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Skin ulcer and immunoblot patterns by inoculation sites in BALB/c mice infected with Leishmania major
M Lee,J K Lee,S B Kim,S H Lee and S T Hong*
Department of Parasitology and Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea.
Received February 17, 1997; Accepted February 25, 1997.
Abstract
The skin ulcer in Leishmania major infection is known to be variable according to the inoculation sites even in a susceptible host. The present study traced the immunoblot patterns by the site of inoculation and duration of infection in BALB/c mice. L. major were subcutaneously inoculated on the nose, footpad, and back of the mice, in a dose of 3 × 106 promastigotes. Sera of the mice were collected every 10 days after inoculation. SDS-PAGE separated soluble protein bands of the promastigotes and immunoblot was carried out with the infection sera. The skin ulcer first appeared on the nose at 15 days, and on the footpad at 17 days after inoculation. The ulcer on the back appeared after 90 days. In the mice with ulcer on the nose or footpad, serum IgG antibody reacted to 202, 139, 98, 83, 81, 67 65, 62, 59, 54, 52, 42, 26 and 23 kDa bands at 20 days after inoculation. In mice inoculated on the back, however, the immunoblot showed visible reactions with 202, 83, 81, 74, 67, 65, 62, 59, 54, 52, 20 and 17 kDa bands at 90 days after inoculation. The present result showed that the antigenic protein bands of L. major promastigotes were differed by the inoculation site and duration of infection. Since the skin ulcer and the serum antibodies to antigenic bands between 67-52 kDa appeared simultaneously, it is suggested that the serum IgG antibodies may play a role in formation of the skin ulcer in BALB/c mice.
Figures
Fig. 1 A. Ulceration on the nose of a BALB/c mouse at 20 days after inoculation of L. major promastigotes. B. Ulceration on the footpad of a BALB/c mouse at 20 days after inoculation of L. major promastigotes. C. Skin lesion on the back of a BALB/c mouse at 90 days after inoculation of L. major promastigotes.
Fig. 2 Crude extract of L. major promastigotes in 12.5% SDS-GAGE.
Fig. 3 Immunoblot patterns of L. major crude antigen with sera of mice with nasal ulceration. Lane 1, 10 days after inoculation; 2, 20 days; 3, 30 days; 4, 40 days after inoculation.
Fig. 4 Immunoblot patterns of L. major crude antigen with sera collected at 40 days after inoculation. lanes 1-5: sera from mice with nasal ulcer; lanes 6-10: sera from mice with pedal ulcer; lanes 11-15: sera fron mice with dorsal inoculation.
Fig. 5 Immunoblot patterns of L. major crude antigen with sera of mice with dorsal ulceration. lanes 1-6, intervals of 10 days after inoculation; 7, 90 days; 8, 100days.
Fig. 6 Summary of immunoblot patterns of IgG antibody to crude antigen of L. major by inoculation sites.
Tables
Table 1 Number of mice with ulceration at the inoculation site of L. major by duration of infection
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