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Systematic evaluation of monoclonal antibodies and immunoassays for the detection of Interferon-gamma and Interleukin-2 in old and new world non-human primates
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Number of Authors: 7
2017 (English)In: JIM - Journal of Immunological Methods, ISSN 0022-1759, E-ISSN 1872-7905, Vol. 441, 39-48 p.Article in journal (Refereed) Published
Abstract [en]

Non-human primates (NHP) provide important animal models for studies on immune responses to infections and vaccines. When assessing cellular immunity in NHP, cytokines are almost exclusively analyzed utilizing cross-reactive anti-human antibodies. The functionality of antibodies has to be empirically established for each assay/application as well as NHP species.

A rational approach was employed to identify monoclonal antibodies (mAb) cross-reactive with many NHP species. Panels of new and established mAbs against human Interferon (IFN)-gamma and Interleukin (IL)-2 were assessed for reactivity with eukaryotically expressed recombinant IFN-gamma and IL-2, respectively, from Old (rhesus, cynomolgus and pigtail macaques, African green monkey, sooty mangabey and baboon) and New World NHP (Ma's night monkey, squirrel monkey and common marmoset).

Pan-reactive mAbs, recognizing cytokines from all NHP species, were further analyzed in capture assays and flow cytometry with NHP peripheral blood mononuclear cells (PBMC). Pan-reactive mAb pairs for IFN-gamma well as IL-2 were identified and used in ELISA to measure IFN-gamma and IL-2, respectively, in Old and New World NHP PBMC supernatants. The same mAb pairs displayed high functionality in ELISpot and FluoroSpot for the measurement of antigen-specific IFN-gamma and IL-2 responses using cynomolgus PBMC Functionality of pan-reactive mAbs in flow cytometry was also verified with cynomolgus PBMC.

The development of well-defined immunoassays functional with a panel of NHP species facilitates improved analyses of cellular immunity and enables inclusion in multiplex cytokine assays intended for a variety of NHP.

Place, publisher, year, edition, pages
2017. Vol. 441, 39-48 p.
Keyword [en]
Antibody, ELISA, ELISpot, Interferon-gamma, Interleukin-2, Non-human primates
National Category
Biological Sciences Immunology in the medical area
Identifiers
URN: urn:nbn:se:su:diva-141377DOI: 10.1016/j.jim.2016.11.011ISI: 000393250400006PubMedID: 27889562OAI: oai:DiVA.org:su-141377DiVA: diva2:1091927
Available from: 2017-04-28 Created: 2017-04-28 Last updated: 2017-04-28Bibliographically approved

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Department of Molecular Biosciences, The Wenner-Gren Institute
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CiteExportLink to record
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Citation style
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