In a previous study, we demonstrated the therapeutic efficacy of a subcutaneous injection of GK1 peptide in a melanoma mouse model, effectively increasing the mean survival time by 42.58%, delaying tumor growth, and increasing intratumoral necrosis compared with the control. As a first approach to investigate the anti-melanoma effect of GK1, this study was carried out to determine the hematological effects along with both serum and lung cytokine profiles in a melanoma lung metastatic model.
Materials and
Methods: Thirteen C57BL6 female mice were transfected in the lateral tail vein with 2×105 B16-F0 melanoma cells. After 7 days, mice were separated in two different groups and treatments were initiated (day 0): The GK1-treated group (seven mice) were injected every 5 days intravenously with GK1 (10 μg) in the lateral tail vein, and the control group (six mice) were injected every 5 days with intravenous saline solution. Blood samples were collected every 5 days from day 0; tumor samples were obtained for cytokine measurements on the day of sacrifice.
Results:
In the peripheral blood, mice treated with GK1 presented a statistically significant decrease in IFN-γ (p<0.05), and lymphocytes tended to be lower compared with the control mice (p=0.06). Lung metastatic analysis demonstrated a significant increase in IFN-γ and IL-12p70 (p<0.05); a significant decrease in IL-17, IL-4, IL-22, IL-23, and IL-12p40 (p<0.05); and a marginal decrease in IL-1β (p=0.07) compared with the control.
Discussion:
Our results suggest that an intratumoral increase of cytokines with antitumor activity along with an intratumoral decrease of cytokines with protumor activity could explain, in part, the anti-melanoma effects of GK1 in a lung metastatic melanoma mouse model. Further studies must be performed to elucidate the precise mechanisms of action for GK1 peptide against melanoma, and their eventual application in humans.
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References
1.
JemalA, et al.Cancer statistics, 2006. CA Cancer J Clin, 2006; 56:106.
2.
BrownCK, KirkwoodJM. Medical management of melanoma. Surg Clin North Am, 2003; 83:283.
3.
American Cancer Society. Cancer Facts & Figures 2014. Atlanta: American Cancer Society, 2014.
4.
KaufmanHL, et al.The society for immunotherapy of cancer consensus statement on tumour immunotherapy for the treatment of cutaneous melanoma. Nat Rev Clin Oncol, 2013; 10:588.
5.
EggermontAM, et al.Adjuvant therapy with pegylated interferon alfa-2b versus observation alone in resected stage III melanoma: Final results of EORTC 18991, a randomised phase III trial. Lancet, 2008; 372:117.
6.
HodiFS, et al.Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med, 2010; 363:711.
7.
ChapmanPB, et al.Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N Engl J Med, 2011; 364:2507.
8.
RosenbergSA, YangJC, RestifoNP. Cancer immunotherapy: Moving beyond current vaccines. Nat Med, 2004; 10:909.
9.
ListT, NeriD. Immunocytokines: A review of molecules in clinical development for cancer therapy. Clin Pharmacol, 2013; 5:29.
10.
ManoutcharianK, et al.Cysticercosis: Identification and cloning of protective recombinant antigens. J Parasitol, 1996; 82:250.
11.
ToledoA, et al.Towards a Taenia solium cysticercosis vaccine: An epitope shared by Taenia crassiceps and Taenia solium protects mice against experimental cysticercosis. Infect Immun, 1999; 67:2522.
12.
Segura-VelazquezR, FragosoG, SciuttoE, et al.Towards identification of the mechanisms of action of parasite-derived peptide GK1 on the immunogenicity of an influenza vaccine. Clin Vaccine Immunol, 2009; 16:1338.
13.
Perez-TorresA, et al.The synthetic parasite-derived peptide GK1 increases survival in a preclinical mouse melanoma model. Cancer Biother Radiopharm, 2013; 28:682.
14.
WorkmanP, et al.UKCCCR guidelines for the welfare of animals in experimental neoplasia. Lab Anim, 1988; 22:195.
15.
NakamuraK, et al.Characterization of mouse melanoma cell lines by their mortal malignancy using an experimental metastatic model. Life Sci, 2002; 70:791.
16.
Salinas-JazminN, et al.Antimetastatic, antineoplastic, and toxic effects of 4-hydroxycoumarin in a preclinical mouse melanoma model. Cancer Chemother Pharmacol, 2010; 65:931.
17.
ErhardtW, HebestedtA, AschenbrennerG, et al.A comparative study with various anesthetics in mice (pentobarbitone, ketamine-xylazine, carfentanyl-etomidate). Res Exp Med (Berl), 1984; 184:159.
HoonDS, BanezM, OkunE, et al.Modulation of human melanoma cells by interleukin-4 and in combination with gamma-interferon or alpha-tumor necrosis factor. Cancer Res, 1991; 51:2002.
20.
HemmerleT, NeriD. The antibody-based targeted delivery of interleukin-4 and 12 to the tumor neovasculature eradicates tumors in three mouse models of cancer. Int J Cancer, 2014; 134:467.
21.
HungK, et al.The central role of CD4(+) T cells in the antitumor immune response. J Exp Med, 1998; 188:2357.
22.
OssendorpF, ToesRE, OffringaR, et al.Importance of CD4(+) T helper cell responses in tumor immunity. Immunol Lett, 2000; 74:75.
23.
KnutsonKL, DisisML. Tumor antigen-specific T helper cells in cancer immunity and immunotherapy. Cancer Immunol Immunother, 2005; 54:721.
24.
BrassardDL, GraceMJ, BordensRW. Interferon-alpha as an immunotherapeutic protein. J Leukoc Biol, 2002; 71:565.
25.
LesinskiGB, et al.The antitumor effects of IFN-alpha are abrogated in a STAT1-deficient mouse. J Clin Invest, 2003; 112:170.
26.
BironCA. Interferons alpha and beta as immune regulators—a new look. Immunity, 2001; 14:661.
27.
Martin-HenaoGA, et al.L-selectin expression is low on CD34+ cells from patients with chronic myeloid leukemia and interferon-a up-regulates this expression. Haematologica, 2000; 85:139.
28.
von StammU, et al.Effects of systemic interferon-alpha (IFN-alpha) on the antigenic phenotype of melanoma metastases. EORTC melanoma group cooperative study No. 18852. Melanoma Res, 1993; 3:173.
29.
BalchCM (ed.). Cutaneous Melanoma. 3rd ed. St. Louis: Quality Medical Publishing, 1998; 89.
GrimmEA, MazumderA, ZhangHZ, et al.Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes. J Exp Med, 1982; 155:1823.
32.
LotzeMT, GrimmEA, MazumderA, et al.Lysis of fresh and cultured autologous tumor by human lymphocytes cultured in T-cell growth factor. Cancer Res, 1981; 41:4420.
33.
AtkinsMB. Interleukin-2 in metastatic melanoma: What is the current role?. Cancer J Sci Am, 2000; 6Suppl 1:S8.
34.
DudleyME, et al.Cancer regression and autoimmunity in patients after clonal repopulation with antitumor lymphocytes. Science, 2002; 298:850.
35.
PalmerDC, et al.Effective tumor treatment targeting a melanoma/melanocyte-associated antigen triggers severe ocular autoimmunity. Proc Natl Acad Sci U S A, 2008; 105:8061.
36.
BoehmU, KlampT, GrootM, et al.Cellular responses to interferon-gamma. Annu Rev Immunol, 1997; 15:749.
37.
FurutaJ, InozumeT, HaradaK, et al.CD271 on melanoma cell is an IFN-gamma-inducible immunosuppressive factor that mediates downregulation of melanoma antigens. J Invest Dermatol, 2014; 134:1369.
38.
ParryRV, et al.CTLA-4 and PD-1 receptors inhibit T-cell activation by distinct mechanisms. Mol Cell Biol, 2005; 25:9543.
39.
BlankC, et al.PD-L1/B7H-1 inhibits the effector phase of tumor rejection by T cell receptor (TCR) transgenic CD8+ T cells. Cancer Res, 2004; 64:1140.
40.
BlankC, GajewskiTF, MackensenA. Interaction of PD-L1 on tumor cells with PD-1 on tumor-specific T cells as a mechanism of immune evasion: Implications for tumor immunotherapy. Cancer Immunol Immunother, 2005; 54:307.
41.
KlemmF, JoyceJA. Microenvironmental regulation of therapeutic response in cancer. Trends Cell Biol, 2015; 25:198.
TrinchieriG. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat Rev Immunol, 2003; 3:133.
44.
PetersonAC, HarlinH, GajewskiTF. Immunization with Melan-A peptide-pulsed peripheral blood mononuclear cells plus recombinant human interleukin-12 induces clinical activity and T-cell responses in advanced melanoma. J Clin Oncol, 2003; 21:2342.
45.
HamidO, et al.Alum with interleukin-12 augments immunity to a melanoma peptide vaccine: Correlation with time to relapse in patients with resected high-risk disease. Clin Cancer Res, 2007; 13:215.
46.
AgarwalP, et al.Gene regulation and chromatin remodeling by IL-12 and type I IFN in programming for CD8 T cell effector function and memory. J Immunol, 2009; 183:1695.
47.
HayesMP, WangJ, NorcrossMA. Regulation of interleukin-12 expression in human monocytes: Selective priming by interferon-gamma of lipopolysaccharide-inducible p35 and p40 genes. Blood, 1995; 86:646.
48.
SnijdersA, KalinskiP, HilkensCM, et al.High-level IL-12 production by human dendritic cells requires two signals. Int Immunol, 1998; 10:1593.
49.
MoscaPJ, et al.A subset of human monocyte-derived dendritic cells expresses high levels of interleukin-12 in response to combined CD40 ligand and interferon-gamma treatment. Blood, 2000; 96:3499.
50.
CarrenoBM, et al.IL-12p70-producing patient DC vaccine elicits Tc1-polarized immunity. J Clin Invest, 2013; 123:3383.
51.
GaffenSL. An overview of IL-17 function and signaling. Cytokine, 2008; 43:402.
52.
BettelliE, KornT, OukkaM, et al.Induction and effector functions of T(H)17 cells. Nature, 2008; 453:1051.
53.
KryczekI, et al.Cutting edge: Opposite effects of IL-1 and IL-2 on the regulation of IL-17+ T cell pool IL-1 subverts IL-2-mediated suppression. J Immunol, 2007; 179:1423.
54.
WilkeCM, et al.Th17 cells in cancer: Help or hindrance?. Carcinogenesis, 2011; 32:643.
55.
NumasakiM, et al.Interleukin-17 promotes angiogenesis and tumor growth. Blood, 2003; 101:2620.
56.
NumasakiM, et al.IL-17 enhances the net angiogenic activity and in vivo growth of human non-small cell lung cancer in SCID mice through promoting CXCR-2-dependent angiogenesis. J Immunol, 2005; 175:6177.
57.
NamJS, et al.Transforming growth factor beta subverts the immune system into directly promoting tumor growth through interleukin-17. Cancer Res, 2008; 68:3915.
58.
XiaoM, et al.IFNgamma promotes papilloma development by up-regulating Th17-associated inflammation. Cancer Res, 2009; 69:2010.
59.
ZouW, RestifoNP. T(H)17 cells in tumour immunity and immunotherapy. Nat Rev Immunol, 2010; 10:248.
60.
WangL, et al.IL-17 can promote tumor growth through an IL-6-Stat3 signaling pathway. J Exp Med, 2009; 206:1457.
61.
TengMW, et al.IL-23 suppresses innate immune response independently of IL-17A during carcinogenesis and metastasis. Proc Natl Acad Sci U S A, 2010; 107:8328.
62.
LangowskiJL, et al.IL-23 promotes tumour incidence and growth. Nature, 2006; 442:461.
63.
TengMW, von ScheidtB, DuretH, et al.Anti-IL-23 monoclonal antibody synergizes in combination with targeted therapies or IL-2 to suppress tumor growth and metastases. Cancer Res, 2011; 71:2077.
64.
ElarajDM, et al.The role of interleukin 1 in growth and metastasis of human cancer xenografts. Clin Cancer Res, 2006; 12:1088.
65.
LaviG, VoronovE, DinarelloCA, et al.Sustained delivery of IL-1 Ra from biodegradable microspheres reduces the number of murine B16 melanoma lung metastases. J Control Release, 2007; 123:123.
66.
ArendWP. The balance between IL-1 and IL-1Ra in disease. Cytokine Growth Factor Rev, 2002; 13:323.
67.
CarmiY, et al.The role of IL-1beta in the early tumor cell-induced angiogenic response. J Immunol, 2013; 190:3500.
68.
HoejbergL, et al.Serum interleukin-6 as a prognostic biomarker in patients with metastatic melanoma. Melanoma Res, 2012; 22:287.
69.
HoejbergL, BastholtL, SchmidtH. Interleukin-6 and melanoma. Melanoma Res, 2012; 22:327.
70.
HodgeDR, HurtEM, FarrarWL. The role of IL-6 and STAT3 in inflammation and cancer. Eur J Cancer, 2005; 41:2502.
71.
BartonBE. Interleukin-6 and new strategies for the treatment of cancer, hyperproliferative diseases and paraneoplastic syndromes. Expert Opin Ther Targets, 2005; 9:737.
72.
Rose-JohnS, WaetzigGH, SchellerJ, et al.The IL-6/sIL-6R complex as a novel target for therapeutic approaches. Expert Opin Ther Targets, 2007; 11:613.
73.
AraT, DeclerckYA. Interleukin-6 in bone metastasis and cancer progression. Eur J Cancer, 2010; 46:1223.
74.
NaYR, LeeJS, LeeSJ, et al.Interleukin-6-induced Twist and N-cadherin enhance melanoma cell metastasis. Melanoma Res, 2013; 23:434.
75.
SharmaP, WagnerK, WolchokJD, et al.Novel cancer immunotherapy agents with survival benefit: Recent successes and next steps. Nat Rev Cancer, 2011; 11:805.
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