RESUMO
Presented are experiences with the determination of immunosuppressive activity of alpha globulin fraction of human blood plasma, crude (Cohn fraction IV), partially purified and so-called low-molecular isolates of these substances. For testing the following techniques were used: the lymphocytotoxic and lymphoagglutination test, rosette-inhibition tests with human lymphocytes and splenocytes of mice immunized with sheep erythrocytes, lymphocyte blastic transformation inhibition test using PHA stimulated cells as well as in the mixed culture, inhibition of haemolytic plaque formation tests, determination of GVH reactivity using regional tests in the popliteal nodes, determination of HVG reactivity by skin graft survival test, determination of the effect on formation of precipitins against soluble antigens. The best results were obtained with the determination of survival times for skin grafts and the regional test in the popliteal nodes Fairly reproducible results were obtained in experiments on influencing the humoral antibody response and inhibition of formation of precipitating antibodies. Of in vitro methods tests of inhibition of lymphocyte blastic transformation both after PHA stimulation and in the mixed culture are applicable. Fully unsuitable proved the rosette, lymphocytotoxic and lymphoagglutination tests. In vitro modification of haemolytic plaque formation tests. In vitro modification of haemolytic plaque formation test is unfit for testing the crude fractions or partially purified drugs, its in vivo modification seems more feasible provided a large panel of animals is examined.
Assuntos
alfa-Globulinas/fisiologia , Terapia de Imunossupressão , Animais , Formação de Anticorpos , Citotoxicidade Imunológica , Sobrevivência de Enxerto , Humanos , Imunidade Celular , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos A , Camundongos Endogâmicos C57BL , Peso Molecular , Coelhos , Imunologia de TransplantesRESUMO
Immunosuppressive activity of alpha-globulin fraction of human blood plasma (Cohn fraction IV) was studied to determine the possible clinical utilization of this fraction as such, purified, or as the starting material for isolation of the active factor. The ethanol fractions IV, isolated and waste in the production of blood derivatives were found to be directly inapplicable because of relatively low activity and poor solubility. Therefore purification experiments were done to remove the denatured proteins and lipids and to increase the specific activity, they were performed on laboratory scale using DEAE ionex chromatography, and on semipilot scale utilizing the principles of ethanol fractionation of plasma. From the viewpoint of technology, the latter procedure seems more feasible; partial purification yielded good solubility and somewhat higher specific activity. Under study are further techniques capable of eliminating the contaminating proteins with do not carry the immunosuppressively active component, as well as the effectivity of pasteurization at 60 degrees C/10 hrs., which seems indispensable with respect to inactivation of infectious hepatitis viruses and the potential clinical application. Experimental isolations of the active component have led immunosuppressively active low-molecular part, which, however, is chemically nonhomogeneous. Its subsequent subfractionation will be necessary.
Assuntos
Formação de Anticorpos , Proteínas Sanguíneas/farmacologia , Reação Enxerto-Hospedeiro , Reação Hospedeiro-Enxerto , Imunossupressores/farmacologia , Animais , Proteínas Sanguíneas/isolamento & purificação , Imunossupressores/isolamento & purificação , Transfusão de Linfócitos , Camundongos , Camundongos Endogâmicos C57BL , Coelhos , Transplante de PeleAssuntos
Proteínas Sanguíneas/isolamento & purificação , Imunidade Celular/efeitos dos fármacos , Animais , Proteínas Sanguíneas/farmacologia , Reação Hospedeiro-Enxerto/efeitos dos fármacos , Ativação Linfocitária/efeitos dos fármacos , Métodos , Camundongos , Camundongos Endogâmicos C57BL , Transplante de PeleRESUMO
The effect of the alpha globulin fractions of human serum administered before the inoculum of tumour cells was studied in two sublines of C3H mice (C3H/VUFB and C3H/Sumice) showing a different resistance to a solid Gardner lymphosarcoma. The survival time of animals was shortened in both sublines, but more markedly in C3H/VUFB mice that were more resistant to the tumour. The survival was found to depend on the dose and the immunosuppressive activity of the alpha globulin fractions as revealed in other in vitro and in vivo tests. We believe that the tumour model using C3H/VUFB mice may be used as a further test for determining the activity of alpha globulin fractions.