Delayed hypersensitivity reactions by the mycobacterial antigen A60 and cutaneous testing in tuberculosis

Antigen A60 has been purified from the cytoplasm of Mycobacterium bovis BCG, and its composition has been determined: it has proved to be able to elicit immune reactions of both humoral and cellular type. Inoculation of A60 into the footpad of mice previously sensitized with the same antigen, or with whole mycobacterial cells produced a footpad swelling showing a peak at 24 h. Similar delayed hypersensitivity reactions were induced in sensitized guinea-pigs by subcutaneous injection of an A60 dose of 0.01 g (minimal revealing dose). A quantity thousandfold higher (15 g A60) was unable to induce in unsensitized guinea pigs the mounting of a cellular immunisation against A60, as shown by negative cutaneous testings 1 month later. Our results show that A60 preparations satisfied the requirements of the European Pharmacopoeia Commission and met the WHO recommandations for new tuberculins. Handicaps of old tuberculin and PPD (heterogeneous mixtures titrated biologically and unstable in solution) can be overcome by A60 preparations (a single antigen spectrophoretically measurable and stable).Abbreviations BCG Mycobacterium bovis, strain Calmette Guérin - A60 Antigen 60 of BCG (classification of Closs et al. [4] - CMN Group of Gram-positive bacteria including genera Corynebacterium, Mycobacterium and Nocardia - TMA (thermostable macromolecular antigens): a group of mycobacterial components including A60 of BCG and A7 of M. leprae - Elisa (enzyme-linked immunosorbent assay) immunoenzymometric method - Ph.Eur.U. European Pharmacopoeia Units of tuberculins