Combination immunotherapy with extract of heated 4T1 and naloxone in mouse model of breast cancer

Combination immunotherapy with extract of heated 4T1 and naloxone in mouse model of breast cancer

Background/aim: This study was designed to investigate the efficacy of a new vaccine against breast cancer, which was made by mixing the extract of heated 4T1 cells and naloxone, as an adjuvant. Materials and methods: Female BALB/c mice of 6 8 weeks old were challenged subcutaneously in the right flanks with 4T1 cells. When all animals developed a palpable tumor, immunotherapy was initiated. Mice in the experimental groups received, twice with a 1-week interval, either the extract of heated 4T1 alone or in combination with naloxone, and mice in the negative control group received phosphate-buffered saline. One week after the last immunotherapy, half of the mice were euthanized in order to determine the immune response profile. The remaining animals were kept until the time when death occurred spontaneously. Results: The combined-treated mice with mammary tumors showed a more favorable survival curve and slower rate of tumor development compared to the mice with tumors that received only heated 4T1 and/or negative control mice. Moreover, the combined immunization significantly amplified the respiratory burst potential and the secretion of IFN-γ, and, conversely, diminished the secretion of IL-4, IL-10, and TGF-β in the splenocyte population compared to splenocytes from other groups. Conclusion: The combined naloxone and heated 4T1 cells promote beneficial outcomes in the mouse model of breast cancer.

___

  • 1. Raval RR, Sharabi AB, Walker AJ, Drake CG, Sharma P. Tumor immunology and cancer immunotherapy: summary of the 2013 SITC primer. J Immunother Cancer 2014; 2: 1–11.
  • 2. Corthay A. Does the immune system naturally protect against cancer? Front Immunol 2014; 5: 1–8.
  • 3. Maueroder C, Munoz LE, Chaurio RA, Herrmann M, Schett G, Berens C. Tumor immunotherapy: lessons from autoimmunity. Front Immunol 2014; 5: 1–5.
  • 4. Wood LM, Paterson Y. Attenuated Listeria monocytogenes: a powerful and versatile vector for the future of tumor immunotherapy. Front Cell Infect Microbiol 2014; 4: 1–22.
  • 5. Reed SG, Bertholet S, Coler RN, Friede M. New horizons in adjuvants for vaccine development. Trends Immunol 2009; 30: 23–32.
  • 6. Lombard M, Pastoret PP, Moulin AM. A brief history of vaccines and vaccination. Rev Sci Tech 2007; 26: 29–48.
  • 7. De Gregorio E, Tritto E, Rappuoli R. Alum adjuvanticity: unraveling a century old mystery. Eur J Immunol 2008; 38: 2068–2071.
  • 8. Claassen E, de Leeuw W, de Greeve P, Hendriksen C, Boersma W. Freund’s complete adjuvant: an effective but disagreeable formula. Res Immunol 1992; 143: 478–483.
  • 9. Jamali A, Mahdavi M, Hassan ZM, Sabahi F, Farsani MJ, Bamdad T, Soleimanjahi H, Motazakker M, Shahabi S. A novel adjuvant, the general opioid antagonist naloxone, elicits a robust cellular immune response for a DNA vaccine. Int Immunol 2009; 21: 217–225.
  • 10. Jazani NH, Parsania S, Sohrabpour M, Mazloomi E, Karimzad M, Shahabi S. Naloxone and alum synergistically augment adjuvant activities of each other in a mouse vaccine model of Salmonella typhimurium infection. Immunobiology 2011; 216: 744–451.
  • 11. Jazani NH, Sohrabpour M, Mazloomi E, Shahabi S. A novel adjuvant, a mixture of alum and the general opioid antagonist naloxone, elicits both humoral and cellular immune responses for heat-killed Salmonella typhimurium vaccine. FEMS Immunol Med Microbiol 2011; 61: 54–62.
  • 12. Motaharinia Y, Rezaee MA, Rashidi A, Jalili A, Rezaie MJ, Shapouri R, Hossieni W, Rahmani MR. Induction of protective immunity against brucellosis in mice by vaccination with a combination of naloxone, alum, and heat-killed Brucella melitensis 16 M. J Microbiol Immunol Infect 2013; 46: 253– 258.
  • 13. Tappeh KH, Khorshidvand Z, Shahabi S, Mohammadzadeh H. A novel adjuvant, mixture of alum and naltrexone, elicits humoral immune responses for excreted/secreted antigens of Toxoplasma gondii tachyzoites vaccine in Balb/c murine model. Turkiye Parazitol Derg 2013; 37: 92–96.
  • 14. Rohan TE, Xue X, Lin HM, D’Alfonso TM, Ginter PS, Oktay MH,  Robinson BD,  Ginsberg M,  Gertler FB,  Glass AG et al. Tumor microenvironment of metastasis and risk of distant metastasis of breast cancer. J Natl Cancer Inst 2014; 3: 106–110.
  • 15. Tao K, Fang M, Alroy J, Sahagian GG. Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer 2008; 8: 1–19.
  • 16. Heppner GH, Miller FR, Shekhar PM. Nontransgenic models of breast cancer. Breast Cancer Res 2000; 2: 331–334.
  • 17. Guo O, Li X, Yang Y, Wei J, Zhao Q, Luo F, Qian Z. Enhanced 4T1 breast carcinoma anticancer activity by co-delivery of doxorubicin and curcumin with core-shell drug-carrier based on heparin modified poly (L-lactide) grafted polyethylenimine cationic nanoparticles. J Biomed Nanotechnol 2014; 10: 227– 237.
  • 18. Abtahi Froushani SM, Delirezh N, Hobbenaghi R, Mosayebi G. Synergistic effects of atorvastatin and all-trans retinoic acid in ameliorating animal model of multiple sclerosis. Immunol Invest 2014; 43: 54–68.
  • 19. Esmaili Gouvarchin Galeh H, Delirezh N, Abtahi Froushani SM, Afzale Ahangaran N. Calcitriol modulates the effects of the supernatants of bone-marrow–derived mesenchymal stem cells on neutrophil functions. Turk J Biol 2014; 38: 365–370.
  • 20. Shilling DA, Smith MJ, Tyther R, Sheehan D, England K, Kavanagh EG, Redmond HP, Shanahan F, O’Mahony L. Salmonella typhimurium stimulation combined with tumourderived heat shock proteins induces potent dendritic cell antitumour responses in a murine model. Clin Exp Immunol 2007; 149: 109–116.
  • 21. Morecki S, Yacovlev E, Gelfand Y, Trembovler V, Shohami E, Slavin S. Induction of antitumor immunity by indomethacin. Cancer Immunol Immunother 2000; 48: 613–620.
  • 22. Mahmood K, Jadoon S. Synergistic effects of toxic elements on heat shock proteins. BioMed Research International 2014; 2014: 564136.
  • 23. Mahmood Q, Irshad M, Hussain J, Ahmed K, Zaidi SF. Treating cancer with heat: hyperthermia as promising strategy to enhance apoptosis. Biomed Res Int 2013; 63: 504–508.
  • 24. Rohrer KM, Haug M, Schworer D, Kalbacher H, Holzer U. Mutations in the substrate binding site of human heat-shock protein 70 indicate specific interaction with HLA-DR outside the peptide binding groove. Immunology 2014; 142: 237–247.
  • 25. Wang L, Yu Y. Dendritic cells primed with protein-protein fusion adjuvant. Methods Mol Biol 2014; 1139: 57–75.
  • 26. Zhou YJ, Messmer MN, Binder RJ. Establishment of tumorassociated immunity requires interaction of heat shock proteins with CD91. Cancer Immunol Res 2014; 2: 217–228.
  • 27. Bausero MA, Page DT, Osinaga E, Asea A. Surface expression of Hsp25 and Hsp72 differentially regulates tumor growth and metastasis. Tumour Biol 2004; 25: 243–251.
  • 28. Procaccini C, Pucino V, De Rosa V, Marone G, Matarese G. Neuro-endocrine networks controlling immune system in health and disease. Front Immunol 2014; 5: 1–10.
  • 29. Singhal G, Jaehne EJ, Corrigan F, Baune BT. Cellular and molecular mechanisms of immunomodulation in the brain through environmental enrichment. Front Cell Neurosci 2014; 8: 1–29.
  • 30. Sacerdote P, Manfredi B, Gaspani L, Panerai AE. The opioid antagonist naloxone induces a shift from type 2 to type 1 cytokine pattern in BALB/cJ mice. Blood 2000; 95: 2031–2036.
  • 31. Al-Hashimi M, Scott SW, Thompson JP, Lambert DG. Opioids and immune modulation: more questions than answers. Br J Anaesth 2013; 111: 80–88.
  • 32. Feng Y, He X, Yang Y, Chao D, Lazarus LH, Xia Y. Current research on opioid receptor function. Curr Drug Targets 2012; 13: 230–246.
  • 33. Stein C, Machelska H. Modulation of peripheral sensory neurons by the immune system: implications for pain therapy. Pharmacol Rev 2011; 63: 860–881.
  • 34. Mathers AR, Tckacheva OA, Janelsins BM, Shufesky WJ, Morelli AE, Larregina AT. In vivo signaling through the neurokinin 1 receptor favors transgene expression by Langerhans cells and promotes the generation of Th1- and Tc1-biased immune responses. J Immunol 2007; 178: 7006–7017.
  • 35. Bileviciute-Ljungar I, Saxne T, Spetea M. Anti-inflammatory effects of contralateral administration of the kappa-opioid agonist U-50,488H in rats with unilaterally induced adjuvant arthritis. Rheumatology (Oxford) 2006; 45: 295–302.
  • 36. Jazani NH, Karimzad M, Mazloomi E, Sohrabpour M, Hassan ZM, Ghasemnejad H, Roshan-Milani S, Shahabi S. Evaluation of the adjuvant activity of naloxone, an opioid receptor antagonist, in combination with heat-killed Listeria monocytogenes vaccine. Microbes Infect 2010; 12: 382–388.
  • 37. Molla Hassan AT, Hassan ZM, Moazzeni SM, Mostafaie A, Shahabi S, Ebtekar M, Hashemi SM. Naloxone can improve the anti-tumor immunity by reducing the CD4+CD25+Foxp3+ regulatory T cells in BALB/c mice. Int Immunopharmacol 2009; 9: 1381–1386.
  • 38. Panerai AE, Manfredi B, Granucci F, Sacerdote P. The betaendorphin inhibition of mitogen-induced splenocytes proliferation is mediated by central and peripheral paracrine/ autocrine effects of the opioid. J Neuroimmunol 1995; 58: 71– 76.
  • 39. Talar B, Czyz M. TGF-beta signaling pathways in cancers. Postepy Hig Med Dosw 2013; 67: 1008–1017.
  • 40. Nagai S, Toi M. Interleukin-4 and breast cancer. Breast Cancer 2000; 7: 181–186.
  • 41. Mantovani A, Biswas SK, Galdiero MR, Sica A, Locati M. Macrophage plasticity and polarization in tissue repair and remodelling. J Pathol 2013; 229: 176–185.
  • 42. Cho HJ, Jung JI, Lim Do Y, Kwon GT, Her S, Park JH, Park JH. Bone marrow-derived, alternatively activated macrophages enhance solid tumor growth and lung metastasis of mammary carcinoma cells in a Balb/C mouse orthotopic model. Breast Cancer Res 2012; 14: 1–12.