An improved microassay for Plasmodium falciparum cytoadherence using stable transformants of Chinese hamster ovary cells expressing CD36 or intercellular adhesion molecule-1.
Stable transformants of Chinese hamster ovary (CHO) cell lines expressing high levels of human CD36 or intercellular adhesion molecule-1 (ICAM-1) have been produced as target cells for cytoadherence of Plasmodium falciparum-infected erythrocytes. An improved adherence microassay has been designed using small sample volumes and allowing convenient and reliable measurements on a large number of samples. The assay can be used both with purified proteins spotted on plastic and with the stably transformed CHO cell lines. The same assay plate can be evaluated either microscopically or by scintillation counting after use of 3H-hypoxanthine-labeled parasites. Using the microassay, functional expression of the transfected receptor molecules on CHO-CD36 and CHO-ICAM was confirmed using parasites with different cytoadherence phenotypes and cytoadherence inhibition experiments with a panel of anti-CD36 antibodies. The use of isolates from The Gambia confirmed the applicability of these assays for laboratory studies of these isolates.
Hasler
T
T
Laboratory of Infectious Disease, DNAX Research Institute, Palo Alto, California.
Albrecht
G R
GR
Van Schravendijk
M R
MR
Aguiar
J C
JC
Morehead
K E
KE
Pasloske
B L
BL
Ma
C
C
Barnwell
J W
JW
Greenwood
B
B
Howard
R J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D015153:Blotting, Western; D018955:CD36 Antigens; D016466:CHO Cells; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000276:immunology*; D002469:Cell Separation; D006224:Cricetinae; D015925:Cryopreservation; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000166:cytology / Q000469:parasitology*; D005434:Flow Cytometry; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D018799:Intercellular Adhesion Molecule-1; D008545:Melanoma; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000166:cytology* / Q000276:immunology; D011233:Precipitin Tests; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018955:CD36 Antigens; D015815:Cell Adhesion Molecules; D007106:Immune Sera; D018799:Intercellular Adhesion Molecule-1