3-Methylindole-induced splenotoxicity: Functional analysis of immune parameters and lymphocyte phenotyping by flow cytometry

Abstract

In this laboratory, 3-methylindole (3-MI), a pneumotoxic metabolite of l-tryptophan that forms in the digestive tract of humans and ruminants, has been demonstrated to be toxic to rat and mouse splenic cells both in vitro and in vivo. The present studies examine whether the reduction in nucleated splenic cells is associated with alterations in: (1) immune functioning (e.g., B and T cell mitogenic responses to lectins), (2) natural resistance (e.g., natural killer (NK) activity and cytokine release from macrophages (MPs)), or (3) the relative percentages of B and T cells in the remaining cells as determined by flow cytometric phenotyping. A dose-dependent decrease in splenic weight (24–46%) and nucleated cell numbers (54–73%) was observed 24 hr after intraperitoneal (ip) administration of 100–300 mg/kg 3-MI to B6C3F1 mice. At a dose of 300 mg/kg, the blastogenic response of splenic lymphocytes to 1 μg/ml phytohemagglutinin, a T cell mitogen, was reduced 37 and 64%, and NK activity was reduced 20 and 60%, in rats and mice, respectively. Following exposure to 400 mg/kg 3-MI, interleukin-1 and tumor necrosis factor production by lipopolysaccharide-stimulated rat splenic MPs was decreased 58 and 38%, respectively. Despite the reduction in total nucleated cell number in 3-MI-treated mice, the percentages of splenic B and T cells remained the same. These findings indicate that, in addition to its toxicity to splenic cells, 3-MI can significantly impair the functioning of the remaining viable cells. The potential importance of these functional changes for alterations in host resistance in rodents exposed to 3-MI or other alkylindoles is unknown.