rapa) and cyclosporine A (CsA) on generation of human tolerogenic DCs (tolDCs) and also to evaluate the ability of these cells to induce mechanisms of immune tolerance. tolDCs were generated in the environment of Rapa or CsA. Next, we evaluated the effects of these agents on surface phenotypes (CD11c, MHC II, CD40, CD80, CD83, CD86, CCR7, TLR2, TLR4), cytokine production (IL-4, IL-6, IL-10, IL-12p70, TGF-ß), phagocytic capacity and resistant to lipopolysaccharide activation of these DCs. Moreover, we assessed ability of such tolDCs to induce T cell activation and apoptosis, Treg differentiation and production of Th1- and Th2-characteristic cytokine profile. Data obtained in this study demonstrate that rapamycin is effective at generating maturation-resistant tolDCs, however, does not change the ability of these cells to induce mechanisms of immune tolerance. In contrast, CsA affects the ability of these cells to induce mechanisms of immune tolerance, but is not efficient at generating maturation-resistant tolDCs.
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Ciclosporina , Sirolimus , Células Dendríticas , Humanos , Tolerancia Inmunológica , InmunosupresoresRESUMEN
Endometriosis is a common gynecological disorder characterized by the presence of endometrial-like tissue outside the uterus. The disease is associated with disturbed local and systemic immunity. It has been reported that the proportion of CD4+CD25highFOXP3+ Treg cells may be significantly increased in the peritoneal fluid of patients with endometriosis. Therefore, the aim of our study was to investigate whether the proportions of Treg cells in the peritoneal cavity of patients with endometriosis are related to the chemotactic and stimulatory activity of the local peritoneal milieu. The peritoneal fluid was collected from 13 women with ovarian endometriosis and 12 control women without the disease. T cell populations were analyzed by flow cytometry, cytokines and chemokines were evaluated using the cytometric bead kit, and cell chemotaxis was studied by cell migration assay. We confirmed that the proportions of Treg cells are increased in the peritoneal fluid of women with endometriosis as compared to the control women. Endometriosis was also associated with elevated concentrations of IL-6, IL-10, and TGF-ß1/2 as well as CCL20, CXCL8, CXCL9, and CXCL10. We did not reveal any changes in the proportion of peritoneal Th17 cells and concentrations of IL-17A. Peritoneal Treg cells positively correlated with concentrations of TGF-ß, IL-10, and CCL20. Endometriotic peritoneal fluid stimulated chemotaxis of both CD4+ and Treg cells. This chemotactic activity positively correlated with concentrations of CCL20. CCL20 stimulated the migration of Treg cells, and the chemotactic activity of the endometriotic peritoneal fluid was inhibited by neutralizing anti-CCL20 antibodies. These results imply that increased proportions of the peritoneal Treg cells in women with endometriosis may result from attraction and activation by local chemokines and cytokines, especially CCL20 and TGF-ß. Since Treg cells contribute to the immunopathogenesis of endometriosis, their chemotaxis and activation may be considered as a target for therapeutic intervention.
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Endometriosis is a common gynaecological disorder characterized by the ectopic growth of endometrial tissue outside the uterine cavity. It is associated with chronic pelvic inflammation and autoimmune reactivity manifesting by autoantibody production and abrogated cellular immune responses. Endometriotic peritoneal fluid contains various infiltrating leucocyte populations and a bulk of proinflammatory and immunoregulatory cytokines. However, the nature and significance of the peritoneal milieu in women with endometriosis still remains obscure. Therefore, the aim of the present study was to investigate the immunoregulatory activity of the peritoneal fluid (PF) from women with endometriosis. The peritoneal fluid samples were collected during laparoscopic surgery from 30 women with and without endometriosis. Immunoregulatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) and chemokines (CCL2, CCL5, CXCL8 and CXCL9) were evaluated in PF and culture supernatants generated by unstimulated and CD3/CD28/IL-2-stimulated CD4+ T cells cultured in the presence of PF. The effect of PF on the generation of Treg and Th17 cells in CD4+ T cell cultures, as well as the natural cytotoxic activity of peripheral blood mononuclear cells, was also investigated. Concentrations of IL-6, IL-10, CCL2, CXCL8 and CXCL9 were significantly upregulated in the PF from women with endometriosis when compared to control women, whereas concentrations of other cytokines and chemokines were unaffected. The culturing of unstimulated and CD3/CD28/IL-2-stimulated CD4+ T cells in the presence of endometriotic PF resulted in the downregulation of their IL-2, IFN-γ, IL-17A and TNF production as compared to culture medium alone. On the other side, endometriotic PF significantly stimulated the production of IL-4 and IL-10. Endometriotic PF also stimulated the release of CCL2 and CXCL8, whereas the production of CCL5 and CXCL9 was downregulated. Endometriotic PF stimulated the generation of Treg cells and had an inhibitory effect on the generation of Th17 cells in cultures of CD4+ T cells. It also inhibited the NK cell cytotoxic activity of the peripheral blood lymphocytes. These results strongly imply that the PF from patients with endometriosis has immunoregulatory/immunosuppressive activity and shifts the Th1/Th2 cytokine balance toward the Th2 response, which may account for deviation of local and systemic immune responses. However, a similar trend, albeit not a statistically significant one, was also observed in case of PF from women without endometriosis, thus suggesting that peritoneal milieu may in general display some immunoregulatory/immunosuppressive properties. It should be stressed, however, that our present observations were made on a relatively small number of PF samples and further studies are needed to reveal possible mechanism(s) responsible for this phenomenon.
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Líquido Ascítico/inmunología , Quimiocinas/metabolismo , Endometriosis/inmunología , Tolerancia Inmunológica , Células Th2/inmunología , Adulto , Líquido Ascítico/metabolismo , Estudios de Casos y Controles , Células Cultivadas , Femenino , Humanos , Persona de Mediana Edad , Linfocitos T Reguladores/inmunología , Células TH1/inmunología , Células Th17/inmunología , Regulación hacia Arriba , Adulto JovenRESUMEN
Atrial fibrillation (AF) and atherosclerotic disease are independent risk factors for acute ischaemic stroke (AIS). The optimal biological marker which could allow differentiation between AF and non-AF AIS patients is still not available. AIM OF THE STUDY: Aim of the present study was to investigate the role of pentosidine as a potential biological marker for AF in an AIS patient group. MATERIALS AND METHODS: Sixty-three acute ischaemic hemispheric stroke patients were recruited and divided into two groups according to the presumed underlying mechanism: with or without atrial rhythm disorders. Ten healthy volunteers were a reference group for serum level of pentosidine. Carotid artery ultrasound was performed, and common carotid artery stiffness and intima-media thickness were measured. Serum levels of pentosidine and selected routine biochemical risk factors for atherosclerosis (cholesterol and its lipoprotein fractions, homocysteine) were examined. RESULTS: A higher serum level of pentosidine was observed in patients without atrial fibrillation (1,509 ± 485.13pmol/ml); a statistically significant difference was observed compared to the reference group (1,041.52 ± 411.17pmol/ml; p = 0.01), but not the AF patients (1,438.19 ± 495.97pmol/ml; p = 0.59). No significant difference in the non-AF group compared to the AF group for carotid intima-media thickness (IMT)/stiffness and pentosidine serum level was recorded. CONCLUSIONS AND CLINICAL IMPLICATIONS: A higher serum level of pentosidine was observed in AIS patients without atrial fibrillation compared to the healthy volunteers. According to the results of the present study, no difference between these patients in the selected risk factors of atherosclerosis were observed. Further studies are needed to identify a reliable marker of AF that would bring added value to the standard diagnostic workup after acute ischaemic stroke.
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Fibrilación Atrial , Isquemia Encefálica , Accidente Cerebrovascular , Arginina/análogos & derivados , Grosor Intima-Media Carotídeo , Humanos , Lisina/análogos & derivados , Factores de RiesgoRESUMEN
The original article has been published without acknowledgment section. The acknowledgement section is given below for your reading.
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Regulation of immune response was found to play an important role in the course of many diseases such as autoimmune diseases, allergy, malignancy, organ transplantation. The studies on immune regulation focus on the role of regulatory cells (Tregs, Bregs, regulatory myeloid cells) in these disorders. The number and function of Tregs may serve as a marker of disease activity. As in allergy, the depletion of Tregs is observed and the results of allergen-specific immunotherapy could be measured by an increase in the population of IL-10+ regulatory cells. On the basis of the knowledge of anti-cancer immune response regulation, new directions in therapy of tumors are introduced. As the proportion of regulatory cells is increased in the course of neoplasm, the therapeutic action is directed at their inhibition. The depletion of Tregs may be also achieved by an anti-check-point blockade, anti-CD25 agents, and inhibition of regulatory cell recruitment to the tumor site by affecting chemokine pathways. However, the possible favorable role of Tregs in cancer development is considered and the plasticity of immune regulation should be taken into account. The new promising direction of the treatment based on regulatory cells is the prevention of transplant rejection. A different way of production and implementation of classic Tregs as well as other cell types such as double-negative cells, Bregs, CD4+ Tr1 cells are tested in ongoing trials. On the basis of the results of current studies, we could show in this review the significance of therapies based on regulatory cells in different disorders.
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Linfocitos B Reguladores/inmunología , Rechazo de Injerto/inmunología , Hipersensibilidad/inmunología , Neoplasias/inmunología , Linfocitos T Reguladores/inmunología , Animales , Autoinmunidad , Humanos , Tolerancia Inmunológica , TrasplanteRESUMEN
Immunosuppressive activity of regulatory T and B cells is critical to limit autoimmunity, excessive inflammation, and pathological immune response to conventional antigens or allergens. Both types of regulatory cells are intensively investigated, however, their development and mechanisms of action are still not completely understood. Both T and B regulatory cells represent highly differentiated populations in terms of phenotypes and origin, however, they use similar mechanisms of action. The most investigated CD4+CD25+ regulatory T cells are characterized by the expression of Foxp3+ transcription factor, which is not sufficient to maintain their lineage stability and suppressive function. Currently, it is considered that specific epigenetic changes are critical for defining regulatory T cell stability in the context of their suppressive function. It is not yet known if similar epigenetic regulation determines development, lineage stability, and function of regulatory B cells. Phenotype diversity, confirmed or hypothetical developmental pathways, multiple mechanisms of action, and role of epigenetic changes in these processes are the subject of this review.
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Linfocitos B Reguladores/inmunología , Inmunidad Celular , Linfocitos T Reguladores/inmunología , Animales , Biodiversidad , Diferenciación Celular , Linaje de la Célula , Epigénesis Genética , Factores de Transcripción Forkhead/metabolismo , Humanos , Terapia de Inmunosupresión , Fenotipo , Receptores de Antígenos de Linfocitos B/metabolismo , Receptores de Antígenos de Linfocitos T/metabolismoRESUMEN
The objective of this study was to evaluate the effects of A3R phage and Staphylococcus aureus lysate obtained after phage infection on neutrophil degranulation. The exocytosis of primary and secondary granules from neutrophils was investigated in vitro in whole blood specimens by flow cytometry based on the expression of specific markers of exocytosis (CD63 for primary granules and CD66b for secondary granules). We found that both A3R and S. aureus lysate had no significant effect on the exocytosis of primary and secondary granules. These data suggest that neither A3R virions nor any products of phage-induced lysis of S. aureus are likely to induce neutrophil degranulation in patients who are treated with phage preparations. Since neutrophil granules contain some potentially toxic proteins, our results provide an important argument for the safety of phage therapy. Moreover, these data indicate that the induction of neutrophil degranulation is not likely to contribute to antibacterial effects of phages.
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Bacteriólisis , Degranulación de la Célula , Neutrófilos/inmunología , Fagos de Staphylococcus/inmunología , Staphylococcus aureus/inmunología , Adulto , Exocitosis , Citometría de Flujo , Voluntarios Sanos , Humanos , Terapia de Fagos/efectos adversosRESUMEN
A growing body of data shows that bacteriophages can interact with different kinds of immune cells. The objective of this study was to investigate whether T4 bacteriophage and T4-generated Escherichia coli lysate affect functions of monocytes, the key population of immune cells involved in antibacterial immunity. To that end, we evaluated how T4 and E. coli lysate influence the expression of main costimulatory molecules including CD40, CD80 and CD86, TLR2, TLR4 on monocytes, as well as the production of IL-6 and IL-12 in cultures of peripheral blood mononuclear cells (PBMCs). Separate experiments were performed on unactivated and LPS-activated PBMCs cultures. Both studied preparations significantly increased the percentage of CD14(+)CD16(-)CD40(+) and CD14(+)CD16(-)CD80(+) monocytes in unactivated PBMCs cultures, as well as the concentration of IL-6 and IL-12 in culture supernates. However, neither purified T4 nor E. coli lysate had any significant effect on monocytes in LPS-activated PBMCs cultures. We conclude that LPS-activated monocytes are unresponsive to phages and products of phage-induced lysis of bacteria. This study is highly relevant to phage therapy because it suggests that in patients with infections caused by Gram-negative bacteria the administration of phage preparations to patients and lysis of bacteria by phages are not likely to overly stimulate monocytes.
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Bacteriophages (phages) are viruses of bacteria. Here we evaluated the effects of T4 and A3/R bacteriophages, as well as phage-generated bacterial lysates, on differentiation of human myeloid dendritic cells (DCs) from monocytes. Neither of the phages significantly reduced the expression of markers associated with differentiation of DCs and their role in the activation of T cells (CD40, CD80, CD83, CD86, CD1c, CD11c, MHC II, PD-L1, PD-L2, TLR2, TLR4, and CCR7) and phagocytosis receptors (CD64 and DEC-205). By contrast, bacterial lysate of T4 phage significantly decreased the percentages of DEC-205- and CD1c-positive cells. The percentage of DEC-205-positive cells was also significantly reduced in DCs differentiated in the presence of lysate of A3/R phage. Thus while bacteriophages do not substantially affect differentiation of DCs, some products of phage-induced lysis of bacterial cells may influence the differentiation and potentially also some functions of DCs. Our results have important implications for phage therapy of bacterial infections because during infections monocytes recruited to the site of inflammation are an important source of inflammatory DCs.
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Human CD8(+)CD28(-) T suppressor cells were previously shown to be involved in the control of the immune response to transplanted allografts. It seems essential to examine how immunosuppressive drugs influence these cells. However, the CD8(+)CD28(-) population contains both suppressor (Ts) and cytotoxic (Tc) T cells, and the phenotype of the Ts subpopulation has not been identified explicitly. It is proposed that the transcription factor FOXP3 may be helpful in distinguishing the Ts and Tc subpopulations. The aim of this study was to evaluate the influence of the immunosuppressive drugs cyclosporine A (CsA) and rapamycin (RAPA) on the level, suppressor properties, and phenotype of human CD8(+)CD28(-) T cells in vitro. The model used was the mixed leukocyte reaction performed with peripheral blood mononuclear cells from healthy volunteers. It was observed that CD8(+)CD28(-) T cells from cultures with CsA or RAPA had similar suppressor properties to cells from control cultures, although the drugs influenced the expression of FOXP3. CsA and RAPA did not interfere with the suppressor properties of human CD8(+)CD28(-) T cells in vitro, although they affected the expression of the FOXP3 molecule.
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Antígenos CD28/metabolismo , Linfocitos T CD8-positivos/citología , Ciclosporina/administración & dosificación , Inmunosupresores/administración & dosificación , Sirolimus/administración & dosificación , Adolescente , Adulto , Linfocitos T CD8-positivos/inmunología , Ciclosporina/química , Factores de Transcripción Forkhead/metabolismo , Rechazo de Injerto/inmunología , Humanos , Inmunosupresores/química , Inmunosupresores/uso terapéutico , Leucocitos Mononucleares/inmunología , Activación de Linfocitos , Prueba de Cultivo Mixto de Linfocitos , Persona de Mediana Edad , Fenotipo , Sirolimus/química , Linfocitos T/inmunología , Adulto JovenRESUMEN
N-glycosylation plays an important role in the majority of physiological and pathological processes occurring in the immune system. Alteration of the type and abundance of glycans is an element of lymphocyte differentiation; it is also common in the development of immune-mediated inflammatory diseases. The N-glycosylation process is very sensitive to different environmental agents, among them the pharmacological environment of immunosuppressive drugs. Some results show that high-mannose oligosaccharides have the ability to suppress different stages of the immune response. We evaluated the effects of cyclosporin A (CsA) and rapamycin (Rapa) on high-mannose/hybrid-type glycosylation in human leukocytes activated in a two-way mixed leukocyte reaction (MLR). CsA significantly reduced the number of leukocytes covered by high-mannose/hybrid N-glycans, and the synergistic action of CsA and Rapa led to an increase of these structures on the remaining leukocytes. This is the first study indicating that ß1 and ß3 integrins bearing high-mannose/hybrid structures are affected by Rapa and CsA. Rapa taken separately and together with CsA changed the expression of ß1 and ß3 integrins and, by regulating the protein amount, increased the oligomannose/hybrid-type N-glycosylation on the leukocyte surface. We suggest that the changes in the glycosylation profile of leukocytes may promote the development of tolerance in transplantation.
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Inmunosupresores/farmacología , Leucocitos/metabolismo , Manosa/metabolismo , Polisacáridos/metabolismo , Recuento de Células , Ciclosporina/farmacología , Sinergismo Farmacológico , Glicoproteínas/metabolismo , Humanos , Integrinas/metabolismo , Leucocitos/efectos de los fármacos , Lectinas de Unión a Manosa/metabolismo , Lectinas de Plantas/metabolismo , Subunidades de Proteína/metabolismo , Sirolimus/farmacología , Espectrometría de Masas en TándemRESUMEN
STUDY QUESTION: Is endometriosis associated with changes in CD4⺠CD25⺠FOXP3⺠regulatory T cells (Treg cells)? SUMMARY ANSWER: Endometriosis is associated with disturbed compartmentalization of CD25(high) FOXP3⺠Treg cells. WHAT IS KNOWN ALREADY: Endometriosis is associated with an abrogated immune response and displays some features of an autoimmune disorder. Treg cells play a part in the development of autoimmune reactions; however, their role in pathogenesis of endometriosis is still poorly recognized. STUDY DESIGN, SIZE AND DURATION: Case-control study comparing 17 women with laparoscopically and histopathologically confirmed ovarian endometriosis with 15 control women without visible endometriosis foci, pelvic inflammation or related pathology who were subjected to laparoscopic surgery between 2010 and 2011. PARTICIPANTS/MATERIALS, SETTING AND METHODS: Peripheral blood and peritoneal fluid were collected during laparoscopy and T cell subpopulations were analysed by flow cytometry using specific monoclonal antibodies recognizing CD4âº, CD25⺠and FOXP3⺠markers. MAIN RESULTS: The percentage of CD25(high) FOXP3⺠Treg cells was significantly decreased in the peripheral blood of women with ovarian endometriosis compared with control women. On the other hand, the proportion of these cells was significantly increased in the peritoneal fluid of women with endometriosis. LIMITATIONS, REASONS FOR CAUTION: The present study is limited to patients with ovarian endometrioma and further investigations are needed, including patients with lower grade of endometriosis. WIDER IMPLICATIONS OF THE FINDINGS: The present results suggest that Treg cells may play a part in immunopathogenesis of endometriosis being responsible for abrogated local cellular immune responses and facilitation and development of autoimmune reactions. Treg cells may be thus a potential target in the treatment of endometriosis. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by 1M15/N/2011 and NK1W grants from the I Faculty of Medicine, Warsaw Medical University. None of the authors has any competing interests to declare.
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Líquido Ascítico/inmunología , Endometriosis/inmunología , Inmunidad Celular , Quistes Ováricos/inmunología , Linfocitos T Reguladores/inmunología , Adulto , Líquido Ascítico/patología , Biomarcadores/metabolismo , Antígenos CD4/metabolismo , Estudios de Casos y Controles , Recuento de Células , Endometriosis/sangre , Endometriosis/metabolismo , Endometriosis/cirugía , Femenino , Factores de Transcripción Forkhead/metabolismo , Humanos , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Laparoscopía , Quistes Ováricos/sangre , Quistes Ováricos/metabolismo , Quistes Ováricos/cirugía , Índice de Severidad de la Enfermedad , Linfocitos T Reguladores/metabolismo , Adulto JovenRESUMEN
Although the natural hosts for bacteriophages are bacteria, a growing body of data shows that phages can also interact with some populations of mammalian cells, especially with cells of the immune system. In general, these interactions include two main aspects. The first is the phage immunogenicity, that is, the capacity of phages to induce specific immune responses, in particular the generation of specific antibodies against phage antigens. The other aspect includes the immunomodulatory activity of phages, that is, the nonspecific effects of phages on different functions of major populations of immune cells involved in both innate and adaptive immune responses. These functions include, among others, phagocytosis and the respiratory burst of phagocytic cells, the production of cytokines, and the generation of antibodies against nonphage antigens. The aim of this chapter is to discuss the interactions between phages and cells of the immune system, along with their implications for phage therapy. These topics are presented based on the results of experimental studies and unique data on immunomodulatory effects found in patients with bacterial infections treated with phage preparations.
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Anticuerpos Antivirales/sangre , Bacteriófagos/inmunología , Productos Biológicos/farmacología , Factores Inmunológicos/farmacología , Animales , Infecciones Bacterianas/terapia , Terapia Biológica/métodos , Terapias Complementarias/métodos , Citocinas/metabolismo , Humanos , Macrófagos/inmunología , Macrófagos/virología , Fagocitosis , Estallido RespiratorioRESUMEN
Endometriosis is a common gynaecological disorder characterized by the presence of endometrial tissue outside the uterine cavity. This disease is associated with pelvic inflammation and displays some features of autoimmune disorder. Human neutrophil peptides 1, 2, and 3 (HNP 1-3) belonging to α-defensin family play a crucial role in innate immunity against infections and may exert immunoregulatory effects. They may play a role in various inflammatory reactions; however, their role in endometriosis has not been studied. Therefore, the aim of the present study was to evaluate HNP 1-3 in the peritoneal fluid of 67 patients with endometriosis and 16 healthy control women in relation to peritoneal leukocyte subpopulations (neutrophils, T cells, and macrophages) and inflammatory cytokines (IL-6 and IL-8). HNP 1-3, IL-6 and IL-8 were evaluated in the peritoneal fluid by specific enzyme-linked immunosorbent assays (ELISA), and peritoneal leukocyte subpopulations were evaluated by flow cytometry. We found that the levels of HNP 1-3 were significantly increased in the peritoneal fluid of endometriosis patients, compared with control women, and correlated with severity of the disease. Endometriosis was also associated with increased concentrations of peritoneal neutrophils. In endometriosis the levels of HNP 1-3 strongly correlated with concentrations of neutrophils, T cells and IL-8. HNP 1-3 levels were not associated with peritoneal IL-6 or macrophages. These data suggest that HNP 1-3 and neutrophils might play a role in immunopathogenesis of endometriosis and may be worth evaluating as targets for anti-endometriosis therapy.
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Líquido Ascítico/metabolismo , Defensinas/metabolismo , Endometriosis/metabolismo , Interleucina-8/metabolismo , Neutrófilos/metabolismo , Linfocitos T/metabolismo , alfa-Defensinas/metabolismo , Adulto , Líquido Ascítico/inmunología , Biomarcadores/metabolismo , Estudios de Casos y Controles , Defensinas/inmunología , Endometriosis/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunidad Innata , Interleucina-8/inmunología , Neutrófilos/inmunología , Linfocitos T/inmunología , alfa-Defensinas/inmunologíaRESUMEN
Immunuosenescence-related dysregulation of cytokine production is not fully understood and the roles of cytokines in organ aging have been insufficiently studied. This work aimed to compare the expression of interleukin-2 (IL-2), IL-4, IL-6, interferon-gamma (IFNγ), and transforming growth factor-beta (TGFß) in lymphoid organs of young (3 months; n = 10), adult (1 year; n = 10), and aged (2 years; n = 7) rats under healthy, steady-state conditions. Cytokine mRNA levels were determined with TaqMan real-time PCR. In the spleen, all cytokine expression gradually declined with age (for representative cytokine IL-2 averages dCt in spleens of 3 months, 1 year and 2 years rats were 13,645, 13,19 and 15,470, respectively). In lymph nodes, all cytokines except TGF-ß showed markedly upregulated expression in adult compared to young rats, but all were expressed at very low levels in aged rats. In bone marrow, adult animals showed enhanced IL-2, IFNγ, and TGFß expression, but similar IL-4 and IL-6 expression, compared to young rats. Bone marrow of aged rats showed very low expression of all the measured cytokines. Our results demonstrated that changes occurred unevenly with aging in different immune system compartments.
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Regulación de la Expresión Génica , Ganglios Linfáticos/patología , Envejecimiento , Animales , Médula Ósea/metabolismo , Citocinas/metabolismo , Femenino , Sistema Inmunológico , Interferón gamma/metabolismo , Interleucina-2/metabolismo , Interleucina-4/metabolismo , Interleucina-6/metabolismo , Reacción en Cadena de la Polimerasa/métodos , Ratas , Ratas Endogámicas Lew , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Bazo/metabolismo , Factores de Tiempo , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Bacteriophages (viruses of bacteria) are currently considered a promising means of treating antibiotic-resistant infections. The main objective of this study was to evaluate the intensity of the whole-blood monocyte and neutrophil respiratory burst induced by purified preparations and lysates of the bacteriophages T4 and A3/R. While A3/R phage preparations did not induce a significant respiratory burst, T4 phage preparations increased the production of reactive oxygen species in a dose-dependent manner. However, the intensity of the phage-induced respiratory burst was much lower than that triggered by heat-inactivated Staphylococcus aureus cells. These results suggest that phage preparations are not likely to induce oxidative stress following their administration to patients.
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Bacteriófagos/inmunología , Monocitos/inmunología , Neutrófilos/inmunología , Estallido Respiratorio , Humanos , Especies Reactivas de Oxígeno/metabolismo , Staphylococcus aureus/inmunologíaRESUMEN
BACKGROUND: A growing body of data shows that CD4(+)CD25(+) regulatory T cells (Tregs) can induce transplantation tolerance by suppressing immune responses to allograft antigens. However, both the generation and the suppressive capacity of CD4(+)CD25(+) Tregs can be substantially affected by different immunosuppressive drugs used in clinical transplantation. The goal of this study was to compare the effects of cyclosporine A and rapamycin on the induction and suppressive functions of human CD4(+)CD25(+) Tregs in vitro. METHODS: CD4(+)CD25(+) Tregs were induced in two-way mixed lymphocyte reaction (MLR) in the presence of rapamycin (Treg-Rapa) or cyclosporine A (Treg-CsA). Tregs were identified in MLR cultures by flow cytometry using anti-CD4, anti-CD25, anti-CTLA-4, anti-CD122, anti-GITR mAbs and ant-PE-FOXP3 staining sets. Suppressive capacity of induced Tregs was evaluated by their capability to inhibit anti-CD3 Ab-triggered proliferation of peripheral blood mononuclear cells (PBMCs), as measured by flow cytometry. The concentration of TGF-beta1 in culture supernatants was measured by enzyme-linked immunosorbent assay. RESULTS: Although both rapamycin and cyclosporine A suppressed the induction of CD4(+)CD25(+) Tregs during MLRs, this effect was significantly more pronounced in cells cultured with cyclosporine. On the other hand, only rapamycin significantly decreased the percentage of CD4(+)CD25(+) Tregs which expressed GITR, a negative regulator of Treg's suppressive capacity. Importantly, Treg-Rapa, unlike Treg-CsA, displayed significant suppressive activity and were capable of inhibiting the proliferation of anti-CD3 Ab-activated PBMCs. This activity was likely mediated by TGF-beta1. CONCLUSIONS: Rapamycin, unlike cyclosporine A, does not inhibit the function of CD4(+)CD25(+) Tregs. This implies that rapamycin could contribute to the development of transplantation tolerance by promoting the induction of functional CD4(+)CD25(+) Tregs. Moreover, our results suggest that rapamycin could be combined with functional Tregs.
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Ciclosporina/farmacología , Inmunosupresores/farmacología , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Sirolimus/farmacología , Linfocitos T Reguladores/efectos de los fármacos , Linfocitos T Reguladores/inmunología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Proteína Relacionada con TNFR Inducida por Glucocorticoide , Humanos , Técnicas In Vitro , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Receptores de Factor de Crecimiento Nervioso/metabolismo , Receptores del Factor de Necrosis Tumoral/metabolismo , Linfocitos T Reguladores/metabolismo , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
INTRODUCTION: Endometriosis is a common, complex and chronic disease related to ectopic implantation and growth of endometrial tissue that may manifest by pelvic inflammatory reactions, chronic pelvic pain and subfertility. Endometriosis may be associated with increased peritoneal fluid leptin levels. Leptin is known to exert immunomodulatory effects; however, an association between leptin and inflammatory reactions in endometriosis has not been documented. Therefore, the aim of this study was to investigate a relationship between leptin concentrations in peritoneal fluid and the levels of peritoneal fluid inflammatory cytokines and mononuclear leukocyte subpopulations. MATERIALS AND METHODS: Peritoneal fluid was aspirated by laparoscopy from 46 women in whom endometriosis had been confirmed by clinical and histopathological examinations and from 10 control women qualified for ART in whom pelvic pathology has been excluded. Concentrations of leptin and inflammatory cytokines (IL-1beta, IL-6, IFN-gamma and TNF) in peritoneal fluid were evaluated by specific ELISAs. Percentage of peritoneal leukocyte subpopulations (CD3+, CD4+, CD8+ and CD14+) was analyzed by FACS using specific monoclonal antibodies. RESULTS: Leptin concentrations in peritoneal fluid correlated negatively with concentrations of IL-1beta and IFN-gamma (r(s)=-0.38, p=0.01 and r(s)=-0.31, p=0.03, respectively) and correlated positively with the percentage of CD3+ pan-T cells (r(s)=0.69, p=0.009) and CD4+ T helper cells (r(s)=0.74, p=0.036). CONCLUSIONS: Increased leptin levels in peritoneal fluid from endometriosis patients may affect local inflammatory/immune reactions, especially infiltration of CD4+ T helper cells. Thus, leptin may play an important role in the immunopathogenesis of endometriosis.
Asunto(s)
Líquido Ascítico/inmunología , Citocinas/fisiología , Endometriosis/etiología , Leptina/fisiología , Subgrupos Linfocitarios/fisiología , Adulto , Citocinas/análisis , Endometriosis/inmunología , Femenino , Humanos , Leptina/análisisRESUMEN
BACKGROUND: The storage of organs for liver transplantation has resulted in expanding the donor pool for the "living-related" organ donors. Hence, understanding the factors that moderate liver regeneration, both in donor and recipient, is of great importance. Liver regeneration is accompanied by an appearance of cytokines, growth factors and hormones which enable hepatocytes acquiring the ability to reenter the cell cycle. Within few hours after partial hepatectomy (PH) increase concentration of a large number of mitogens for hepatocytes. Among them essential function in recovering after PH plays interleukin 6 (IL-6). However the gene expression of this factor after the reconstruction of the liver is completed is still uncertain. We have assessed the gene expression of IL-6 during liver regeneration in tissue of regenerating liver in tenth day after PH. MATERIAL/METHODS: Materials for the research were obtained from 12 rats. PH surgeries were performed on 7 rats (study group), on the 5 left laparotomy was performed (control group). In the tenth day after surgery on whole 12 animals hepatectomy was performed. RNA was isolated from tissues and then copied to cDNA. Using real-time polymerase chain reaction (real-time PCR) the gene expression of IL-6 was defined. The Mann-Whitney-Wilcoxon Test was performed for the statistic analysis. The p<0,05 was considered as statistically significant. RESULTS: mRNA transcripts for IL-6 in study group compared to control group were 6-fold elevated 10 days following PH (p=0,03). CONCLUSIONS: These results indicate that IL-6 plays a pivotal role in liver regeneration. Elevated level of mRNA transcripts for IL-6 is sustained even after the operation and rebuild of the organ size.