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Simultaneous Intranasal/Intravascular Antibody Labeling of CD4+ T Cells in Mouse Lungs

Featured protocol,  Authors: Yanqun Wang*
Yanqun WangAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3996
Jing Sun*
Jing SunAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3997
Rudragouda Channappanavar
Rudragouda ChannappanavarAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
Bio-protocol author page: a3999
Jingxian Zhao
Jingxian ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3998
Stanley Perlman
Stanley PerlmanAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
For correspondence: stanley-perlman@uiowa.edu
Bio-protocol author page: a1251
 and Jincun Zhao
Jincun ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou
For correspondence: zhaojincun@gird.cn
Bio-protocol author page: a1250
 (*contributed equally to this work) date: 1/5/2017, 90 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2099.

Brief version appeared in Immunity, Jun 2016
CD4+ T cell responses have been shown to be protective in many respiratory virus infections. In the respiratory tract, CD4+ T cells include cells in the airway and parenchyma and cells adhering to the pulmonary vasculature. Here we discuss in detail the methods that are useful for characterizing CD4+ T cells in different anatomic locations in mouse lungs.

In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor

Featured protocol,  Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 73 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2097.

Brief version appeared in Nat Immunol, Jun 2016
Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger et al., 2014). This protocol is designed for treatment with Kyn or FICZ in mouse embryonic fibroblasts (MEFs) or primary peripheral monocytes.

FICZ Exposure and Viral Infection in Mice

Featured protocol,  Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 79 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2096.

Brief version appeared in Nat Immunol, Jun 2016
The aryl hydrocarbon receptor (AHR) is known as a sensor for dioxins that mediates their toxicity, and also has important biophysiological roles such as circadian rhythms, cell differentiation and immune responses. 6-formylindolo(3,2-b)carbazole (FICZ), which is derived through the metabolism of L-tryptophan by ultraviolet B irradiation, is one of putative physiological ligands for AHR (Smirnova et al., 2016). It has recently been shown that endogenously-activated AHR signaling modulates innate immune response during viral infection (Yamada et al., 2016). This section describes how to treat mice with FICZ and to infect them with virus.

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Featured protocol,  Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 97 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

Brief version appeared in Cell Death Differ, Apr 2016
Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and -10 are fully activated by several proteolytic cleavage steps and induce the caspase cascade leading to apoptotic cell death. Analysing the processing of procaspases-8 and -10 by Western blot is a commonly used method to study the induction of apoptosis by death receptor stimulation. To analyse procaspase-8 and -10 cleavage, cells are stimulated with a death ligand for different time intervals, lysed and subjected to Western blot analysis using anti-caspase-8 and anti-caspase-10 antibodies. This allows monitoring the caspase cleavage products and thereby induction of apoptosis.

Simultaneous Intranasal/Intravascular Antibody Labeling of CD4+ T Cells in Mouse Lungs

Authors: Yanqun Wang*
Yanqun WangAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3996
Jing Sun*
Jing SunAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3997
Rudragouda Channappanavar
Rudragouda ChannappanavarAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
Bio-protocol author page: a3999
Jingxian Zhao
Jingxian ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
Bio-protocol author page: a3998
Stanley Perlman
Stanley PerlmanAffiliation: Departments of Microbiology, University of Iowa, Iowa, USA
For correspondence: stanley-perlman@uiowa.edu
Bio-protocol author page: a1251
 and Jincun Zhao
Jincun ZhaoAffiliation: State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou
For correspondence: zhaojincun@gird.cn
Bio-protocol author page: a1250
 (*contributed equally to this work) date: 1/5/2017, 90 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2099.

[Abstract] CD4+ T cell responses have been shown to be protective in many respiratory virus infections. In the respiratory tract, CD4+ T cells include cells in the airway and parenchyma and cells adhering to the pulmonary vasculature. Here we discuss in detail the methods that are useful for characterizing CD4+ T cells in different anatomic locations in mouse ...

In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor

Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 73 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2097.

[Abstract] Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger ...

FICZ Exposure and Viral Infection in Mice

Authors: Taisho Yamada
Taisho YamadaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
Bio-protocol author page: a3976
 and Akinori Takaoka
Akinori TakaokaAffiliation: Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan
For correspondence: takaoka@igm.hokudai.ac.jp
Bio-protocol author page: a2879
date: 1/5/2017, 79 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2096.

[Abstract] The aryl hydrocarbon receptor (AHR) is known as a sensor for dioxins that mediates their toxicity, and also has important biophysiological roles such as circadian rhythms, cell differentiation and immune responses. 6-formylindolo(3,2-b)carbazole (FICZ), which is derived through the metabolism of L-tryptophan by ultraviolet B irradiation, is one of ...

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 97 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

[Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and ...

Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)

Authors: Mickaël M. Ménager
Mickaël M. MénagerAffiliation: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
For correspondence: mickael.menager@med.nyu.edu
Bio-protocol author page: a3703
 and Dan R. Littman
Dan R. LittmanAffiliation 1: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
Affiliation 2: Howard Hughes Medical Institute, Washington, USA
Bio-protocol author page: a3704
date: 11/20/2016, 301 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2004.

[Abstract] The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy....

Evaluation of Cross-presentation in Bone Marrow-derived Dendritic Cells in vitro and Splenic Dendritic Cells ex vivo Using Antigen-coated Beads

Authors: Andrés Alloatti
Andrés AlloattiAffiliation: Institute Curie, PSL Research University, INSERM U932, Paris, France
For correspondence: andres.alloatti@curie.fr
Bio-protocol author page: a3727
Fiorella Kotsias
Fiorella KotsiasAffiliation 1: Institute Curie, PSL Research University, INSERM U932, Paris, France
Affiliation 2: Department of Virology, University of Buenos Aires, Buenos Aires, Argentina
Bio-protocol author page: a3728
Eik Hoffmann
Eik HoffmannAffiliation 1: Institute Curie, PSL Research University, INSERM U932, Paris, France
Affiliation 2: VIB Inflammation Research Center, Unit of Molecular Signal Transduction in Inflammation, Ghent, Belgium
Affiliation 3: Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Bio-protocol author page: a3725
 and Sebastian Amigorena
Sebastian AmigorenaAffiliation: Institute Curie, PSL Research University, INSERM U932, Paris, France
Bio-protocol author page: a3730
date: 11/20/2016, 303 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2015.

[Abstract] Antigen presentation by MHC class I molecules, also referred to as cross-presentation, elicits cytotoxic immune responses. In particular, dendritic cells (DC) are the most proficient cross-presenting cells, since they have developed unique means to control phagocytic and degradative pathways.
   This protocol allows the evaluation of antigen cross-presentation ...

Analysis of Phagosomal Antigen Degradation by Flow Organellocytometry

Authors: Eik Hoffmann
Eik HoffmannAffiliation 1: Institute Curie, INSERM U932, PSL Research University, Paris, France
Affiliation 2: VIB Inflammation Research Center, Unit of Molecular Signal Transduction in Inflammation, Ghent, Belgium
Affiliation 3: Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
For correspondence: eik.hoffmann@irc.vib-ugent.be
Bio-protocol author page: a3725
Anne-Marie Pauwels
Anne-Marie PauwelsAffiliation 1: VIB Inflammation Research Center, Unit of Molecular Signal Transduction in Inflammation, Ghent, Belgium
Affiliation 2: Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Bio-protocol author page: a3726
Andrés Alloatti
Andrés AlloattiAffiliation: Institute Curie, INSERM U932, PSL Research University, Paris, France
Bio-protocol author page: a3727
Fiorella Kotsias
Fiorella KotsiasAffiliation 1: Institute Curie, INSERM U932, PSL Research University, Paris, France
Affiliation 2: Department of Virology, University of Buenos Aires, Buenos Aires, Argentina
Bio-protocol author page: a3728
 and Sebastian Amigorena
Sebastian AmigorenaAffiliation: Institute Curie, INSERM U932, PSL Research University, Paris, France
Bio-protocol author page: a3730
date: 11/20/2016, 246 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2014.

[Abstract] Professional phagocytes internalize self and non-self particles by phagocytosis to initiate innate immune responses. After internalization, the formed phagosome matures through fusion and fission events with endosomes and lysosomes to obtain a more acidic, oxidative and hydrolytic environment for the degradation of its cargo. Interestingly, phagosome ...

Determination of H2O2 Generation by pHPA Assay

Authors: Jennifer L. Larson-Casey
Jennifer L. Larson-CaseyAffiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
Bio-protocol author page: a3716
 and A. Brent Carter
A. Brent CarterAffiliation 1: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
Affiliation 2: Birmingham Veterans Administration Medical Center, Birmingham, AL, USA
For correspondence: bcarter1@uab.edu
Bio-protocol author page: a3717
date: 11/20/2016, 264 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2010.

[Abstract] The production of reactive oxygen species, including H2O2, is a process that can be used in signaling, cell death, or immune response. To quantify oxidative stress in cells, a fluorescence technique has been modified from a previously described method to measure H2O2 release from cells (Panus et al., 1993; Murthy et al., 2010; Larson-Casey et al., ...

Assay to Evaluate BAL Fluid Regulation of Fibroblast α-SMA Expression

Authors: Jennifer L. Larson-Casey
Jennifer L. Larson-CaseyAffiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
Bio-protocol author page: a3716
 and A. Brent Carter
A. Brent CarterAffiliation 1: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
Affiliation 2: Birmingham Veterans Administration Medical Center, Birmingham, AL, USA
For correspondence: bcarter1@uab.edu
Bio-protocol author page: a3717
date: 11/20/2016, 307 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2009.

[Abstract] Because transforming growth factor-β (TGF-β1) induces differentiation of fibroblasts to myofibroblasts, we developed a protocol to evaluate alveolar macrophage-derived TGF-β1 regulation of lung fibroblast differentiation (Larson-Casey et al., 2016). The protocol evaluates the ability of mouse bronchoalveolar lavage (BAL) fluid to alter fibroblast differentiation. ...

Establishment of Patient-Derived Xenografts in Mice

Authors: Dongkyoo Park
Dongkyoo ParkAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3712
Dongsheng Wang
Dongsheng WangAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3713
Guo Chen
Guo ChenAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
Bio-protocol author page: a3714
 and Xingming Deng
Xingming DengAffiliation: Division of Cancer Biology, Department of Radiation Oncology, Emory University School of Medicine and Winship Cancer Institute of Emory University, Atlanta, GA, USA
For correspondence: xdeng4@emory.edu
Bio-protocol author page: a3715
date: 11/20/2016, 352 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2008.

[Abstract] Patient-derived xenograft (PDX) models for cancer research have recently attracted considerable attention in both the academy and industry (Hidalgo et al., 2014; Wilding and Bodmer, 2014). PDX models have been developed from different tumor types including lung cancer to improve the drug development process. These models are used for pre-clinical drug ...
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In vitro Culture of Human PBMCs

Authors: Santosh K Panda
Santosh K PandaAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
Bio-protocol author page: a221
 and Balachandran Ravindran
Balachandran RavindranAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
For correspondence: ravindran8@gmail.com
Bio-protocol author page: a222
date: 2/5/2013, 37570 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.322.

[Abstract] Peripheral blood mononuclear cells (PBMCs) consist of chiefly of lymphocytes and monocytes. Purified PBMCs are used in vitro to evaluate a variety of functions of lymphocytes viz; a) proliferation to mitogenic (PHA, Con-A) stimulation, b) monitoring of prior sensitisation in antigen recall assays by ...

Isolation and Culture of Mouse Bone Marrow-derived Macrophages (BMM’phi’)

Author: Ran Chen
Ran ChenAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: rcchen@jfkbio.com
Bio-protocol author page: a34
date: 2/5/2012, 33935 views, 15 Q&A
DOI: https://doi.org/10.21769/BioProtoc.68.

[Abstract] Bone marrow derived macrophages are a type of white blood cell that can be isolated from mammalian bone marrow. In this protocol, a method is described in which bone marrow cells are isolated from mouse leg bones (femur and tibia), and then differentiated to bone marrow-derived macrophages in approximately ...

[Bio101] In vitro Differentiation of Mouse Th0, Th1 and Th2 from Naïve CD4 T Cells

Author: Jia Li
Jia LiAffiliation: Department of Immunology, Medical Center, Duke University, Durham, North Carolina, USA
For correspondence: jiali.email@gmail.com
Bio-protocol author page: a16
date: 11/20/2011, 29201 views, 18 Q&A
DOI: https://doi.org/10.21769/BioProtoc.157.

[Abstract] In vitro differentiation of helper T cells of various lineages is frequently used in T helper cell study. Naïve CD4 T cells can differentiate into certain lineage of T help cells in vitro in the presence of specific stimulatory cytokines and inhibition of cytokines that are essential for the differentiation ...

[Bio101] Whole Blood Staining of Human Monocyte Subsets for Flow Cytometry

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 5/20/2011, 20806 views, 10 Q&A
DOI: https://doi.org/10.21769/BioProtoc.69.

[Abstract] This is a general protocol to stain whole human blood for flow analysis with minimal spontaneous activation of monocytes. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research....

[Bio101] In vivo BrdU Incorporation and Detection in Mouse

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 6/5/2011, 18610 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.81.

[Abstract] BrdU (Bromodeoxyuridine or 5-bromo-2’-deoxyuridine) is a synthetic nucleoside that is incorporated into DNA by proliferating cells. This protocol is to be used to incorporate and detect BrdU in murine plasma cells. The plasma cells described in this protocol are formed spontaneously in autoimmune mice ...

Isolation of Human PBMCs

Authors: Santosh K Panda
Santosh K PandaAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
Bio-protocol author page: a221
 and Balachandran Ravindran
Balachandran RavindranAffiliation: Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, India
For correspondence: ravindran8@gmail.com
Bio-protocol author page: a222
date: 2/5/2013, 18077 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.323.

[Abstract] Peripheral blood mononuclear cells (PBMCs) are chiefly lymphocytes and monocytes. PBMCs are separated from the whole blood by a density gradient centrifugation method using Ficoll Histopaque....

[Bio101] Thioglycollate Induced Peritonitis

Author: Zheng Liu
Zheng LiuAffiliation: The Feinstein Institute for Medical Research, Manhasset, NY, USA
For correspondence: zl2119@caa.columbia.edu
Bio-protocol author page: a12
Anne Davidson Lab, date: 6/20/2011, 16627 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.84.

[Abstract] Intraperitoneal (i.p.) injection of thioglycollate elicits a robust influx of neutrophils into peritoneal cavity. The trafficking of the cells is believed to be mediated by chemokines CXCL1, CXCL2, and CXCL8 (Call et al., 2001; Cacalano et al., 1994). Thus this model can be used to test the ability ...

Intracellular Cytokine (INF-gamma) Staining Assay

Author: Huagang Zhang
Huagang ZhangAffiliation: Albert Einstein College of Medicine, Yeshiva University, New York City, USA
For correspondence: huagangzhang@gmail.com
Bio-protocol author page: a21
date: 4/5/2012, 16507 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.122.

[Abstract] An intracellular cytokine (INF-gamma) staining assay is used to analyze the function of lymphocytes at the single cell level. By combining surface staining and intracellular cytokine staining, this assay can reveal the percentage of cytokine-releasing cells in a particular population, which cannot be ...

Isolation of Dendritic Cells and Macrophages from the Murine Kidneys of Lupus by Cell Sorter

Authors: Ramalingam Bethunaickan
Ramalingam BethunaickanAffiliation: Center for Autoimmunity and Musculoskeletal Diseases, Feinstein Institute for Medical Research, New York, USA
For correspondence: bramalingam@gmail.com
Bio-protocol author page: a24
 and Anne Davidson
Anne DavidsonAffiliation: Center for Autoimmunity and Musculoskeletal Diseases, Feinstein Institute for Medical Research, New York, USA
Bio-protocol author page: a1712
Anne Davidson Lab, date: 4/20/2012, 12204 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.168.

[Abstract] Methods for the isolation and characterization of mononuclear phagocytes from the kidneys of mice with SLE are essential to understand the patho-physiology of the disease. Activation of these cells is associated with the onset of clinical disease in mice and infiltration with these cells is associated ...

Isolation, Purification, and Culture of Primary Murine Microglia Cells

Authors: Xuqin Chen
Xuqin ChenAffiliation: Department of Neurology, Affiliated Children's Hospital, Soochow University, Suzhou, China
For correspondence: xuqinlili@yahoo.com
Bio-protocol author page: a202
Ye Zhang
Ye ZhangAffiliation: Department of Neurology, Affiliated Children's Hospital, Soochow University, Suzhou, China
Bio-protocol author page: a203
Gaitri Sadadcharam
Gaitri SadadcharamAffiliation: Department of Neurology, Affiliated Children's Hospital, Soochow University, Suzhou, China
Bio-protocol author page: a204
Weili Cui
Weili CuiAffiliation: Department of Neurology, Affiliated Children's Hospital, Soochow University, Suzhou, China
Bio-protocol author page: a205
 and Jiang Huai Wang
Jiang Huai WangAffiliation: Department of Neurology, Affiliated Children's Hospital, Soochow University, Suzhou, China
For correspondence: jh.wang@ucc.ie
Bio-protocol author page: a206
date: 1/5/2013, 11171 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.314.

[Abstract] The following is a detailed protocol for the isolation, purification and culture of murine brain microglia cells using neutral enzyme digestion and shaking. The protocol below is designed to isolate and culture a large number of purified inactivated microglia cells. Neutral enzyme digestion allows ...
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