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

Frank Liang, Aurélie Ploquin, José Dela O. Hernández, Hugues Fausther-Bovendo, Gustaf Lindgren, Daphne Stanley, Aiala Salvador Martinez, Jason M. Brenchley, Richard A. Koup, Karin Loré, Nancy J. Sullivan

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

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 classic 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 provide a basis for defining immediate post-injection vaccine responses in primates.

Original languageEnglish (US)
Pages (from-to)69-78
Number of pages10
JournalJournal of Immunological Methods
Volume425
DOIs
StatePublished - Oct 1 2015
Externally publishedYes

Keywords

  • Flow cytometry
  • Nonhuman primate
  • Skeletal muscle
  • Vaccine administration

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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