Delayed-type hypersensitivity reaction induced in broilers via trachea inoculation of killed Staphylococcus aureus

Abstract

A study was conducted to determine whether the delayed-type hypersensitivity (DTH) reaction to killed Staphylococcus aureus antigen in chickens could be induced through multiple intratracheal inoculations. Three criteria were used to assess DTH: 1) delayed footpad reaction (DFR) with a peak response at 24 to 48 h postchallenge, 2) inhibition of monocyte/macrophage migration, and 3) mononuclear cell infiltration at the challenge site. Broilers were sensitized three times with a s.c. injection in the neck or intratracheal inoculation of killed S. aureus in polyethylene glycol at 2, 3, and 4 wk of age. Controls were given polyethylene glycol with a s.c. injection in the neck or intratracheal inoculation. Migration inhibition tests were conducted at 6 wk of age. At 7 wk of age, all birds were challenged intradermally with S. aureus antigen in PBS in the right footpad. The left footpad was injected with PBS. The thickness of the footpad was measured at 0, 4, 24, and 48 h postchallenge to evaluate the DFR. Birds were euthanatized, and both footpads were removed for histopathological examination. Subcutaneously or intratracheally sensitized birds showed significant DFR compared with nonsensitized birds (P < 0.0001), which reached maximum response at 24 h postchallenge. The s.c. sensitization resulted in an inhibition of the in vitro migration of monocytes/macrophages (P < 0.0001), whereas intratracheally sensitized birds did not show migration inhibition of monocytes/macrophages. Histological examination showed typical perivascular infiltration of small lymphocytes in S. aureus-injected footpads from s.c. and intratracheally sensitized birds. These results indicate that multiple intratracheal inoculation, as well as s.c. injection of killed S. aureus antigen, can be used to induce a cell-mediated DTH reaction in chickens.