Inhibitory activity on bacterial motility and in vivo protective activity of human monoclonal antibodies against flagella of Pseudomonas aeruginosa.
Three stable hybridoma cell lines, IN-2A8, IN-5D6, and ZI-3A8, that secrete human monoclonal antibodies (MAbs) specific for b-type flagella of Pseudomonas aeruginosa were established by fusing peripheral blood lymphocytes from healthy volunteers with murine myeloma P3X63-Ag8.653 cells. The immunoglobulin M MAbs reacted specifically with flagellin (Mr, 52,000) by Western blotting (immunoblotting) analysis and bound specifically to clinical isolates belonging to Homma serotypes A, B, H, I, and M at frequencies of 58, 50, 46, 30, and 35%, respectively, but did not bind to any serotype E or G isolates. Overall, the MAbs bound to 31% of the clinical isolates. MAb IN-2A8 strongly protected burned mice challenged with P. aeruginosa bearing b-type flagella from death following parenteral administration of 0.1 microgram per mouse. This MAb also inhibited P. aeruginosa colony spreading in soft agar at a concentration of more than 1 microgram/ml but only slightly enhanced opsonophagocytosis by human polymorphonuclear leukocytes. A line of evidence suggests that the potent in vivo activity of MAb IN-2A8 in the burned-mouse model is likely to be caused by its inhibition of bacterial motility after binding to flagella.
Ochi
H
H
Laboratory of Biotechnology, Takarazuka Research Center, Sumitomo Chemical Co., Ltd., Hyogo, Japan.
Ohtsuka
H
H
Yokota
S
S
Uezumi
I
I
Terashima
M
M
Irie
K
K
Noguchi
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial / Q000032:analysis; D002056:Burns / Q000382:microbiology; D002465:Cell Movement; D005407:Flagella / Q000276:immunology*; D006801:Humans; D051379:Mice; D009068:Movement; D010587:Phagocytosis; D011550:Pseudomonas aeruginosa / Q000276:immunology* / Q000472:pathogenicity
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial