Extracorporeal photochemotherapy (ECP) is a procedure effective in the treatment of several human T-cell mediated diseases. During ECP treatment the patient’s blood is processed by means of a cell separator to collect leukocytes (leukapheresis), mostly lymphocytes and monocytes (PBMC), which are then added with the photoactive drug 8-methoxypsoralen (8-MOP), exposed to ultraviolet-A light (UV-A) and reinfused into the patient. Even if the mechanisms of action of ECP remain elusive, it has been shown that it has an in-vivo immunomodulatory effect in experimental ellergic encephalomyelitis (EAE) and in a small pilot study in multiple sclerosis (MS) patients. It has been suggested that during ECP not only UV-A irradiation but also the environmental condition changes may be relevant, an aspect which has never been investigated in detail. Therefore, we developed a new bench device which reliably mimics ex-vivo the complete ECP cycle and we investigated the effect of 8-MOP, UV-A and their combination on the synthesis of IFN-gamma, IL-2 and TNF-alpha on healthy controls and a MS patient. PBMC were collected and treated with 8-MOP and/or UV-A under the same conditions used for the ECP therapy. PBMC were polyclonally stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of Brefeldin A (BFA) to prevent newly synthesized cytokine egression. Intracellular pro-inflammatory cytokines produced by activated T-lymphocytes were evaluated. We observed a significant decrease in activated CD4+ and CD8+ T-lymphocytes producing cytokines after UV-A irradiation and a further decrease in the presence of 8-MOP + UV-A. The decrease in cytokines production seemed to be both cytokine- and cell type-related. In fact TNF-alpha production was reduced to a lesser extent than IFN-gamma and IL-2 ones by both UV-A and the co-treatment, while CD4+ T-cells seemed to be more sensitive than CD8+ lymphocytes when IFN-gamma and IL-2 production was considered. Both T-cell population showed similar behaviour when TNF-alpha production was evaluated. 8-MOP and UV-A co-treatment affected T-lymphocytes activation after polyclonal stimulation and. reduced IFN-gamma, IL-2 and TNF-alpha production with different extent among the cytokines considered. Following this preliminary experience this ex-vivo protocol will be used to extend the determinations in the EAE model and to reproduce these observations in a larger series of healthy controls and MS patients.

Rigolio, R., Perseghin, P., Petersson, J., Jonsson, S., Biffi, A., Cavaletti, G., et al. (2006). Ex-vivo study of extracorporeal photochemotherapy effects on immune system: early experience in healthy controls and a multiple sclerosis patient.. In Abstract Book (pp.63-63).

Ex-vivo study of extracorporeal photochemotherapy effects on immune system: early experience in healthy controls and a multiple sclerosis patient.

RIGOLIO, ROBERTA
Primo
;
CAVALETTI, GUIDO ANGELO
Penultimo
;
2006

Abstract

Extracorporeal photochemotherapy (ECP) is a procedure effective in the treatment of several human T-cell mediated diseases. During ECP treatment the patient’s blood is processed by means of a cell separator to collect leukocytes (leukapheresis), mostly lymphocytes and monocytes (PBMC), which are then added with the photoactive drug 8-methoxypsoralen (8-MOP), exposed to ultraviolet-A light (UV-A) and reinfused into the patient. Even if the mechanisms of action of ECP remain elusive, it has been shown that it has an in-vivo immunomodulatory effect in experimental ellergic encephalomyelitis (EAE) and in a small pilot study in multiple sclerosis (MS) patients. It has been suggested that during ECP not only UV-A irradiation but also the environmental condition changes may be relevant, an aspect which has never been investigated in detail. Therefore, we developed a new bench device which reliably mimics ex-vivo the complete ECP cycle and we investigated the effect of 8-MOP, UV-A and their combination on the synthesis of IFN-gamma, IL-2 and TNF-alpha on healthy controls and a MS patient. PBMC were collected and treated with 8-MOP and/or UV-A under the same conditions used for the ECP therapy. PBMC were polyclonally stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of Brefeldin A (BFA) to prevent newly synthesized cytokine egression. Intracellular pro-inflammatory cytokines produced by activated T-lymphocytes were evaluated. We observed a significant decrease in activated CD4+ and CD8+ T-lymphocytes producing cytokines after UV-A irradiation and a further decrease in the presence of 8-MOP + UV-A. The decrease in cytokines production seemed to be both cytokine- and cell type-related. In fact TNF-alpha production was reduced to a lesser extent than IFN-gamma and IL-2 ones by both UV-A and the co-treatment, while CD4+ T-cells seemed to be more sensitive than CD8+ lymphocytes when IFN-gamma and IL-2 production was considered. Both T-cell population showed similar behaviour when TNF-alpha production was evaluated. 8-MOP and UV-A co-treatment affected T-lymphocytes activation after polyclonal stimulation and. reduced IFN-gamma, IL-2 and TNF-alpha production with different extent among the cytokines considered. Following this preliminary experience this ex-vivo protocol will be used to extend the determinations in the EAE model and to reproduce these observations in a larger series of healthy controls and MS patients.
abstract + poster
Extracorporeal Photochemotherapy, multiple sclerosis, T lymphocyte cytokine production
English
XVI Associazione Italiana Neuroimmunologia (AINI) Congress
2006
Abstract Book
2006
63
63
open
Rigolio, R., Perseghin, P., Petersson, J., Jonsson, S., Biffi, A., Cavaletti, G., et al. (2006). Ex-vivo study of extracorporeal photochemotherapy effects on immune system: early experience in healthy controls and a multiple sclerosis patient.. In Abstract Book (pp.63-63).
File in questo prodotto:
File Dimensione Formato  
2006 AINI Ostuni Rigolio.PDF

accesso aperto

Descrizione: Abtract
Dimensione 17.44 MB
Formato Adobe PDF
17.44 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/60864
Citazioni
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
Social impact