Active human grB results in tumor regression without the induction of adverse effects. Article Swipe
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· 2015
· Open Access
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· DOI: https://doi.org/10.1371/journal.pone.0041341.g006
· OA: W2595194801
<p>(<b>A</b>) CT26 tumor-bearing BALB/c mice were injected (i.p.) either with PBS (ctrl), inactive grB (inact grB) or enzymatically active grB (grB; 20 µg/g mouse body weight in 200 µl PBS) on days 6, 7, 13, 14 after i.p. injection of CT26 tumor spheroids. Mice were sacrificed on day 21. The weights of tumors in mice treated with grB were significantly lower than those in the control group (n = 15 per group; <i>p</i>*<0.05). The weight of tumors in mice treated with active or inactive grB were also significantly different (<i>p</i>*<0.05), whereas there was no significant difference in the weights of tumors in control mice or mice that had been treated with inactive grB (<i>p</i> = 0.49). (<b>B</b>) Comparison of CT26 tumors derived from control mice (ctrl), active and inactive grB-treated mice (see above). Representative sections of formalin-fixed paraffin-embedded tumors stained with H&E. R1 marks necrotic/apoptotic areas in the centre of the tumor of ctrl animals; R2 marks necrotic/apoptotic areas in the border area of the tumor of active grB-treated mice; vessels are marked with a circle. (<b>C</b>) Comparison of sections from formalin-fixed paraffin-embedded liver, kidney, heart, lung and spleen derived from control mice and mice treated with grB (see above) stained with H&E were analyzed using light microscopy (20x objective, indicated scale bar 100 µm). (<b>D</b>) Intraoperative detection of Cy5.5-labeled cmHsp70.1 mAb in BALB/c mice bearing CT26 tumors. Antibody (100 µg) was injected (i.v.) into the tail vein on day 6 after tumor injection. Representative false color images and views of the Cy5.5 fluorescence of the neck part of three mice were taken 24 hours after i.v. injection. The circled area indicates the histologically proven tumor tissue; arrows indicate superficial blood vessels in the normal tissue (upper arrow) and the tumor (lower arrow). (<b>E</b>) Relative binding of cmHsp70.1 mAb to isolated CT26 tumor cells (tumor) and endothelial (normal) cells. The capacity of cells to bind cmHsp70.1-FITC is presented as mean equivalents of soluble fluorochrome (MESF) divided by the antibody binding capacity (ABC) and was determined using the Quantum Simply Cellular anti-Mouse IgG according to the manufacturer’s protocol (Bangs Laboratories, Inc., Fishers, IN, USA). The Y-axis is a logarithmic scale.</p>
