Production and characterization of monoclonal antibodies against specific serotypes of Mycobacterium avium and the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex

Abstract

Serotype-specific and Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex (MAIS complex)-specific monoclonal antibodies (MAbs) were prepared. A series of MAbs were obtained, five specific for serotype 2, three specific for serotype 4, eight against a strain of serotype 19, two specific for the MAIS complex, and two against a common glycolipid shared by all the mycobacteria tested so far. The serotype-specific and the MAIS complex-specific MAbs reacted in immunofluorescence with intact mycobacteria and in enzyme-linked immunosorbent assay and immuno-thin-layer chromatography with lipid extracts of mycobacteria. The two MAbs against a common mycobacterial glycolipid reacted only in lipid enzyme-linked immunosorbent assay and immuno-thin-layer chromatography. All MAbs were directed against glycopeptidolipids (GPLs), except for four MAbs against proteins of serotype 19. The serotype-and MAIS complexspecific epitopes on GPLs are exposed on the mycobacterial cell wall, in contrast with the common mycobacterial glycolipid, which is probably located inside the cell wall. The serotype-specific MAbs reacted with native as well as deacetylated GPLs, in contrast with the MAIS complex-specific MAbs, which reacted only with native GPLs. The MAbs will be useful for the identification of MAIS complex and M. avium serotypes 2 and 4 and a strain of serotype 19, GPL analyses with immuno-thin-layer chromatography, and the localization of GPL epitopes in mycobacteria.

Detection and identification of mycobacteria by DNA amplification and oligonucleotide-specific capture plate hybridization

“…However, because 16S rDNA is perfectly conserved, no differentiation between members of the complex can be made. For strains belonging to M. avium-M. intracellulare, the determination made by OSCPH correlated with what had been previously described (2,19,27).…”

“…Strains and clinical specimens. Mycobacterial reference strains (n ϭ 12), strains belonging to the different serovars (1 to 20) of M. avium-M. intracellulare (n ϭ 20) (19), and 38 well-characterized strains isolated in our institute were used for testing (Table 1). In addition, 15 clinical specimens (3 sputum, 8 biopsy, and 2 pus samples, 1 blood sample, and 1 peritoneal effusion sample) from patients with clinically suspected infection by NTM (AIDS patients and patients with previous NTM isolation) were investigated in parallel by PCR-OSCPH and culture.…”

Serovars of Mycobacterium avium complex isolated from AIDS and non-AIDS patients in Spain

“…On the other hand, anti-Ser·8 presented positive reactivity for all antigens tested. Thus, murine Mabs as generated by Rivoire et al (1989) and colleagues (Kolk et al 1989), which have exquisite specificities for the sugar epitope of GPLs, were a benefit to this type of work. These antibodies, which have no appreciable crossreactivities, have increased the specificity and sensitivity of the test and, above all, have allowed the reaction of whole cells with antibody with no misinterpretations.…”

Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

“…1A). In some cases, the 6-deoxytalose moiety may be further extended with haptenic oligosaccharides resulting in highly antigenic serovar specific GPLs (Kolk et al ., 1989;Daffe and Lemmassu, 2000). They have been studied from a structural point of view during the last 50 years (Smith et al ., 1960a;Barrow and Brennan, 1982; for review Billman-Jacobe, 2004), but the first molecular structure of a GPL was solved in the late 60s (Smith et al ., 1960b).…”