Figure 1

Protein expression in NHP lung and spleen tissue was quantified using in situ computerized image analysis of immunohistochemistry data. Immunohistochemistry images (left panel) illustrate positive immunostaining for CD8α⁺ T cells or CD3⁺ T cells (brown) in Mtb-infected tissue obtained from an rBCG/rAd35-vaccinated animal. The threshold values for the positive staining (green contour line) and the total cell area (red/orange contour line), illustrated in the overlay image (right panel), was set using a digital image analysis system including a highly sensitive Qwin 550 software. The data are presented in field statistics as the total area measured, intensity, positive area and cellular area. A mean value of the percent positive area of the total cell area in the tissue was quantified in 10–40 high-power fields and plotted in separate graphs. Magnification 125×.