Dissociation of skeletal muscle for flow cytometric characterization of immune cells in macaques

Autor: Aurélie Ploquin, Karin Loré, Gustaf Lindgren, José DelaO Hernández, Daphne A. Stanley, Hugues Fausther-Bovendo, Frank Liang, Richard A. Koup, Aiala Salvador Martinez, Nancy J. Sullivan, Jason M. Brenchley
Rok vydání: 2015
Předmět:
Zdroj: Journal of Immunological Methods. 425:69-78
ISSN: 0022-1759
DOI: 10.1016/j.jim.2015.06.011
Popis: The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. While methods for obtaining and characterizing murine muscle cell suspensions have been reported, protocols for nonhuman primates (NHPs) have not been well defined. NHPs comprise important in vivo models for studies of immune cell function due to their high degree of resemblance with humans. In this study, we developed and systematically compared methods to collect vaccine-injected muscle tissue to be processed into single cell suspensions for flow cytometric characterization of immune cells. We found that muscle tissue processed by mechanical disruption alone resulted in significantly lower immune cell yields compared to enzymatic digestion using Liberase. Dendritic cell subsets, monocytes, macrophages, neutrophils, B cells, T cells and NK cells were readily detected in the muscle by the classical human markers. The methods for obtaining skeletal muscle cell suspension established here offer opportunities to increase the understanding of immune responses in the muscle, and provides a basis for defining immediate post-injection vaccine responses in primates.
Databáze: OpenAIRE
Externí odkaz: