La Jolla Institute for Immunology

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells

Yanfang Peipei Zhu¹, Lindsey Padgett¹, Huy Q. Dinh¹, Paola Marcovecchio¹, Runpei Wu¹, Denise Hinz², Cheryl Kim², Catherine C. Hedrick¹

¹Division of Inflammation Biology, La Jolla Institute for Immunology, ²Flow Cytometry Core Facility, La Jolla Institute for Immunology

Here, we present a protocol to process fresh bone marrow (BM) isolated from mouse or human for high-dimensional mass cytometry (Cytometry by Time-Of-Flight, CyTOF) analysis of neutrophil-lineage cells.

Biology

A Semiautomated ChIP-Seq Procedure for Large-scale Epigenetic Studies

Justin Cayford¹, Sara Herrera-de la Mata¹, Benjamin Joachim Schmiedel¹, Vivek Chandra¹, Pandurangan Vijayanad¹, Grégory Seumois¹

¹Division of Vaccine Discovery, La Jolla Institute for Immunology

This paper describes a semiautomated, microscaled ChIP-Seq protocol of DNA regions associated with a specific histone modification (H3K27ac) using a ChIP liquid-handler platform, followed by library preparation using tagmentation. The procedure includes control assessment for quality and quantity purposes and can be adopted to other histone modifications or transcription factors.

JoVE Core

Experimental Melanoma Immunotherapy Model Using Tumor Vaccination with a Hematopoietic Cytokine

Hsin Yu Liu¹, Amnon Altman¹, Ann J. Canonigo-Balancio¹, Michael Croft¹

¹La Jolla Institute for Immunology

The protocol presents a cancer immunotherapy model using cell-based tumor vaccination with Flt3L-expressing B16-F10 melanoma. This protocol demonstrates the procedures, including preparation of cultured tumor cells, tumor implantation, cell irradiation, measurement of tumor growth, isolation of intratumoral immune cells, and flow cytometry analysis.