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ELISPOT 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.
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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
NP_000567 |
IL-2
NP_000577 |
IL-4
NP_000580 |
IL-6
NP_000591 |
IL-13
NP_002179 |
TNF-alpha
NP_000585 |
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% |
Mangabey
(Cercocebus torquatus) |
91% |
94% |
92% |
95% |
NA |
NA |
Baboon
(Papio hamadryas) |
NA |
90% |
NA |
NA |
NA |
95% |
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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 |
Notes:
- 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.
- NCR - No cross-reactivity.
- Confluent spots - unable to analyze.
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