f Amylosin, a heat-stable channel-forming non-ribosomally synthesized peptide toxin produced by strains of Bacillus amyloliquefaciens isolated from moisture-damaged buildings, is shown in this paper to have immunotoxic and cytotoxic effects on human cells as well as antagonistic effects on microbes. Human macrophages exposed to 50 ng of amylosin ml ؊1 secreted high levels of cytokines interleukin-1 (IL-1) and IL-18 within 2 h, indicating activation of the NLRP3 inflammasome, an integral part of the innate immune system. At the same exposure level, expression of IL-1 and IL-18 mRNA increased. Amylosin caused dose-dependent potassium ion efflux from all tested mammalian cells (human monocytes and keratinocytes and porcine sperm cells) at 1 to 2 M exposure. Amylosin also inhibited the motility of porcine sperm cells and depolarized the mitochondria of human keratinocytes. Amylosin may thus trigger the activation of the NLRP3 inflammasome and subsequently cytokine release by causing potassium efflux from exposed cells. The results of this study indicate that exposure to amylosin activates the innate immune system, which could offer an explanation for the inflammatory symptoms experienced by occupants of moisture-damaged buildings. In addition, the amylosin-producing B. amyloliquefaciens inhibited the growth of both prokaryotic and eukaryotic indoor microbes, and purified amylosin also had an antimicrobial effect. These antimicrobial effects could make amylosin producers dominant and therefore significant causal agents of health problems in some moisture-damaged sites.A mylosin is a heat-stable 1,197-Da peptide toxin originally found to be produced by strains of Bacillus amyloliquefaciens isolated from moisture-damaged buildings (1, 2). B. amyloliquefaciens belongs to the Bacillus subtilis group, members of which have been found in high numbers in moisture-damaged buildings where occupants have experienced building-related ill health symptoms (3, 4). Amylosin forms cation-permeant ion channels with high selectivity for K ϩ in lipid membranes (5) and thus depolarizes the plasma membrane and the mitochondria inside live mammalian cells, disrupting cellular ion homeostasis, mitochondrial functions, and energy metabolism (1, 2). It has been suggested that amylosin causes health risks for people living or working in moisture-damaged buildings contaminated with amylosin-producing bacteria (1). Amylosin has also been shown to be produced by B. subtilis and Bacillus mojavensis strains connected to foodborne illness (6).Moisture-damaged buildings harbor many kinds of fungi and bacteria, including producers of bioactive compounds (7, 8; reviewed in reference 9). The symptoms experienced by people living or working in moisture-damaged buildings vary (10), but they could at least partly result from activation of the innate immune system (11,12). Symptoms such as acute inflammatory responses and pseudoallergic reactions may be a result of the activation of inflammasomes (12,13). Inflammasomes are components of the innate immun...