title: Assessment of human natural killer and lymphokine-activated killer cell cytotoxicity against Toxoplasma gondii trophozoites and brain cysts.
abstract: Because previous work has suggested that NK cells may be important in host resistance against the intracellular parasite Toxoplasma gondii we examined whether human NK cells and lymphokine-activated killer (LAK) cells have activity against trophozoites and cysts of this organism in vitro. A method to radiolabel Toxoplasma trophozoites with 51Cr was developed and direct cytotoxic activity was determined by using modifications of the standard 51Cr release assay. Viability of 51Cr-labeled trophozoites assessed by both methylene blue staining and trypan blue exclusion was greater than 90%. Significantly more 51Cr was released by anti-Toxoplasma antibody and C than by antibody in the absence of C. Incubation of trophozoites with freshly isolated human NK cells or NK cells activated with either rIL-2 or rIFN-alpha did not result in significant release of 51Cr (specific lysis was 0 to 2.3%). In contrast, the average specific lysis of radiolabeled trophozoites by LAK cells was significant (specific lysis was 7.8% +/- 1.1, p less than 0.01). In a series of separate experiments, preincubation of radiolabeled trophozoites with heat-inactivated normal or Toxoplasma antibody-positive human serum increased the cytotoxicity of LAK cells from a mean specific lysis of 15% +/- 4.5 to 39% +/- 8.5, respectively (p less than 0.05), as assessed by 51Cr release. Because previous work has shown that radioisotope release from parasites may be nonspecific, separate experiments were performed to determine the cytotoxicity of LAK cells against antibody-coated trophozoites by using ethidium bromide-acridine orange staining to assess effector cell damage. LAK cells had a mean specific lysis of 51% against antibody-coated trophozoites by ethidium bromide-acridine orange staining. Preincubation with heat-inactivated Toxoplasma-antibody positive human serum did not increase activity of rIL-2-activated NK cells against 51CR-labeled trophozoites. Neither human NK cells (freshly isolated or activated by rIL-2 or rIFN-alpha) nor LAK cells were cytotoxic for purified preparations of cysts of Toxoplasma isolated from the brains of chronically infected mice.
authors: Department of Immunology and Infectious Diseases, Palo Alto Medical Foundation, CA 94301.
authors: B R
authors: BR
authors: Dannemann
authors: V A
authors: VA
authors: Morris
authors: F G
authors: FG
authors: Araujo
authors: J S
authors: JS
authors: Remington
mesh_terms: D000165:Acridine Orange; D000818:Animals; D000913:Antibodies, Protozoan / Q000502:physiology; D000953:Antigens, Protozoan / Q000502:physiology; D001927:Brain Diseases / Q000276:immunology* / Q000401:mortality; D002860:Chromium Radioisotopes / Q000378:metabolism; D003165:Complement System Proteins / Q000502:physiology; D003560:Cysts / Q000276:immunology* / Q000401:mortality; D003602:Cytotoxicity, Immunologic; D004996:Ethidium; D005260:Female; D006801:Humans; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology*; D007694:Killer Cells, Natural / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D013194:Staining and Labeling; D014122:Toxoplasma / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism; D014124:Toxoplasmosis, Animal / Q000276:immunology / Q000401:mortality
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D002860:Chromium Radioisotopes; D003165:Complement System Proteins; D004996:Ethidium; D000165:Acridine Orange