RESUMEN
Endogenous histamine regulates the haematopoiesis. Histidine decarboxylase inhibitor decreases the histamine level, and its intracellular antagonist decreases the histamine effect. The effect of histidine decarboxylase inhibitor (alpha-fluoromethyl histidine) and the intracellular antagonist of histamine [N'N-diethyl-2-4-(phenylmethyl) phenoxyethan-amine-HCl] was investigated on the colony-forming ability of human peripheral progenitor cells. Semi-solid culture medium was used both in the presence and in the absence of 3 U/ml erythropoietin. alpha-Fluoromethyl histidine was used in the range of 50 through 150 micro Mol/ml, the concentration of N'N-diethyl-2-4-(phenylmethyl) phenoxyethanamine-HCl was between 5 and 25 micro Mol/ml. The number of both the erythroide and the granulocyte macrophage colony was significantly decreased in a concentration dependent manner by the presence of both N'N-diethyl-2-4-(phenylmethyl) phenoxyethanamine-HCl (in all concentrations used) and &alpha-fluoromethyl histidine (at higher concentration). The inhibitory effect was decreased by erythropoietin.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Metilhistidinas/farmacología , Éteres Fenílicos/farmacología , Femenino , Citometría de Flujo , Células Madre Hematopoyéticas/citología , Histamina/metabolismo , Antagonistas de los Receptores Histamínicos/farmacología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
The activity of monocyte/lymphocyte system and the splanchnic circulation was investigated in acute pancreatitis. The splanchnic blood flow was characterised indirectly by gastric intramucosal pH changes, which strongly correlated with APACHE-II score, and predicted the bad prognosis. The high CD14/HLA-DR and CD14/CD16 coexpression, the low TNF-alpha production proved to be unfavourable prognostic factor in the early phase of the disease. The expression of IL-2, IL-10 and moderately of IL-4 was elevated, but the expression of INF gamma did not show significant alteration. The thrombocyte aggregation decreased in the early phase. There are bad prognostic signs if its level remains constantly low and ATP-release increases. The activity and index of phagocytosis were higher in comparison with controls, but these parameters were not increasable with higher cell concentration. The ROI production increased, and the increase in the fMLP and o'zymosan indicated LDCL seems to be unfavourable prognostic sign, which well correlated with the APACHE-II score.
Asunto(s)
Citocinas/metabolismo , Linfocitos/metabolismo , Monocitos/metabolismo , Pancreatitis/complicaciones , Pancreatitis/inmunología , Fagocitosis , Enfermedad Aguda , Adulto , Anciano , Estudios de Casos y Controles , Femenino , Citometría de Flujo , Humanos , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-2/sangre , Interleucina-4/sangre , Masculino , Persona de Mediana Edad , Pronóstico , Factores de Riesgo , Índice de Severidad de la Enfermedad , Factor de Necrosis Tumoral alfa/metabolismoAsunto(s)
Células de la Médula Ósea/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Histamina/fisiología , Animales , Células de la Médula Ósea/citología , Recuento de Células , Cimetidina/farmacología , Ensayo de Unidades Formadoras de Colonias , Inhibidores Enzimáticos/farmacología , Células Madre Hematopoyéticas/citología , Antagonistas de los Receptores Histamínicos/farmacología , Histidina Descarboxilasa/antagonistas & inhibidores , Leucemia Mielomonocítica Aguda , Metilhistidinas/farmacología , Ratones , Triprolidina/farmacología , Células Tumorales CultivadasRESUMEN
Two murine hybridoma cell lines named 1D12 and 3B1 secreting antibody against human blood group B were developed. Effect of medium composition and hyperosmotic stress on cell growth, viability and monoclonal antibody (MAb) production was examined during down-stream process in suspension culture. Both of the cell lines secreted MAb of higher titre if the basal medium was supplemented with 2.5 g/l L-glucose and 419 mg/l L-glutamine. Additional nutrients like transferrine, amino acids, ethanolamine, selenium were added to promote cell growth and viability. In case of cell line 1D12 enhancement of MAb production was observed using hyper-osmotic medium as well. Having data of these preliminary experiments we optimized the large scale production of MAbs in bioreactor. These MAbs serve as a base of the anti-B blood group reagent registered in Hungary.
Asunto(s)
Sistema del Grupo Sanguíneo ABO/inmunología , Anticuerpos Monoclonales/biosíntesis , Animales , División Celular , Supervivencia Celular , Medios de Cultivo , Humanos , Hibridomas , Ratones , Células Tumorales CultivadasRESUMEN
A human/mouse heterohybridoma cell line secreting anti-Rh(D) antibody was established by the fusion of Epstein-Barr Virus (EBV) transformed human B lymphocytes and human/mouse heteromyelomas. During scaling-up the cell line was adapted to suspension culture, then medium composition was gradually altered into a more economical one. The adapted cell line has kept its multiplication and antibody secreting characteristics in the new medium. Parameters of large scale batch cultivation in bioreactors were optimized in working volume of 3.6 litre and production was carried out under these parameters in working volume of 24 litre. Optimized parameters were the follows: (i) the cell line is highly sensitive to dissolved oxygen (DO) level, its optimal DO is 10%; (ii) optimal inoculum cell count is 3 x 10(5) cell/ml; (iii) Marine-blade impeller was chosen for supporting the requested oxygen transfer. Using the optimized parameters HUMAN Co. Ltd. produces the Monoclonal Anti-D (IgM) reagent, which has been registered in 1995 in Hungary and now it is on the market.