RESUMEN
In cultures of peritoneal macrophages (MP) of male BALB/c mice infected with Mycobacterium tuberculosis from the BCG vaccine, the expression of CD1, CD14, CD25, CD30, CD35, and CD95 receptors was studied in vitro 3 months after infection. In MP cultures from intact and infected mice, mononuclear MP predominated (96 and 92%, respectively). Bi- and trinuclear MP in MP cultures from control and infected mice constituted 4 and 8.3% of all MP, respectively. In the cultures of both groups, no obvious correlations between the number of MP expressing CD-receptors and number of nuclei in these cells were found, but the expression of CD14 receptor was more often noted. In cultures from infected animals, hypertrophied MP and enhanced (by several times) expression of all CD-receptors were observed. The increase in the expression of CD-receptor can be determined by activation of plastic processes in hypertrophied MP (in epithelioid and in numerically insignificant polynuclear MP), which is due to the phenomenon of prolonged M. tuberculosis persistence in the vacuolar apparatus of these cells.
Asunto(s)
Antígenos CD1/biosíntesis , Macrófagos Peritoneales/inmunología , Mycobacterium tuberculosis/inmunología , Receptores de Citocinas/biosíntesis , Tuberculosis/inmunología , Animales , Vacuna BCG/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Tuberculosis/patologíaRESUMEN
Cultured peritoneal macrophages from intact (control) and BCG-infected (experiment) male BALB/c mice were studied 90 days after infection. Polarization of macrophages by M1 (expression of GM-CSF, IFNγ, and CD16/32) and M2 (expression of bFGF and CD36) differentiation pathways was studied with consideration for their the nuclearity class. Mononuclear cells predominated (90% and higher) in macrophage cultures of both groups and presumably, were presented by mainly epithelioid cells. The results indicated polarization of mononuclear and multinuclear macrophages in the M2 direction under conditions of BCG granulomatosis and a higher initial M2 polarization of binuclear macrophages. In control cultures, the ratio of M2 to M1 macrophages was 0.57, in experimental cultures this ratio was 1.6. It seems that long persistence of Mycobacterium tuberculosis in macrophages served as a factor stimulating the plastic processes and transformation of macrophages into epithelioid cells that form the "core" of granulomas and their enlargement upon incorporation of macrophages.
Asunto(s)
Células Epitelioides/patología , Regulación de la Expresión Génica/inmunología , Macrófagos Peritoneales/patología , Mycobacterium bovis/crecimiento & desarrollo , Tuberculosis/patología , Animales , Antígenos CD36/genética , Antígenos CD36/inmunología , Diferenciación Celular , Transdiferenciación Celular/genética , Transdiferenciación Celular/inmunología , Células Epitelioides/inmunología , Células Epitelioides/microbiología , Factor 2 de Crecimiento de Fibroblastos/genética , Factor 2 de Crecimiento de Fibroblastos/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Interferón gamma/genética , Interferón gamma/inmunología , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/microbiología , Masculino , Ratones , Ratones Endogámicos BALB C , Mycobacterium bovis/patogenicidad , Cultivo Primario de Células , Receptores de IgG/genética , Receptores de IgG/inmunología , Tuberculosis/genética , Tuberculosis/inmunología , Tuberculosis/microbiologíaRESUMEN
Expression of CD11, CD29, CD36, and DC-STAMP molecules by macrophages was analyzed in in vitro experiments. These molecules mediate cell fusion, one of the mechanisms underlying the formation of multinuclear macrophages. Macrophages were obtained from intact and BCG-infected male BALB/c mice. In intact cultures, multinuclear macrophages appeared primarily due to amitotic division of cell nuclei, while in macrophage cultures from infected mice, the process of cell fusion predominated. In intact macrophage cultures, bi- and multinuclear cells expressed primarily CD29 and CD36. In cultures from infected mice, macrophages expressing CD29 and DC-STAMP predominated, but bi- and multinuclear macrophages expressing CD11 and CD36 predominated over mononuclear ones. The study of macrophage fusion mechanism can be useful for understanding of this biological phenomenon as the mechanisms of delivery of M. tuberculosis and lysosomotropic anti-tuberculosis drugs into tuberculous granulomas to suppress M. tuberculosis persisting in macrophages and reduce the destructive potential of granulomas.
Asunto(s)
Vacuna BCG/administración & dosificación , Antígenos CD11/genética , Antígenos CD36/genética , Integrina beta1/genética , Macrófagos/microbiología , Proteínas de la Membrana/genética , Proteínas del Tejido Nervioso/genética , Animales , Antígenos CD11/inmunología , Antígenos CD36/inmunología , Fusión Celular , Núcleo Celular/ultraestructura , Expresión Génica , Integrina beta1/inmunología , Macrófagos/inmunología , Macrófagos/patología , Masculino , Proteínas de la Membrana/inmunología , Ratones , Ratones Endogámicos BALB C , Proteínas del Tejido Nervioso/inmunología , Cultivo Primario de CélulasRESUMEN
Intraperitoneal infection of Mycobacterium tuberculosis from BCG vaccine to male BALB/c mice provoked the formation of multinuclear macrophages in the peritoneal transudate. In 2 and 3 months after infection, their number increased respectively by 34.4 and 66.7% in comparison with intact mice. At all postinfection terms, far greater amounts of caspase 3, p53, Bad, and TNFα were expressed by multinuclear macrophages than by the mononuclear ones. At these terms, the content of Bcl-2-expressing macrophages with different numbers of nuclei was pronouncedly high; of them, the number of multinuclear macrophages was maximum. In 3 months after infection, the number of macrophages expressing Bcl-2 significantly decreased irrespective of the number of their nuclei. At this term, the number of macrophages expressing caspase 3, and TNFα also decreased. In parallel, the number of macrophages expressing p53 and Bad somewhat increased. These data attest to potentiality ("readiness") of macrophages infected with Mycobacterium tuberculosis to apoptotic self-elimination.
Asunto(s)
Macrófagos/metabolismo , Mycobacterium bovis/patogenicidad , Tuberculosis/metabolismo , Animales , Apoptosis/fisiología , Vacuna BCG/uso terapéutico , Caspasa 3/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The incidence of mono- and multinuclear cells and their expression of pro- and antifibrotic factors were studied in cultured peritoneal macrophages from intact and BCG-infected mice. Generally, the expression of factors increased with an increase in the number of nuclei per cell. However, the expression was higher in macrophages from BCG infected mice, except the cells with 3 and more nuclei, extremely rarely expressing IL-1α in cultures from intact and BCG-infected animals. The number of macrophages with 3 and more nuclei, expressing CatD, was comparable with the number of mono- and binuclear macrophages. Presumably, this was determined by various mechanisms of formation of multinuclear (3-5 and more nuclei) macrophages, for example, by amitosis.
Asunto(s)
Catepsina D/inmunología , Factor 2 de Crecimiento de Fibroblastos/inmunología , Interferón gamma/inmunología , Interleucina-1alfa/inmunología , Macrófagos Peritoneales/inmunología , Metaloproteinasa 13 de la Matriz/inmunología , Infecciones por Mycobacterium/inmunología , Factor de Crecimiento Transformador beta1/inmunología , Animales , Catepsina D/genética , Núcleo Celular , Factor 2 de Crecimiento de Fibroblastos/genética , Regulación de la Expresión Génica , Interferón gamma/genética , Interleucina-1alfa/genética , Macrófagos Peritoneales/microbiología , Macrófagos Peritoneales/patología , Masculino , Metaloproteinasa 13 de la Matriz/genética , Ratones , Ratones Endogámicos BALB C , Infecciones por Mycobacterium/microbiología , Infecciones por Mycobacterium/patología , Mycobacterium bovis/inmunología , Cultivo Primario de Células , Transducción de Señal , Factor de Crecimiento Transformador beta1/genéticaRESUMEN
Peritoneal macrophages were isolated from intact and BCG-infected BALB/c mice and explanted in vitro. Multinuclear macrophages formed in these cultures differed by the number of nuclei, expression of apoptosis inductors and regulators (TNF-α, p53 protein, caspase 3, and Bcl-2 protein), and cytophysiological characteristics (phagocytic activity, ROS generation, and antimycobacterial properties). Our results indicate that the formation of multinuclear macrophages is accompanied by induction of apoptosis (p53 signaling pathway) and appearance of multinuclear macrophage-derived cells characterized by high phagocytic and antimycobacterial activity.
Asunto(s)
Apoptosis/inmunología , Macrófagos Peritoneales/inmunología , Mycobacterium bovis/inmunología , Fagocitosis/inmunología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Tuberculosis/inmunología , Animales , Caspasa 3/metabolismo , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/microbiología , Masculino , Ratones , Ratones Endogámicos BALB C , Tuberculosis/microbiología , Factor de Necrosis Tumoral alfa/metabolismo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
The review examines the current data on the cellular and tissue mechanisms of scar formation in the liver. On the basis of the analysis of the literature, authors conclude that to minimize the destructive processes in the liver, the most advisable and promising method during the operations on this organ is the use of laser surgical devices. However, the problems of the selection of the optimal regimes of laser radiation and of the methods of conducting these operations to promote reliable and rapid healing of postoperative wound of the liver.