Journal: Annals of Medicine and Surgery

Article Title: Da-yuan-yin decoction ameliorates inflammatory injury in severe pancreatitis by protecting intestinal mucosal barrier and immune function and preventing intestinal dysbiosis

doi: 10.1097/MS9.0000000000004873

Figure Lengend Snippet: Serum interleukin (IL)-1β and IL-10 expression levels (pg/ml) and cluster of differentiation (CD)3 + , CD4 + , and CD8 + cells from five ileal Peyer’s patches in the sham operation (SO), severe acute pancreatitis (SAP), and da-yuan-yin (DYY) + SAP groups, respectively. Serum IL-1β and IL-10 levels in the SAP and DYY + SAP groups were significantly higher than those in the SO group at 24, 48, and 72 h after surgery (all P < 0.05). Serum IL-1β and IL-10 levels in the DYY + SAP group were significantly lower than those in the SAP group 72 h postoperatively ( t = 4.37, P = 0.003; t = 2.41, P = 0.04, respectively). Levels of CD3 + , CD4 + , and CD8 + cells from ileal Peyer’s patches in the SAP and DYY + SAP groups decreased with time post-surgery. At 72 h postoperatively, the DYY + SAP group had significantly higher levels of CD3 + , CD4 + , and CD8 + cells than the SAP group ( t = 2.58, P = 0.036; t = 2.78, P = 0.027; t = 2.77, P = 0.028, respectively). There were no significant differences in early or late apoptosis in ileal Peyer’s patches 24, 48, and 72 h postoperatively in the SO, SAP, and DYY + SAP groups.

Article Snippet: Single-cell suspension is stained with the following antibodies: Live/Dead Fixable Violet Dead Cell (Cat# 2549272, Invitrogen), anti-Rat CD3 (Cat# G4.18, MultiSciences Biotech Co., Ltd), and anti-Rat CD4 (Cat# OX35, MultiSciences).

Techniques: Expressing

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 1. T cells are present in white matter (WM), gray-white junction (GWJ), gray matter (GM), cerebellar granular layer (GL), and molecular layer (ML) areas harboring JCV-infected cells. Triple (A) and quadruple (BYD) immunostaining assays for CD8 (brown), CD3 (gray blue), T antigen (T Ag, purple), and VP1 (cyan-blue) show that numerous CD8-positive T cells (brown, filled circle arrow) are present in the vicinity of JC virus (JCV)Yinfected cells expressing T Ag (purple, arrowheads) in partially burnt out lesions located in the GWJ (A) square 1, magnified in inset 1 of an HIV-seropositive patient with progressive multifocal leukoencephalopathy. Rare presumed CD4-positive T cells (CD3 positive/CD8 negative, gray blue, asterisk in inset 1) are also present. The PML lesions extend partially in the GM (A, square 2, magnified in inset 2) where cells expressing T Ag can be seen (purple, arrowheads) as well as a few CD8-positive T cells located in the vessels (brown, empty circle arrows, inset 2). In B, the presence of CD8-positive and frequent presumed CD4-positive T cells in the GM of a JCV encephalopathy (JCVE) patient harboring numerous JCV-infected cells expressing T Ag and VP1. Numerous CD8-positive T cells in the parenchyma and blood vessels (brown, filled and empty circle arrows, respectively), presumed CD4-positive T cells (gray blue, asterisk), and JCV-infected cells expressing T Ag (purple, arrowhead) or VP1 (cyan-blue, arrow) are visible in the inset. An isolated cerebellar lesion at the border of the WM and GL of a JCV granule cell neuronopathy (JCV GCN) patient with chronic lymphocytic leukemia (C) and the magni- fication of the square in D show that CD8-positive T cells (brown, filled circle arrows) are present in the parenchyma in areas with JCV-infected cells expressing T Ag (purple, arrowheads) and/or VP1 (cyan-blue, arrows). Aggregates of CD8 on JCV-infected cells expressing VP1 are also evident. Scale bars = (AYC) 500 Km; (D) 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Infection, Immunostaining, Virus, Expressing, Isolation

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 3. CD8-positive T cells aggregate mostly around cells with intact nuclei, expressing JC virus (JCV) T antigen (T Ag) and VP1. (A, B) Quadruple immunostaining for CD8 (brown), CD3 (gray blue), T Ag (purple), and VP1 (cyan-blue) done on an HIV- seronegative PML patient with chronic lymphocytic leukemia (A) and an HIV-seropositive PML patient (B). In PML lesions of the cerebral gray-white junction (GWJ) and gray matter (GM) (A, inset), where JCV-infected cells expressing VP1 are present (cyan-blue arrows), there are numerous parenchymal CD8-positive T cells (brown, black circle arrow). They are located mostly at the leading edge of infection where newly infected cells have still intact nuclei and express T Ag alone (purple, arrowhead) or T Ag and VP1 (cyan-blue arrows). In JCV-infected cerebellum (B), only rare CD8-positive T cells (brown, filled circle arrow) colocalize in the GL with hundreds of JCV-infected cells expressing T Ag (inset, purple, arrowheads), whereas more numerous CD8-positive T cells and some CD4-positive T cells (gray-blue, asterisk) colocalize in the contiguous WM with a few JCV-infected cells expressing VP1 (cyan-blue, arrow). (CYE) Triple immunofluorescence (IFA) staining for CD3, T Ag, and VP1 done in the cerebrum of an HIV-seronegative PML patient (C), an HIV-seropositive PML patient (D), and a JCV encephalopathy (JCVE) patient (E). The IFA immunostains confirm that T cells are absent in an area with T Ag only (C), whereas T cells are mostly present in areas where JCV-infected cells have intact nuclei with coexpression of T Ag and VP1 (pink, arrows in squares in D and E). Separate color channels are shown in insets. Scale bars = (A, B) 250 Km; (CYE) 100 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Expressing, Virus, Immunostaining, Infection, Staining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 5. CD8-positive T cells colocalize with JC virus (JCV)Yinfected glia rather than neurons in mixed areas. (A) Triple immu- nostaining for CD8 (brown), microtubule-associated protein 2 ([MAP-2] gray blue), and T antigen (T Ag) (purple) of the cerebrum of a HIV-seropositive patient with progressive multifocal leukoencephalopathy (PML).CD8-positive T cells (brown, black circle arrow) are common in the gray-white junction (GWJ) area (inset 1) in the vicinity of JCV-infected glia (MAP-2 negative) expressing T Ag (purple, arrowhead). Inset 2 shows that the isolated JCV-infected cells, which express T Ag alone (purple, arrowheads) and are located in the area free of CD8-positive T cells of the gray matter (GM), are in fact neurons (gray blue, arrowheads, MAP-2 positive)., (B) Triple immunofluorescence (IFA) staining for VP1 (inset 1), MAP-2 (inset 2), and granzyme B (inset 3) in the cerebrum of an HIV-seropositive PML patient shows that the CD8-positive T cells of the GWJ produce granzyme B when they are in contact with productively JCV-infected glia (MAP-2 negative, arrows). (C) Triple immunostaining for granzyme B (brown), MAP-2 (gray blue), and T Ag (purple) of the cerebrum of a JCV encephalopathy (JCVE) patient showing a T cell (black circle arrow) releasing its granzyme B (double arrow) in a JCV-infected neuron. (D) Quadruple IFA for CD3, CD8, granzyme B, and VP1 done on the same patient shown in C show numerous CD3-positive/CD8-positive T cells (yellow-red, filled circle arrows) producing granzyme B (white plus sign) as well as some presumed CD4-positive T cells (CD3 positive/CD8 negative, asterisk) devoid of granzyme B near VP1- expressing cells. A granzyme-producing CD3-positive/CD8-positive T cell (full circle arrow) apposed to a JCV-infected cell of neuronal phenotype expressing VP1 (arrow) is visible at the right of the inset. WM, white matter. Scale bars = (A) 1 mm; (BYD) 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Virus, Infection, Expressing, Isolation, Staining, Triple Immunostaining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 6. Major histocompatibility complex class I (MHC I) is upregulated in infiltrates and in parenchymal cells in JC virus (JCV)Yinfected areas. (AYF) Multiple immunofluorescence (IFA) staining images of CD3, MHC I, and VP1 (A, D, E), microtubule- associated protein 2 (MAP-2), MHC I, CD8, and VP1 (B, C), and CD68 and MHC I (F). A typical uninfected area in the temporal lobe of an HIV-seropositive patient with progressive multifocal leukoencephalopathy (PML) are shown in A and B. In those areas, cells expressing high level of MHC I are rare and restricted to vessels, parenchymal CD3-positive (A) and CD8-positive T cells (B) are absent and only rare CD8-positive T cells (B, empty circle arrow) are seen in blood vessels. Conversely, numerous cells expressing high levels of MHC I (red) are present in lesions of an HIV-seronegative patient with miliary PML (C) and a JCV encephalopathy (JCVE) patient (D) that harbor parenchymal CD8-positive (C) or CD3-positive (D) T cells (filled circle arrows) close to JCV-infected cells expressing VP1 (arrow and inset). Some of the cells expressing a high level of MHC I are infiltrates in the parenchyma and perivascular cuffs as shown by double positive for CD3-positive T cells and MHC I in a JCVE patient (white, E, inset) and for CD68-positive macrophages and MHC I in a pediatric HIV-seropositive PML patient (white, F, inset). Scale bar = 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Immunopeptidomics, Virus, Staining, Expressing, Infection

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Claudin-2 upregulation enhances intestinal permeability, immune activation, dysbiosis, and mortality in sepsis.

doi: 10.1073/pnas.2217877121

Figure Lengend Snippet: Fig. 4. Claudin-2 expression augments mucosal immune activation. (A) Representative immunofluorescent staining of myeloperoxidase-positive neutrophils (green) and F4/80-positive macrophages (red). The graphs show numbers of neutrophils and macrophages within mucosa of sham WT (green) and claudin-2 KO (light green) and CLP WT (red) and claudin-2 KO (pink) mice. (B) Mucosal CD3+CD4- (green) and CD3+CD4+ (yellow) T cell infiltration is similar across all conditions. (C) Flow cytometric plot and graph of CD8αβ IEL numbers 24 h after CLP. (D) Flow cytometric plot and graph of IL-17 producing TCRγδ IELs after CLP. (E) Cytokine mRNA expression shows that claudin-2 KO mice have reduced IL-1β and IL-6 transcription, relative to WT, at 24 h after CLP. (F) Within Peyer's patches, the fractions of CD4+CD69+, CD4+CD25+, and CD8+CD25+ lymphocytes among all CD3+ cells were significantly greater in WT, relative to claudin-2 KO, mice. n = 5 to 12 for each condition. *P < 0.05; **P < 0.01; ***P < 0.001. NaK ATPase (blue) is shown for reference (A and B). Scale bars, 100 μm, 20 μm (Insets).

Article Snippet: Primary antibodies used for immunohisto chemistry Antigen Host Source Clone/Catalog RRID Concentration Claudin- 2 Rabbit Turner lab Rb188- 1/2 AB_2916077 1 μg/mL (IF) Claudin- 3 Rabbit Thermo fisher scientific 34- 1700 AB_86804 1 μ g/mL (IF) Claudin- 4 Rabbit Abcam ab210796 AB_2732879 0.1 μg/mL (IF) Claudin- 5 Rabbit BiCell 205 1 μ g/mL (IF) Claudin- 7 Rabbit Abcam ab207300 AB_2783812 0.5 μ g/mL (IF) Claudin- 15 Rabbit BiCell 215 1 μ g/mL (IF) pMLC Rabbit Turner lab 6889+6890 AB_2916078 1 μg/mL (IF) Occludin Rat Turner lab Clone 6B8A3 AB_2819194 1 μ g/mL (IF) Occludin Rat Turner lab Clone 5E5A6 AB_2819196 1 μg/mL (IF) E- cadherin Mouse Abnova MAB1388 AB_1671631 1 μ g/mL (IF) NaKATPase Mouse SantaCruz sc- 48345 AB_626712 0.5 μg/mL (IF) ZO- 1 Rat Turner lab Clone 6B6E4 AB_2783858 0.5 μ g/mL (IF) ZO- 1 Rat Turner lab Clone R40.76 AB_2783859 0.5 μ g/mL (IF) CD3 Rabbit Abcam Ab16669 AB_443425 5 μg/mL (IF) CD4 Rat eBioscience 13- 9766- 82 AB_2572833 5 μg/mL (IF) CD8 Rat eBioscience 13- 0808- 82 AB_2572771 5 μ g/mL (IF) CD25 Mouse Novus biologicals NB600- 564 AB_10002565 5 μg/mL (IF) CD68 Mouse SantaCruz sc- 20060 AB_2891106 1 μg/mL (IF) F4/80 Rabbit Cell signaling 70076T AB_2799771 1 μ g/mL (IF) MPO Rabbit Abcam Ab9535 AB_307322 1 μ g/mL (IF) JAMA Rabbit Thermo fisher scientific 36- 1700 AB_148483 0.5 μg/mL (IF) Ki67 Rabbit Novus biologicals NB600- 1252 AB_2142376 5 μ g/mL (IF) CD3 Rat BioRad MCA1477A488 AB_321245 5 μg/mL (IF) CD8 Rat Biolegend 372902 AB_2650657 10 μ g/mL (IF)

Techniques: Expressing, Activation Assay, Staining

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Exploring the Mechanisms of Lijin Fang on Treg/Th17 Cell Imbalance in COPD Based on Network Pharmacology

doi: 10.2147/COPD.S512469

Figure Lengend Snippet: Flow Cytometry Results for Treg (CD3+CD4+FOXP3+) / Th17 (CD3+CD4+IL17) Detection. LJF can reverse the decrease in Treg cells and the upregulation of Th17 cells induced by CSE. ( A ). Percentage of Treg cells in the Control group, CSE group, CSE+LJF-L group, and CSE+LJF-H group. ( B ). Percentage of Th17 cells in the Control group, CSE group, CSE+LJF-L group, and CSE+LJF-H group. (N=3. ***p<0.001, **p<0.01.).

Article Snippet: Following the manufacturer’s instructions, the appropriate volumes of antibodies against CD3 (E-AB-F1228J, Elabscience, China), CD4 (E-AB-F1105C, Elabscience, China), FOXP3 (B369751, BioLegend, China), and IL-17 (17–7177-81, eBioscience, USA) were added and mixed thoroughly.

Techniques: Flow Cytometry, Control