Figure 4.

Human leukocytes contain the Tag7–Mts1 complex. (A) Tag7 and Mts1 are contained in the medium conditioned by PBMCs. The PBMC cells were incubated for 18 hours in sterile conditions, than conditioned medium was collected and passed through an anti-Tag7 Sepharose, and the absorbed proteins were eluted and analyzed by immunoblotting with antibodies to Tag7 (on the left) and Mts1 (on the right). (B) Tag7, Mts1, and the Tag7–Mts1 complex are localized in the cytoplasmic granular inclusions in CD4⁺ and CD8⁺ lymphocytes and neutrophils. Confocal micrographs of (a–c) neutrophil, (d–f) CD8⁺ lymphocyte, (g–i) CD4⁺ lymphocyte, and (j–l) monocyte double-immunostained with (a, d, g, j) rabbit anti-Mst1 antibodies and FITC-conjugated anti-rabbit IgG (green fluorescence) and (b, e, h, k) mouse anti-Tag7 antibodies and phycoerythrin-conjugated anti-mouse IgG (red fluorescence), and (c, f, i, l) merged images of the same cells. Images were taken at ×100 magnification. (C) The effect of Tag7–Mts1 complex (TM) from different sources on the migration of CD56⁺ cells. Isolated CD56⁺ cells in 200 µL of RPMI were placed in the upper compartment of the Transwell system while the lower compartment contained immune-adsorbed Tag7–Mts1 from the medium conditioned by neutrophils, CD4⁺ or CD8⁺ lymphocytes, monocytes, or RPMI (control). The initial number of cells in the upper compartment was taken as 100%, and the relative number of cells that migrated to the lower compartment was estimated after 1 h using the MTT test.