Simultaneous detection of multiple cytokines and chemokines from nonhuman primates using luminex technology

Luis D. Giavedoni

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

Cytokines and chemokines are soluble mediators of the immune system that play a crucial role in intercellular signaling, and in the recruitment of cells to inflammation sites. Identification of these molecules in nonhuman primates (NHP) is crucial for the understanding of complex physiological and pathological mechanisms that occur in these species, and to demonstrate whether these mechanisms function similarly in humans. The Luminex100 system is a bench-top flow cytometer that allows the user to perform up to 100 tests simultaneously in a single tube. Recently, a significant number of commercial vendors have developed kits for the simultaneous detection of multiple cytokines and chemokines of human origin with the Luminex system. These kits were tested for their capacity to recognize chemokines and cytokines of nonhuman primate origin. ELISA and ELISPOT assays were also adapted to the Luminex format, and novel assays based on new combinations of antibodies were developed. PBMC were isolated from blood from chimpanzees, rhesus macaques, baboons, cynomolgus macaques, pig-tailed macaques, and African green monkeys; these cells were stimulated in vitro and culture supernatants were used for the determination of cytokines and chemokines. Crossreactivity tables were prepared based on the ability of the reagents to detect cytokines and chemokines in NHP samples with similar intensity to the ones observed in human samples. By mixing commercially available reagents and newly developed ones, a combination has been created that allows for the detection of 20 NHP chemokines and cytokines in a single sample, including G-CSF, GM-CSF, IFN-γ, IL-1β, IL-1Ra, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p40), IL-17, IL-18, MCP-1, MIP-1α, MIP-1β, RANTES, TNF-α, and TNF-β. These reagents may become a very useful resource for scientists working with these NHP species, which are relevant pre-clinical models for human diseases and transplantation because they approximate humans in physiology and genetics more closely than any other animal.

Original languageEnglish (US)
Pages (from-to)89-101
Number of pages13
JournalJournal of Immunological Methods
Volume301
Issue number1-2
DOIs
StatePublished - Jun 2005
Externally publishedYes

Fingerprint

Chemokines
Primates
Cytokines
Technology
Macaca
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin 1 Receptor Antagonist Protein
Enzyme-Linked Immunospot Assay
Cercopithecus aethiops
Chemokine CCL5
Interleukin-18
Pan troglodytes
Interleukin-17
Papio
Interleukin-5
Granulocyte Colony-Stimulating Factor
Interleukin-12
Macaca mulatta
Interleukin-8
Interleukin-1

Keywords

  • Chemokine
  • Cytokine
  • Luminex
  • Multiplexing
  • Nonhuman primate

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Simultaneous detection of multiple cytokines and chemokines from nonhuman primates using luminex technology. / Giavedoni, Luis D.

In: Journal of Immunological Methods, Vol. 301, No. 1-2, 06.2005, p. 89-101.

Research output: Contribution to journalArticle

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