Human monoclonal anti-Plasmodium falciparum antibodies produced by stable EBV-transformed lymphocytes from patients with falciparum malaria.
Peripheral blood lymphocytes (PBL) from 10 persons living in a malaria endemic area and 18 patients recovered from falciparum malaria were studied, nine of whom were admitted to the Hospital for Tropical Diseases and the remaining nine patients were from Trad District Hospital. PBL were divided into two portions, one of which was transformed directly by EBV in the presence of cyclosporin A to eliminate T cell suppression and the other was pre-incubated before transformation with the extract of ultrasonically disrupted, schizont-enriched P. falciparum parasites from in vitro culture. The products of transformed cells were tested for antibodies against blood stages and sporozoites and cells from positive wells were cloned and propagated. With antigen pre-stimulation, cells from 212 of 317 wells (64.5%) were transformed, and this level of transformation was not significantly different from that in the absence of antigen stimulation in which 193 of 311 wells (62.5%) showed transformation (p greater than 0.05). In contrast, 85 of 212 (40.2%) clones from antigen prestimulated wells secreted antibodies whereas 18 of 193 (9.3%) wells without prior antigen stimulation did (p less than 0.0001). Only 44 of 103 antibody-positive clones were subjected to further analysis, of which 42 had activities against blood stages and two against sporozoites. Based on indirect immunofluorescent reactivities, our anti-blood stage monoclonal antibodies (MABs) were conformed to group I (21 clones), III (11 clones) and V (5 clones) and group VI (5 clones).(ABSTRACT TRUNCATED AT 250 WORDS)
Khusmith
S
S
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Tharavanij
S
S
Chongsa-Nguan
M
M
Tapchaisri
P
P
Sabchareon
A
A
Bunnag
D
D
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology; D002472:Cell Transformation, Viral; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human; D006801:Humans; D007132:Immunoglobulin Isotypes / Q000032:analysis; D008288:Malaria / Q000276:immunology*; D010963:Plasmodium falciparum / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes