Scientific Publications by FDA Staff

J Interferon Cytokine Res 2012 Sep;32(9):407-15

High-Throughput Quantitative Real-Time Polymerase Chain Reaction Array for Absolute and Relative Quantification of Rhesus Macaque Types I, II, and III Interferon and Their Subtypes.

Schramm LM, Kirschman KD, Heuer M, Chen AA, Verthelyi D, Puig M, Rabin RL

Rhesus macaques provide a valuable research and preclinical model for cancer and infectious diseases, as nonhuman primates share immune pathways with humans. Interferons (IFNs) are key cytokines in both innate and adaptive immunity, so a detailed analysis of gene expression in peripheral blood and tissues may shed insight into immune responses. Macaques have 18 IFN genes, of which 14 encode for 13 distinct IFN-alpha subtypes, and one for IFN-beta. Here, we developed a high-throughput array to evaluate each of the IFN-alpha subtypes, as well as IFN-beta, IFN-gamma and 2 subtypes of IFN-lambda. With this array, expression of each IFN species may be quantified as relative to a reference (housekeeping) gene (DeltaCq) or fitted to its own 4-point standard curve for absolute quantification (copy number per mass unit RNA). After validating the assay with IFN complementary DNA, we determined the IFN expression profile of peripheral blood mononuclear cells from 3 rhesus macaques in response to TLR agonists, and demonstrated that the profiles are consistent among animals. Furthermore, because the IFN expression profiles differ depending on the TLR stimuli, they suggest different biological functions for many of the IFN species measured, including individual subtypes of IFN-alpha.