Monoclonal antibody E8-18 identifies an integral membrane surface protein unique to Mycoplasma capricolum subsp. capripneumoniae

Abstract

Monoclonal antibody (MAb) E8-18 reacted with four isolates of Mycoplasma capricolum subsp. capripneumoniae in Western blots identifying an epitope on a 24 kDa antigen (p24). MAb E8-18 did not react with 11 isolates belonging to four other Mycoplasma species or subspecies closely related to M. capricolum subsp. capripneumoniae. A combination of trypsin treatment of intact organisms and detergent-phase partitioning revealed p24 to be an integral M. capricolum subsp. capripneumoniae surface membrane protein. Mycoplasma capricolum subsp. capripneumoniae, formerly designated Mycoplasma strain F38, is the causative agent of classical contagious caprine pleuropneumonia (CCPP) (31). The disease is of major economic importance in Africa and Asia and poses a major constraint to goat production because of high mortalities. M. capricolum subsp. capripneumoniae was originally isolated from a goat with pleuropneumonia in Kenya (30) and produces classical CCPP, which involves transmission by contact, high mortality, and a fibrinous pleuropneumonia characterized by massive hepatization and pleurisy (31). Whereas M. capricolum subsp. capripneumoniae has been isolated from goats in only seven other countries, including Sudan (19), Tunisia (33), Ethiopia (45), Chad (28), Oman (23), Turkey (47), and Uganda (4), it is not known whether the pleuropneumonia observed in other parts of the world is due to M. capricolum subsp. capripneumoniae or other mycoplasmas of the Mycoplasma mycoides cluster (9, 10, 14). The M. mycoides cluster includes seven Mycoplasma species: M. mycoides (small colony), M. mycoides (large colony), Mycoplasma capricolum subsp. capricolum, M. primatum, M. equigenitalium, Mycoplasma bovine group 7 described by Leach (26a), M. capri, and M. capricolum subsp. capripneumoniae (9, 10). All these organisms have extensive serological cross-reactivity (12-14, 24, 26) and biochemical similarities (1, 9, 10, 24, 26). The serological cross-reactivities have made it difficult to develop simple and yet specific diagnostic tests for the diseases caused by the different organisms. Various diagnostic tests have been reported, including latex agglutination (36), tests for monoclonal antibody (MAb) blocking and/or MAb reactivity with antigen in body fluids (44), dot blot immunobinding assay (18), and a combination of PCR and enzyme restriction analysis (5). Latex agglutination to detect serum antibodies to M. capricolum subsp. capripneumoniae is simple and can be performed in the field (36). The specificity of this test for field use is adequate; however, rabbit antisera to some other Mycoplasma species cause agglutination of M. capricolum subsp. capripneumoniae polysaccharide latex-coated beads, although these organisms

Development and Evaluation of an Immuno-Capture Enzyme-Linked Immunosorbent Assay to Quantify the Mycoplasma capricolum subsp. capripneumoniae (Mccp) Protein in Contagious Caprine Pleuropneumonia (CCPP) Vaccine

“…Eight mAbs were generated and evaluated to design the ICE. e number of mAbs obtained are slightly higher than those reported [28]. ese mAbs were selected based on the optical density on the antigen obtained in iELISA.…”

Novel Candidates for Vaccine Development Against Mycoplasma Capricolum Subspecies Capripneumoniae (Mccp)—Current Knowledge and Future Prospects

“…However, the immunogenic potential of these antigens as vaccines is yet to be evaluated. Rurangirwa et al [55] identified a specific integral membrane surface protein of Mccp using the mAb E8-18. The inhibitory effects of vaccines containing subunit fractions of Mccp were studied by March and Jones [56].…”

“…These six members of the mycoides cluster are antigenically related, as they share some common antigens [21,42,67,85,86,87]. Hence, cross-reactions are frequently noted in diagnostic tests [54,55]. In other words, antigens identified or evaluated in other members of the mycoides cluster can be explored in Mccp also.…”