Fig. 6
CA2-2 increases the amount of activated M1 macrophages inside tumor tissue after macrophage injection in the near tumor region. (A) General scheme of macrophage administration. (B) Immunohistochemical detection of BALB/c mouse peritoneal macrophages in cryosections of mouse solid Ehrlich carcinoma tissue 24 h after subcutaneous injection of macrophages at a concentration of 1 × 105 cells/mouse. Macrophages were stained with antibodies to the F4/80 marker conjugated with FITC (green fluorescence), to CD86 marker conjugated with FITC (green fluorescence), to Iba-1 marker conjugated with FITC (green fluorescence), to IL-1β cytokine conjugated with TRITC (red fluorescence); NIR-NPs have a red fluorescence; the nuclei were stained with Hoechst 33,342 (blue fluorescence). The figure shows sections of a tumor from 3 groups of mice: the control group, the group receiving non-activated M0 peritoneal macrophages and the group receiving CA2-2 pre-treated (0.02 µM, 2 h) M1 peritoneal macrophages. Quantitative analysis of the content of peritoneal macrophages in tumor tissue. (C) Amount of F4/80 + cells. (D) Amount of CD86 + cells. (E) Amount of Iba-1 + cells; (F) amount of IL-1β producing cells; and (G) number of macrophages loaded with NIR-NPs in cryosections of Ehrlich carcinoma tissue 1 and 24 h after subcutaneous injection of macrophages. Groups of mice: control group; a group that received non-treated M0 peritoneal macrophages (1 and 24 h after the injection); the group that received CA2-2 pre-treated (0.02 µM, 2 h) M1 peritoneal macrophages (1 and 24 h after injection). Fluorescent images and images in transmitted light were obtained using a LSM510 META confocal microscope (Carl Zeiss, Germany). Image processing was carried out using the software ZEN 2012 lite (Carl Zeiss, Germany). Data are presented as mean ± SE (n = 8). *p < 0.05 compared to control