Quantitation of CXCL9/Mig mRNA expression in spleen tissues from rhesus macaques by image capture and analysis and real-time RT-PCR.

(A) Quantitative image capture and analysis was used to determine the signal intensities after ISH for CXCL9/Mig mRNA in spleen tissue sections from rhesus macaques that were either uninfected or had the indicated stage of SIV disease. Each data point represents the surface area of reflected light determined for an individual 20 × microscopic field from the same tissue section after subtraction of the surface area of reflected light determined for a tissue section hybridized in parallel with the corresponding sense control probe. The values in each vertical collection of data points are from an individual animal. The differences in CXCL9/Mig expression were significant for both the acute infection (P = .005) and AIDS (P < .001) spleen measurements compared with uninfected spleen measurements. (B) Real-time RT-PCR was used to determine the relative levels of expression of CXCL9/Mig and IFN-γ mRNA in snap-frozen spleen tissue specimens, normalized against an endogenous control, β-GUS. The data were further normalized by using values from an uninfected macaque (M6600) for calibration.