TY - JOUR
T1 - Dissociation of skeletal muscle for flow cytometric characterization of immune cells in macaques
AU - Liang, Frank
AU - Ploquin, Aurélie
AU - Hernández, José Dela O.
AU - Fausther-Bovendo, Hugues
AU - Lindgren, Gustaf
AU - Stanley, Daphne
AU - Martinez, Aiala Salvador
AU - Brenchley, Jason M.
AU - Koup, Richard A.
AU - Loré, Karin
AU - Sullivan, Nancy J.
N1 - Publisher Copyright:
© 2015 Elsevier B.V.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - 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.
AB - 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.
KW - Flow cytometry
KW - Nonhuman primate
KW - Skeletal muscle
KW - Vaccine administration
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U2 - 10.1016/j.jim.2015.06.011
DO - 10.1016/j.jim.2015.06.011
M3 - Article
C2 - 26099800
AN - SCOPUS:84943661565
SN - 0022-1759
VL - 425
SP - 69
EP - 78
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
ER -