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1.
Cell Biol Int ; 43(5): 574-579, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30761646

RESUMEN

Although interleukin-24 (IL-24) has been extensively explored in the immunopathologies of autoimmune diseases, neoplasms, and infections, its role in HIV-1 infection has not been thoroughly elucidated to date. Therefore, the objective of this study was to evaluate the gene and protein expressions of IL-24 at the initial moments of HIV infection in PBMCs. Due to the pro-apoptotic role of IL-24, we evaluated the protein expression of caspase-3, as well as Annexin V/Propidium Iodide flow cytometry and phosphorylation of ERK, which may induce an apoptotic signal block when phosphorylated. The results of this study demonstrated that HIV-1 infection had an impact on the gene and protein expressions of IL-24 and ERK. Annexin V/Propidium Iodide assay demonstrated decrease in the mechanisms of apoptosis in infected cells after incubation of IL-24 neutralizing antibody. Studies on how HIV-1 regulates IL-24 expression may play a role in characterizing viral persistence mechanisms and designing antiretroviral strategies.


Asunto(s)
Infecciones por VIH/inmunología , VIH-1/inmunología , Interleucinas/inmunología , Anexina A5/inmunología , Apoptosis/fisiología , Células Sanguíneas/inmunología , Caspasa 3/inmunología , Humanos , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Cultivo Primario de Células
2.
J Immunol Res ; 2015: 120348, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26634216

RESUMEN

N-Formyl-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) induce similar intracellular signalling profiles; but only fMLP induces interleukin-8 (IL-8) release and nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase activity in neutrophils. Because the role of ROS on IL-8 release in neutrophils is until now controversial, we assessed if NADPH oxidase is involved in the IL-8 secretions and PI3K/Akt, MAPK, and NF-κB pathways activity induced by fMLP. Neutrophils were obtained from healthy volunteers. IL-8 was measured by ELISA, IL-8 mRNA by qPCR, and ROS production by luminol-amplified chemiluminescence, reduction of ferricytochrome c, and FACS. Intracellular pH changes were detected by spectrofluorescence. ERK1/2, p38 MAPK, and Akt phosphorylation were analysed by immunoblotting and NF-κB was analysed by immunocytochemistry. Hydroxy-3-methoxyaceto-phenone (HMAP), diphenyleneiodonium (DPI), and siRNA Nox2 reduced the ROS and IL-8 release in neutrophils treated with fMLP. HMAP, DPI, and amiloride (a Na(+)/H(+) exchanger inhibitor) inhibited the Akt phosphorylation and did not affect the p38 MAPK and ERK1/2 activity. DPI and HMAP reduced NF-κB translocation induced by fMLP. We showed that IL-8 release induced by fMLP is dependent on NADPH oxidase, and ROS could play a redundant role in cell signalling, ultimately activating the PI3K/Akt and NF-κB pathways in neutrophils.


Asunto(s)
Interleucina-8/metabolismo , Glicoproteínas de Membrana/genética , N-Formilmetionina Leucil-Fenilalanina/farmacología , NADPH Oxidasas/genética , Neutrófilos/efectos de los fármacos , Especies Reactivas de Oxígeno/inmunología , Transducción de Señal/efectos de los fármacos , Acetofenonas/farmacología , Amilorida/farmacología , Bloqueadores del Canal de Sodio Epitelial/farmacología , Regulación de la Expresión Génica , Humanos , Interleucina-8/genética , Interleucina-8/inmunología , Glicoproteínas de Membrana/antagonistas & inhibidores , Glicoproteínas de Membrana/inmunología , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , NADPH Oxidasa 2 , NADPH Oxidasas/antagonistas & inhibidores , NADPH Oxidasas/inmunología , FN-kappa B/genética , FN-kappa B/inmunología , Neutrófilos/citología , Neutrófilos/inmunología , Compuestos Onio/farmacología , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/inmunología , Fosforilación/efectos de los fármacos , Cultivo Primario de Células , Transporte de Proteínas/efectos de los fármacos , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/inmunología
3.
Appl Microbiol Biotechnol ; 99(3): 1323-36, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25359475

RESUMEN

The correct and early identification of humans and dogs infected with Leishmania are key steps in the control of leishmaniasis. Additionally, a method with high sensitivity and specificity at low cost that allows the screening of a large number of samples would be extremely valuable. In this study, we analyzed the potential of mitogen-activated protein kinase 3 (MAPK3) and mitogen-activated protein kinase 4 (MAPK4) proteins from Leishmania braziliensis to serve as antigen candidates for the serodiagnosis of human visceral and tegumentary leishmaniasis, as well as canine visceral disease. Moreover, we mapped linear B-cell epitopes in these proteins and selected those epitopes with sequences that were divergent in the corresponding orthologs in Homo sapiens, in Canis familiaris, and in Trypanosoma cruzi. We compared the performance of these peptides with the recombinant protein using ELISA. Both MAPK3 and MAPK4 recombinant proteins showed better specificity in the immunodiagnosis of human and canine leishmaniasis than soluble parasite antigens and the EIE-leishmaniose-visceral-canina-bio-manguinhos (EIE-LVC) kit. Furthermore, the performance of this serodiagnosis assay was improved using synthetic peptides corresponding to B-cell epitopes derived from both proteins.


Asunto(s)
Enfermedades de los Perros/parasitología , Epítopos de Linfocito B/química , Leishmania braziliensis/enzimología , Leishmaniasis/parasitología , Leishmaniasis/veterinaria , Proteína Quinasa 3 Activada por Mitógenos/química , Proteínas Quinasas Activadas por Mitógenos/química , Proteínas Protozoarias/química , Secuencia de Aminoácidos , Animales , Anticuerpos/análisis , Anticuerpos/inmunología , Línea Celular , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/inmunología , Perros , Mapeo Epitopo , Epítopos de Linfocito B/genética , Epítopos de Linfocito B/inmunología , Humanos , Leishmania braziliensis/química , Leishmania braziliensis/genética , Leishmania braziliensis/inmunología , Leishmaniasis/diagnóstico , Leishmaniasis/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/inmunología , Modelos Moleculares , Datos de Secuencia Molecular , Proteínas Protozoarias/genética , Proteínas Protozoarias/inmunología , Alineación de Secuencia , Pruebas Serológicas
4.
Cell Biol Toxicol ; 29(3): 175-87, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23605514

RESUMEN

Immunomodulatory actions exerted by some classes of tryptamines, such as benzoyltryptamine analogues, suggest these molecules as promising candidates to develop new therapies to treat conditions associated to acute and chronic pain and inflammation. N-salicyloyltryptamine (STP) was observed to act as an anticonvulsive agent and exert antinociceptive effects in mouse. In the present work, we performed a screening of cytotoxic, cytoprotective, immunomodulatory, and redox properties of STP in RAW 264.7 macrophages challenged with hydrogen peroxide and LPS. Our results show that STP presents no cytotoxicity in the range of 0.001 to 1 µg/mL, but doses of 50 and 100 µg/mL caused loss of cell viability (IC(50) = 22.75 µg/mL). Similarly, STP at 0.001 to 1 µg/mL did not cause oxidative stress to RAW 264.7 cells, although it did not prevent cell death induced by H(2)O(2) 0.5 mM. At 1 µg/mL, STP reversed some redox and inflammatory parameters induced by LPS. These include thiol (sulfhydryl) oxidation, superoxide dismutase activation, and morphological changes associated to macrophage activation. Besides, STP significantly inhibited LPS-induced TNF-α and IL-1ß release, as well as CD40 and TNF-α protein upregulation. Signaling events induced by LPS, such as phosphorylation of ERK 1/2 and IκBα and p65 nuclear translocation (NF-kB activation) were also inhibited by STP. These data indicate that STP is able to modulate inflammatory parameters at doses that do not interfere in cell viability.


Asunto(s)
Inmunomodulación , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Salicilatos/farmacología , Triptaminas/farmacología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Regulación de la Expresión Génica , Peróxido de Hidrógeno/farmacología , Proteínas I-kappa B/genética , Proteínas I-kappa B/inmunología , Concentración 50 Inhibidora , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/inmunología , Ratones , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Inhibidor NF-kappaB alfa , FN-kappa B/genética , FN-kappa B/inmunología , Estrés Oxidativo , Transducción de Señal , Superóxido Dismutasa/genética , Superóxido Dismutasa/inmunología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/metabolismo
5.
Scand J Immunol ; 69(6): 521-8, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19439013

RESUMEN

RATIONALE: Patients with chronic Schistosoma mansoni infection show lower anti-soluble egg antigen (SEA) proliferation responses and higher responses to soluble worm antigen preparation (SWAP). OBJECTIVE: To compare the activation status and proliferation response of peripheral blood mononuclear cells (PBMC) of infected (XTO) and egg-negative individuals (NI) living in the same endemic area. METHODS: XTO (n = 51) and NI individuals from the same geographical area (n = 37) and healthy blood donors (n = 22) were evaluated before and after stimulation with SEA and SWAP. The expression of activation markers (CD4(+) HLADR(+), CD8(high+)HLA-DR(+) and CD8(+) CD28(+)) and proliferation assay was assessed by flow cytometry. FINDINGS: PBMC from infected patients showed lower frequency of CD4(+) but no change in CD8(+) T cells when compared with the healthy donor group. The ratio CD4(+)/CD8(+) was 1.3, 0.6 and 0.5 in healthy donors, infected and non-infected individuals, respectively. The HLA-DR(+) expression on CD8(+) was higher in PBMC from infected and non-infected individuals than from healthy donors, but similar in both total lymphocytes and CD4(+) populations. No intergroup proliferation response differences were observed in CD4(+) and CD8(+) PBMC unstimulated and stimulated with SEA and SWAP. The SEA but not SWAP-stimulated cells showed a decrease in the expression of phosphorylated extracellular signal-regulated kinase (ERK1/2). CONCLUSIONS: XTO and NI individuals living in the same area presented a smaller per cent of CD4(+) and a higher per cent of CD8(+) cells. The activation by either CD8(high+)HLA-DR(+) or CD8(high+)HLA-DR(+)/CD8(+) was enhanced and decreased in XTO and NI by CD8(+) CD28(+) and CD8(+) CD28(+)/CD8(+) when compared with healthy donor. ERK phosphorylation was attenuated in XTO and NI individuals when stimulated with SEA but not SWAP.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Esquistosomiasis mansoni/inmunología , Subgrupos de Linfocitos T/inmunología , Adolescente , Adulto , Animales , Antígenos Helmínticos/inmunología , Antígenos CD28/inmunología , Citometría de Flujo , Antígenos HLA-DR/inmunología , Antígenos HLA-DR/metabolismo , Humanos , Persona de Mediana Edad , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Fosforilación , Schistosoma mansoni/inmunología , Transducción de Señal/inmunología
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