The pathogenesis of leprosy in the nine-banded armadillo and the significance of IgM antibodies to PGL-1.

Twenty-seven nine-banded armadillos captured from the wild and tested free of wild M. leprae infection were distributed into four groups. They were injected at the right hind footpad with saline suspensions of M. Leprae at doses of 10(3), 10(4), 10(5) and 10(6). PGL-1 antibody levels were estimated using an ELISA test, twice during six months before the infection and every two months after the infection. One animal from each group was sacrificed at 6, 12, 18, 24, and 30-month intervals and another eight at unspecified intervals. A thorough autopsy and histopathological examination were conducted on all of them. Of the 27 animals, 18 developed the infection. In 10, there were granulomas at the site of inoculation and in 17 the regional lymph nodes were infected. The disease spread extensively to other lymph nodes and to the liver and spleen and then to the other organs. Peripheral nerves were invaded by M. leprae in only five animals. PGL-1 antibody levels registered a positive reading in only six of the 18 animals with the infection. In armadillo leprosy, the lesions did not persist at the site of entry in all animals M. leprae multiplied in the macrophages at the site of inoculation and the reticuloendothelial cells of the lymph nodes before they spread to other organs. There was evidence of invasion of endothelial cells of capillaries and possible bacteraemia even at an early phase of the infection. Peripheral nerves were not the preferred sites of entry or multiplication of M. leprae. Progressive increase in PGL-1 antibodies was recorded in five lepromatous armadillos with disseminated infection and high bacterial load. However, PGL-1 antibodies response was not sensitive enough to detect early disease.

An attempt to produce experimental tuberculoid leprosy in the nine-banded armadillo.

In an attempt to produce experimental tuberculoid leprosy, three nine-banded armadillos, two with borderline tuberculoid lepromin reaction, and one with tuberculoid lepromin reaction, were chosen. They were injected subcutaneously in a four square centimetre area in the abdominal skin with saline suspension of 6.5 x 10(7) M. leprae. Induration of skin at the injected site appeared in 24 hours and persisted for 6 months in one and for 18 months in the other two animals. Histopathological examination of the infected site at 6 weeks, 18 and 20 months showed progressively decreasing granulomatous inflammation; but the cutaneous nerves were uninvolved. Autopsy examination of the three animals failed to show disseminated disease. Since there was no evidence of nerve involvement, experimental transmission of tuberculoid leprosy to armadillos could not be established in this study.

Early infection with M. leprae and antibodies to phenolic glycolipid-I in the nine-banded armadillo.

Nine-banded armadillos were intravenously infected with 10(9) M. leprae. IgM antibodies to PGL-I were evaluated three times during the six months before and every two months after the infection. A thorough autopsy examination was done on animals that died or were sacrificed at intervals of 3, 4, 6, 12, 15 and 18 months after the infection. Three animals which had acquired the infection in the wild and one experimentally infected animal showed significant increases in antibody levels corresponding to their high bacterial load. In the other five experimentally infected animals, M. leprae infection was established in the cells of the reticulo endothelial system (RES) long before the IgM antibody levels to PGL-I became positive. It is possible that in human leprosy also M. leprae may enter and multiply in the RES initiating antibody production during the incubation period before clinical disease with neuritis becomes manifest.

Phagocytosis of Mycobacterium leprae by cardiac muscle cells--a preliminary report.

Fetal cardiac muscle cells were shown to ingest M. leprae easily within 20 minutes of exposure in vitro. This phagocytosis is considered nonspecific and facilitated by the lipid coat of the mycobacteria. The presence of M. leprae free in the cytoplasm of the muscle cells did not seriously affect the morphology or rhythmic contractions of the cells. The significance of the presence of M. leprae in somatic cells needs further study.

Our experiences with breeding of nine-banded armadillos (Dasypus novemcinctus) in captivity.

The nine-banded armadillo is considered the best animal model in the study of leprosy. Armadillos have never been successfully bred in the laboratory and therefore animals required for the experiments are captured from the wild and are likely to carry many diseases including leprosy. An attempt was made to breed them in captivity. Our attempt to house them in various combinations in their natural environment in the farmlands of Louisiana was successful. Although a significant increase in conception and delivery was recorded, consistent breeding of the animals was not obtained.

Effect of repeated lepromin testing on experimental nine-banded armadillo leprosy.

Twenty-eight armadillos were lepromin tested and infected with M. leprae; 18 intravenously and 10 intradermally. The lepromin test was repeated after 3 months and at intervals of 6 months thereafter until their death or sacrifice up to 30 months. The one animal with tuberculoid lepromin was resistant and 14 of the 16 with lepromatous lepromin developed generalized disease. Of the 11 with borderline lepromin, 6 developed disseminated disease and 5 were resistant. There is a definite relationship between resistance and tuberculoid lepromin in the armadillo. Repeated lepromin testing had no effect in the rate of infection and the course of the disease in animals infected intravenously. In the intradermally infected animals the results were inconclusive. Whereas all the 20 animals with disseminated disease showed lesions in the liver, spleen and lymph nodes, only 4 animals had sciatic nerve involvement. Peripheral nerve trunk is not necessarily the preferred site in the armadillo. Lung lesions were an important cause of death in lepromatous armadillos.

First lesion in experimental armadillo leprosy.

Eighteen armadillos were infected intravenously with 10(8) M. leprae and 10 intracutaneously with 10(7) M. leprae. Among those which developed disseminated disease, a nodule at the site of inoculation was the first lesion noticed in 14 of the 16 infected intravenously and 4 of the 4 infected intradermally. It is possible that in human leprosy the first sign of infection is localized proliferation of M. leprae at the site of entry, and even nodule formation in lepromatous patients. It may be important to search for asymptomatic swelling or keloid-like lesions in skin or in nasal mucosa while screening a population for early leprosy.