Mechanisms for enhanced antitumor immune responses induced by irradiated hepatocellular carcinoma cells engineered to express hepatitis B virus X protein

Huang,Anliang; Ma,Jinhu; Huang,Liyan; Yang,Fan; Cheng,Ping

doi:10.3892/ol.2018.8430

Mechanisms for enhanced antitumor immune responses induced by irradiated hepatocellular carcinoma cells engineered to express hepatitis B virus X protein

Authors:
- Anliang Huang
- Jinhu Ma
- Liyan Huang
- Fan Yang
- Ping Cheng
View Affiliations

Affiliations: Department of Pathology, Key Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Department of Abdominal Oncology, Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: April 4, 2018 https://doi.org/10.3892/ol.2018.8430
Pages: 8505-8515
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Tumor associated antigen (TAA) induces both humoral immunity and cellular immunity. The T cell-mediated immune response has an important role in the immune response induced by TAA. The hepatitis B virus X protein (HBx) sequence is mapped with Custer of differentiation (CD)8+ T cell (CTL) epitopes, while a large number of studies have indicated that HBx may enhance the autophagy. In our previous study, a novel hepatocellular carcinoma vaccine was designed that was an irradiated HBx modified hepatocellular carcinoma cell vaccine in autophagic form, which significantly induced antitumor immune responses in vivo. However, the mechanism by which this vaccine contributes to enhancing antitumor immune responses have yet to be fully elucidated. In the present study, we examined how autophagy was induced by this vaccine's influence on the generation of the ‘danger signal’ by hepatoma tumor cells and the subsequent activation of the immunoresponse. The data showed that the vaccine induced phenotypic maturation of DCs, which leads to efficient cross-presentation and a specific response. Both CD8+ and CD4+ T lymphocytes were involved in the antitumor immune response, as reflected by IFN-γ secretion. In addition, damage-associated molecular pattern molecules (DAMPs) were significantly elevated in the vaccine, and the elevation of DAMPs was autophagy-dependent. Furthermore, the antitumor activity was achieved by adoptive transfer of lymphocytes but not serum. The present findings indicated that this vaccine enhanced antitumor immune responses, which was in accordance with our previous study.

View Figures

View References

1	Steinman RM: The dendritic cell system and its role in immunogenicity. Annu Rev Immunol. 9:271–296. 1991. View Article : Google Scholar : PubMed/NCBI
2	Hart DN: Dendritic cells: Unique leukocyte populations which control the primary immune response. Blood. 90:3245–3287. 1997.PubMed/NCBI
3	Rock KL: A new foreign policy: MHC class I molecules monitor the outside world. Immunol Today. 17:131–137. 1996. View Article : Google Scholar : PubMed/NCBI
4	Fields RC, Shimizu K and Mulè JJ: Murine dendritic cells pulsed with whole tumor lysates mediate potent antitumor immune responses in vitro and in vivo. Proc Natl Acad Sci USA. 95:9482–9487. 1998. View Article : Google Scholar : PubMed/NCBI
5	Perez CA, Fu A, Onishko H, Hallahan DE and Geng L: Radiation induces an antitumour immune response to mouse melanoma. Int J Radiat Biol. 85:1126–1136. 2009. View Article : Google Scholar : PubMed/NCBI
6	Weiss EM, Frey B, Rödel F, Herrmann M, Schlücker E, Voll RE, Fietkau R and Gaipl US: Ex vivo- and in vivo-induced dead tumor cells as modulators of antitumor responses. Ann NY Acad Sci. 1209:109–117. 2010. View Article : Google Scholar : PubMed/NCBI
7	Das A and Ali N: Vaccine prospects of killed but metabolically active Leishmania against visceral leishmaniasis. Expert Rev Vaccines. 11:783–785. 2012. View Article : Google Scholar : PubMed/NCBI
8	Hoshimoto S, Faries MB, Morton DL, Shingai T, Kuo C, Wang HJ, Elashoff R, Mozzillo N, Kelley MC, Thompson JF, et al: Assessment of prognostic circulating tumor cells in a phase III trial of adjuvant immunotherapy after complete resection of stage IV melanoma. Ann Surg. 255:357–362. 2012. View Article : Google Scholar : PubMed/NCBI
9	Miguel A, Herrero MJ, Sendra L, Botella R, Algás R, Sánchez M and Aliño SF: Comparative antitumor effect among GM-CSF, IL-12 and GM-CSF+IL-12 genetically modified tumor cell vaccines. Cancer Gene Ther. 20:576–581. 2013. View Article : Google Scholar : PubMed/NCBI
10	Yan HX, Cheng P, Wei HY, Shen GB, Fu LX, Ni J, Wu Y and Wei YQ: Active immunotherapy for mouse breast cancer with irradiated whole-cell vaccine expressing VEGFR2. Oncol Rep. 29:1510–1516. 2013. View Article : Google Scholar : PubMed/NCBI
11	Meijer SL, Dols A, Jensen SM, Hu HM, Miller W, Walker E, Romero P, Fox BA and Urba WJ: Induction of circulating tumor-reactive CD8⁺ T cells after vaccination of melanoma patients with the gp100 209-2M peptide. J Immunother. 30:533–543. 2007. View Article : Google Scholar : PubMed/NCBI
12	Zhang X, Mei W, Zhang L, Yu H, Zhao X, Fan X, Qian G and Ge S: Co-expression of P1A_35–43/β2m fusion protein and co-stimulatory molecule CD80 elicits effective anti-tumor immunity in the P815 mouse mastocytoma tumor model. Oncol Rep. 22:1213–1220. 2009.PubMed/NCBI
13	Iero M, Filipazzi P, Castelli C, Belli F, Valdagni R, Parmiani G, Patuzzo R, Santinami M and Rivoltini L: Modified peptides in anti-cancer vaccines: Are we eventually improving anti-tumour immunity? Cancer Immunol Immunother. 58:1159–1167. 2009. View Article : Google Scholar : PubMed/NCBI
14	Yan Y, Liu N, Lu L, Zang CM, Shao B, Li Y, Wen Y, Wei Y and Cheng P: Autophagy enhances antitumor immune responses induced by irradiated hepatocellular carcinoma cells engineered to express hepatitis B virus X protein. Oncol Rep. 30:993–999. 2013. View Article : Google Scholar : PubMed/NCBI
15	Luo C, Shen G, Liu N, Gong F, Wei X, Yao S, Liu D, Teng X, Ye N, Zhang N, et al: Ammonia drives dendritic cells into dysfunction. J Immunol. 193:1080–1089. 2014. View Article : Google Scholar : PubMed/NCBI
16	Norman JM, Cohen GM and Bampton ET: The in vitro cleavage of the hAtg proteins by cell death proteases. Autophagy. 6:1042–1056. 2010. View Article : Google Scholar : PubMed/NCBI
17	Bevan MJ: Cross-priming for a secondary cytotoxic response to minor H antigens with H-2 congenic cells which do not cross-react in the cytotoxic assay. J Exp Med. 143:1283–1288. 1976. View Article : Google Scholar : PubMed/NCBI
18	Huang AY, Golumbek P, Ahmadzadeh M, Jaffee E, Pardoll D and Levitsky H: Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science. 264:961–965. 1994. View Article : Google Scholar : PubMed/NCBI
19	Rehermann B and Nascimbeni M: Immunology of hepatitis B virus and hepatitis C virus infection. Nat Rev Immunol. 5:215–229. 2005. View Article : Google Scholar : PubMed/NCBI
20	Malmassari S, Lone YC, Zhang M, Transy C and Michel ML: In vivo hierarchy of immunodominant and subdominant HLA-A*0201-restricted T-cell epitopes of HBx antigen of hepatitis B virus. Microbes Infect. 7:626–634. 2005. View Article : Google Scholar : PubMed/NCBI
21	Ding FX, Wang F, Lu YM, Li K, Wang KH, He XW and Sun SH: Multiepitope peptide-loaded virus-like particles as a vaccine against hepatitis B virus-related hepatocellular carcinoma. Hepatology. 49:1492–1502. 2009. View Article : Google Scholar : PubMed/NCBI
22	Li Y, Cheng P, Wen Y, Chen P, Yang L, Zhao X, Lv H, Quan Q, Wu Y, Yang H, et al: T lymphocyte responses against hepatitis B virus-related hepatocellular carcinoma induced by adenovirus vaccine encoding HBx. Int J Mol Med. 26:869–876. 2010.PubMed/NCBI
23	Podojil JR and Miller SD: Targeting the B7 family of co-stimulatory molecules: Successes and challenges. BioDrugs. 27:1–13. 2013. View Article : Google Scholar : PubMed/NCBI
24	Townsend SE and Allison JP: Tumor rejection after direct costimulation of CD8+ T cells by B7-transfected melanoma cells. Science. 259:368–370. 1993. View Article : Google Scholar : PubMed/NCBI
25	Kim R, Emi M and Tanabe K: Cancer immunoediting from immune surveillance to immune escape. Immunology. 121:1–14. 2007. View Article : Google Scholar : PubMed/NCBI
26	Beatty GL and Gladney WL: Immune escape mechanisms as a guide for cancer immunotherapy. Clin Cancer Res. 21:687–692. 2015. View Article : Google Scholar : PubMed/NCBI
27	Rabinovich GA, Gabrilovich D and Sotomayor EM: Immunosuppressive strategies that are mediated by tumor cells. Annu Rev Immunol. 25:267–296. 2007. View Article : Google Scholar : PubMed/NCBI
28	Mizushima N, Yoshimori T and Ohsumi Y: The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol. 27:107–132. 2011. View Article : Google Scholar : PubMed/NCBI
29	Virgin HW and Levine B: Autophagy genes in immunity. Nat Immunol. 10:461–470. 2009. View Article : Google Scholar : PubMed/NCBI
30	Mizushima N and Levine B: Autophagy in mammalian development and differentiation. Nat Cell Biol. 12:823–830. 2010. View Article : Google Scholar : PubMed/NCBI
31	Li Y, Wang LX, Yang G, Hao F, Urba WJ and Hu HM: Efficient cross-presentation depends on autophagy in tumor cells. Cancer Res. 68:6889–6895. 2008. View Article : Google Scholar : PubMed/NCBI
32	Uhl M, Kepp O, Jusforgues-Saklani H, Vicencio JM, Kroemer G and Albert ML: Autophagy within the antigen donor cell facilitates efficient antigen cross-priming of virus-specific CD8⁺ T cells. Cell Death Differ. 16:991–1005. 2009. View Article : Google Scholar : PubMed/NCBI
33	Tang H, Da L, Mao Y, Li Y, Li D, Xu Z, Li F, Wang Y, Tiollais P, Li T and Zhao M: Hepatitis B virus X protein sensitizes cells to starvation-induced autophagy via up-regulation of beclin 1 expression. Hepatology. 49:60–71. 2009. View Article : Google Scholar : PubMed/NCBI
34	Sir D, Tian Y, Chen WL, Ann DK, Yen TS and Ou JH: The early autophagic pathway is activated by hepatitis B virus and required for viral DNA replication. Proc Natl Acad Sci USA. 107:4383–4388. 2010. View Article : Google Scholar : PubMed/NCBI
35	Esensten JH, Helou YA, Chopra G, Weiss A and Bluestone JA: CD28 costimulation: From mechanism to therapy. Immunity. 44:973–988. 2016. View Article : Google Scholar : PubMed/NCBI
36	Keir ME, Butte MJ, Freeman GJ and Sharpe AH: PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol. 26:677–704. 2008. View Article : Google Scholar : PubMed/NCBI
37	Bennett SR, Carbone FR, Karamalis F, Miller JF and Heath WR: Induction of a CD8⁺ cytotoxic T lymphocyte response by cross-priming requires cognate CD4⁺ T cell help. J Exp Med. 186:65–70. 1997. View Article : Google Scholar : PubMed/NCBI
38	Cassell D and Forman J: Linked recognition of helper and cytotoxic antigenic determinants for the generation of cytotoxic T lymphocytes. Ann NY Acad Sci. 532:51–60. 1988. View Article : Google Scholar : PubMed/NCBI
39	Matzinger P: Tolerance, danger, and the extended family. Annu Rev Immunol. 12:991–1045. 1994. View Article : Google Scholar : PubMed/NCBI
40	Nickel W and Rabouille C: Mechanisms of regulated unconventional protein secretion. Nat Rev Mol Cell Biol. 10:148–155. 2009. View Article : Google Scholar : PubMed/NCBI
41	Deretic V, Jiang S and Dupont N: Autophagy intersections with conventional and unconventional secretion in tissue development, remodeling and inflammation. Trends Cell Biol. 22:397–406. 2012. View Article : Google Scholar : PubMed/NCBI
42	Tian J, Avalos AM, Mao SY, Chen B, Senthil K, Wu H, Parroche P, Drabic S, Golenbock D, Sirois C, et al: Toll-like receptor 9-dependent activation by DNA-containing immune complexes is mediated by HMGB1 and RAGE. Nat Immunol. 8:487–496. 2007. View Article : Google Scholar : PubMed/NCBI
43	Garg AD, Krysko DV, Vandenabeele P and Agostinis P: Hypericin-based photodynamic therapy induces surface exposure of damage-associated molecular patterns like HSP70 and calreticulin. Cancer Immunol Immunother. 61:215–221. 2012. View Article : Google Scholar : PubMed/NCBI
44	Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini JL, Castedo M, Mignot G, Panaretakis T, Casares N, et al: Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med. 13:54–61. 2007. View Article : Google Scholar : PubMed/NCBI