RESUMO
Food allergy is an increasing health problem in Western countries. Previously, it has been shown that the intensity of food allergic reactions can be regulated by regulatory T (T(reg)) cells. In addition, it has been shown that activation of the aryl hydrocarbon receptor (AhR) regulates T-cell responses by induction of T(reg) cells. Therefore, we hypothesized that activation of the AhR pathway can suppress development of food allergic responses through the induction of T(reg) cells. This was investigated by using a mouse model for peanut allergy. C3H/HeOuJ mice (AhR(b)(-2)) were sensitized to peanut by administering peanut extract (PE) by gavage in the presence of cholera toxin and were treated with the prototypical AhR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (0.6, 1.7, 5, and 15 µg/kg body weight) on days 3 and 11 orally. The functional role of CD4(+)CD25(+)Foxp3(+) T(reg) cells was investigated by depleting these cells with anti-CD25 mAb during sensitization to PE. TCDD treatment dose dependently suppressed sensitization to peanut (PE-specific IgE, IgG1, and IgG2a and PE-induced IL-5, IL-10, and IL-13, respectively). The percentage, but not the number, of CD4(+)CD25(+)Foxp3(+) T(reg) cells dose dependently increased by AhR activation in both spleen and mesenteric lymph nodes. Depletion of CD4(+)CD25(+)Foxp3(+) T(reg) cells markedly reversed the suppressive effect of TCDD on PE-specific antibody levels and PE-induced IL-5, IL-10, and IL-13 cytokine production. Present data demonstrate for the first time that activation of the AhR by TCDD suppressed the development of Th2-mediated food allergic responses. A functional shift within the CD4(+) cell population toward CD4(+)CD25(+)Foxp3(+) T(reg) cells appeared to underlie this effect. This suggests that the AhR pathway might provide potential therapeutic targets to treat food allergic diseases.
Assuntos
Alérgenos/imunologia , Arachis/imunologia , Tolerância Imunológica/imunologia , Hipersensibilidade a Amendoim/imunologia , Receptores de Hidrocarboneto Arílico/biossíntese , Animais , Anticorpos Bloqueadores/farmacologia , Arachis/química , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Antagonismo de Drogas , Fatores de Transcrição Forkhead/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Hipersensibilidade a Amendoim/metabolismo , Extratos Vegetais/imunologia , Extratos Vegetais/toxicidade , Dibenzodioxinas Policloradas/farmacologia , Linfócitos T Reguladores/imunologiaRESUMO
BACKGROUND: Food allergy affects approximately 6% of children and is the leading cause of hospitalization for anaphylactic reactions in westernized countries. Crucial in the establishment of allergy is the activation of dendritic cells (DC) leading to T helper 2-mediated responses. OBJECTIVE: We, therefore, investigated whether changes in DC subsets precede the establishment of food allergy, and which DC subsets have functional relevance during allergic sensitization in a mouse model. METHODS: Changes in DC populations in the intestine were analysed after exposure to cholera toxin alone and in combination with peanut extract (PE) as an allergen. To study the functional role of DC subsets in relation to food allergy, we used expansion of DC in vivo by treatment with Flt3L. RESULTS: Sensitization to PE in this mouse model was accompanied by a shift in DC subsets in intestinal tissues towards more CD11b(+) DC and less CD103(+) DC. No significant changes in the plasmacytoid DC (pDC) numbers were observed. Flt3L treatment, resulting in the expansion of all DC subtypes, inhibited allergic manifestations in our model, including Th2 cytokine production, PE-specific IgE and PE-induced mast cell degranulation. pDC depletion reversed Flt3L-induced inhibition of IgE responses and mast cell degranulation. conclusions and clinical relevance: The establishment of food allergy is accompanied by profound changes in DC subsets in the intestine towards more inflammatory CD11b(+) DC. In addition, expansion of DC numbers by Flt3L, in particular pDC, inhibits the establishment of allergic manifestations in the intestine. These findings are of relevance for understanding the role of DC subsets early during the process of allergic sensitization, and may lead to new therapeutic or prophylactic opportunities to prevent food allergy.
Assuntos
Arachis/química , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/metabolismo , Intestinos/citologia , Intestinos/imunologia , Extratos Vegetais/imunologia , Adjuvantes Imunológicos/farmacologia , Animais , Antígenos CD/imunologia , Antígeno CD11b/imunologia , Células Dendríticas/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Cadeias alfa de Integrinas/imunologia , Intestinos/efeitos dos fármacos , Linfonodos/citologia , Proteínas de Membrana/farmacologia , Camundongos , Camundongos Endogâmicos C3H , Organismos Livres de Patógenos EspecíficosRESUMO
BACKGROUND: Recent studies have implicated CD4(+) CD25(+) regulatory T cells (nTregs) in the maintenance of tolerance to oral antigens and in the regulation of the food allergic IgE response. OBJECTIVE: The objective was to assess if nTregs can transfer allergen-specific oral tolerance to naïve, non-TCR transgenic mice and regulate peanut extract (PE)-specific hypersensitivity responses. Additionally, the role of the regulatory cytokines IL-10 and TGF-ß in the modulation of peanut-allergic sensitization was studied. METHODS: CD25-enriched T cells from PE-tolerant mice were adoptively transferred to recipient mice, which were subsequently sensitized to PE. Depletion of CD25(+) cells and neutralization of IL-10 and TGF-ß were compared in a CH3/HeOuJ mouse model of peanut-allergic sensitization. RESULTS: Transfer of CD25(+) Tregs-enriched cell populations did not affect the PE-specific cytokine production or PE-specific antibody levels compared with control mice but interestingly resulted in a decrease of mast cell responsiveness. On the contrary, transfer of CD25(+) Tregs-depleted cells caused an increase in non-specific cytokine production, in the absence of changes in PE-specific responses. TGF-ß neutralization resulted even in a larger increase in spontaneous release of all cytokines measured (IL-4, IL-5, IL-10, IL-13, and IFN-γ), but surprisingly also to a higher PE-specific Th2-associated (IL-4, IL-5, IL-13) cytokine production compared with depletion of CD25 cells or neutralization of IL-10. Similarly, depletion of CD25 cells and TGF-ß neutralization but not of IL-10 neutralization lead to an increase in PE-specific antibody levels and elevated mast cell degranulation following a PE challenge. CONCLUSIONS AND CLINICAL RELEVANCE: We conclude that CD4(+) CD25(+) Tregs from non-transgenic-tolerant mice cannot transfer specific oral tolerance of exogenous antigens to naïve mice and are more involved in general immune suppressive mechanisms. However, we found evidence that TGF-ß secreting Tregs (Th3) may play an important role.
Assuntos
Alérgenos/imunologia , Arachis/imunologia , Tolerância Imunológica/imunologia , Hipersensibilidade a Amendoim/imunologia , Linfócitos T Reguladores/imunologia , Transferência Adotiva , Alérgenos/administração & dosagem , Animais , Anticorpos/sangue , Anticorpos/imunologia , Quimiocina CCL2/metabolismo , Citocinas/biossíntese , Citocinas/imunologia , Modelos Animais de Doenças , Feminino , Fatores de Transcrição Forkhead/imunologia , Fatores de Transcrição Forkhead/metabolismo , Subunidade alfa de Receptor de Interleucina-2/imunologia , Linfonodos/imunologia , Linfonodos/metabolismo , Depleção Linfocítica , Mastócitos/imunologia , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos C3H , Baço/imunologia , Baço/metabolismoRESUMO
BACKGROUND: Diclofenac and other non-steroidal anti-inflammatory drugs (NSAIDs) interfere with cyclo-oxygenase-mediated synthesis of prostaglandins, resulting in the inhibition of inflammatory immune responses. In contrast, it is known that NSAIDs are able to induce gastrointestinal damage. OBJECTIVE: Our aim was to investigate whether NSAIDs are able to enhance sensitization or abrogate tolerance to food antigens. METHODS: Mice were exposed to diclofenac and sensitized to peanut using cholera toxin as a mucosal adjuvant. In a tolerance model, oral tolerance was induced via feeding of peanut 3 weeks before sensitization with peanut. Diclofenac was administered before peanut feeding. After 4 weeks, peanut-specific antibodies in the serum and cytokine production in the spleen were measured. Induction of intestinal damage after oral exposure with diclofenac and peanut + cholera toxin was examined microscopically. RESULTS: Diclofenac-exposed animals showed increased levels of peanut-specific IgG1, IgG2a and IgE in the serum compared with vehicle-treated animals. Furthermore, peanut-induced cytokine production in the spleen was elevated upon diclofenac treatment. Importantly, diclofenac did not induce peanut-allergic responses in the absence of the cholera toxin, although exposure to diclofenac and peanut + cholera toxin resulted in intestinal epithelial damage. Reduced peanut-specific antibody production in the case of oral tolerance was not reversed after diclofenac exposure. However, oral tolerance, as measured by inhibition of peanut-specific cytokine responses, was reverted by diclofenac. CONCLUSIONS: These data point towards an increased risk for induction of food-allergic responses by diclofenac, when other circumstances are also in favour of induction of allergy.
Assuntos
Anti-Inflamatórios não Esteroides/efeitos adversos , Diclofenaco/efeitos adversos , Tolerância Imunológica/efeitos dos fármacos , Tolerância Imunológica/imunologia , Hipersensibilidade a Amendoim/imunologia , Animais , Modelos Animais de Doenças , Feminino , CamundongosRESUMO
BACKGROUND: Food allergy affects approximately 5% of children and is the leading cause of hospitalization for anaphylactic reactions in westernized countries. The mucosal adjuvant cholera toxin induces allergic sensitization to co-administered proteins in mice, while feeding the protein alone induces oral tolerance. Intestinal γδ T cells could be of importance in the induction of oral tolerance. This study aims to investigate whether γδ T cells have functional relevance in food allergic sensitization. METHODS: Changes in γδ T cells on days 1, 2, 3, and 7 after initiation of food allergy were evaluated using flowcytometry. Furthermore, the anti-γδ T-cell receptor (TCR) antibody UC7 was used to block the γδ TCR in mice in vivo, followed by sensitization to peanut. After 4 weeks, peanut-specific antibodies in serum and cytokine production in spleen were measured. RESULTS: Induction of food allergy resulted in a profound decrease in the percentage of γδ T cells in intestinal tissues and Peyer's Patches, but not in mesenteric lymph nodes or spleen. This decrease could be detected from days 1 to 2 after the initiation of food allergy and the number of γδ T cells returned to normal on day 7. Blockade of the γδ TCR resulted in elevated food allergic responses upon sensitization with peanut characterized by increased IgE and Th2 cytokine production in splenocytes. CONCLUSION: These results demonstrate a unique regulatory role of γδ T cells, suggesting that targeting γδ T cells in the intestine may contribute to strategies to prevent and possibly treat food allergy.
Assuntos
Hipersensibilidade Alimentar/imunologia , Imunização , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Linfócitos T/imunologia , Adjuvantes Imunológicos/farmacologia , Alérgenos/imunologia , Animais , Anticorpos/farmacologia , Arachis/imunologia , Contagem de Células , Células Cultivadas , Toxina da Cólera/farmacologia , Feminino , Intestinos/citologia , Intestinos/imunologia , Linfonodos/citologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Receptores de Antígenos de Linfócitos T gama-delta/antagonistas & inibidores , Baço/citologia , Baço/imunologia , Linfócitos T/efeitos dos fármacosRESUMO
BACKGROUND: Airborne particulate matter (PM) is an important factor associated with the enhanced prevalence of respiratory allergy. The PM adjuvant activity on allergic sensitization is a possible mechanism of action involved, and the induction of airway inflammation is suggested to be of importance in PM-induced adjuvant activity. OBJECTIVE: Because differently sized PM have different toxic potentials, we studied the role of particle size in the induction of airway inflammation and allergic sensitization. This was done using fine (0.250 and 0.260 micro m) and ultrafine (0.029 and 0.014 micro m) titanium dioxide (TiO(2)) and carbon black particles (CBP) with known differences in airway toxicity. METHODS: Mice were intranasally exposed to ovalbumin (OVA) alone or in combination with one of the different particles. The induction of airway inflammation and the immune adjuvant activity were studied in the lungs and lung-draining peribronchial lymph nodes (PBLN) at day 8. OVA-specific antibodies were measured at day 21, and the development of allergic airway inflammation was studied after OVA challenges (day 28). RESULTS: When administered at the same total particle mass (200 micro g), exposure to ultrafine TiO(2) and CBP-induced airway inflammation, and had immune adjuvant activity. The latter was shown by increasing both the PBLN cell numbers and the production of OVA-specific T-helper type 2 (Th2) cytokines (IL-4, IL-5, IL-10 and IL-13). Whereas OVA-specific IgE and IgG1 levels in serum were only increased in animals exposed to the ultrafine TiO(2), allergic airway inflammation could be detected in both ultrafine TiO(2)-and CBP-treated groups after challenges with OVA. CONCLUSION: Our data show that only the ultrafine particles, with a small diameter and a large total surface area/mass, cause airway inflammation and have immune adjuvant activity in the current model supporting the hypothesis that particle toxicity is site-dependent and related to adjuvant activity.
Assuntos
Antígenos/administração & dosagem , Material Particulado , Hipersensibilidade Respiratória/imunologia , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Feminino , Citometria de Fluxo , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Ovalbumina , Tamanho da Partícula , Hipersensibilidade Respiratória/patologia , Fuligem , TitânioRESUMO
Many chemicals, in particular drugs, cause systemic allergy or autoimmune-like disorders. Due to complex pathogenesis and strong dependence on genetic make-up, these immunotoxicological effects are usually missed in standard toxicity testing. Besides, animal studies that demonstrate chemically induced systemic allergy or autoimmune-like disorders are scarce. Here, animal models are presented that would fit into a predictive two-tiered strategy, designed to allow screening for immunostimulatory potential in the first tier, and more elaborate testing for allergenic or autoimmunogenic potential of selected chemicals in the second tier. The popliteal lymph node assay (PLNA), with or without reporter antigens, would fit in the first tier, and relevant route of exposure protocols with selected strains of mice or rats may be further developed to compose the second tier. To date, the relevant route of exposure models mentioned here (with 'normal' inbred mice and/or Brown Norway rats) has been tested with only a few chemicals, and the PLNA, although tested with over 100 chemicals, is not validated as yet. Conceivably, a major challenge in immunotoxicology is to incorporate the present knowledge on chemical-induced systemic allergy and autoimmunity in further development and validation of predictive models and strategies.
Assuntos
Doenças Autoimunes/imunologia , Autoimunidade/imunologia , Hipersensibilidade a Drogas/imunologia , Animais , Doenças Autoimunes/induzido quimicamente , Hipersensibilidade a Drogas/etiologia , Humanos , Imunidade/imunologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Valor Preditivo dos Testes , Xenobióticos/efeitos adversos , Xenobióticos/toxicidadeRESUMO
HgCl(2) and diphenylhydantoin (DPH) are prototype chemicals associated with diverse (auto)immune effects in genetically susceptible individuals. Both chemicals activate T cells, and the balance of Th1 versus Th2 activation may influence the clinical outcome of exposure. It is unknown which chemically created neoantigens are responsible for Th activation. We therefore investigated the effect of DPH and HgCl(2) on specific responses to TNP-ovalbumin, in mouse strains with varying sensitivity for the adverse effects. HgCl(2) was found to enhance Th2-driven antibody responses in susceptible B10.s, but protective type 1 responses in resistant B10.d2 mice. This was chemical-specific, as DPH enhanced type 2 responses in both strains. DBA/2 mice were relatively unresponsive to HgCl(2), whereas DPH stimulated type 1 responses in these mice. Interestingly, prior exposure to HgCl(2), but not DPH, facilitated IC deposition in B10.s mice only. Thus, we demonstrate that, depending on MHC-II and background genes, HgCl(2) and DPH preferentially adjuvate type 1 or type 2 responses. In case of HgCl(2), the type of response corresponds with susceptibility to antibody-mediated autoimmunity induced by this chemical. In addition, we demonstrate that, within one strain, different autoimmunogenic chemicals can enhance distinct responses to the same antigen.
Assuntos
Adjuvantes Imunológicos/farmacologia , Camundongos Endogâmicos/genética , Ovalbumina/administração & dosagem , Xenobióticos/farmacologia , Animais , Complexo Antígeno-Anticorpo/análise , Autoimunidade , Complemento C3/análise , Feminino , Hipersensibilidade Tardia/etiologia , Imunização , Isotipos de Imunoglobulinas/sangue , Túbulos Renais/imunologia , Cloreto de Mercúrio/farmacologia , Camundongos , Camundongos Endogâmicos/sangue , Camundongos Endogâmicos/imunologia , Fenitoína/farmacologia , Organismos Livres de Patógenos Específicos , Fatores de TempoRESUMO
Exposure to certain drugs and environmental chemicals can provoke the onset of autoimmune disease in susceptible individuals by releasing (self) epitopes for which tolerance has not been established, while simultaneously providing the necessary adjuvant activity. The resulting response type is influenced by the genotype of exposed individuals and relates to susceptibility to the adverse immune effects of the chemicals. Here, we assessed the modulatory role of the chemical compounds themselves. A single injection of streptozotocin (STZ) increased the number of CD8+ cells, macrophages, apoptotic cells, and IFN-gamma-producing T helper and T cytotoxic cells, whereas the number of CD4+ cells and B cells was reduced in the draining lymph node. Coinjection with the reporter antigen TNP-OVA resulted in primary and secondary production of TNP-specific antibodies that were predominantly of IgG2a and IgG2b isotype, whereas STZ did not enhance priming for delayed-type hypersensitivity (DTH) responses to TNP-OVA. Injection of HgCl2 on the other hand, reduced the number of IFN-gamma-producing cells, induced accumulation of B cells and CD4+ and CD8+ T cells, enhanced IgG1 and IgE production to TNP-OVA, and primed for secondary IgG1 and IgE production as well as for DTH reactions. Together these results indicate that a single injection of STZ stimulates type-1 responses, whereas HgCl2 enhanced mixed type-1 and -2 responses in BALB/c mice. These response types match the (auto)immune effects elicited to unknown (auto)antigens following multiple injections of these chemicals.
Assuntos
Autoimunidade , Linfócitos B/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Cloreto de Mercúrio/imunologia , Estreptozocina/imunologia , Xenobióticos/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Feminino , Ficoll/administração & dosagem , Ficoll/análogos & derivados , Ficoll/imunologia , Haptenos/administração & dosagem , Haptenos/imunologia , Cloreto de Mercúrio/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Estreptozocina/administração & dosagem , Trinitrobenzenos/administração & dosagem , Trinitrobenzenos/imunologia , Xenobióticos/administração & dosagemRESUMO
The effects of the presumed autoimmunogenic chemical hexachlorobenzene (HCB), and the closely related non-autoimmunogenic pentachlorobenzene (PCB) in the local popliteal lymph node assay (PLNA) were investigated. To that end 1-5 mg of HCB, equimolar amounts of PCB or the vehicle only, were injected into the hind footpads of rats or mice, and the reaction in the draining lymph node was evaluated on days 7 and 21 after injection. PLN were isolated, weighed, and cell suspensions were prepared to determine PLN cell numbers, and antibody production of PLN cells with an ELISPOT assay or a line immunoassay. The extent of the lymphoproliferative effect was examined by detection of proliferating cells with the BrdU method, and by measurement of paracortex and follicle areas, by combined immunohistochemistry and morphometry of PLN cryosections. We demonstrate here that HCB elevated PLN weights and cell numbers of the rat PLN, by day 7 after injection, but no elevation of antibody production in the PLN. Moreover, HCB caused an enlargement of both the PLN paracortical and follicular areas, and an elevation of proliferating paracortical T cells. None of the HCB-induced effects were found on day 21. HCB caused the same effects in the mouse PLNA, but they tended to sustain at least until day 21. Hardly any of the HCB-induced changes were found when PCB was injected. Previously, we have shown that oral exposure of Wistar rats to HCB elevated the number of splenic T cells and B cells, but also the serum levels of (auto-)antibodies and the production of these antibodies in the spleen, which is thus only partly in accordance with the results of the local reaction to HCB described in this study. This seeming contradiction is discussed.
Assuntos
Clorobenzenos/toxicidade , Fungicidas Industriais/toxicidade , Hexaclorobenzeno/toxicidade , Linfonodos/efeitos dos fármacos , Animais , Linfócitos B/efeitos dos fármacos , Linfócitos B/metabolismo , Edema/etiologia , Edema/imunologia , Feminino , Pé , Imunoensaio , Imunoglobulina G/biossíntese , Imunoglobulina G/efeitos dos fármacos , Imunoglobulina M/biossíntese , Imunoglobulina M/efeitos dos fármacos , Linfonodos/patologia , Ativação Linfocitária/efeitos dos fármacos , Contagem de Linfócitos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar , Linfócitos T/efeitos dos fármacosRESUMO
In this study, it is examined whether the organotin compound di-n-butyltindichloride (DBTC), which has been shown to inhibit immature thymocyte proliferation, is able to disturb the binding between thymocytes and thymic epithelial cells (TEC). To that end, an enzyme-linked binding assay was developed in which the amount of binding of Thy-1+ (mAb ER4)-thymocytes to the rat-derived TEC-line IT45R1 (IT45R1-TEC) could be detected. It was found that preincubation of thymocytes with 3-5 microM DBTC for 30 min inhibited the binding by 50-60% during a 1 h adhesion period. By extending the preincubation period to 1 h and the adhesion period to 22 h, 0.1 microM DBTC was already sufficient to reduce the binding with 60-80%. Further characterization of the binding revealed that splenic lymphocytes were unable to bind to the MHC class II-negative IT45R1-TEC. Since dextran sulfate inhibited the binding as well, sulfated polysaccharide-binding molecules such as Thy-1 and CD2 are likely to be involved in the binding. Electron microscopy showed filament-containing microvilli at the site of interaction. The results are discussed in relation to the mechanism of DBTC-induced thymus atrophy.
Assuntos
Compostos Orgânicos de Estanho/farmacologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Timo/efeitos dos fármacos , Animais , Atrofia , Antígenos CD2/metabolismo , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Depressão Química , Sulfato de Dextrana/farmacologia , Epitélio/metabolismo , Epitélio/ultraestrutura , Antígenos de Histocompatibilidade Classe II/imunologia , Microscopia Eletrônica , Compostos Orgânicos de Estanho/toxicidade , Ratos , Subpopulações de Linfócitos T/metabolismo , Antígenos Thy-1/metabolismo , Timo/patologiaRESUMO
The peroxisome-proliferating effects of clofibric acid and beclobric acid were studied in primary cultures of hepatocytes derived from rat, monkey (Macaca fascicularis) and human liver. Determination of peroxisomal fatty acid beta-oxidation and morphometrical analysis of the peroxisomal compartment were performed after incubation of 1-day-old hepatocyte cultures for 3 days with either compound. In rat liver cell cultures both compounds gave a 10-fold increase in peroxisomal beta-oxidation, a 3-fold increase in the relative number of peroxisomes and a 1.5-fold increase in the mean size of peroxisomes. Beclobric acid gave its maximal effect at a concentration of 10 microM, which is at least one order of magnitude lower than the maximum-effect concentration of clofibric acid. At concentrations greater than 300 microM beclobric acid was cytotoxic. No stimulation of peroxisomal fatty acid beta-oxidation was found in either monkey or human hepatocyte cultures. Morphometrical analysis also showed no increase in the peroxisomal compartment in cultures derived from these species, as indicated by the lack of increase in both relative number and size of peroxisomes. In all three species tested beclobric acid was equally cytotoxic for hepatocytes in vitro. These results are of relevance for the interpretation of the peroxisome-proliferating effects of clofibrate and similar compounds in rats. Since peroxisome proliferation may be correlated to increased hepatic tumour incidences in the rat, the absence of peroxisome proliferation in primates suggests the absence of tumourogenic activity by hypolipidemic compounds in these species.