lps group contained lps pg standard  (InvivoGen)

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: Chronic kidney disease (CKD) affects colonization and survival of P. gingivalis strain ATCC 32277 in solid organs upon ligation model. (A) Body mass was monitored for 60 days in all the experimental groups. (B) Survival of P. gingivalis was assessed by counting the number of colony-forming units in lung, liver, spleen, heart, and kidney homogenates collected from the control group, CKD group (AAI), P. gingivalis -infected group (P.g.), and CKD- P. gingivalis -infected group (AAI + P.g.) at day 60 upon CKD induction followed by chronic infection ( n = 6 per each group). The bacteria grew only in the AAI + P.g. group. The radar chart represents the percentage of mice that tested positive for P. gingivalis as assessed by colony counting in the indicated organs. (C) The radar chart shows the number of colonies that grew in mice whose organs tested positive for P. gingivalis upon infection: lung (210 colonies), liver (23 colonies), spleen (10 colonies), heart (0 colonies), and kidney (80 colonies). (D) The number of bacteria in gingival tissues and in tissues was assessed by RT-PCR and presented as the number of copies of the detected nucleic acid (based on standard curve). (E) The levels of several innate immunity-related genes were investigated in the spleens of mice. A heat map shows altered genes from expression analysis of preselected transcripts. Genes indicated in green are upregulated, and genes indicated in pink are downregulated to highlight differences between the samples. The rows are Z -score scaled. (F) The levels of the indicated cytokines and chemokines was investigated in the blood of mice. (G) The mRNA expression levels of genes during CKD and P. gingivalis infection. A heat map shows altered genes from expression analysis of preselected transcripts. Genes indicated in green are upregulated, and genes indicated in pink are downregulated to highlight differences between the samples. The rows are Z -score scaled. (H) The levels of several adaptive immunity-related genes were investigated in the spleens of mice. We observed reduction of several gene expressions within the AAI + P.g. group. Data are shown as means ± SD. ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Ligation, Control, Infection, Bacteria, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: We investigated the serum IgG levels (A) and kidney IgG deposition (B) in the control group, CKD group (AAI), P. gingivalis -infected group (P.g.), and CKD- P. gingivalis -infected group (AAI + P.g.) at day 60 upon CKD induction followed by chronic infection. Our data indicate that adaptive immune responses (including antibody production) might be affected by chronic kidney disease. (C) Serum creatinine levels, (D) blood urea nitrogen, (E) proteinuria, and (F) ACR were assessed. Data are shown as means ± SD. ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Control, Infection

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: Differences between the control groups, CKD group (AAI), P. gingivalis -infected group (P.g.), and CKD- P. gingivalis -infected group (AAI + P.g.) in (A) kidney weight and (B) fibrosis scored based on Sirius red staining and (C) collagen within the kidney tissue. (D) The treated and control kidneys from all groups were stained for interstitial macrophages (F4/80), proliferation marker (Ki67), and proximal tubules to illustrate kidney inflammation and regeneration. Quantification was performed in Photoshop as a percentage of positively stained high-power field (data not shown). Differences between groups of mice in F4/80+ macrophages, Ki67, and proximal tubules staining. Scale bar = 50 µm. Data are shown as means ± SD. ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Control, Infection, Staining, Marker

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: (A) Expression levels of several adaptive immunity-related genes were investigated in the A20 lymphocytes stimulated with indoxyl sulfate (IS), LPS from P. gingivalis , or LPS from E. coli . We observed reduction of several gene expressions within upon IS-stimulated group. A heat map shows altered genes from expression analysis of preselected transcripts. Genes indicated in green are upregulated, and genes indicated in pink are downregulated to highlight differences between the samples. The rows are Z -score scaled. (B) Expression levels of selected adaptive immunity-related genes were investigated in the A20 lymphocytes stimulated with IS. Ahr expression was used as a control. Data are shown as means ± SD. ⁣ ∗ p < 0.05 and ⁣ ∗∗ p < 0.01.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Expressing, Control

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: B lymphocyte cell antibody production during uremia condition and infection. B lymphocyte cells were stimulated with IS toxin in the presence/absence of (A) LPS P. gingivalis or (B) P. gingivalis ATCC 33277 for 48 h. For evaluation of the antibody production, cells were stained for CD138, Ig, and MHCII and were analyzed by flow cytometry. (A) CD138 expression did not change between the groups after stimulation with IS, P. gingivalis lipopolysaccharide (LPS P. gingivalis ), IS + LPS P. gingivalis , and the control group. Ig production was reduced in the presence of IS and even more reduced in the presence of IS + LPS P. gingivalis . The results demonstrated that there were no significant differences observed among the IS + LPS P. gingivalis and LPS P. gingivalis groups in MHCII expression. (B) CD138 expression decreased after P. gingivalis and IS+ P. gingivalis stimulation, the same as Ig antibody production. The expression level of MHCII increased after P. gingivalis stimulation compared to the IS and IS + P. gingivalis . All data represent mean values ± SD. ⁣ ∗ p < 0.05, ⁣ ∗∗∗ p < 0.001, and ⁣ ∗∗∗∗ p < 0.0001.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Infection, Staining, Flow Cytometry, Expressing, Control

Journal: Journal of Immunology Research

Article Title: Bidirectional Interaction Between Chronic Kidney Disease and Porphyromonas gingivalis Infection Drives Inflammation and Immune Dysfunction

doi: 10.1155/jimr/8355738

Figure Lengend Snippet: Bone loss following CKD and P. gingivalis infection ATCC 32277. Quantification of P. gingivalis induced bone loss in wild-type mice with CKD. Exemplary images of sectional micro-computed tomography measurements of the distances from the cementoenamel junction to the alveolar bone crest (CEJ–ABC) on the buccal side at day 60. Results are the mean ± SD. ⁣ ∗∗ p < 0.01.

Article Snippet: Cells (RPMI medium including 10% FBS and 1% P/S) were seeded with 1% FBS at 0.5 million cells/well and stimulated with IS (60 μg/mL) (Sigma-Aldrich) for 2 h. Next, cells were stimulated with LPS P. gingivalis standard (10 µg/mL) (InvivoGen) and P. gingivalis ATCC 33277 (MOI 1:10) for 48 h. The following antibodies were used for flow cytometric analysis: FITC antimouse MHC class II (I-A/I-E) (eBioscience), PE rat antimouse Ig k light chain (BD Pharmingen), and APC rat antimouse CD138 (BD Bioscience).

Techniques: Infection, Micro-CT

Journal: Scientific Reports

Article Title: Porphyromonas gingivalis lipopolysaccharides act exclusively through TLR4 with a resilience between mouse and human

doi: 10.1038/s41598-017-16190-y

Figure Lengend Snippet: ESI-MS analysis of lipid A from Pg LPS. ( A ) Structure and molecular mass of penta-acylated lipid A (1690) of P. gingivalis . ( B ) Structure and molecular mass of penta-acylated lipid A was modified during electrospray ionization to generated a deacylated lipid A. Negative ions analysis of deacylated lipid A from STD Pg LPS ( C ) or UP Pg LPS ( D ) by ESI-MS.

Article Snippet: Ultrapure P. gingivalis LPS (UP Pg LPS) (Invivogen, cat: tlrl-ppglps, lot: PPG-3701 and 3801), Standard P. gingivalis LPS (STD Pg LPS) (cat: tlrl-pglps, lot: LPG 37-01 and 38-01) were from InvivoGen (France).

Techniques: Modification, Generated

Journal: Scientific Reports

Article Title: Porphyromonas gingivalis lipopolysaccharides act exclusively through TLR4 with a resilience between mouse and human

doi: 10.1038/s41598-017-16190-y

Figure Lengend Snippet: Differential recognition of lipid A on Pg LPS and Ec LPS. LPS recognition by TLR4 is based on the lipid A structure. Lipid A is different between Ec LPS and Pg LPS. Lipid A binding to MD2 induces dimerization of TLR4, activates the downstream signaling pathways and then leads to the secretion of pro-inflammatory cytokines. Our result suggests that recognition of lipid A structures by mouse and human TLR4 is different. Hexa-acylated E. coli lipid A is recognized as a strong agonist by both mouse and human TLR4, and induced an important production of pro-inflammatory cytokines. Penta-acylated P. gingivalis lipid A is recognized as an agonist by human TLR4 through MD2, and induces a production of proinflammatory cytokines, whereas it was weakly recognized by mouse MD2 and induces a weak production of pro-inflammatory cytokines. Penta-acylated P. gingivalis lipid A does not interact with TLR2.

Article Snippet: Ultrapure P. gingivalis LPS (UP Pg LPS) (Invivogen, cat: tlrl-ppglps, lot: PPG-3701 and 3801), Standard P. gingivalis LPS (STD Pg LPS) (cat: tlrl-pglps, lot: LPG 37-01 and 38-01) were from InvivoGen (France).

Techniques: Binding Assay