Interaction of bacterial toxin with leukocytes measured by flow cytometry.
Enterobacter cloacae toxin was purified in the form of monomer and polymer. Both forms stimulated the generation of reactive oxygen species (ROS) at sublytic concentration; the oxidative stress produced was studied by using chemiluminescence (CL). The alteration generated caused death of leukocytes, especially at high toxin concentration. Polymeric toxin produced more oxidative stress than the monomeric one. Cytometry allowed the detection of more toxin binding to neutrophils rather than to monocytes or lymphocytes. There was binding at 4 degrees C, and the amount of toxin in the cells increased at 37 degrees C. The interaction of toxin with leukocytes was evident even after 100 degrees C treatment of toxin during 5 min. The incubation with 2-mercaptoethanol was not necessary for toxin binding.
Departamento de Farmacia, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Ciudad Universitaria, Haya de la Torre y Medina Allende 5000 Córdoba, Argentina.
María Gabriela
MG
Paraje
Pablo
P
Iribarren
Inés
I
Albesa
D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D016972:Enterobacter cloacae / Q000472:pathogenicity; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008163:Luminescent Measurements; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress; D011108:Polymers / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D017382:Reactive Oxygen Species / Q000378:metabolism
D001427:Bacterial Toxins; D011108:Polymers; D017382:Reactive Oxygen Species