title: Intracellular chemiluminescence activates targeted photodynamic destruction of leukaemic cells.
abstract: Photodynamic therapy (PDT) involves a two-stage process. A light-absorbing photosensitiser (Ps) is endocytosed and then stimulated by light, inducing transfer of energy to a cytoplasmic acceptor molecule and the generation of reactive oxygen species that initiate damage to cellular membrane components and cytolysis. The expanded use of PDT in the clinic is hindered by the lack of Ps target-cell specificity and the limited tissue penetration by external light radiation. This study demonstrates that bioconjugates composed of transferrin and haematoporphyrin (Tf-Hp), significantly improve the specificity and efficiency of PDT for erythroleukemic cells by a factor of almost seven-fold. Fluorescence microscopy showed that the conjugates accumulate in intracellular vesicles whereas free Hp was mostly membrane bound. Experiments with cells deliberately exposed to Tf-Hp at <LD(100) doses showed that surviving cells did not develop resistance to subsequent treatments with the conjugate. Furthermore, we show that the compound luminol induces intracellular chemiluminescence. This strategy was then used to obviate the use of external radiation for Ps activation by incubating the cells with luminol either before or together with Tf-Hp. This novel chemical means of PDT activation induced cytotoxicity in 95% of cells. These combined approaches provide an opportunity to develop broader and more effective applications of PDT.
authors.affiliation: Department of Chemical Engineering and Biotechnology, College of Judea and Samaria, Ariel 44837, Israel.
authors.forename: R
authors.initials: R
authors.lastname: Laptev
authors.forename: M
authors.initials: M
authors.lastname: Nisnevitch
authors.forename: G
authors.initials: G
authors.lastname: Siboni
authors.forename: Z
authors.initials: Z
authors.lastname: Malik
authors.forename: M A
authors.initials: MA
authors.lastname: Firer
mesh_terms: D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D022162:Cytoplasmic Vesicles / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D006415:Hematoporphyrins / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D008163:Luminescent Measurements; D008165:Luminol / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000302:isolation & purification / Q000494:pharmacology*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014168:Transferrin / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D006415:Hematoporphyrins; D017319:Photosensitizing Agents; D014168:Transferrin; D008165:Luminol