Method description
We performed effusion of 20-30 ml of the patient’s whole blood in a syringe (50ml) with 5ml of anticoagulant (ACD-A). The volume of effusion depended on the level of hematocrit. This was the most important limiting factor, as the maximum dilution was allowed up to 300ml as required by the ECP system manufacturer. The obtained whole blood was injected into a photopheresis bag (MacoPharma, France) adding 5 ml of 8-MOP (Metoxsalene S.A.L.F, Italy) and incubated in the dark at room temperature for 1 hour. Then sterile NaCl 0.9% solution was added into the bag to 300 ml. The resulting cell product was irradiated with ultraviolet light in the MacoGenic G2 (MacoPharma, France). After phototreatment, the cell product was reinfused to the patient.