IL-23- and Imiquimod-Induced Models of Experimental Psoriasis in Mice.
This article provides detailed methodologies for creating and evaluating the IL-23- and Aldara/IMQ-induced mouse models of psoriasis. The article also provides a protocol for analyzing skin leukocytes by flow cytometry. © 2019 by John Wiley & Sons, Inc.
PMID: 30615272 [PubMed - as supplied by publisher] (Source: Current Protocols in Immunology)
Source: Current Protocols in Immunology - January 8, 2019 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Isolation and Culture of Microglia.
This article describes approaches for purification of microglia by fluorescence-activated cell sorting using microglia-specific surface markers and for enrichment of microglia by magnetic sorting and immunopanning. Detailed information about culturing primary microglia at various developmental stages is also provided. Throughout, we focus on special considerations for handling microglia and compare the relative strengths or disadvantages of different protocols. © 2018 by John Wiley & Sons, Inc.
PMID: 30414379 [PubMed - as supplied by publisher] (Source: Current Protocols in Immunology)
Source: Current Protocols in Immunology - November 12, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Statistics for Immunologists
Authors:
PMID: 30400720 [PubMed - in process] (Source: Current Protocols in Immunology)
Source: Current Protocols in Immunology - November 9, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Ribonucleoprotein Transfection for CRISPR/Cas9-Mediated Gene Knockout in Primary T Cells.
Authors: Oh SA, Seki A, Rutz S
Abstract
CRISPR/Cas9 has enabled the rapid and efficient generation of gene knockouts across various cell types of several species. T cells are central players in adaptive immune responses. Gene editing in primary T cells not only represents a valuable research tool, but is also critical for next generation immunotherapies, such as CAR T cells. Broad application of CRIPSR/Cas9 for gene editing in primary T cells has been hampered by limitations in transfection efficiency and the requirement for TCR stimulation. In this article, we provide a detailed protocol for Cas9/gRNA rib...
Source: Current Protocols in Immunology - October 20, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Methods to Measure MDSC Immune Suppressive Activity In Vitro and In Vivo.
Authors: Solito S, Pinton L, De Sanctis F, Ugel S, Bronte V, Mandruzzato S, Marigo I
Abstract
This unit presents methods to assess the immunosuppressive properties of immunoregulatory cells of myeloid origin, such as myeloid-derived suppressor cells (MDSCs), both in vitro and in vivo in mice, as well as in biological samples from cancer patients. These methods could be adapted to test the impact of different suppressive populations on T cell activation, proliferation, and cytotoxic activity; moreover, they could be useful to assess the influence exerted by genetic modifications, chemical inhibitors, and dr...
Source: Current Protocols in Immunology - October 13, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Organotypic Brain Slice Cultures.
Authors: Humpel C
Abstract
Ex vivo cell culture models are of particular interest for neurobiologists, as these allow the study of brain cells in an isolated environment. Organotypic brain slice cultures allow growth of three-dimensional brain tissue that conserves the cellular architecture. This unit describes the preparation and culturing of organotypic brain slices from mice. In three basic protocols, the general procedure, the specific preparation of chopper slices, and slicing of whole-brain vibratome sections are described. Support protocols explain the use of postnatal or adult mice, the preparation...
Source: Current Protocols in Immunology - October 13, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Efficient CRISPR/Cas9-Mediated Mutagenesis in Primary Murine T Lymphocytes.
This article outlines a protocol for CRISPR/Cas9-mediated mutagenesis in primary T lymphocytes from Cas9 transgenic mice using retroviral delivery of guide RNAs. © 2018 by John Wiley & Sons, Inc.
PMID: 30312021 [PubMed - as supplied by publisher] (Source: Current Protocols in Immunology)
Source: Current Protocols in Immunology - October 13, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Neuroinflammation Quantification for Spinal Cord Injury.
Authors: Amo-Aparicio J, Martínez-Muriana A, Sánchez-Fernández A, López-Vales R
Abstract
Spinal cord injury (SCI) leads to irreversible devastating neurological disabilities. Accumulated evidence in the literature indicates that the inflammatory response that occurs in the spinal cord following injury contributes importantly to spread tissue damage to healthy regions adjacent to the lesion site, and consequently, to increase neurological deficits. Therefore, targeting inflammation could lead to the development of new therapies to prevent tissue damage and neurological impairments after SCI. Inflammatio...
Source: Current Protocols in Immunology - September 27, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Mouse Model for Human Vitiligo.
This article discusses the first mouse model of vitiligo that develops epidermal depigmentation, similar to disease in human patients. To achieve epidermal depigmentation, mice are genetically engineered to retain melanocytes in the skin epidermis. Induction of disease occurs by adoptive transfer of melanocyte-specific CD8+ T cells into recipient mice and the subsequent activation of these T cells using a viral vector. Depigmentation of the epidermis occurs within 5 to 7 weeks in a patchy pattern similar to patients with vitiligo. This article describes the methods of vitiligo induction, quantification of lesion progressio...
Source: Current Protocols in Immunology - September 27, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
LC3-Associated Phagocytosis and Antigen Presentation.
Authors: Ligeon LA, Loi M, Münz C
Abstract
LC3-associated phagocytosis (LAP) is an unconventional form of autophagy that relies on parts of the canonical autophagy machinery for its function. LAP is triggered upon receptor-mediated phagocytosis and is characterized by the formation of a single-membrane vesicle decorated with the autophagy protein LC3. In professional phagocytic cells, such as macrophages, the role of LAP in immune processes has been characterized, although how LAP functions at the molecular level remains poorly defined. It is important to point out that as for all autophagic pathways, the...
Source: Current Protocols in Immunology - September 27, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Proximity Ligation Assay (PLA).
Authors: Alam MS
Abstract
Proximity ligation assay (PLA), also referred to as Duolink® PLA technology, permits detection of protein-protein interactions in situ (at distances <40 nm) at endogenous protein levels. It exploits specific antibodies identifying (either directly or indirectly) the two proteins of interest and utilizes specific DNA primers covalently linked to the antibodies. A hybridization step followed by a PCR amplification with fluorescent probes permit visualization of spots of proximity by fluorescence microscopy. Since the development of PLA in 2002, it has been increasingly used to ...
Source: Current Protocols in Immunology - September 23, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Bone Marrow Chimeras to Study Neuroinflammation.
Authors: Laflamme N, Préfontaine P, Lampron A, Rivest S
Abstract
Bone marrow transplantation is the standard of care for a host of diseases such as leukemia and multiple myeloma, as well as genetically inherited metabolic diseases affecting the central nervous system. In mouse models, bone marrow transplantation has proven a valuable tool for understanding the hematopoietic system and the homing of hematopoietic cells to their target organs. Many techniques have been developed to create chimeric mice, animals with a hematopoietic system derived from a genetic background that differs from the rest of the b...
Source: Current Protocols in Immunology - September 19, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Toll Receptors.
Authors: Takeda K, Akira S
Abstract
This has led to the identification of several important roles for Toll receptors in mammals. This unit discusses mammalian Toll receptors (TLR1-10) that have an essential role in the innate immune recognition of microorganisms. Also discussed are TLR-mediated signaling pathways and antibodies that are available to detect specific TLRs.
PMID: 30153720 [PubMed] (Source: Current Protocols in Immunology)
Source: Current Protocols in Immunology - August 31, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Cytomegalovirus Infection: Mouse Model.
Authors: Brizić I, Lisnić B, Brune W, Hengel H, Jonjić S
Abstract
This unit describes procedures for infecting newborn and adult mice with murine cytomegalovirus (MCMV). Methods are included for propagating MCMV in cell cultures and for preparing a more virulent form of MCMV from salivary glands of infected mice. A plaque assay is provided for determining MCMV titers of infected tissues or virus stocks. Also, a method is described for preparing the murine embryonic fibroblasts used for propagating MCMV and for the plaque assay. © 2018 by John Wiley & Sons, Inc.
PMID: 30044539 [PubMed - as s...
Source: Current Protocols in Immunology - July 28, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
Leukocyte Isolation from Brain, Spinal Cord, and Meninges for Flow Cytometric Analysis.
Authors: Manglani M, Gossa S, McGavern DB
Abstract
A continual dialogue exists between the central nervous system (CNS) and immune system that contributes to neural homeostasis as well as protection from microbes, repair following damage, autoimmune disease, and neurodegeneration. Characterization of resident and peripherally derived leukocyte populations within the central nervous system can provide valuable information regarding how these cells contribute to steady-state and inflammatory conditions. Flow cytometry provides a method to conduct detailed multi-parameter analyses of immune cells isolated fro...
Source: Current Protocols in Immunology - July 25, 2018 Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research
