tff2 protein  (Proteintech)

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Postoperative sleep-restriction (SR) increased myeloid-derived suppressor cells (MDSCs) expansion and decreased splenic CD8 + cells activity via inhibiting splenic trefoil factor 2 (TFF2). ( A ) The expression of TFF2 in the spleen of SR mice with or without dexmedetomidine (Dex) treatment was analysed by real-time PCR (RT-PCR and western blotting. ( B ) Spleen weight in each group after 7 days of SR. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in SR mice with or without Dex treatment. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from SR mice with or without Dex treatment. ( E ) The mRNA and protein expression of TFF2 in the spleen were analysed by RT-PCR and western blotting in SR mice with or without vagus nerve stimulation (VNS). ( F ) Spleen weight in SR mice with the treatment of recombinant human TFF2 protein (rTFF2) or PBS. ( G ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in SR mice with the treatment of rTFF2 or PBS. ( H ) ELISPOT assay of the levels of IFN-γ and GrB in splenic CD8 + T cells from SR mice with the treatment of rTFF2 or PBS. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Derivative Assay, Activity Assay, Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Enzyme-linked Immunospot, Recombinant

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Subdiaphragmatic vagus nerve mediated the attenuated effects of dexmedetomidine (Dex) on sleep-restriction (SR)-induced decrease in splenic trefoil factor 2 (TFF2) expression, increase in the expansion of myeloid-derived suppressor cells (MDSCs) and decrease in CD8 + T cells activity. ( A ) The expression of TFF2 in the spleen of dexmedetomidine (Dex)-treated SR mice with or without bilateral sub-diaphragmatic vagotomy (SDV) was analysed by western blotting. ( B ) spleen weight in Dex-treated SR mice with or without SDV. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in Dex-treated SR mice with or without SDV. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from Dex-treated SR mice with or without SDV. All data represent mean ± SEM, n = 5; # P < 0.05, N.S., not significant.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Expressing, Derivative Assay, Activity Assay, Western Blot, Flow Cytometry, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Dexmedetomidine (Dex) attenuated postoperative sleep-restriction (SR)-induced decrease in splenic trefoil factor 2 (TFF2) expression, increase in myeloid-derived suppressor cells (MDSCs) expansion and decrease in splenic CD8 + cells activity via improving gut microbiota disturbance. ( A ) Western blotting analysis of splenic TFF2 expression in sham-operated mice and pseudo-germ-free mouse received fecal microbiota transplantation (FMT). ( B ) Spleen weight in sham-operated mice and pseudo-germ-free mouse received FMT. ( C , D ) Flow cytometry analysis of spleen for CD11b + Gr-1 + MDSCs in sham-operated mice and pseudo-germ-free mouse received FMT. ( E ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from sham-operated mice and pseudo-germ-free mouse received FMT. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Expressing, Derivative Assay, Activity Assay, Western Blot, Transplantation Assay, Flow Cytometry, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Subdiaphragmatic vagus nerve served as a bridge between gut microbiota and spleen after postoperative sleep-restriction (SR). ( A ) Western blotting analysis of splenic trefoil factor 2 (TFF2) expression in pseudo-germ-free mouse received fecal microbiota transplantation (FMT) with feces of SR mice, dexmedetomidine (Dex)-treated SR mice or Dex-treated SR mice with sub-diaphragmatic vagotomy (SDV). ( B ) Spleen weight in pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. ( C ) Flow cytometry analysis of spleen for CD11b + Gr-1 + myeloid-derived suppressor cells (MDSCs) in pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. ( D ) The enzyme-linked immunospot (ELISPOT) assay of the levels of interferon-γ (IFN-γ) and Granzyme B (GrB) in splenic CD8 + T cells from pseudo-germ-free mouse received FMT with feces of SR mice, Dex-treated SR mice or Dex-treated SR mice with SDV. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Western Blot, Expressing, Transplantation Assay, Flow Cytometry, Derivative Assay, Enzyme-linked Immunospot

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Splenic trefoil factor 2 (TFF2) was essential to attenuate SR-induced reduced protective ability against Escherichia coli ( E. coli ) pneumonia, increased expression of IL-4 and IL-13 in the lung and M2 polarization of alveolar macrophages (AMs), and decreased phagocytic activity of AMs. ( A ) Enumeration of colony-forming units (CFU) per milliliter of bronchoalveolar lavage analyzed one day after E. coli pneumonia in postoperative SR mice treated with recombinant human TFF2 protein (rTFF2) or PBS. ( B ) ELISA determination of the concentrations of IL-4 and IL-13 in the lungs of postoperative SR mice treated with rTFF2 or PBS one day after E. coli pneumonia. ( C ) The phagocytic activity of alveolar macrophages from postoperative SR mice treated with rTFF2 or PBS. ( D ) Real-time PCR (RT-PCR) analysis of the mRNA expression of Arg1, YM and iNOS in alveolar macrophages from postoperative SR mice treated with rTFF2 or PBS. All data represent mean ± SEM, n = 5; # P < 0.05, ## P < 0.01.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Expressing, Activity Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: Schematics illustrating the signaling mechanisms of sleep-restriction in postoperative immunosuppression and its treatment by dexmedetomidine. Sleep-restriction exaggerates intestinal flora disorder, furtherly decreases splenic trefoil factor 2 (TFF2) expression, which subsequently leads to the increase in myeloid-derived suppressor cells (MDSCs) numbers and decrease in splenic CD8 + T cells activity. Subdiaphragmatic vagus nerve (SVN) served as an important conduit of gut microbiota-spleen communication. SR-induced exaggeration of postoperative immunosuppression was characterized by increased expression of IL-4 and IL-13 in the lung, increased M2 polarization of alveolar macrophages (AMs), decreased phagocytic activity of AMs and thus decreased antimicrobial activity in E. coli pneumonia. Dexmedetomidine treatment during SR alleviated SR-induced decrease in postoperative immunosuppression through gut microbiota and SVN.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques: Expressing, Derivative Assay, Activity Assay

Journal: Aging (Albany NY)

Article Title: Dexmedetomidine alleviates sleep-restriction-mediated exaggeration of postoperative immunosuppression via splenic TFF2 in aged mice

doi: 10.18632/aging.102952

Figure Lengend Snippet: The sequences of primers for real-time PCR.

Article Snippet: After 7 days of SR, recombinant human TFF2 protein (rTFF2; 0.2 mg/kg; R&D Systems, Minneapolis, MN, USA) was injected intravenously via the tail vein.

Techniques:

Journal: Cancer Gene Therapy

Article Title: Therapeutic potential of adenovirus-mediated TFF2-CTP-Flag peptide for treatment of colorectal cancer

doi: 10.1038/s41417-018-0036-z

Figure Lengend Snippet: Identification of wild-type TFF2 and fusion TFF2-CTP-Flag protein delivered by Ad- Tff2 and Ad- Tff2 -CTP-Flag in the blood. a TFF2 level assayed by ELISA in the blood from naive wild-type and CD2- Tff2 transgenic mice and mice at time point 6 months after induction of tumorigenesis with AOM/DSS treatment. Dunn’s multiple comparisons test after one-way ANOVA test, ns, non-significant, * p < 0.05, **** p < 0.001. b Amino acid sequence of fusion protein TFF2-CTP-Flag: TFF2 amino acid sequence—bold regular, CTP—regular, S —underlined, original cysteine substituted for serine, additional amino acids residues -bold italic shadow, Flag-underlined regular, Stop—stop codon. c Schematic presentation of fusion construct Tff2 -2CTP-3Flag. d Map of GV314 vector with inserted TFF2 gene. e – j Time course of TFF2 and TFF2-CTP-Flag level after single administration of adenoviruses Ad- Tff2 and Tff2 -CTP-Flag. Tff2 -null mice were injected with 5 × 10 8 pfu of Ad- Tff2 or Ad- Tff2 -CTP-Flag and sacrificed at indicated times post infection. Time-course of Tff2 mRNA ( e ) and Tff2 -CTP-Flag mRNA ( h ) expression in the liver. Total mRNA was isolated from the liver and then Tff2 mRNA was detected by qPCR, fold of change normalized on housekeeper Hprt mRNA. For each time point 3 mice were used, graphed as mean ± SD. f , i Kinetic of TFF2 protein and TFF2-CTP-Flag fusion protein in the blood assessed by ELISA and expressed as mean ± SD at each time point, three animals per group. g , j western blot for TFF2 and TFF2-CTP-Flag in mouse serum. Five microliter of mouse serum were loaded in each lane and western blot was developed with antibody produced against C-end of TFF2 molecule, arrows indicate the position of TFF2 and TFF2-CTP-Flag with calculated sizes 12 and 25–26 kDa accordingly. k Western blot of blood sample (5 µl) from Tff2 -null mouse taken on day 7 after Ad- Tff2 -CTP-Flag administration. Western blot was developed with anti-Flag antibody; arrows indicate the positions of TFF2-CTP-Flag fusion under reduced and non-reduced conditions

Article Snippet: Fusion TFF2-CTP-Flag protein from this diluted soup was purified using anion-exchange chromatography (Capto Q, GE life sciences).

Techniques: Enzyme-linked Immunosorbent Assay, Transgenic Assay, Sequencing, Construct, Plasmid Preparation, Injection, Infection, Expressing, Isolation, Western Blot, Produced

Journal: Cancer Gene Therapy

Article Title: Therapeutic potential of adenovirus-mediated TFF2-CTP-Flag peptide for treatment of colorectal cancer

doi: 10.1038/s41417-018-0036-z

Figure Lengend Snippet: Ad- Tff2 or Ad- Tff2 -CTP-Flag reduces splenomegaly and proportion of myeloid cells in the spleen of wild-type and Tff2 -null mice. a – f Wild-type mice were treated 2% DSS for 5 consecutive days, then on day 12 post treatment with DSS single injection of Ad- Tff2 ( a – c ) or Ad- Tff2 -CTP ( d – f ) and Ad-Fc or Ad-Fc-CTP-Flag as controls. Dose 5 × 10 8 pfu was administered in tail vein. Mice were sacrificed on day 19. Spleen appearance ( a , d ), spleen size mass ( b , e ) and proportion of CD11b + Gr-1 + cells ( c, f ) in spleen, unpaired t -test, * p < 0.05. Two experiments with three mice in each group have been done. g – i Ad- Tff2- CTP-Flag reduces splenomegaly ( g , h ) and proportion of CD11b + Gr-1 + cells ( i ) in the spleen of Tff2 -null mice, unpaired t -test, * p < 0.05. One experiment with three mice in each group has been done. j Administration of Ad- Tff2 -CTP-Flag decreases granulocytes/macrophages colonies-forming units in the spleen of wild-type mice treated with DSS. Data obtained from mice treated with 2% DSS for 5 consecutive days and sacrificed on day 19, unpaired t -test, * p < 0.05. k Immunostaining for Gr-1 in the red pulp area in spleens of Tff2 -null mice treated DSS water and received Ad-Fc-CTP (upper raw) or Ad- Tff2 -CTP (lower raw), day 19. Bar size is 100 µm

Article Snippet: Fusion TFF2-CTP-Flag protein from this diluted soup was purified using anion-exchange chromatography (Capto Q, GE life sciences).

Techniques: Injection, Immunostaining

Journal: Cancer Gene Therapy

Article Title: Therapeutic potential of adenovirus-mediated TFF2-CTP-Flag peptide for treatment of colorectal cancer

doi: 10.1038/s41417-018-0036-z

Figure Lengend Snippet: Validation of Ad- Tff2 -CTP-Flag activity in AOM/DSS-induced colon cancer model. a , b Ad- Tff2 suppresses colon tumorigenesis in wild-type mice treated with AOM/DSS. a Colon appearance (left), tumor number (right), b number of splenic CD11b + Gr-1 + cells. Two experiments have been done with 3–4 mice in each group. c , d Ad Tff2 -CTP-Flag suppresses colon tumorigenesis in AOM/DSS-induced colon cancer model. c Colon appearance and tumor number, d proportion CD11b + Gr-1 + cells in the spleen from wild-type mice treated with Ad- Tff2 -CTP versus Ad-Fc-CTP, unpaired t -test, * p < 0.05. Two experiments have been done with 3–4 mice in each group. e Overlapping staining for Flag and Gr-1 in the spleen of Tff2 -null mice injected with Ad- Tff2 -CTP-Flag. Mice were given 2% DSS for 5 consecutive days, then seven days later adenovirus was administrated via tail vein and mice were sacrificed in one week after adenovirus administration. Bar size is 50 µm. f BrdU incorporation in splenic CD11b + Gr-1 + cells of wild-type mice injected with Ad-Fc-CTP-Flag compare with Ad- Tff2 -CTP-Flag, unpaired t-test. Wild-type mice were treated 2% DSS for 5 consecutive days, then on day 7 post treatment with DSS single injection of Ad- Tff2 -CTP or Ad-Fc-CTP-Flag (both 5 × 10 8 pfu) was administered via tail vein injection. Mice were sacrificed on day 19 after start of DSS treatment, BrdU was injected three hours before sacrifice

Article Snippet: Fusion TFF2-CTP-Flag protein from this diluted soup was purified using anion-exchange chromatography (Capto Q, GE life sciences).

Techniques: Activity Assay, Staining, Injection, BrdU Incorporation Assay

Journal: Cancer Gene Therapy

Article Title: Therapeutic potential of adenovirus-mediated TFF2-CTP-Flag peptide for treatment of colorectal cancer

doi: 10.1038/s41417-018-0036-z

Figure Lengend Snippet: Clearance of recombinant mouse TFF2 and fusion TFF2-CTP-Flag from the blood. a – c Tff2- null mice were tail vein injected with equal molar amount of purified recombinant TFF2 and TFF2-CTP-Flag per kg of mouse weight. At indicated time points blood was taken and assayed for TFF2 and TFF2-CTP-Flag by western blot with antibodies produced against C-terminus of TFF2 molecule ( a , b ) and ELISA ( c ). Excretion of TFF2 with the urine was analyzed by western blot ( a ). ELISA data are plotted as a mean of value and standard deviation at each time point. d Dose-dependent downregulation of CCND1 mRNA in CD11b + Gr-1 + cells upon administration of recombinant wild-type TFF2 and fusion TFF2-CTP-Flag protein. Tff2 -null mice were given 2.5% DSS for 5 days, then CD11b + Gr-1 + were sorted and cultured with recombinant TFF2 and TFF2-CTP-Flag in indicated concentrations for a 7 days

Article Snippet: Fusion TFF2-CTP-Flag protein from this diluted soup was purified using anion-exchange chromatography (Capto Q, GE life sciences).

Techniques: Recombinant, Injection, Purification, Western Blot, Produced, Enzyme-linked Immunosorbent Assay, Standard Deviation, Cell Culture

Journal: The Journal of Physiology

Article Title: Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

doi: 10.1113/JP277259

Figure Lengend Snippet: Results from WT and TFF2 KO gastric organoids imaging over time, measuring the movement of fluorescent nuclei (Hoechst 33342 stain) after PD. PD occurred at t = 0 min. WT and TFF2 KO gastric organoids were treated with AMD3100 (1 μm) for 1 h or BAPTA/AM (50 μm) for 30 min before PD as indicated. rTFF2 was microinjected into the lumen of organoids 30 min before the study (see Methods). Exfoliation was determined based on the maximum distance of damaged nuclei into gastric organoid lumen over 20 min. Vehicle (WT control, n = 7; TFF2 KO control, n = 10; TFF2 + rTFF2 Control, n = 10); AMD3100 (WT, n = 6; TFF2 KO, n = 6; TFF2 KO + rTFF2, n = 5); BAPTA/AM (WT, n = 4; TFF2 KO, n = 4; TFF2 KO + rTFF2, n = 8). * P < 0.05 vs. WT vehicle, #P < 0.05 vs. rTFF2 treatment in TFF2 KO.

Article Snippet: Microinjection For rescue experiments in TFF2 and NHE2 KO gastric organoids, recombinant human TFF2 (rTFF2; 40 μ m stock; R&D Systems, Minneapolis, MN, USA) was microinjected as described previously (Engevik et al . 2018 ).

Techniques: Imaging, Staining, Control

Journal: The Journal of Physiology

Article Title: Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

doi: 10.1113/JP277259

Figure Lengend Snippet: Results from imaging of WT and TFF2 KO organoids over time; measuring the movement of fluorescent nuclei (Hoechst 33342 stain) after PD. Some organoids were treated with AG1478 (200 nm) as indicated. rTFF2 was microinjected into the lumen of organoids before the study. Exfoliation was determined based on the maximum distance of damaged nuclei into gastric organoid lumen over 20 min. Vehicle (WT control, n = 6; TFF2 KO control, n = 8; TFF2 KO + rTFF2, n = 8); AG1478 (WT, n = 6; TFF2 KO, n = 5; TFF2 KO + rTFF2, n = 5). * P < 0.05 vs. WT vehicle, #P < 0.05 vs. rTFF2 treatment in TFF2 KO.

Article Snippet: Microinjection For rescue experiments in TFF2 and NHE2 KO gastric organoids, recombinant human TFF2 (rTFF2; 40 μ m stock; R&D Systems, Minneapolis, MN, USA) was microinjected as described previously (Engevik et al . 2018 ).

Techniques: Imaging, Staining, Control

Journal: The Journal of Physiology

Article Title: Trefoil factor 2 activation of CXCR4 requires calcium mobilization to drive epithelial repair in gastric organoids

doi: 10.1113/JP277259

Figure Lengend Snippet: Fluorescence of YC‐Nano gastric organoids imaged over time in (A) to (D) and cell exfoliation measured over time in (E) to (F). Where indicated, Hoe 694 (100 μm) was added to organoid medium 1 h prior to experimentation. In time courses, PD occurred at t = 0 min. A, damage area measured in YC‐Nano control (black) and Hoe 694 supplemented gastric organoids (white) (n = 4). B, comparison of rate of repair between YC‐Nano control (black) and Hoe 694 supplemented gastric organoids (white) (* P < 0.05). C, measurement of the normalized FRET/CFP ratio of the lateral membrane region of cells adjacent to the damage site comparing control (black) and Hoe 694 supplemented gastric organoids (white). D, comparison of the maximum FRET/CFP ratio from (C) between control (black) and Hoe 694 supplemented gastric organoids (white) (n = 4, * P < 0.05). E, comparison of exfoliation in WT (n = 5) and NHE2 KO vehicle (n = 5) and rTFF2 injected organoids (n = 6) (* P < 0.05). F, comparison of exfoliation in WT and TFF2 KO gastric organoids treated with Hoe 694 and/or microinjection of rTFF2. Vehicle (WT Control, n = 5; TFF2 KO, n = 6; TFF2 KO + rTFF2, n = 4); Hoe 694 (WT, n = 5; TFF2 KO, n = 4; TFF2 KO + rTFF2, n = 4). * P < 0.05 vs. WT vehicle, #P < 0.05 vs. rTFF2 treatment in TFF2 KO.

Article Snippet: Microinjection For rescue experiments in TFF2 and NHE2 KO gastric organoids, recombinant human TFF2 (rTFF2; 40 μ m stock; R&D Systems, Minneapolis, MN, USA) was microinjected as described previously (Engevik et al . 2018 ).

Techniques: Fluorescence, Control, Comparison, Membrane, Injection, Microinjection