Detail of multiwell plate containing PANC-1 cells stained with crystal violet showing Newcastle disease virus Ulster strain (NDV-U) 0.16 PFU

Two Avirulent, Lentogenic Strains of Newcastle Disease Virus Are Cytotoxic for Some Human Pancreatic Tumor Lines In Vitro

Robert J Walter, Bashar M Attar, Asad Rafiq, Megan Delimata, Sooraj Tejaswi


Context Pancreatic cancer is the fourth leading cause of cancer death in the U.S. Highly infectious Newcastle disease virus (NDV) strains are known to be very cytotoxic for an array of human tumor cell types in vitro and in vivo but the effects of these and avirulent NDV strains on pancreatic neoplasms are little known. Objective Here, the direct cytolytic effects of the avirulent Hitchner-B1 (B1) and Ulster (U) NDV strains on 7 human pancreatic tumor cell lines and 4 normal human cell lines were studied. Methods Cytotoxicity assays used serially diluted NDV to determine minimum cytotoxic plaque forming unit (PFU) doses. Results For NDV-B1, normal human cells were killed only by relatively high doses (range: 471-3,724 PFU) whereas NDV-U killed these cells at low PFU (range: 0.32-1.60 PFU). Most pancreatic cancer cell types were killed by much lower NDV-B1 doses (range: 0.40-2.60 PFU) while NDV-U killed Capan-1 and SU.86.86 cultures at very low doses (0.00041 PFU and 0.0034 PFU, respectively). Conclusions On average, 1,555 times more NDV-B1 was needed to kill normal cells than most pancreatic tumor cells and 558 times more NDV-U to kill the two most sensitive pancreatic cancer lines. These innately-targeted lentogenic viruses may have meaningful potential in treating pancreatic cancer.

Image: Detail of multiwell plate containing PANC-1 cells stained with crystal violet showing Newcastle disease virus Ulster strain (NDV-U) 0.16 PFU.


Antineoplastic Agents; Newcastle disease virus /classification; Oncolytic Virotherapy; Pancreatic Neoplasms; Viral Vaccines /administration and dosage

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American Cancer Society. Cancer Facts and Figures 2010. Atlanta, Ga: American Cancer Society. 2010.

Greenlee RT, Murray T, Bolden S, Wingo PA. Cancer statistics, 2000. CA Cancer J Clin 2000; 50:7-33. [PMID 10735013]

Oettle H, Post S, Neuhaus P, Gellert K, Langrehr J, Ridwelski K, Schramm H, Fahlke J, Zuelke C, Burkart C, Gutberlet K, Kettner E, Schmalenberg H, Weigang-Koehler K, Bechstein WO, Niedergethmann M, Schmidt-Wolf I, Roll L, Doerken B, Riess H. Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: a randomized controlled trial. JAMA 2007; 297:267-277. [PMID 17227978]

Regine WF, Winter KA, Abrams RA, Safran H, Hoffman JP, Konski A, Benson AB, Macdonald JS, Kudrimoti MR, Fromm ML, Haddock MG, Schaefer P, Willett CG, Rich TA. Fluorouracil vs gemcitabine chemotherapy before and after fluorouracil-based chemoradiation following resection of pancreatic adenocarcinoma: a randomized controlled trial. JAMA 2008; 299:1019-1026. [PMID 18319412]

National Cancer Institute. Pancreatic Cancer Treatment.

Stathis A, Moore MJ. Advanced pancreatic carcinoma: current treatment and future challenges. Nat Rev Clin Oncol 2010; 7:163-172. [PMID 20101258]

Lorence RM, Katubig BB, Reichard KW, Reyes HM, Phuangsab A, Sassetti MD, Walter RJ, Peeples ME. Complete regression of human fibrosarcoma xenografts after local Newcastle disease virus therapy. Cancer Res 1994; 54:6017-6021. [PMID 7954437]

Lorence RM, Reichard KW, Katubig BB, Reyes HM, Phuangsab A, Mitchell BR, Cascino CJ, Walter RJ, Peeples ME. Complete regression of human neuroblastoma xenografts in athymic mice after local Newcastle disease virus therapy. J Natl Cancer Inst 1994; 86:1228-1233. [PMID 8040891]

Phuangsab A, Lorence RM, Reichard KW, Peeples ME, Walter RJ. Newcastle disease virus therapy of human tumor xenografts: antitumor effects of local or systemic administration. Cancer Lett 2001; 172:27-36. [PMID 11595126]

Reichard KW, Lorence RM, Katubig BB, Peeples ME, Reyes HM. Retinoic acid enhances killing of neuroblastoma cells by Newcastle disease virus. J Pediatr Surg 1993; 28:1221-1225. [PMID 8263678]

Reichard KW, Lorence RM, Cascino CJ, Peeples ME, Walter RJ, Fernando MB, Reyes HM, Greager JA. Newcastle disease virus selectively kills human tumor cells. J Surg Res 1992; 52:448-453. [PMID 1619912]

de Leeuw OS, Koch G, Hartog L, Ravenshorst N, Peeters BP. Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein. J Gen Virol 2005; 86:1759-1769. [PMID 15914855]

Steward M, Vipond IB, Millar NS, Emmerson PT. RNA editing in Newcastle disease virus. J Gen Virol 1993; 74 ( Pt 12):2539-2547. [PMID 8277263]

Stojdl DF, Lichty B, Knowles S, Marius R, Atkins H, Sonenberg N, Bell JC. Exploiting tumor-specific defects in the interferon pathway with a previously unknown oncolytic virus. Nat Med 2000; 6:821-825. [PMID 10888934]

Lorence RM, Roberts MS, O'Neil JD, Groene WS, Miller JA, Mueller SN, Bamat MK. Phase 1 clinical experience using intravenous administration of PV701, an oncolytic Newcastle disease virus. Curr Cancer Drug Targets 2007; 7:157-167. [PMID 17346107]

Toyoda T, Sakaguchi T, Hirota H, Gotoh B, Kuma K, Miyata T, Nagai Y. Newcastle disease virus evolution. II. Lack of gene recombination in generating virulent and avirulent strains. Virology 1989; 169:273-282. [PMID 2705298]

Sakaguchi T, Toyoda T, Gotoh B, Inocencio NM, Kuma K, Miyata T, Nagai Y. Newcastle disease virus evolution. I. Multiple lineages defined by sequence variability of the hemagglutinin-neuraminidase gene. Virology 1989; 169:260-272. [PMID 2705297]

Krishnamurthy S, Takimoto T, Scroggs RA, Portner A. Differentially regulated interferon response determines the outcome of Newcastle disease virus infection in normal and tumor cell lines. J Virol 2006; 80:5145-5155. [PMID 16698995]

Apostolidis L, Schirrmacher V, Fournier P. Host mediated anti-tumor effect of oncolytic Newcastle disease virus after locoregional application. Int J Oncol 2007; 31:1009-1019. [PMID 17912426]

Yaacov B, Eliahoo E, Lazar I, Ben-Shlomo M, Greenbaum I, Panet A, Zakay-Rones Z. Selective oncolytic effect of an attenuated Newcastle disease virus (NDV-HUJ) in lung tumors. Cancer Gene Ther 2008; 15:795-807. [PMID 18535620]

Schirrmacher V, Haas C, Bonifer R, Ahlert T, Gerhards R, Ertel C. Human tumor cell modification by virus infection: an efficient and safe way to produce cancer vaccine with pleiotropic immune stimulatory properties when using Newcastle disease virus. Gene Ther 1999; 6:63-73. [PMID 10341877]

Schirrmacher V, Griesbach A, Ahlert T. Antitumor effects of Newcastle Disease Virus in vivo: local versus systemic effects. Int J Oncol 2001; 18:945-952. [PMID 11295039]

Sergel T, McGinnes LW, Morrison TG. The fusion promotion activity of the NDV HN protein does not correlate with neuraminidase activity. Virology 1993; 196:831-834. [PMID 8372451]

Sergel T, McGinnes LW, Peeples ME, Morrison TG. The attachment function of the Newcastle disease virus hemagglutinin-neuraminidase protein can be separated from fusion promotion by mutation. Virology 1993; 193:717-726. [PMID 8384752]

Sergel TA, McGinnes LW, Morrison TG. A single amino acid change in the Newcastle disease virus fusion protein alters the requirement for HN protein in fusion. J Virol 2000; 74:5101-5107. [PMID 10799584]

HANSON RP, BRANDLY CA. Identification of vaccine strains of Newcastle disease virus. Science 1955; 122:156-157. [PMID 14396376]

Fabian Z, Csatary CM, Szeberenyi J, Csatary LK. p53-independent endoplasmic reticulum stress-mediated cytotoxicity of a Newcastle disease virus strain in tumor cell lines. J Virol 2007; 81:2817-2830. [PMID 17215292]

Zamarin D, Martinez-Sobrido L, Kelly K, Mansour M, Sheng G, Vigil A, Garcia-Sastre A, Palese P, Fong Y. Enhancement of oncolytic properties of recombinant newcastle disease virus through antagonism of cellular innate immune responses. Mol Ther 2009; 17:697-706. [PMID 19209145]

Jarahian M, Watzl C, Fournier P, Arnold A, Djandji D, Zahedi S, Cerwenka A, Paschen A, Schirrmacher V, Momburg F. Activation of natural killer cells by newcastle disease virus hemagglutinin-neuraminidase. J Virol 2009; 83:8108-8121. [PMID 19515783]

Pecora AL, Rizvi N, Cohen GI, Meropol NJ, Sterman D, Marshall JL, Goldberg S, Gross P, O'Neil JD, Groene WS, Roberts MS, Rabin H, Bamat MK, Lorence RM. Phase I trial of intravenous administration of PV701, an oncolytic virus, in patients with advanced solid cancers. J Clin Oncol 2002; 20:2251-2266. [PMID 11980996]

Liang W, Wang H, Sun TM, Yao WQ, Chen LL, Jin Y, Li CL, Meng FJ. Application of autologous tumor cell vaccine and NDV vaccine in treatment of tumors of digestive tract. World J Gastroenterol 2003; 9:495-498. [PMID 12632504]

Walter RJ, Attar BM, Rafiq A, Tejaswi S, Delimata M. Newcastle disease virus LaSota strain kills human pancreatic cancer cells in vitro with high selectivity. JOP 2012; 13:45-53. [PMID 22233946]

McGinnes LW, Pantua H, Reitter J, Morrison TG. Newcastle disease virus: propagation, quantification, and storage. Curr Protoc Microbiol 2006; Chapter 15:Unit. [PMID 18770579]

McGinnes LW, Pantua H, Laliberte JP, Gravel KA, Jain S, Morrison TG. Assembly and biological and immunological properties of Newcastle disease virus-like particles. J Virol 2010; 84:4513-4523. [PMID 20181713]

Furukawa T, Duguid WP, Rosenberg L, Viallet J, Galloway DA, Tsao MS. Long-term culture and immortalization of epithelial cells from normal adult human pancreatic ducts transfected by the E6E7 gene of human papilloma virus 16. Am J Pathol 1996; 148:1763-1770. [PMID 8669463]

Ouyang H, Mou L, Luk C, Liu N, Karaskova J, Squire J, Tsao MS. Immortal human pancreatic duct epithelial cell lines with near normal genotype and phenotype. Am J Pathol 2000; 157:1623-1631. [PMID 11073822]

Davis PB, Drumm M, Konstan MW. Cystic fibrosis. Am J Respir Crit Care Med 1996; 154:1229-1256. [PMID 8912731]

Sheppard DN, Welsh MJ. Structure and function of the CFTR chloride channel. Physiol Rev 1999; 79:S23-S45. [PMID 9922375]

Shumaker H, Amlal H, Frizzell R, Ulrich CD, Soleimani M. CFTR drives Na+-nHCO-3 cotransport in pancreatic duct cells: a basis for defective HCO-3 secretion in CF. Am J Physiol 1999; 276:C16-C25. [PMID 9886916]

Schirrmacher V, Jurianz K, Roth C, Griesbach A, Bonifer R, Zawatzky R. Tumor stimulator cell modification by infection with Newcastle Disease Virus: analysis of effects and mechanism in MLTC-CML cultures. Int J Oncol 1999; 14:205-215. [PMID 9917494]

Csatary LK, Eckhardt S, Bukosza I, Czegledi F, Fenyvesi C, Gergely P, Bodey B, Csatary CM. Attenuated veterinary virus vaccine for the treatment of cancer. Cancer Detect Prev 1993; 17:619-627. [PMID 8275514]

Csatary LK, Bakacs T. Use of Newcastle disease virus vaccine (MTH-68/H) in a patient with high-grade glioblastoma. JAMA 1999; 281:1588-1589. [PMID 10235150]

Csatary LK, Moss RW, Beuth J, Torocsik B, Szeberenyi J, Bakacs T. Beneficial treatment of patients with advanced cancer using a Newcastle disease virus vaccine (MTH-68/H). Anticancer Res 1999; 19:635-638. [PMID 10216468]

Zorn U, Dallmann I, Grosse J, Kirchner H, Poliwoda H, Atzpodien J. Induction of cytokines and cytotoxicity against tumor cells by Newcastle disease virus. Cancer Biother 1994; 9:225-235. [PMID 7820184]

Cassel WA, Garrett RE. Newcastle disease virus as an antineoplastic agent. Cancer 1965; 18:863-868. [PMID 14308233]

Murray DR, Cassel WA, Torbin AH, Olkowski ZL, Moore ME. Viral oncolysate in the management of malignant melanoma. II. Clinical studies. Cancer 1977; 40:680-686. [PMID 196740]

Cassel WA, Murray DR. A ten-year follow-up on stage II malignant melanoma patients treated postsurgically with Newcastle disease virus oncolysate. Med Oncol Tumor Pharmacother 1992; 9:169-171. [PMID 1342060]

Lorence RM, Pecora AL, Major PP, Hotte SJ, Laurie SA, Roberts MS, Groene WS, Bamat MK. Overview of phase I studies of intravenous administration of PV701, an oncolytic virus. Curr Opin Mol Ther 2003; 5:618-624. [PMID 14755888]

Roberts MS, Lorence RM, Groene WS, Bamat MK. Naturally oncolytic viruses. Curr Opin Mol Ther 2006; 8:314-321. [PMID 16955694]

Freeman AI, Zakay-Rones Z, Gomori JM, Linetsky E, Rasooly L, Greenbaum E, Rozenman-Yair S, Panet A, Libson E, Irving CS, Galun E, Siegal T. Phase I/II trial of intravenous NDV-HUJ oncolytic virus in recurrent glioblastoma multiforme. Mol Ther 2006; 13:221-228. [PMID 16257582]

Lazar I, Yaacov B, Shiloach T, Eliahoo E, Kadouri L, Lotem M, Perlman R, Zakay-Rones Z, Panet A, Ben-Yehuda D. The oncolytic activity of Newcastle disease virus NDV-HUJ on chemoresistant primary melanoma cells is dependent on the proapoptotic activity of the inhibitor of apoptosis protein Livin. J Virol 2010; 84:639-646. [PMID 19864394]

Liebrich W, Schlag P, Manasterski M, Lehner B, Stohr M, Moller P, Schirrmacher V. In vitro and clinical characterisation of a Newcastle disease virus-modified autologous tumour cell vaccine for treatment of colorectal cancer patients. Eur J Cancer 1991; 27:703-710. [PMID 1829908]

Ockert D, Schirrmacher V, Beck N, Stoelben E, Ahlert T, Flechtenmacher J, Hagmuller E, Buchcik R, Nagel M, Saeger HD. Newcastle disease virus-infected intact autologous tumor cell vaccine for adjuvant active specific immunotherapy of resected colorectal carcinoma. Clin Cancer Res 1996; 2:21-28. [PMID 9816085]

Ahlert T, Sauerbrei W, Bastert G, Ruhland S, Bartik B, Simiantonaki N, Schumacher J, Hacker B, Schumacher M, Schirrmacher V. Tumor-cell number and viability as quality and efficacy parameters of autologous virus-modified cancer vaccines in patients with breast or ovarian cancer. J Clin Oncol 1997; 15:1354-1366. [PMID 9193327]

Pomer S, Schirrmacher V, Thiele R, Lohrke H, Brkovic D, Staehler G. Tumor response and 4 year survival-data of patients with advanced renal-cell carcinoma treated with autologous tumor vaccine and subcutaneous R-IL-2 and IFN-alpha(2b). Int J Oncol 1995; 6:947-954. [PMID 21556623]

Steiner HH, Bonsanto MM, Beckhove P, Brysch M, Geletneky K, Ahmadi R, Schuele-Freyer R, Kremer P, Ranaie G, Matejic D, Bauer H, Kiessling M, Kunze S, Schirrmacher V, Herold-Mende C. Antitumor vaccination of patients with glioblastoma multiforme: a pilot study to assess feasibility, safety, and clinical benefit. J Clin Oncol 2004; 22:4272-4281. [PMID 15452186]

Cassel WA, Murray DR, Phillips HS. A phase II study on the postsurgical management of Stage II malignant melanoma with a Newcastle disease virus oncolysate. Cancer 1983; 52:856-860. [PMID 6871827]

Csatary LK, Telegdy L, Gergely P, Bodey B, Bakacs T. Preliminary report of a controlled trial of MTH-68/B virus vaccine treatment in acute B and C hepatitis: a phase II study. Anticancer Res 1998; 18:1279-1282. [PMID 9615801]

Lorence RM, Rood PA, Kelley KW. Newcastle disease virus as an antineoplastic agent: induction of tumor necrosis factor-alpha and augmentation of its cytotoxicity. J Natl Cancer Inst 1988; 80:1305-1312. [PMID 2459402]

Cassel WA, Garrett RE. Tumor immunity after viral oncolysis. J Bacteriol 1966; 92:792. [PMID 4288498]

Fabian Z, Torocsik B, Kiss K, Csatary LK, Bodey B, Tigyi J, Csatary C, Szeberenyi J. Induction of apoptosis by a Newcastle disease virus vaccine (MTH-68/H) in PC12 rat phaeochromocytoma cells. Anticancer Res 2001; 21:125-135. [PMID 11299726]

Csatary LK, Gosztonyi G, Szeberenyi J, Fabian Z, Liszka V, Bodey B, Csatary CM. MTH-68/H oncolytic viral treatment in human high-grade gliomas. J Neurooncol 2004; 67:83-93. [PMID 15072452]

Hrabak A, Csuka I, Bajor T, Csatary LK. The cytotoxic anti-tumor effect of MTH-68/H, a live attenuated Newcastle disease virus is mediated by the induction of nitric oxide synthesis in rat peritoneal macrophages in vitro. Cancer Lett 2006; 231:279-289. [PMID 16399229]

Bohle W, Schlag P, Liebrich W, Hohenberger P, Manasterski M, Moller P, Schirrmacher V. Postoperative active specific immunization in colorectal cancer patients with virus-modified autologous tumor-cell vaccine. First clinical results with tumor-cell vaccines modified with live but avirulent Newcastle disease virus. Cancer 1990; 66:1517-1523. [PMID 2208003]

Schlag P, Manasterski M, Gerneth T, Hohenberger P, Dueck M, Herfarth C, Liebrich W, Schirrmacher V. Active specific immunotherapy with Newcastle-disease-virus-modified autologous tumor cells following resection of liver metastases in colorectal cancer. First evaluation of clinical response of a phase II-trial. Cancer Immunol Immunother 1992; 35:325-330. [PMID 1394336]

Schirrmacher V, Fournier P. Newcastle disease virus: a promising vector for viral therapy, immune therapy, and gene therapy of cancer. Methods Mol Biol 2009; 542:565-605. [PMID 19565923]

Alexander DJ, Allan WH. Newcastle disease virus pathotypes. Avian Pathol 1974; 3:269-278. [PMID 18777282]

Ananth R, Kirubaharian J, Priyadarshini MLM, Albert A. Isolation of Newcastle Disease Viruses of High Virulence in Unvaccinated Healthy Village Chickens in South India. Intl J Poult Sci 2008; 7:368-373.

Orsi M, Doretto JL, Reischak D, Silva LHA, Spilki FR, Buzinaro MG, Arns CW. Newcastle disease virus vaccine strains: immunogenicity is not influenced by ICPI. 11 ed. 2009:129-133.

Nagai Y. Virus activation by host proteinases. A pivotal role in the spread of infection, tissue tropism and pathogenicity. Microbiol Immunol 1995; 39:1-9. [PMID 7783672]

Nagai Y. Protease-dependent virus tropism and pathogenicity. Trends Microbiol 1993; 1:81-87. [PMID 8143121]

Gotoh B, Ogasawara T, Toyoda T, Inocencio NM, Hamaguchi M, Nagai Y. An endoprotease homologous to the blood clotting factor X as a determinant of viral tropism in chick embryo. EMBO J 1990; 9:4189-4195. [PMID 2174359]

Alexander DJ. Gordon Memorial Lecture. Newcastle disease. Br Poult Sci 2001; 42:5-22. [PMID 11337967]

Gotoh B, Ohnishi Y, Inocencio NM, Esaki E, Nakayama K, Barr PJ, Thomas G, Nagai Y. Mammalian subtilisin-related proteinases in cleavage activation of the paramyxovirus fusion glycoprotein: superiority of furin/PACE to PC2 or PC1/PC3. J Virol 1992; 66:6391-6397. [PMID 1404596]

Ogasawara T, Gotoh B, Suzuki H, Asaka J, Shimokata K, Rott R, Nagai Y. Expression of factor X and its significance for the determination of paramyxovirus tropism in the chick embryo. EMBO J 1992; 11:467-472. [PMID 1371460]

Swayne DE, King DJ. Avian influenza and Newcastle disease. J Am Vet Med Assoc 2003; 222:1534-1540. [PMID 12784958]

Fujii Y, Sakaguchi T, Kiyotani K, Yoshida T. Comparison of substrate specificities against the fusion glycoprotein of virulent Newcastle disease virus between a chick embryo fibroblast processing protease and mammalian subtilisin-like proteases. Microbiol Immunol 1999; 43:133-140. [PMID 10229267]

Walter RJ, Attar BM, Rafiq A, Delimata M, Tejaswi SLK. Three avirulent, lentogenic strains of Newcastle disease virus are cytotoxic for human pancreatic tumor cells in vitro. Gastroenterology 2010; 138(Suppl 1):s452.

Walter RJ, Attar BM, Tejaswi SLK, Rafiq A, Delimata M. Newcastle disease virus LaSota strain kills human pancreatic cancer tumor cells in vitro with high selectivity. Gastroenterology 2010; 138(Suppl 1):s453.



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