Delayed hypersensitivity responses in mice and guinea pigs to Mycobacterium leprae, Mycobacterium vaccae, and Mycobacterium nonchromogenicum cytoplasmic proteins

Abstract

Antigenic relationships between Mycobacterium vaccae, M. nonchromogenicum, and M. leprae were examined in mice and guinea pigs injected with M. vaccae or M. nonchromogenicum suspensions. The growth of both organisms in outbred ICR and four inbred mouse strains was followed up to 30 days. M. nonchromogenicum persisted in the livers and spleens of the inbred mice substantially better than did the M. vaccae population in the same mouse strains. A translucent colony variant of M. vaccae isolated from the opossum survived in vivo better than the opaque colony isolated from opossums and cattle. Persistence of M. vaccae and M. nonchromogenicum was not markedly increased in T-celldepleted (nude) mice. Normal mice infected with increasing numbers of M. vaccae did not develop delayed-type hypersensitivity to the homologous M. vaccae cytoplasmic protein antigen. When heat-killed M. vaccae were incorporated into Freund adjuvant, both mice and guinea pigs developed delayed hypersensitivity to cytoplasmic antigens prepared from M. vaccae, M. nonchromogenicum, and M. keprae, but not to purified protein derivative. Both M. nonchromogenicum and M. vaccae vaccines cross-sensitized guinea pigs to the M. leprae cytoplasmic antigens.

T-cell immune responses in Mycobacterium avium-infected mice

“…The suspension was checked for sterility and stored at -20°C until required. The cell suspension was subjected to sonic disruption at 0°C with a Braun-Sonic probe as described previously (19). The residual whole cells were removed by centrifugation at 10,000 x g at 4°C for 20 min followed by ultracentrifugation at 100,000 x g for 2 h. The clear supernatant fluid was sterilized by passage through a 0.45-,um-pore-size membrane filter.…”

“…Delayed-hypersensitivity determinations. Groups of infected mice were injected in the left hind footpad with 10 ,ug of purified protein derivative (Evans Medical, Langhurst, United Kingdom) or 10 jig of MASE dissolved in 25 ,ll of pyrogen-free saline (19). Saline was injected into the contralateral footpad, and the increase in foot thickness was compared by using dial gauge calipers at 3, 24, and 48 h. An increase of 0.18 mm (1.8 U) or more was considered to be significant at the 1% level (6).…”

Systemic Mycobacterium kansasii infection and regulation of the alloantigenic response

“…Animals. Specific-pathogen-free inbred C3H(He) and B6D2F1 hybrid mice were obtained from the Animal Breeding Facility at the Trudeau Institute and kept under conditions described previously (7,25).…”

“…Footpad hypersensitivity determinations. Groups of five infected and five normal mice were footpad tested after 2 or 4 weeks by injecting 5 l&g of M. kansasii 1203 cytoplasmic protein antigen prepared by sonic disruption of whole cells, as described previously (25). Increases in foot thickness were measured with a Schnelltister after 3, 6, and 24 h (6), the normal saline control values were subtracted, and the mean values were calculated.…”

Induction of immunity against experimental tuberculosis with mycobacterial mannophosphoinositides encapsulated in liposomes containing lipid A

“…In humans a typical delayed reaction usually peaks between 48 and 72 h [19], whereas in the present study PIM-based antigens elicited a maximum reaction at 24 h in mice. Earlier observations [20,21] have also demonstrated that in guinea-pigs and mice the skin reaction reaches a maximum earlier (24 h) than in humans. These differences in the rates of hypersensitivity reaction may be because of difference in host species.…”