Determination of the preformed immune status in the case of active diseases with intracellular causative agents for the reconstruction of infection because

摘要

A process for the determination of the preformed immune status in the case of active diseases with intracellular causative agents for the reconstruction of infection because by means of flow cytometry analysis of cytokines,characterized by the,that the origin of an active disease, namely primarily occurrence of an infection, or reactivation, by means of a specific cytokine displacement, comprising the steps of:– Stimulation of heparinised whole blood with the pathogen - specific antigens, with staphylococcus enterotoxin b (seb) as a positive control, as well as with solvent as a negative control with or without the addition of co - stimulatory antibodies anti - cd49d and anti - cd28.– Incubation for 30 min to 4 h hours at 35 to 39°C. and 3 (5) to 10% co₂.– The addition of a secretion inhibitor, for example, brefeldin a– It was incubated for 2 to 12 hours at 35 to 39°C. and 3 (5) to 10% co₂– Lysis of erythrocytes and fixing of leukocytes– Multiple washing steps for the isolation of the leukocytes– Conveyance of the cell membrane, by means of resistance of the present invention is that by the addition of saponin or another, the cell membrane - permeability which.– Incubation with specific fluorochrome - labelled antibodies against cd4, cd69, ifnγ - γ and il - 2 for at least 10 minutes on ice or at room temperature - or in the case of 35 to 39°C.– Renewed washing steps for the removal of antibodies unbonded– Analysis of the cells to a flow cytometerThe evaluation of the component and / or of the ratio of ifnγ -, il - 2 - or both cytokines producing t - cells for the determination of the infection origin in the sense of a primary infection or a reactivation in patients with an active infection disease.