LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion

Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10−6 M (mTORC2/mTORC1 inhibitor) and 10−6 M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10−6 M U0126, an MEK/ERK inhibitor, and 10−6 M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases

Soroepidemiologia da toxoplasmose em caprinos e ovinos de três municípios do estado do Rio de Janeiro

A toxoplasmose é uma zoonose de ampla distribuição mundial, causada pelo Toxoplasma gondii. O estudo da prevalência desta infecção em animais produtores de carne e leite é de interesse à saúde pública, devido ao fato desses produtos oriundos de animais infectados serem importantes vias de transmissão para o homem, quando consumidos in natura. Além disso, há o aspecto econômico, uma vez que pode causar aborto, retardo no crescimento e animais debilitados, levando prejuízos ao pecuarista. Este trabalho objetivou estimar a soroprevalência da infecção por T. gondii, por meio da reação de imunofluorescência indireta (RIFI) em caprinos e ovinos de três municípios do estado do Rio de Janeiro, provenientes de 10 propriedades. A prevalência de anticorpos IgG anti-T.gondii foi de 29,12% (60/206) nos caprinos e de 38,05% (137/360) nos ovinos, sendo observada nessa última espécie associação (p<0,05) entre sexo (fêmeas), idade adulta, sistema de criação extensivo, dieta de pastagem e água de beber de açude com a soropositividade. Os títulos variaram de 64 a 256, podendo ser sugestivos de infecção crônica. Melhorias nas técnicas de criação podem reduzir as fontes de infecção por T. gondii nos rebanhos

Seroepidemiology of toxoplasmosis in goats and sheep from three counties of Rio de Janeiro state, Brazil

Submitted by Sandra Infurna ([email protected]) on 2018-03-27T11:47:05Z No. of bitstreams: 1 magyda_duharoug_etal_IOC_2011.pdf: 167561 bytes, checksum: 27921249420884bbe655f43b68e22dbf (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-03-27T11:54:46Z (GMT) No. of bitstreams: 1 magyda_duharoug_etal_IOC_2011.pdf: 167561 bytes, checksum: 27921249420884bbe655f43b68e22dbf (MD5)Made available in DSpace on 2018-03-27T11:54:47Z (GMT). No. of bitstreams: 1 magyda_duharoug_etal_IOC_2011.pdf: 167561 bytes, checksum: 27921249420884bbe655f43b68e22dbf (MD5) Previous issue date: 2011Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Serviço de Zoonoses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Serviço de Zoonoses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório deToxoplasmose. Rio de Janeiro, RJ. Brasil.Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii. Epidemiological surveys of T. gondii infection among livestock have great economical importance since this infection may cause abortion, growth retardation and neonatal mortality, with significant losses to breeders. In regard of public health, human infection can be acquired by ingestion of meat or milk in natura from infected livestock. The aim of this study was to assess the toxoplasmosis seroprevalence by indirect immunofluorescence antibody test (IFAT) in goats and sheep, in three counties of Rio de Janeiro state, from 10 different farms. The seroprevalences of anti-T.gondii IgG antibodies were 29.12% (60/206) in goats and 38.05% (137/360) in sheep, with titers ranging from 64 to 256, suggesting chronic infection. Association of each of the following epidemiological factors: female gender, adult age, extensive management system, grazing pasture and drinking lake water with seropositivity was observed only in sheep (p≤0.05). Improvement in breeding conditions may reduce the sources of infection in herds

Data_Sheet_4_LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion.ZIP

<p>Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10⁻⁶ M (mTORC2/mTORC1 inhibitor) and 10⁻⁶ M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10⁻⁶ M U0126, an MEK/ERK inhibitor, and 10⁻⁶ M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases.</p

Data_Sheet_2_LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion.ZIP

<p>Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10⁻⁶ M (mTORC2/mTORC1 inhibitor) and 10⁻⁶ M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10⁻⁶ M U0126, an MEK/ERK inhibitor, and 10⁻⁶ M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases.</p

Data_Sheet_3_LPS Induces mTORC1 and mTORC2 Activation During Monocyte Adhesion.ZIP

<p>Monocyte adhesion is a crucial step in transmigration and can be induced by lipopolysaccharide (LPS). Here, we studied the role of mammalian target of rapamycin (mTOR) complexes, mTORC1 and mTORC2, and PKC in this process. We used THP-1 cells, a human monocytic cell line, to investigate monocyte adhesion under static and flow conditions. We observed that 1.0 μg/mL LPS increased PI3K/mTORC2 pathway and PKC activity after 1 h of incubation. WYE-354 10⁻⁶ M (mTORC2/mTORC1 inhibitor) and 10⁻⁶ M wortmannin avoided monocyte adhesion in culture plates. In addition, WYE also blocked LPS-induced CD11a expression. Interestingly, rapamycin and WYE-354 blocked both LPS-induced monocyte adhesion in a cell monolayer and actin cytoskeleton rearrangement, confirming mTORC1 involvement in this process. Once activated, PKC activates mTORC1/S6K pathway in a similar effect observed to LPS. Activation of the mTORC1/S6K pathway was attenuated by 10⁻⁶ M U0126, an MEK/ERK inhibitor, and 10⁻⁶ M calphostin C, a PKC inhibitor, indicating that the MEK/ERK/TSC2 axis acts as a mediator. In agreement, 80 nM PMA (a PKC activator) mimicked the effect of LPS on the activation of the MEK/ERK/TSC2/mTORC1/S6K pathway, monocyte adhesion to ECV cells and actin cytoskeleton rearrangement. Our findings show that LPS induces activation of mTOR complexes. This signaling pathway led to integrin expression and cytoskeleton rearrangement resulting in monocyte adhesion. These results describe a new molecular mechanism involved in monocyte adhesion in immune-based diseases.</p