TAX and HBZ: hFc Ɣ 1 proteins as targets for passive immunotherapy

Document Type : Original Article

Authors

1 Inflammation and Inflammatory Diseases Research Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Cancer and Immunology Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

4 Animal Laboratory, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Blood Borne Infections Research Center, Academic Center for Education, Culture, and Research (ACECR), Razavi Khorasan, Mashhad, Iran

10.22038/ijbms.2022.64787.14266

Abstract

Objective(s): Human T leukemia virus type one (HTLV-1) causes two life-threatening diseases in around five percent of infected subjects, a T cell malignancy and a neurodegenerative disease. TAX and HBZ are the main virulence agents implicated in the manifestation of HTLV-1–associated diseases. Therefore, this study aims to produce these HTLV-1 factors as recombinant Fc fusion proteins to study the structures, their immunogenic properties as vaccines, and their capability to produce specific neutralization antibodies.
Materials and Methods: TAX and HBZ sequences were chosen from the NCBI-nucleotide database, then designed as human Fc chimers and cloned into Pichia pastoris. Produced proteins were purified by HiTrap affinity chromatography and subcutaneously injected into rabbits. Rabbit Abs were purified by batch chromatography, and their neutralization activities for the HTLV-1-infected MT-2 cell line were assessed. Furthermore, the protective abilities of recombinant proteins were evaluated in Tax or HBZ immunized rabbits by MT-2 cell line inoculation and measurement of HTLV-1-proviral load.
Results: Specific Abs against Tax and HBZ can eliminate 2 million MT-2 cells in 1/1000 dilution in vitro. In challenging assays, the immunization of the animals using Tax or HBZ had no protective activity as HTLV-1 PVL was still positive.
Conclusion: The result suggests that recombinant TAX and HBZ: hFcγ1 proteins can produce a proper humoral immune response. Therefore, they could be considered a passive immunotherapy source for HTLV-1-associated diseases, while total TAX and HBZ proteins are unsuitable as HTLV-1 vaccine candidates. 

Keywords


1. Eusebio-Ponce E, Anguita E, Paulino-Ramirez R, Candel FJ. HTLV-1 infection: an emerging risk. pathogenesis, epidemiology, diagnosis and associated diseases. Rev Esp Quimioter 2019; 32:485-496.
2. Khan MY, Khan IN, Farman M, Al Karim S, Qadri I, Kamal MA, et al. HTLV-1 associated neurological disorders. Curr Top Med Chem 2017; 17:1320-1330. 
3. Iwanaga M. Epidemiology of HTLV-1 infection and ATL in Japan: an update. Front Microbiol 2020; 11:1124-1134.
4. Taylor GP. The epidemiology of HTLV-I in Europe. J Acquir Immune Defic Syndr Hum Retrovirol 1996; 1:8-14. 
5. Rafatpanah H, Hedayati-Moghaddam MR, Fathimoghadam F, Bidkhori HR, Shamsian SK, Ahmadi S, et al. High prevalence of HTLV-I infection in Mashhad, Northeast Iran: a population-based seroepidemiology survey. J Clin Virol 2011; 52:172-176.
6. Araujo AQ, Silva MT. The HTLV-1 neurological complex. Lancet Neurol 2006; 5:1068-1076. 
7. Mahieux R, Gessain A. HTLV-1 and associated adult T-cell leukemia/lymphoma. Rev clin Exp Hematol 2003; 7:336-361.
8. Garcia I, Hennington É A. HTLV: a stigmatizing infection? Cad Saude Publica 2019; 35:1-14.
9.Watanabe T. HTLV-1-associated diseases. Int J Hematol 1997; 66:257-278.
10.    Rodríguez-Zúñiga MJM, Cortez-Franco F, Qujiano-Gomero E. Adult T-cell leukemia/lymphoma. Review of the literature. Actas Dermosifiliogr 2018; 109:399-407.
11.    Phillips AA, Harewood JCK. Adult T Cell leukemia-lymphoma (ATL): state of the art. Curr Hematol Malig Rep 2018; 13:300-307.
12.    Mehta-Shah N, Ratner L, Horwitz SM. Adult T-Cell leukemia/lymphoma. J Oncol Pract 2017; 13:487-492. 
13.    Mohanty S, Harhaj EW. Mechanisms of oncogenesis by HTLV-1 tax. Pathogens 2020; 9:543-570. 
14.    Martin JL, Maldonado JO, Mueller JD, Zhang W, Mansky LM. Molecular studies of HTLV-1 replication: an update. Viruses 2016; 8:31-52.
15.    Mirhosseini A, Mohareri M, Arab R, Rezaee SA, Shirdel A, Koshyar MM, et al. Complete sequence of human T cell leukemia virus type 1 in ATLL patients from Northeast Iran, Mashhad revealed a prematurely terminated protease and an elongated pX open reading frame III. Infect Genet Evol 2019; 73:460-469. 
16.    Tagaya Y, Gallo RC. The exceptional oncogenicity of HTLV-1. Front Microbiol 2017; 8:1425-1428. 
17.    Akbarin MM, Shirdel A, Bari A, Mohaddes ST, Rafatpanah H, Karimani EG, et al. Evaluation of the role of TAX, HBZ, and HTLV-1 proviral load on the survival of ATLL patients. Blood Res 2017; 52:106-111.
18.    Tarokhian H, Rahimi H, Mosavat A, Shirdel A, Rafatpanah H, Akbarin MM, et al. HTLV-1-host interactions on the development of adult T cell leukemia/lymphoma: virus and host gene expressions. BMC Cancer 2018; 18:1287-1298.
19.    Matsuoka M, Mesnard JM. HTLV-1 bZIP factor: the key viral gene for pathogenesis. Retrovirology 2020; 17:2-9.
20.    Al-Saleem J DW, Martinez MP, Shkriabai N, Kvaratskhelia M, Ratner L, et al. HTLV-1 tax-1 interacts with SNX27 to regulate cellular localization of the HTLV-1 receptor molecule, GLUT1. PLoS One 2019; 14:e0214059.
21.    Fochi S, Mutascio S, Bertazzoni U, Zipeto D, Romanelli MG. HTLV deregulation of the NF-κB pathway: an update on tax and antisense proteins role. Front Microbiol  2018; 9:285-292.
22.    Rauch DA, Ratner L. Targeting HTLV-1 activation of NFκB in mouse models and ATLL patients. Viruses 2011; 3:886-900.
23.    Wurm T, Wright DG, Polakowski N, Mesnard J-M, Lemasson I. The HTLV-1-encoded protein HBZ directly inhibits the acetyl transferase activity of p300/CBP. Nucleic Acids Res 2012; 40:5910-5925.
24.    Mozhgani SH, Jaberi N, Rezaee SA, Bustani R, Jazayeri SM, Akbarin MM, et al. Evaluation of HTLV-1 HBZ and proviral load, together with host IFN λ3, in pathogenesis of HAM/TSP. J Med Virol 2017; 89:1102-1107.
25.    Zhao T. The role of HBZ in HTLV-1-induced oncogenesis. Viruses 2016; 8:34-45.
26.    Katsuya H, Ishitsuka K. Treatment advances and prognosis for patients with adult T-cell leukemia-lymphoma. J Clin Exp Hematopathol 2017; 57:87-97. 
27.    Boostani R, Vakili R, Hosseiny SS, Shoeibi A, Fazeli B, Etemadi MM, et al. Triple therapy with prednisolone, pegylated interferon and sodium valproate improves clinical outcome and reduces human T-Cell leukemia virus type 1 (HTLV-1) proviral load, tax and HBZ mRNA expression in patients with HTLV-1-associated myelopathy/tropical spastic paraparesis. Neurotherapeutics 2015; 12:887-895.
28.    Torshizi R, Ghayour Karimani E, Etminani K, Akbarin MM, Jamialahmadi K, Shirdel A, et al. Altered expression of cell cycle regulators in adult T-cell leukemia/ lymphoma patients. Rep Biochem Mol Biol 2017; 6:88-94.
29.    Marino-Merlo F, Balestrieri E, Matteucci C, Mastino A, Grelli S, Macchi B. Antiretroviral therapy in HTLV-1 infection: an updated overview. Pathogens 2020; 9:342-355.
30.    de Thé G, Bomford R. An HTLV-I vaccine: why, how, for whom? AIDS Res Hum Retroviruses 1993; 9:381-386. 
31.    Fields PA, Taylor GP. “Antivirals” in the treatment of adult T cell leukaemia- lymphoma (ATLL). Curr Hematol Malig Rep 2012; 7:267-275. 
32.    Théry C, Amigorena S. The cell biology of antigen presentation in dendritic cells. Curr Opin Immunol 2001; 13:45-51. 
33.    Guyre PM, Graziano RF, Goldstein J, Wallace PK, Morganelli PM, Wardwell K, et al. Increased potency of Fc-receptor-targeted antigens. Cancer Immunol Immunother 1997; 45:146-148.
34.    Arina A, Tirapu I, Alfaro C, Rodríguez-Calvillo M, Mazzolini G, Inogés S, et al. Clinical implications of antigen transfer mechanisms from malignant to dendritic cells. exploiting cross-priming. Exp Hematol 2002; 30:1355-1364. 
35.    Krieger E, Joo K, Lee J, Raman S, Thompson J, Tyka M, et al. Improving physical realism, stereochemistry, and side-chain accuracy in homology modeling: four approaches that performed well in CASP8. Proteins 2009; 77:114-122.
36.    Colovos C, Yeates TO. Verification of protein structures: patterns of nonbonded atomic interactions. Protein Sci 1993; 2:1511-1519.
37.    Lovell SC, Davis IW, Arendall WB 3rd, de Bakker PI, Word JM, Prisant MG, et al. Structure validation by Calpha geometry: phi,psi and Cbeta deviation. Proteins 2003; 50:437-450.
38.    Gupta R, Brunak S. Prediction of glycosylation across the human proteome and the correlation to protein function. Pac Symp Biocomput 2002: 310-322. 
39.    Steentoft C, Vakhrushev SY, Joshi HJ, Kong Y, Vester-Christensen MB, Schjoldager KT, et al. Precision mapping of the human O-GalNAc glycoproteome through simpleCell technology. EMBO J 2013; 32:1478-1488.
40.    Karbalaei Zadeh Babaki M, Taghiabadi M, Soleimanpour S, Saleh Moghadam M, Mosavat A, Amini AA, et al. Mycobacterium tuberculosis Ag85b:hfcγ1 recombinant fusion protein as a selective receptor-dependent delivery system for antigen presentation. Microb Pathog 2019; 129:68-73.
41.    Baghani AA, Soleimanpour S, Farsiani H, Mosavat A, Yousefi M, Meshkat Z, et al. CFP10: mFcγ2 as a novel tuberculosis vaccine candidate increases immune response in mouse. Iran J Basic Med Sci 2017; 20:122-130.
42.    Shuh M, Beilke M. The human T-cell leukemia virus type 1 (HTLV-1): new insights into the clinical aspects and molecular pathogenesis of adult T-cell leukemia/lymphoma (ATLL) and tropical spastic paraparesis/HTLV-associated myelopathy (TSP/HAM). Microsc Res Tech 2005; 68:176-196.
43.    Kannagi M, Hasegawa A, Nagano Y, Iino T, Okamura J, Suehiro Y. Maintenance of long remission in adult T-cell leukemia by Tax-targeted vaccine: a hope for disease-preventive therapy. Cancer Sci 2019; 110:849-857. 
44.    Sugata K, Yasunaga J-i, Mitobe Y, Miura M, Miyazato P, Kohara M, et al. Protective effect of cytotoxic T lymphocytes targeting HTLV-1 bZIP factor. Blood 2015; 126:1095-1105.
45.    Martin F, Taylor GP, Jacobson S. Inflammatory manifestations of HTLV-1 and their therapeutic options. Expert Rev Clin Immunol 2014; 10:1531-1546.
46.    Moreno-Ajona D, Yuste JR, Martín P, Gállego Pérez-Larraya J. HTLV-1 myelopathy after renal transplant and antiviral prophylaxis: the need for screening. J Neurovirol 2018; 24:523-525.
47.    Olindo S, Belrose G, Gillet N, Rodriguez S, Boxus M, Verlaeten O, et al. Safety of long-term treatment of HAM/TSP patients with valproic acid. Blood 2011; 118:6306-6309. 
48.    de Thé G, Kazanji M. An HTLV-I/II vaccine: from animal models to clinical trials? J Acquir Immune Defici Syndr Hum Retrovirol 1996; 1:191-198.
49.    Lynch MP, Kaumaya PT. Advances in HTLV-1 peptide vaccines and therapeutics. Curr Protein  Pept Sci 2006; 7:137-145.
50.    Datta-Mannan A, Witcher DR, Lu J, Wroblewski VJ. Influence of improved FcRn binding on the subcutaneous bioavailability of monoclonal antibodies in cynomolgus monkeys. MAbs 2012; 4:267-273.
51.    Peter H-H, Ochs HD, Cunningham-Rundles C, Vinh DC, Kiessling P, Greve B, et al. Targeting FcRn for immunomodulation: benefits, risks, and practical considerations. J Allergy Clin Immunol 2020; 146:479-491.
52.    Kannian P, Yin H, Doueiri R, Lairmore M, Fernandez S, Green P. Distinct transformation tropism exhibited by human T lymphotropic virus type 1 (HTLV-1) and HTLV-2 is the result of postinfection T cell clonal expansion. J Virol 2012; 86:3757-3766.
53.    Begum N, Horiuchi S, Tanaka Y, Yamamoto N, Ichiyama K. New approach for generation of neutralizing antibody against human T-cell leukaemia virus type-I (HTLV-I) using phage clones. Vaccine 2002; 20:1281-1289.
54. Fujii H, Shimizu M, Miyagi T, Kunihiro M, Tanaka R, Takahashi Y, et al. A potential of an anti-HTLV-I gp46 neutralizing monoclonal antibody (LAT-27) for passive immunization against both horizontal and mother-to-child vertical infection with human T cell leukemia virus type-I. Viruses 2016; 8:41-50.