Production and partial characterization of antibody to cord factor (trehalose 6,6'-dimycolate) in mice.
Antibody production against the trehalose 6,6'-dimycolate (TDM, cord factor) of Rhodococcus ruber, a non-pathogenic species of the Actinomycetales group, was investigated in mice by repeated intraperitoneal injection of TDM in water-in-oil-in-water micelles without carrier protein. The antigenic TDM was isolated and purified chromatographically from the chloroform-methanol extractable lipids of R. ruber. The hydrophobic moiety of this TDM was composed of two molecules of monoenoic or dienoic alpha-mycolic acids with a carbon chain length ranging from C44 to C48 centering at C46. To detect the antibody, an enzyme-linked immunosorbent assay (ELISA) system was employed using plastic plates coated with TDM. The antibody reacted against the TDM of R. ruber. The antibody was reactive in similar fashion against glycosyl monomycolates differing in the carbohydrate moiety, such as that of glucose mycolate (GM) and mannose mycolate (MM), obtained from R. ruber. Moreover, the antibody reacted against mycolic acid methyl ester itself when it was used as the antigen in ELISA, and trehalose did not absorb the antibody to TDM or inhibit the reaction. These results indicate that the epitope of TDM recognized by the antibody is mycolic acid, an extremely hydrophobic part of the molecule. Next, we prepared monoclonal anti-TDM antibody (moAb) in mice myeloma cells to examine its biological activities and the role of humoral immunity in mycobacterial infection. MoAb reacted against the TDM, glycosyl mycolate, and mycolic acid methyl ester in ELISA in the same manner as our polyclonal antibody did. The administration of moAb suppressed granuloma formation in the lungs, spleen, and liver induced by TDM and inhibited the production of interleukin-1 (IL-1) and chemotactic factor, which is reported to precede granuloma formation.
Fujiwara
N
N
Department of Bacteriology, Osaka City University Medical School, Osaka, Japan.
Oka
S
S
Ide
M
M
Kashima
K
K
Honda
T
T
Yano
I
I
D000042:Absorption; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D002241:Carbohydrates / Q000276:immunology; D002630:Chemotactic Factors / Q000096:biosynthesis; D003311:Cord Factors / Q000009:adverse effects / Q000276:immunology*; D006017:Glycolipids / Q000276:immunology; D006099:Granuloma / Q000209:etiology / Q000276:immunology; D007375:Interleukin-1 / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009171:Mycolic Acids / Q000276:immunology; D012240:Rhodococcus / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D002630:Chemotactic Factors; D003311:Cord Factors; D006017:Glycolipids; D007375:Interleukin-1; D009171:Mycolic Acids