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Bacterial Survival in Dictyostelium

Authors: Regin Rønn*
Regin RønnAffiliation 1: Department of Biology, University of Copenhagen, Copenhagen, Denmark
Affiliation 2: Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
Bio-protocol author page: a4771
, Xiuli Hao*
Xiuli HaoAffiliation: Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
Bio-protocol author page: a4772
, Freja Lüthje
Freja LüthjeAffiliation: Department of Biology, University of Copenhagen, Copenhagen, Denmark
Bio-protocol author page: a4773
, Nadezhda A. German
Nadezhda A. GermanAffiliation: Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, Amarillo, TX, USA
Bio-protocol author page: a4774
, Xuanji Li
Xuanji LiAffiliation: Department of Biology, University of Copenhagen, Copenhagen, Denmark
Bio-protocol author page: a4775
, Fuyi Huang
Fuyi HuangAffiliation: Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
Bio-protocol author page: a4776
, Javan Kisaka
Javan KisakaAffiliation: Department of Chemistry, Western Michigan University, Kalamazoo, MI, USA
Present address: Division of Infectious Diseases, Washington University School of Medicine, St. Louis, MO, USA
Bio-protocol author page: a4777
, David Huffman
David HuffmanAffiliation 1: Department of Chemistry, Western Michigan University, Kalamazoo, MI, USA
Affiliation 2: Department of Plant and Environmental Sciences, University of Copenhagen, MO, USA
Bio-protocol author page: a4778
, Hend A. Alwathnani
Hend A. AlwathnaniAffiliation: Department of Botany and Microbiology, King Saud University, Riyadh, Saudi Arabia
Bio-protocol author page: a4779
, Yong-Guan Zhu
Yong-Guan ZhuAffiliation: Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
Bio-protocol author page: a4780
 and Christopher Rensing
Christopher RensingAffiliation 1: Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China
Affiliation 2: J. Craig Venter Institute, La Jolla, CA, USA
Affiliation 3: Fujian Provincial Key Laboratory of Soil Environmental Health and Regulation, College of Resources and Environment, Fujian Agriculture & Forestry University, Fuzhou, China
For correspondence: rensing@iue.ac.cn
Bio-protocol author page: a4781
 (*contributed equally to this work),  date: 7/5/2017, 118 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2376.

[Abstract] We performed an assay to test the ability of different E. coli strains to survive inside amoebal cells after ingestion. In the assay we incubated bacteria together with cells of Dictyostelium discoideum for six hours. After co-incubation most of the uningested bacteria were removed by centrifugation and the remaining uningested bacteria were killed ...

Culturing Bacteria from Caenorhabditis elegans Gut to Assess Colonization Proficiency

Authors: Facundo Rodriguez Ayala
Facundo Rodriguez AyalaAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
For correspondence: facundoayala.foncyt@gmail.com
Bio-protocol author page: a4690
, Sebastián Cogliati
Sebastián CogliatiAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4691
, Carlos Bauman
Carlos BaumanAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4692
, Cecilia Leñini
Cecilia LeñiniAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4693
, Marco Bartolini
Marco BartoliniAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4694
, Juan Manuel Villalba
Juan Manuel VillalbaAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4695
, Federico Argañaraz
Federico ArgañarazAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4696
 and Roberto Grau
Roberto GrauAffiliation: Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, CONICET, Rosario, Argentina
Bio-protocol author page: a4697
,  date: 6/20/2017, 264 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2345.

[Abstract] Determining an accurate count of intestinal bacteria from Caenorhabditis elegans is one critical way to assess colonization proficiency by a given bacteria. This can be accomplished by culturing appropriate dilutions of worm gut bacteria on selective or differential agarized media. Because of the high concentration of bacteria in gut worm, dilution ...

Isolation and Infection of Drosophila Primary Hemocytes

Authors: Charles Tracy
Charles TracyAffiliation: Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
Bio-protocol author page: a4450
 and Helmut Krämer
Helmut KrämerAffiliation 1: Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
Affiliation 2: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
Affiliation 3: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA
For correspondence: helmut.kramer@utsouthwestern.edu
Bio-protocol author page: a4451
,  date: 6/5/2017, 317 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2300.

[Abstract] Phagocytosis of invading pathogens and their subsequent clearance in lysosomes is important for organismal fitness. We have devised the following protocol to extract phagocytic hemocytes from wild-type and mutant Drosophila larvae and infect the isolated hemocytes with GFP-labeled E. coli to measure the rate of phagocytosis and degradation within individual ...

In vitro Antigen-presentation Assay for Self- and Microbial-derived Antigens

Author: Laura Campisi
Laura CampisiAffiliation: Department of Microbiology & Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA
For correspondence: laura.campisi@mssm.edu
Bio-protocol author page: a4584
,  date: 6/5/2017, 341 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2307.

[Abstract] Antigen presenting cells (APC) are able to process and present to T cells antigens from different origins. This mechanism is highly regulated, in particular by Patter Recognition Receptor (PRR) signals. Here, I detail a protocol designed to assess in vitro the capacity of APC to present antigens derived from bacteria, apoptotic and infected apoptotic ...

Ex vivo Model of Human Aortic Valve Bacterial Colonization

Authors: Alejandro Avilés-Reyes
Alejandro Avilés-ReyesAffiliation: Department of Oral Biology, Piracicaba Dental School, University of Campinas, Gainesville, USA
For correspondence: aaviles-reyes@dental.ufl.edu
Bio-protocol author page: a2083
, Irlan Almeida Freires
Irlan Almeida FreiresAffiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, Brazil
Bio-protocol author page: a4608
, Pedro Luiz Rosalen
Pedro Luiz RosalenAffiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, Brazil
Bio-protocol author page: a4609
, José A. Lemos
José A. LemosAffiliation: Department of Oral Biology, University of Florida, College of Dentistry, Gainesville, USA
Bio-protocol author page: a2084
 and Jacqueline Abranches
Jacqueline AbranchesAffiliation: Department of Oral Biology, University of Florida, College of Dentistry, Gainesville, USA
Bio-protocol author page: a2085
,  date: 6/5/2017, 424 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2316.

[Abstract] The interaction of pathogens with host tissues is a key step towards successful colonization and establishment of an infection. During bacteremia, pathogens can virtually reach all organs in the human body (e.g., heart, kidney, spleen) but host immunity, blood flow and tissue integrity generally prevents bacterial colonization. Yet, patients with cardiac ...

A Reliable Assay to Evaluate the Virulence of Aspergillus nidulans Using the Alternative Animal Model Galleria mellonella (Lepidoptera)

Authors: Caroline Mota Fernandes
Caroline Mota FernandesAffiliation: Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
For correspondence: cmota@biof.ufrj.br
Bio-protocol author page: a4626
, Fernanda Lopes Fonseca
Fernanda Lopes FonsecaAffiliation: Centro de Desenvolvimento Tecnológico em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil
Bio-protocol author page: a4627
, Gustavo Henrique Goldman
Gustavo Henrique GoldmanAffiliation: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil
Bio-protocol author page: a4628
, Marcos Dias Pereira*
Marcos Dias PereiraAffiliation: Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
For correspondence: marcosdp@iq.ufrj.br
Bio-protocol author page: a4629
 and Eleonora Kurtenbach*
Eleonora KurtenbachAffiliation: Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Bio-protocol author page: a4630
 (*contributed equally to this work),  date: 6/5/2017, 282 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2329.

[Abstract] The greater wax moth Galleria mellonella has emerged as an effective heterologous host to study fungal pathogenesis and the efficacy of promising antifungal drugs (Mylonakis et al., 2005; Li et al., 2013). Here, a methodology describing the Aspergillus nidulans infection in G. mellonella larvae, along with insect survival analysis, is reported. This ...

Spore Preparation Protocol for Enrichment of Clostridia from Murine Intestine

Authors: Eric M. Velazquez
Eric M. VelazquezAffiliation: Department of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, One Shields Avenue, Davis, CA 95616, USA
Bio-protocol author page: a4551
, Fabian Rivera-Chávez
Fabian Rivera-ChávezAffiliation: Department of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, One Shields Avenue, Davis, CA 95616, USA
Bio-protocol author page: a4552
 and Andreas J. Bäumler
Andreas J. BäumlerAffiliation: Department of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, One Shields Avenue, Davis, CA 95616, USA
For correspondence: ajbaumler@ucdavis.edu
Bio-protocol author page: a4553
,  date: 5/20/2017, 406 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2296.

[Abstract] In recent years, many spore-forming commensal Clostridia found in the gut have been discovered to promote host physiology, immune development, and protection against infections. We provide a detailed protocol for rapid enrichment of spore-forming bacteria from murine intestine. Briefly, contents from the intestinal cecum are collected aerobically, ...

Escherichia coli Infection of Drosophila

Authors: Charles Tracy
Charles TracyAffiliation: Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, USA
Bio-protocol author page: a4450
 and Helmut Krämer
Helmut KrämerAffiliation 1: Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, USA
Affiliation 2: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, USA
For correspondence: helmut.kramer@utsouthwestern.edu
Bio-protocol author page: a4451
,  date: 5/5/2017, 429 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2256.

[Abstract] Following septic insults, healthy insects, just like vertebrates, mount a complex immune response to contain and destroy pathogens. The failure to efficiently clear bacterial infections in immuno-compromised fly mutants leads to higher mortality rates which provide a powerful indicator for genes with important roles in innate immunity. The following ...

Pathogenicity Assay of Penicillium expansum on Apple Fruits

Authors: Yong Chen
Yong ChenAffiliation: Key Laboratory of Plant Resources, Institute of Botany, the Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a4471
, Boqiang Li
Boqiang LiAffiliation: Key Laboratory of Plant Resources, Institute of Botany, the Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a1847
, Zhanquan Zhang
Zhanquan ZhangAffiliation: Key Laboratory of Plant Resources, Institute of Botany, the Chinese Academy of Sciences, Beijing, China
Bio-protocol author page: a1845
 and Shiping Tian
Shiping TianAffiliation: Key Laboratory of Plant Resources, Institute of Botany, the Chinese Academy of Sciences, Beijing, China
For correspondence: tsp@ibcas.ac.cn
Bio-protocol author page: a1848
,  date: 5/5/2017, 460 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2264.

[Abstract] Penicillium expansum, a widespread filamentous fungus, is a major causative agent of fruit decay and leads to huge economic losses during postharvest storage and shipping. Furthermore, it produces mycotoxin on the infected fruits that may cause harmful effects to human health. This pathogenicity assay involves a stab inoculation procedure of P. expansum ...

Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes

Authors: Swetha Reddy
Swetha ReddyAffiliation: College of Veterinary Science, Mississippi State University, Starkville, USA
For correspondence: mswethareddy@gmail.com
Bio-protocol author page: a4476
 and Frank Austin
Frank AustinAffiliation: College of Veterinary Science, Mississippi State University, Starkville, USA
Bio-protocol author page: a4477
,  date: 5/5/2017, 488 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2267.

[Abstract] Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like cervical, liver, ...
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[Bio101] Purification of Adenovirus by Cesium Chloride Density Gradients

Author: Huan Pang
Huan PangAffiliation: Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, USA
For correspondence: pang_huan@hotmail.com
Bio-protocol author page: a48
,  date: 4/5/2012, 18558 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.201.

[Abstract] Adenovirus are efficient gene delivery systems. The standard method for purification of adenoviral vectors is based on using a cesium chloride (CsCl) density gradient combined with ultracentrifugation. This method is suitable for small-scale purification and is less expensive than column chromatography ...

Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses

Authors: Anna Sloutskin
Anna SloutskinAffiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
Bio-protocol author page: a1808
 and Ronald S. Goldstein
Ronald S. GoldsteinAffiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
For correspondence: ron.goldstein@biu.ac.il
Bio-protocol author page: a1809
,  date: 11/20/2014, 11605 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1295.

[Abstract] Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for fluorescently labeled viruses, based on the original plaque assay. In addition, the calculation of multiplicity ...

Assay to Evaluate Vascular Permeability Induction in Mice

Authors: Henry Puerta-Guardo
Henry Puerta-GuardoAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a980
, Arturo Raya-Sandino
Arturo Raya-SandinoAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a981
, Lorenza González-Mariscal
Lorenza González-MariscalAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a982
, Victor H. Rosales
Victor H. RosalesAffiliation: General Services Laboratory, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a983
, José Ayala-Dávila
José Ayala-DávilaAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a984
, Bibiana Chávez-Mungía
Bibiana Chávez-MungíaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a985
, Daniel Martínez-Fong
Daniel Martínez-FongAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a986
, Fernando Medina
Fernando MedinaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a987
, Juan E. Ludert
Juan E. LudertAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: jludert@cinvestav.mx
Bio-protocol author page: a988
 and Rosa María del Angel
Rosa María del AngelAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: rmangel@cinvestav.mx
Bio-protocol author page: a989
,  date: 11/20/2013, 9120 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.977.

[Abstract] Dengue virus infection usually courses as a benign self-limited fever, called dengue fever. However, on occasions it can progress to a life-threatening complication known as severe dengue (SD). A hallmark of SD is a sharp increase in vascular permeability. Secondary infections are considered a risk ...

Bacterial Counts in Spleen

Authors: Elías Barquero-Calvo
Elías Barquero-CalvoAffiliation: Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica
Bio-protocol author page: a954
, Carlos Chacón-Díaz
Carlos Chacón-DíazAffiliation: Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
Bio-protocol author page: a958
, Esteban Chaves-Olarte
Esteban Chaves-OlarteAffiliation: Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
Bio-protocol author page: a960
 and Edgardo Moreno
Edgardo MorenoAffiliation: Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica
For correspondence: emoreno@racsa.co.cr
Bio-protocol author page: a961
,  date: 11/5/2013, 8057 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.954.

[Abstract] Bacterial loads can be determined as colony forming units (CFU) at any point of the infection by culturing spleen homogenates on agar plates. This is a reliable method for comparing the kinetics of infection in various mouse strains, estimating the virulence of different bacterial mutants or isolates ...

Gentamicin Protection Assay to Determine Bacterial Survival within Macrophages

Authors: Sargurunathan Subashchandrabose
Sargurunathan SubashchandraboseAffiliation: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, USA
For correspondence: sarguru@umich.edu
Bio-protocol author page: a1651
 and Harry L.T. Mobley
Harry L.T. MobleyAffiliation: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, USA
Bio-protocol author page: a1652
,  date: 9/20/2014, 7361 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1235.

[Abstract] Macrophages are key cells involved in orchestrating host defense against infections. Here, we describe the protocol for a bacterial killing assay in macrophages that can be adapted to any bacterial pathogen. Using this assay, we analyzed the survival of wild-type and mutant strains of Escherichia coli ...

Intracellular Macrophage Infections with E. coli under Nitrosative Stress

Authors: Stacey L. Bateman
Stacey L. Bateman Affiliation: Department of Molecular Genetics and Microbiology Center for Microbial Pathogenesis, Department of Pediatrics, Duke University School of Medicine, Durham, NC, USA
Bio-protocol author page: a129
 and Patrick Seed
Patrick SeedAffiliation: Department of Molecular Genetics and Microbiology Center for Microbial Pathogenesis, Department of Pediatrics, Duke University School of Medicine, Durham, NC, USA
For correspondence: patrick.seed@duke.edu
Bio-protocol author page: a130
,  date: 10/20/2012, 6594 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.275.

[Abstract] Escherichia coli (E. coli) produces disseminated infections of the urinary tract, blood, and central nervous system where it encounters professional phagocytes such as macrophages, which utilize reactive nitrogen intermediates (RNI) to arrest bacteria. In vitro, extraintestinal pathogenic E. coli (ExPEC) ...

Amplification of HIV-1 Infectious Virus in BL3 Lab

Author: Xin Wang
Xin WangAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: xinwang73@hotmail.com
Bio-protocol author page: a26
,  date: 3/5/2012, 6403 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.102.

[Abstract] This method is used for making high titer human immunodeficiency virus type-1 (HIV-1) virus stock for subsequent infection assays. The amplification of T-tropic HIV-1 virus (IIIB strain) uses the CD4+ T cell line H9....

Murine in vivo CD8+ T Cell Killing Assay

Authors: Myoungjoo V. Kim
Myoungjoo V. KimAffiliation: Department of Immunobiology, Yale University School of Medicine, New Haven, USA
For correspondence: myoungjoo.kim@yale.edu
Bio-protocol author page: a1487
, Weiming Ouyang
Weiming OuyangAffiliation: Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Bethesda, USA
Bio-protocol author page: a1460
, Will Liao
Will LiaoAffiliation: Genomics, New York Genome Center, New York, USA
Bio-protocol author page: a1461
, Michael Q. Zhang
Michael Q. ZhangAffiliation: Molecular and Cellular Biology Department, University of Texas at Dallas, Richardson, USA
Bio-protocol author page: a1462
 and Ming O. Li
Ming O. LiAffiliation: Department of Immunology, Memorial Sloan Kettering Cancer Center (MSKCC), New York, USA
Bio-protocol author page: a1463
,  date: 7/5/2014, 6134 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1172.

[Abstract] Antigen-specific killing ability of effector CD8+ T cells is critical for protective immunity against infection. Here, we describe in vivo cytotoxic T cell assay to examine effector function of antigen-specific CD8+ T cells. Mice infected with Listeria monocytogenes (L. monocytogenes) expressing chicken ...

Neutral Red Assay for Murine Norovirus Replication and Detection in a Mouse

Authors: Mariam Bernadette González-Hernández
Mariam Bernadette González-HernándezAffiliation: Department of Microbiology and Immunology, Graduate Program of Immunology, University of Michigan, Ann Arbor, MI, USA
For correspondence: gonmaria@umich.edu
Bio-protocol author page: a312
, Jeffrey William Perry
Jeffrey William PerryAffiliation: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, USA
Bio-protocol author page: a528
 and Christiane E. Wobus
Christiane E. WobusAffiliation: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, USA
For correspondence: cwobus@umich.edu
Bio-protocol author page: a311
,  date: 4/5/2013, 6067 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.415.

[Abstract] Neutral red (NR) is a dye that must be actively imported into the cell, and, therefore, the dye has been used for decades to selectively stain living cells. In addition, NR can also be incorporated into virus particles, although the mechanism behind this is poorly understood. Once encapsulated into ...

Plasmodium falciparum Rosette Formation Assay

Authors: Inès Vigan-Womas
Inès Vigan-WomasAffiliation: Parasite Molecular Immunology Unit, Institut Pasteur, Paris, France
For correspondence: ines.vigan-womas@pasteur.fr
Bio-protocol author page: a577
, Micheline Guillotte
Micheline GuillotteAffiliation: Parasite Molecular Immunology Unit, Institut Pasteur, Paris, France
Bio-protocol author page: a576
 and Odile Mercereau-Puijalon
Odile Mercereau-PuijalonAffiliation: Parasite Molecular Immunology Unit, Institut Pasteur, Paris, France
Bio-protocol author page: a308
,  date: 4/20/2013, 5742 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.412.

[Abstract] Rosetting, i.e. the capacity of red blood cells (iRBCs) infected with mature parasite stages to bind two or more uninfected red blood cells (RBCs) is a virulence factor of Plasmodium falciparum. This protocol describes an in vitro assay to monitor rosette formation by P. falciparum-infected red blood ...
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