Photoimmunotherapy Using Cationic and Anionic Photosensitizer-Antibody Conjugates against HIV Env-Expressing Cells

• Published in: International journal of molecular sciences 2020-12, Vol.21 (23), p.9151
• Main Authors: Sadraeian, Mohammad, Bahou, Calise, da Cruz, Edgar Ferreira, Janini, Luíz Mário Ramos, Sobhie Diaz, Ricardo, Boyle, Ross W, Chudasama, Vijay, Eduardo Gontijo Guimarães, Francisco
• Format: Article
• Language: English
• Subjects: Anions - chemistry; Anti-HIV Agents - chemistry; Anti-HIV Agents - pharmacology; Antibodies, Monoclonal; Apoptosis; Apoptosis - drug effects; Cations - chemistry; Cell Line, Tumor; Cell membranes; Cells, Cultured; Confocal microscopy; Conjugation; Cytotoxicity; Disulfide bonds; Dosimeters; Dosimetry; env Gene Products, Human Immunodeficiency Virus - antagonists & inhibitors; env Gene Products, Human Immunodeficiency Virus - genetics; env Gene Products, Human Immunodeficiency Virus - metabolism; Flow Cytometry; Fluorescent Antibody Technique; Glycoprotein gp41; HIV; HIV - drug effects; HIV - genetics; HIV Infections - drug therapy; HIV Infections - virology; Human immunodeficiency virus; Humans; Immunoconjugates; Immunoconjugates - chemistry; Immunoconjugates - pharmacology; Immunology; Immunotherapy; Lasers; Light; Lysine; Medical research; Monoclonal antibodies; Photochemotherapy; Photodynamic therapy; Photosensitizing Agents - chemistry; Photosensitizing Agents - pharmacology; Phototoxicity; Porphyrins; Reactive oxygen species; Reactive Oxygen Species - metabolism; Virus Replication - drug effects
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21239151

Different therapeutic strategies have been investigated to target and eliminate HIV-1-infected cells by using armed antibodies specific to viral proteins, with varying degrees of success. Herein, we propose a new strategy by combining photodynamic therapy (PDT) with HIV Env-targeted immunotherapy, and refer to it as HIV photoimmunotherapy (PIT). A human anti-gp41 antibody (7B2) was conjugated to two photosensitizers (PSs) with different charges through different linking strategies; "Click" conjugation by using an azide-bearing porphyrin attached via a disulfide bridge linker with a drug-to-antibody ratio (DAR) of exactly 4, and "Lysine" conjugation by using phthalocyanine IRDye 700DX dye with average DARs of 2.1, 3.0 and 4.4. These photo-immunoconjugates (PICs) were compared via biochemical and immunological characterizations regarding the dosimetry, solubility, and cell targeting. Photo-induced cytotoxicity of the PICs were compared using assays for apoptosis, reactive oxygen species (ROS), photo-cytotoxicity, and confocal microscopy. Targeted phototoxicity seems to be primarily dependent on the binding of PS-antibody to the HIV antigen on the cell membrane, whilst being independent of the PS type. This is the first report of the application of PIT for HIV immunotherapy by killing HIV Env-expressing cells.