Transgene delivery and gelonin cytotoxicity enhanced by photochemical internalization in fibroblast-like synoviocytes (FLS) from rheumatoid arthritis patients.
The objective of this study was to determine if photochemical internalization (PCI) of gelonin can improve the treatment outcome as compared to photodynamic therapy (PDT) and gene transduction of fibroblast-like synoviocytes (FLS)in vitro. For this purpose synovial tissue was obtained under synovectomy of rheumatoid arthritis (RA) patients. Primary single cell suspensions were treated with the photosensitizer meso-tetraphenylporphine (TPPS2a) and light exposure (PDT) followed by evaluation of the cell survival by flow cytometry. PCI of gelonin was performed on FLS in passages 4 and 5 after removal from patients followed by measurements of protein synthesis 24 h after treatment. Additionally FLS were transduced with an adenovirus encoding the E.coli. lacZ gene and treated with PCI to evaluate the effect on the transduction rate. As a result all the cells in the primary cell suspension were susceptible to PDT but CD 106- (FLS) and CD14-positive (monocytes) cells were more sensitive to inactivation by PDT than CD2- (T-cells) and CD19-positive (B-cells) cells. With respect to protein synthesis FLS became up to 4-fold more sensitive to light when combining the photochemical treatment with the gelonin incubation. The fraction of virally transduced FLS was approximately doubled by means of PCI. In conclusion our experiments showed that PCI increased the cytotoxic effect of gelonin and adenoviral transduction of FLS derived from RA patients.
Dietze
Andreas
A
Department of Rheumatoid Arthritis Surgery, Betanien Hospital, N-3722, Skien, Norway. adietze@online.no
Engesaeter
B
B
Berg
K
K
D001172:Arthritis, Rheumatoid / Q000188:drug therapy / Q000473:pathology*; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D018014:Gene Transfer Techniques; D006801:Humans; D066298:In Vitro Techniques; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents; D010940:Plant Proteins / Q000493:pharmacokinetics* / Q000494:pharmacology; D011166:Porphyrins; D011500:Protein Synthesis Inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D054789:Ribosome Inactivating Proteins, Type 1; D013583:Synovial Membrane / Q000166:cytology*; D019076:Transgenes
D017319:Photosensitizing Agents; D010940:Plant Proteins; D011166:Porphyrins; D011500:Protein Synthesis Inhibitors; D054789:Ribosome Inactivating Proteins, Type 1; C425082:di(4-sulfonatophenyl)diphenylporphine; C025674:GEL protein, Gelonium multiflorum