The effect of extracorporeal photopheresis on intracellular cytokine expression in chronic cutaneous graft-versus-host disease

BACKGROUND: Cytokines derived from T helper (Th)1 lymphocytes are thought to be involved in the pathogenesis of graft-versus-host disease (GVHD) and extracorporeal photopheresis (ECP) has been reported to affect Th1/Th2 lymphocyte ratios. It may also influence the balance of cytotoxic Tcells (Tc1/Tc2). OBJECTIVES: This study was formulated to assess the effect of ECP on the cytokine profiles of peripheral blood (PB) lymphocytes from patients with chronic GVHD. PATIENTS AND METHODS: Nine patients were studied. Peripheral blood was sampled at baseline and between 3 and 4 months of therapy when clinical effects are demonstrable. Intracellular cytokine production was assessed in vitro by stimulating PB lymphocytes with phorbol-12-myristate 13-acetate (PMA), inhibiting cytokine release and staining with fluorescein-labelled monoclonal antibodies to interleukin (IL)-2, interferon gamma (IFN-gamma) and IL-4. Flow cytometry analysis gave the absolute number and the percentage of cells expressing a particular cytokine within each lymphocyte subset. RESULTS: Absolute counts of CD3, CD4, CD8, CD19 and CD16+ cells per microlitre were recorded before and after ECP. There was a small but non-significant reduction in all subsets after 3 months of ECP. The percentage of cells expressing IL-2 and IFN-gamma rose following ECP in both the CD4 and CD8 subsets. However, only the percentage of CD4 cells expressing IFN-gamma reached statistical significance (P = 0.02; 95% confidence interval, CI 0.6-15.6). There were no significant changes in the percentage of CD4 cells expressing IL-4. CONCLUSIONS: Our findings appear to be inconsistent with current theories regarding the pathogenesis of GVHD as increased production of Th1 or Tc1 cytokines might be expected to exacerbate GVHD. However, chronic GVHD is characterized by a relative deficiency of IL-2 and IFN-gamma producing cells compared with other patients post-bone marrow transplantation (BMT). This indicates that Th1 and Tc1 cytokines are depleted in chronic GVHD. Thus, by reducing disease activity, ECP could allow cytokine production by these cells to recover. This indicates that the therapeutic effect of ECP is mediated by a different mechanism, and that the changes observed in this study are epiphenomena.