A comparative study on the Mitsuda type response to antigens of chemoautotrophic nocardioform bacteria and to standard lepromin in leprosy patients.

Anergy, or contrarily, Mitsuda-type responses towards 4 chemoautotrophic nocardioform antigens (CAN-Ags) and a control standard lepromin were tested in 73 LL, TT and borderline cases of leprosy. The antigens injected per patient varied from a maximum of 5 to a minimum of 2. Complete anergy to CAN-Ags was seen in 92/92 instances tested on 24 LL cases. The anergy was weakly modified or unmodified in 3 other LL cases which had been vaccinated before. Concurrent studies with the same antigens tested on 33 TT cases showed clear-cut, dose-dependent, Mitsuda-type late responses in 80/81 instances. The CAN bacteria, therefore, despite their origin from different unrelated leprous human, mouse footpad (MFP) and armadillo tissues, appeared to be identical with each other and also probably related to the leprosy bacillus, on the basis of these parameters.

Culture of nocardioform bacilli from leprosy patients & clinical evaluation of nocardioform bacilli derived antigen.

An antigen derived from cultured nocardioform bacilli was compared with Mitsuda lepromin in intradermal skin test reactions. Nocardioform bacilli were cultured in gelatin minimal medium from the tissue fluid of 85 lepromatous patients (56 M, 29 F). Of these, 65 samples showed uncontaminated growth of the organism, which were pooled for the manufacture of the test antigen. This antigen was intradermally tested in 50 untreated leprosy patients irrespective of the type, together with Mitsuda lepromin and sterile gelatin minimal media, which served as a control. No early reaction was observed at 72 h, while the late reaction at 28 days was positive in all patients in the Tuberculoid (TT) group with both antigens. Eighteen patients (81.8%) in the Borderline tuberculoid (BT) group reacted strongly to Mitsuda lepromin at 28 days, while 21 patients (95.5%) in this group showed a strong late reaction with the test antigen. The lepromatous (LL) group did not show any reaction with the two antigens. It is inferred that nocardioform bacilli are easy to cultivate, and that the test antigen compares well with Mitsuda lepromin.

Crossreactivity between Mycobacterium leprae and various actinomycetes and related organisms.

Serological crossreactivity was analyzed between M. leprae and strains of various species of Corynebacterium, Mycobacterium, Nocardia, Rhodococcus, Streptomyces, and related organisms. M. leprae shares antigens with most of these organisms, and sera from patients with lepromatous leprosy contain antibodies against them. The results demonstrate that M. leprae shares more antigens with the mycobacteria than with strains of the other tested genera, thus supporting the view that the leprosy organism belongs to the genus Mycobacterium. One precipitinogen (designated p beta) was found to be common to M. leprae and the streptomycetes, and sera from patients with lepromatous leprosy contain antibodies against this antigen.

Immunochemical analysis of soluble antigens of Mycobacterium lepraemurium.

Soluble antigens obtained from purified M. lepraemurium were identified by SDS polyacrylamide gel electrophoresis as three different proteins, M-W 1 X 10(4), 1 X 10(5), and 4 X 10(6), and two polysaccharide-polypeptides. Three antigens were found to react with anti-M. leprae antiserum; a single antigen proved to be immunologically related to M. tuberculosis, M. similae, M. balnei and a few other mycobacteria, but not to Eubacteriales and fungi, as examined by immunoelectrophoresis. In the dermal hypersensitivity assay, the guinea pigs sensitized with soluble antigens of M. lepraemurium only responded to the antigens of M. lepraemurium, M. leprae and partly to M. scrofulaceum, but not to other antigens, including lepromin and leprous serum globulins.

Immunological determination of Mycobacterium leprae by means of cytoplasmic antigens.

Mycobacterium leprae was isolated and purified from lepromas, the spleen, and the liver of leprosy patients. An immunodiffusion analysis of the cytoplasms obtained from four lots of M. leprae and M. lepraemurium, 295 strains of different actinomycetales, and 12 other bacteria was performed with the use of the cytoplasm antisera. Immunological relationships were revealed between the cytoplasms of M. leprae, M. lepraemurium, M. avium, M. gallinarum, M. tuberculosis, M. simiae, M. kansasii, M. chitae, M. capsulatum, Actinomyces israelii, A. naeslundii, and some strains of saprophytic mycobacteria. These studies led to the proposed concept of the immunological evolution of M. leprae and M. lepraemurium and an Actinomyces-like progenitor through M. avium-M. gallinarum and to a proposal for the polyvalent vaccine currently being developed by this research group.