Use of monoclonal antibodies for flow cytometric detection of intracellular Toxoplasma gondii in murine splenic lymphocytes.
Various monoclonal antibodies (mAbs) against Toxoplasma gondii RH tachyzoites were used for flow cytometric detection of intracellular parasites in murine splenic lymphocytes. Tg110 and Tg563 (reacting with the major surface protein SAG1), Tg505 (with another surface protein SAG2), Tg695 and Tg786 (with rhoptry proteins), Tg507, Tg621, and Tg317 (with dense granule proteins), Tg536 (with a microneme protein), and Tg685 (with a cytosol antigen) were the mAbs used. After an in vitro infection of lymphocytes with tachyzoites and reactions with the different mAbs, flow cytometry was performed using an indirect immunofluorescent technique. The proportions of whole infected lymphocytes and of each infected lymphocyte phenotype, CD4+ T cells, CD8+ T cells, and B cells, were determined, and their fluorescent intensities were quantified. The best reaction was seen when Tg110 or Tg695 was used as the mAbs. The results suggest that mAbs against surface or rhoptry proteins are highly useful for the flow cytometric detection of intracellular T. gondii in host cells.
Shin
Eun-Hee
EH
Department of Parasitology, Seoul National University College of Medicine, and Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul 110-799, South Korea.
Kim
Sung-Bum
SB
Nam
Ho-Woo
HW
Han
Eun-Taek
ET
Park
Jae-Hwan
JH
Ahn
Hye-Jin
HJ
Chai
Jong-Yil
JY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D016130:Immunophenotyping; D008214:Lymphocytes / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013047:Specific Pathogen-Free Organisms; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014122:Toxoplasma / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal