ELISpot Homology and Cross-Reactivity Results

Due to the high degree of similarity observed between human and non-human primate cytokine protein sequences (See Table 1), R&D Systems has evaluated the potential use of our human ELISpot kits to detect cytokines from non-human primate cells. Cells from three different species of monkey were tested with each of our human ELISpot kits and found to cross-react with select analytes (See Table 2). Not all species represented in the table have been tested at this time.

Human Protein
Accession Number
Table 1. Amino acid sequences (note: protein accession number) for several human cytokines were BLASTed for homology to non-human primate protein sequences.
Species IL-1 beta
Rhesus monkey
(Macaca mulatta)
91% 94% 93% 96% NA 95%
Pigtail monkey
(Macaca nemestrina)
91% 94% NA NA NA NA
Cynomolgus monkey
(Macaca fasicularis)
90% 93% 92% 96% NA 94%
Common squirrel monkey
(Saimiri sciureus)
89% 89% NA 87% NA 86%
(Cercocebus torquatus)
91% 94% 92% 95% NA NA
(Papio hamadryas)
NA 90% NA NA NA 95%



ELISpot Kita
Table 2. Peripheral blood mononuclear cells from each of the listed species were stimulated with 50 ng/mL of phorbol 12-myristate-13-acetate and 0.5 µg/mL of calcium ionomycin (IL-2, IL-13) or 3 µg/mL phytohemagglutinin (PHA) (IL-4, IL-6) overnight at 37° C in a 5% CO2 incubator. The data shown is representative data from four independent experiments. Due to the inherent variability observed in immune responses within a group of subjects, assay results may vary.
Species Human IL-2
(5x105 cells/mL)
Human IL-4
(1x106 cells/mL)
Human IL-6
(1x105 cells/mL)
Human IL-13
(5x106 cells/mL)
  Spots/well Spots/well Spots/well Spots/well
Rhesus monkey
(Macaca mulatta)
400 NCRb 195 31
417 NCR 165 15
412 NCR 209 29
425 NCR 176 15
Pigtail monkey
(Macaca nemestrina)
412 36 52 NCR
402 29 36 NCR
430 28 30 NCR
462 42 27 NCR
Cynomolgus monkey
(Macaca fasicularis)
Confluentc 36 42 73
Confluent 28 37 61
Confluent 40 30 62
Confluent 25 46 63
  1. R&D Systems Human IFN-gamma, TNF-alpha, and IL-1 beta ELISpot kits were also tested and did not cross-react with any of the monkey species listed above.
  2. NCR - No cross-reactivity.
  3. Confluent spots - unable to analyze.