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ELISpot and ELISA analysis of spontaneous, mitogen-induced and antigen-specific cytokine production in cynomolgus and rhesus macaques.
- Source :
-
Journal of immunological methods [J Immunol Methods] 2002 Dec 01; Vol. 270 (1), pp. 85-97. - Publication Year :
- 2002
-
Abstract
- Evaluation of cytokine production in macaques has been hampered by a lack of availability of optimized and standardized immunoassays such as ELISA and enzyme-linked immune spot assay (ELISpot); only a limited number of macaque cytokines have been assessed by ELISpot. Using monoclonal antibodies (mAb) to human cytokines that cross-react with cynomolgus and rhesus macaque interferon-gamma (IFN-gamma), interleukin (IL)-2, IL-4, IL-5, IL-6, IL-12, IL-13 and granulocyte monocyte colony-stimulating factor, we measured macaque cytokine production by ELISA and ELISpot. Quantitation of spontaneous as well as phytohemagglutinin (PHA)-induced cytokine production in peripheral blood mononuclear cells (PBMC) from rhesus and cynomolgus macaques and humans were compared. The proportional distribution of the different cytokines, in terms of PBMC synthesizing different cytokines as well as the levels of the different cytokines produced, were similar in all species. Spontaneous- and PHA-induced cytokine productions thus appear to be similarly regulated in macaques and man. ELISpot and ELISA assays for macaque IFN-gamma were further used to measure antigen-specific immune responses of PBMC from cynomolgus macaques exposed to, or vaccinated against, simian immunodeficiency virus (SIV). The establishment of reliable immunoassays for detection of macaque cytokines is of importance for future progress of research utilizing macaques as experimental animals.
- Subjects :
- Animals
Antibodies, Monoclonal immunology
Antigens, Viral
Cell Division
Cells, Cultured
Cytokines immunology
Granulocyte-Macrophage Colony-Stimulating Factor biosynthesis
Granulocyte-Macrophage Colony-Stimulating Factor immunology
Humans
Interferon-gamma biosynthesis
Interleukin-12 biosynthesis
Interleukin-12 immunology
Interleukin-13 biosynthesis
Interleukin-13 immunology
Interleukin-2 biosynthesis
Interleukin-2 immunology
Interleukin-4 biosynthesis
Interleukin-4 immunology
Interleukin-5 biosynthesis
Interleukin-5 immunology
Interleukin-6 biosynthesis
Interleukin-6 immunology
Leukocytes, Mononuclear cytology
Leukocytes, Mononuclear drug effects
Leukocytes, Mononuclear immunology
Macaca fascicularis
Macaca mulatta
Mitogens pharmacology
Phytohemagglutinins pharmacology
Simian Immunodeficiency Virus immunology
T-Lymphocytes, Cytotoxic cytology
T-Lymphocytes, Cytotoxic immunology
Tumor Necrosis Factor-alpha biosynthesis
Tumor Necrosis Factor-alpha immunology
Cytokines biosynthesis
Enzyme-Linked Immunosorbent Assay methods
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1759
- Volume :
- 270
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of immunological methods
- Publication Type :
- Academic Journal
- Accession number :
- 12379341
- Full Text :
- https://doi.org/10.1016/s0022-1759(02)00274-0