title: Systematic evaluation of monoclonal antibodies and immunoassays for the detection of Interferon-γ and Interleukin-2 in old and new world non-human primates.
abstract: 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)-γ and Interleukin (IL)-2 were assessed for reactivity with eukaryotically expressed recombinant IFN-γ 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-γ well as IL-2 were identified and used in ELISA to measure IFN-γ 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-γ 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.
authors.affiliation: Mabtech, Box 1233, SE-131 28, Nacka Strand, Sweden.
authors.forename: Ankie
authors.initials: A
authors.lastname: Höglind
authors.affiliation: Mabtech, Box 1233, SE-131 28, Nacka Strand, Sweden.
authors.forename: Irene
authors.initials: I
authors.lastname: Areström
authors.affiliation: Mabtech, Box 1233, SE-131 28, Nacka Strand, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Ehrnfelt
authors.affiliation: Mabtech, Box 1233, SE-131 28, Nacka Strand, Sweden.
authors.forename: Khosro
authors.initials: K
authors.lastname: Masjedi
authors.affiliation: Swedish Orphan Biovitrum AB, SE-112 76 Stockholm, Sweden.
authors.forename: Bartek
authors.initials: B
authors.lastname: Zuber
authors.affiliation: Departments of Virology and Immunology and Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX 78245-0549, USA.
authors.forename: Luis
authors.initials: L
authors.lastname: Giavedoni
authors.affiliation: Mabtech, Box 1233, SE-131 28, Nacka Strand, Sweden; Department of Immunology, Stockholm University, SE-106 91 Stockholm, Sweden. Electronic address: niklas@mabtech.com.
authors.forename: Niklas
authors.initials: N
authors.lastname: Ahlborg
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D002522:Chlorocebus aethiops; D003429:Cross Reactions; D058501:Enzyme-Linked Immunospot Assay; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007371:Interferon-gamma / Q000032:analysis* / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology; D007376:Interleukin-2 / Q000032:analysis* / Q000276:immunology; D008252:Macaca fascicularis; D008253:Macaca mulatta
chemical_list: D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D016753:Interleukin-10; D007371:Interferon-gamma
keywords: Antibody; ELISA; ELISpot; Interferon-γ; Interleukin-2; Non-human primates