Malathion, Lindane, and Piperonyl Butoxide, Individually or in Combined Mixtures, Induce Immunotoxicity via Apoptosis in Murine Splenocytes In Vitro

Abstract

Lindane, malathion, and piperonyl butoxide were cultured singly or as mixtures with murine splenocytes to evaluate changes in cell death and caused cytotoxicity in a concentration- and time-dependent manner. Pesticide mixture studies were then performed based on minimum cytotoxicity concentrations (<LC(25)). Cytologic analysis and the alamarBlue assay revealed that individual pesticides and mixtures of malathion/lindane and malathion/piperonyl butoxide prompted cytotoxicity, which was supported by DNA ladder analysis. Using 7-aminoactinomycin D, apoptosis was quantified at 6.5%, 12.0%, 13.2%, 19.3%, and 23.4% for malathion, lindane, piperonyl butoxide, malathion-lindane, and malathion-piperonyl butoxide, respectively. Staining with 7-aminoactinomycin D and B- or T-cell-specific fluorescent-labeled monoclonal antibodies showed B cells to be more susceptible to malathion and piperonyl butoxide treatments than T cells. Treatment of murine splenocytes in vitro with minimum cytotoxic concentrations of lindane, malathion, and piperonyl butoxide and their mixtures induced apoptosis, the effect elicited by the mixtures being additive compared with the individual pesticide effect.

Estudo dos aspectos toxicológicos à saúde humana em relação ao praguicida organofosforado metamidofós e sua situação regulatória no Brasil e no mundo

“…Apenas alguns organofosforados têm sido sistematicamente investigados quanto a seu potencial imunotóxico (BATTAGLIA et al, 2010;GALLOWAY, HANDY, 2003;HERMANOWICZ, KOSSMAN, 1984;KOVTYUKH, 1995;NEWCOMBE, 1992;REPETTO, BALIGA, 1996;RODGERS et al, 1985a;QING, 2007;VOCCIA et al, 1999) O sistema imunológico é importante para a defesa do organismo contra porém, ocorreu diminuição da proliferação de linfócitos T para a formação de anticorpos. O autor conduziu um outro estudo in vivo, em 1990, com camundongos machos, através da administração intraperitoneal de 0,6; 1,2 e 6 mg/kg pc de metamidofós, e com ratos através de adrenalectomia após a administração de uma dose única intraperitoneal de metamidofós na concentração de 1,2 mg/kg pc.…”

Immunotoxicity in mice induced by short-term exposure to methoxychlor, parathion, or piperonyl butoxide

“…Based on the EPA guideline (1998), mice are a model species recommended for use in immunotoxicity studies that test effects of agricultural chemicals (see as was done in Casale et al, 1984;Diel et al, 1999;Battaglia et al, 2010). Furthermore, for the immunotoxicity study, only one gender needed to be evaluated; in general, female animals are considered to yield more consistent outcomes than are male counterparts during evaluation of effects from test articles on host humoral immune responses.…”

“…Piperonyl butoxide is an agricultural insecticide synergist used mainly with pyrethroids (Carson et al, 1988;Mitsumori et al, 1996;Emerson et al, 2001). Recent evidence suggests that piperonyl butoxide administration depletes T-lymphocytes in the spleen and thymus, induces hypoplasia of the bone marrow, and inhibits T-lymphocyte proliferation in lymphoid tissues (Mitsumori et al, 1996;Diel et al, 1999;Battaglia et al, 2010). For in vitro studies, parathion was dissolved in DMSO to 0.1% (w/v).…”

Effects of short-term oral combined exposure to environmental immunotoxic chemicals in mice

“…Female mice were selected as the model for this study because the EPA Immunotoxicity Guidelines (EPA, 1998) consider the mouse a model species for use in immunotoxicity studies that examine effects of agricultural chemicals (see Casale et al, 1984;Diel et al, 1999;Battaglia et al, 2010). The guideline indicates that either rats or mice may be used.…”

“…methoxychlor (MXC) -an organochlorine compound, parathion (PARA) -an organophosphate compound, and piperonyl butoxide (PBO) -an agricultural insecticide synergist. These three chemicals were selected on the basis of previous studies: MXC exposure causes atrophy of CD4 þ CD8 þ T-cells in the thymus (Takeuchi et al, 2002a,b); PARA markedly inhibits antigen-specific IgM production (Casale et al, 1984); and PBO depletes T-cells in the spleen and thymus, induces bone marrow hypoplasia, and inhibits T-cell proliferation in lymphoid tissues (Diel et al, 1999;Battaglia et al, 2010;Mitsumori et al, 1996). We also previously showed that MXC, PARA, and PBO exposure results in increased thymocyte apoptosis, markedly inhibited sheep red blood cell (SRBC)-specific IgM production, and aggravation of immune disorders such as atopic dermatitis and allergic airway inflammation (Fukuyama et al, 2011;Nishino et al, 2013).…”