Evaluating the Anti-Tumor Efficacy of a Bacterial Immunomodulatory Protein in a Murine Model

Abstract

Source: Codolo, G. et al. Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model. J. Vis. Exp. (2015)

This video demonstrates a technique to assess the anti-tumor activity of Helicobacter pylori neutrophil-activating protein, or HP-NAP, in a mouse bladder cancer model. HP-NAP interacts with tumor-cell neutrophils to produce pro-inflammatory cytokines, inducing the differentiation of naïve T cells into type 1 T helper (Th1) and type 1 cytotoxic (Tc1) T cell phenotypes. The Th1 and Tc1 cells secrete interferon-gamma, inhibiting tumor angiogenesis and promoting cytotoxic T cell-mediated tumor destruction, reducing tumor volume.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Animals

1. Grow C57BL6/J female mice to 8 weeks of age in individually ventilated cages with microisolation filters.
   NOTE: Females are preferred because the anatomic conformation of their external urogenital apparatus renders the catheterization easier than in males.

2. Cell Culture

1. Maintain the mouse urothelial carcinoma cell line MB49 in RPMI 1640 medium supplemented with 10% heat-....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
C57BL/6J female mice	Harlan Italy (Udine, Italy)
MB49 Cells			Obtained from Prof. O'Donnel, University of Iowa Carver College of Medicine, Iowa, USA
RPMI	Sigma-Aldrich (St. Louis, Missouri, USA)	R8758
FBS	Sigma-Aldrich	F7524
PBS	Sigma-Aldrich	D1408	10X, to be diluted in apyrogenic water
Flask	Becton Dickinson (Franklin Lakes, New Jersey, USA)	353135
Syringe 1ml	Becton Dickinson	301358
Trypsin	Life Technologies (Waltham, Massachusetts, USA)
Gentamycin	Life Technologies	15710-049
Xilor	Bio 98 s.r.l. (Milano, Italy)		2% Xylazin
Zoletil	Virbac (Carros, France)	3.59713E+11	5% Zolazepam + 5% Tiletamine
24G Catheter	Terumo (Rome, Italy)	SR+DM2419PX
HCl	Carlo Erba Reagents (Milano, Italy)	403871	Liquid
NaOH	JT Baker (Center Valley, Pennsylvania, USA)	10095011	Powder