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Dissociation between cytokine mRNA expression and protein production in shigellosis

In our study, infection with Shigella dysenteriae type 1 (n = 16) or Shigella flexneri in adults (n = 5) was associated with a gradual accumulation of mRNA for interleukin (IL)‐1β, tumor necrosis factor (TNF)‐α, IL‐6, transforming growth factor‐β, IL‐10, IL‐4, TNF‐β, interferon (IFN)‐γ and perforin in the rectal biopsy samples during the convalescent stage of the disease demonstrated by in situ hybridization. In contrast, immunohistochemical staining in rectal tissues of cytokine protein‐producing cells at the single‐cell level exhibited a steady‐state expression during 2–36 days after the onset of the disease. The frequency of cytokine mRNA‐expressing cells varied in the range of 3–100‐fold higher than that of the corresponding protein‐synthesizing cells. The accumulation of cytokine mRNA in vivo during shigellosis represented a long‐lasting phenomenon throughout the disease course, and may be linked to its immunopathogenesis. The results also indicate that assessment of both protein and mRNA in vivo may provide complementary information. Stimulation in vitro of peripheral blood mononuclear cells from normal healthy donors with Shigella‐derived lipopoly‐saccharide or shiga toxin was carried out to elucidate the role of Shigella antigens in the regulation of translation of cytokine‐specific mRNA. The incidence of cytokine (IFN‐γ, IL‐6 and TNF‐α) mRNA‐ and cytokine protein‐expressing cells was very similar and congruent after both these Shigella‐derived stimuli. We could, thus, not find evidence for shiga toxin‐induced down‐regulation of cytokine mRNA translation as the explanation for the observed discrepancy between cytokine mRNA and protein levels in the tissue biopsies.