Optimierung der extrakorporalen Photopherese

Zusammenfassung

Die Extrakorporale Photopherese ist ein etabliertes Therapieverfahren für Patienten mit T-Zell vermittelten Erkrankungen. Dabei besteht das Verfahren aus der Gewinnung autologer mononukleärer Zellen, deren Behandlung mit 8-Methoxypsoralen und UVA-Licht und die Retransfusion der behandelten Zellen.

Die Wirkmechanismen der Photopherese sind zwar noch nicht vollständig geklärt, ein zentraler Mechanismus stellt jedoch die Apoptose mononukleärer Zellen dar. Das Ziel der Studie war eine Optimierung der Photopherese im Hinblick auf die Behandlung der Zellen mit 8-Methoxypsoralen/UVA und der daraus induzierten verstärkten Apoptose der Lymphozyten. Dabei sind einige Faktoren bekannt, welche die Effektivität der 8-Methoxypsoralen/UVA-Behandlung der Zellen beeinflussen können, wie z.B. der Hämatokrit oder die UVA-Dosis. Unser Fokus lag auf der Verfügbarkeit der photoaktiven Substanz und dem Einfluss der Zellsuspensionsmatrix auf die Apoptose der Lymphozyten.

Die Verfügbarkeit von 8-Methoxypsoralen für die Aufnahme in die Zellen reduzierte sich durch Absorption an Kunststoffe der Bestrahlungssysteme sowie durch Bindung an Proteine bei der Verwendung von autologem Plasma bei der Suspendierung der Zellen. Eine Steigerung der Zugabe von 8-Methoxypsoralen auf 340 ng/mL anstelle von 200 ng/mL führte zu einem Anstieg der T-Zell Apoptose, die sich unter Verwendung von physiologischer Kochsalzlösung als Zellsuspensionsmatrix weiter erhöhte. Eine Anpassung des Verfahrens mit NaCl anstelle von Plasma und die Verwendung einer höheren 8-Methoxypsoralen Konzentration führte zu einer gesteigerten Apoptoseinduktion der T-Zellen. Inwiefern sich eine Erhöhung der Apoptose auf die klinische Wirksamkeit auswirkt, bedarf hingegen noch weiterer klinischen Untersuchungen.

Abstract

Extracorporeal photopheresis is a well-established therapy for patients with T-cell mediated diseases. It is based on collection of autologous mononuclear cells, treatment of cell suspension with 8-methoxypsoralen and UVA light, and subsequent transfusion of treated cells to the patient.

Although the mechanisms of action are not fully understood, a central effect is apoptosis of mononuclear cells. The object of this study was the optimization of the process regarding 8-methoxypsoralen/UVA treatment of the cells and thus resulting lymphocyte apoptosis. In this regard, there are some parameters known which could influence the efficacy of 8-methoxypsoralen/UVA treatment, such as hematocrit or UVA dose. Concerning process optimization, we focused on the availability of the photosensitive substance and on the influence of the cell suspension matrix on the apoptosis of lymphocytes.

Absorption to plastic material of the irradiation systems as well as protein binding when using autologous plasma for cell suspension reduced availability of 8-methoxypsoralen for uptake into the cells. There was an increase in T-cell apoptosis after application of 340 ng/mL 8-methoxypsoralen instead of 200 ng/mL. Apoptosis increased further by using physiological saline for cell suspension. In summary, adapting photopheresis with NaCl instead of plasma and using a higher 8-methoxypsoralen concentration led to an increase of apoptosis induction in the T-cells. To what extent this effect of an apoptosis increase has a clinical consequence has to be clarified.

Schlüsselwörter

Extrakorporale Photopherese - ECP - Apoptose - 8-Methoxypsoralen

Key words

extracorporeal photopheresis - ECP - apoptosis - 8-methoxypsoralen

Publication History

Article published online:
11 August 2022

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