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The cell-mediated immune response against infection with Sporothrix schenckii was investigated in vivo and in vitro. Following primary infection, mice were protected against a secondary subcutaneous inoculation with S. schenckii as determined by the local growth of fungi, and subcutaneously immunized mice were able to survive a lethal intravenous infection. Protection could be transferred to naive congenitally athymic nude mice using lymph node cells taken from immune mice. When immune lymph node cells had CD4+ cells depleted by antibody plus complement, the ability to transfer protection was significantly reduced. Treatment of mice with carrageenan, a macrophage blocker, before and after secondary inoculation abolished protection. An in vitro system, using either immune lymph node cells alone or macrophages alone, failed to kill the organism. However, inhibition of fungal growth was observed when both immune lymph node cells and macrophages were combined. Growth inhibition in vitro was reduced after depletion of CD4+ cells but not CD8+ cells. Addition of anti-interferon (IFN)-gamma monoclonal antibodies (MAb) also reduced growth inhibition. By using reverse transcription-polymerase chain reaction (RT-PCR) analysis, it was shown that immune lymph node cells expressed message for IFN-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-10 after stimulation with heat-killed S. schenckii. These results suggest that acquired immunity against S. schenckii is expressed mainly by macrophages activated by CD4+ T cells.

Involvement of CD4+ T cells and macrophages in acquired protection against infection with Sporothrix schenckii in mice