Figure 1.
Examples of immunostains used to identify M1 and M2 macrophages.
Expression of CD68 or CD168 is indicated by blue cytoplasmic/membraneous staining. The expression of transcription factors pSTA1, RBP-J and CMAF is indicated by brown nuclear staining. (A) Infectious mononucleosis, a Th1 disease, reveals high numbers of CD68+pSTAT1+ macrophages as well as (B) of CD68+RBP-J+ macrophages. By contrast (C) infectious mononucleosis shows, in this field, absence of CD68+CMAF+ macrophages. Similarly, in Crohn’s disease, another Th1-associated condition (D) large numbers of CD163+pSTAT1+ macrophages as well as (E) of CD163+RBP-J+ macrophages are seen. In contrast (F) Crohn´s disease reveals, in this image, absence of CD163+CMAF+ macrophages. (G) An allergic nasal polyp, a Th2-associated disorder, shows, in this field absence of CD163+pSTAT1+ macrophages and (H) rare CD163+RBP-J+ macrophages, while (I) high numbers of CD163+CMAF+ macrophages are present. Similarly, in an allergic nasal polyp (J) in this field there is absence of CD68+pSTAT1+ macrophages and (K) only few CD68+RBP-J+ macrophages are seen. By contrast, (L) high numbers of CD68+CMAF+ macrophages are observed. (original magnification: 400x).
Figure 2.
Dendrogram using average linkage obtained by hierarchical cluster analysis.
Two main identified clusters (I and II) are identified by brackets. Each cell population is stratified by 50th percentile, indicating high number of this cell in the tissue microenvironment of Th1 and Th2 model diseases. Num: order of variable input.
Figure 3.
Box-plot graphs showing the numerical distribution of M1 (CD163+pSTAT1+ cells [A], CD163+RBP-J+ cells [B], CD68+pSTAT1+ cells [D] and CD68+RBP-J+ cells [E]) and M2 macrophages (CD163+CMAF+ cells [C] and CD68+CMAF+ cells [F]) according to Th model diaseases.
The P-values are from Mann-Whitney tests.
Figure 4.
Scatter plots showing the correlation between the numbers of CD163+ and CD68+ macrophages (A), CD163+pSTAT1+ and CD68+pSTAT1+ macrophages (B), CD163+RBP-J+ and CD68+RBP-J macrophages (C), and CD163+CMAF+ and CD68+CMAF+ macrophages (D).
R2 is the constant correlation.