Neutralizing Potential of Fab IgG Hybrid Antibody Against Dengue Virus (DENV-1,2,3,4) Expressed on Mesenchymal Stem Cells

Dengue is an acute mosquito born viral infections in tropical and subtropical countries, that has been steadily increasing every year. Dengue haemorrhagic fever in Indonesia is caused by all 4 viral serotypes termed as DENV-1, DENV-2, DENV-3, and DENV-4. It is very difficult to treat the disease and there is no effective vaccine to eradicate Dengue virus infections .The aim of this research is to express the antibody fragment gene coding VL and VH of Fab IgG antibody in rat bone marrow mesenchymal stem cells (rat BM-MSCs). The genes were isolated from immunized mice using inactive virus of all dengue serotypes. Then genes were hybridized through ligation and insertion into plasmid pBR322, and transfected into rat BMMSCs. To analytically characterize Fab IgG hybrid binding ability, capacity and specificity, we used immune precipitation, western blotting, neutralization assay, and ELISA. All assays suggested that hybrid Fab IgG antibody have high reactivity and affinity, to efficiently neutralize all Dengue virus serotypes 1,2,3,4. Results of this study show that hybrid Fab IgG antibody can be used as neutralizing agent for the treatment of dengue infections in the future.

Mezenkimal Kök Hücreler Üzerinde Dengue Virüsüne Karşı (DENV-1,2,3,4) Fab IgG Hibrit Antikorunun Nötralize Edici Potansiyeli

Dengue humması, tropik ve subtropikal ülkelerde sivrisineklerden kaynaklanan akut viral bir enfeksiyon olup, her yıl giderek artmaktadır. Endonezya’daki Dang kanamalı ateşine DENV-1, DENV-2, DENV-3 ve DENV-4 adı verilen 4 viral serotip neden olmaktadır. Hastalığın tedavisi çok zordur ve Dang virüsü enfeksiyonlarını yok etmek için etkili bir aşı yoktur. Bu araştırmanın amacı, sıçan kemik iliği mezenkimal kök hücrelerinde (sıçan BM-MSCs) bulunan Fab IgG antikorunun VL ve VH antikor fragmentini kodlayan geninin açığa çıkarılmasıdır. Genler, tüm dang serotiplerinin inaktif virüsü kullanılarak aşılanmış farelerden izole edilmiştir. Sonrasında genler, ligasyon ve plasmid pBR322 içine gömülmesi yoluyla hibridize edilmiş ve sıçan BMMSC’lerine nakledilmiştir. Analitik olarak Fab IgG’nin hibrid bağlama yeteneği, kapasitesi ve özgünlüğünü karakterize etmek için; bağışıklık çökeltmesi, western blotting, nötralizasyon testi ve ELISA kullanılmıştır. Bütün deneyler, hibrid Fab IgG antikorunun tüm Dang virüsü serotipleri 1,2,3,4’ü etkili bir şekilde nötrleştirmek için yüksek reaktiviteye ve afiniteye sahip olduğunu ortaya koymaktadır. Bu çalışmanın sonuçları, melez Fab IgG antikorunun gelecekte dang enfeksiyonlarının tedavisinde nötralize edici ajan olarak kullanılabileceğini göstermektedir.

___

Abaitua F., Rodríguez, J.R., Garzón, A., Rodríguez, D., Esteban, M. 2006. Improving recombinant MVA immune responses: potentiation of the immune responses to HIV-1 with MVA and DNA vectors expressing Env and the cytokines IL-12 and IFNgamma. Virus Res 116, 11-20. [CrossRef]

Bielefeldt-Ohmann, H., Meyer, M., Fitzpatrick, D.R., Mackenzie, J.S. 2001. Dengue virus binding to human leukocyte cell lines: receptor usage differs between cell types and virus strains. Virus Res 73(1), 81–9. [CrossRef]

Champion, J.M., Kean, R.B., Rupprecht, C.E., Notkins, A.L., Koprowski, H., Dietzschold, B., Hooper, D.C. 2000. The development of monoclonal human rabies virus-neutralizing antibodies as a substitute for pooled human immune globulin in the prophylactic treatment of rabies virus exposure. J. Immunol. Methods 235, 81–90. [CrossRef]

Chu., Y.K., Rossy, C., LeDuc, J.W., Lee, H.W., Schmaljohn, C.S., Dalrymple. 1994. Serological relationship among virus in the hanta virus genus family bunyaviridae virology 198, 196-204

Crameri, R., 2007. Allergy vaccines: dreams and reality. Expert Rev Vaccines 2007 Dec; 6(6), 991-9. [CrossRef]

de Carvalho Nicacio, C., Lundkvist, A., Sjölander, K.B., Plyusnin, A., Salonen, E.M,. Björling, E. 2000. A neutralizing recombinant human antibody Fab fragment against Puumala hantavirus. J Med Virol 60, 446-454. [CrossRef]

Fried, J.R., Gibbons, R.V., Kalayanarooj, S., Thomas, S.J., Srikiatkhachorn, A., Yoon, I.K., Jarman, R.G., Green, S., Rothman, A.L., Cummings, D.A. 2010. Serotype-specific differences in the risk of dengue hemorrhagic fever: an analysis of data collected in Bangkok, Thailand from 1994 to 2006. PLoS Negl Trop Dis 4(3), e617. [CrossRef]

Gigler A., Dorsch, S., Hemauer, A., Williams, C., Kim, S., Young,N.S., Zolla-Pazner, S., Wolf, H., Gorny M.K., Modrow, S. 1999. Generation of neutralizing human monoclonal antibodies against parvovirus B19 proteins. J Virol 73, 1974-1979.

Gould, L.H., Sui, J,. Foellmer, H., Oliphant, T., Wang, T., Ledizet, M., Murakami, A., Noonan, K., Lambeth, C., Kar, K., Anderson, J.F., de Silva, A.M., Diamond, M.S., Koski, R.A., Marasco, W.A., Fikrig, E. 2005. Protective and therapeutic capacity of human single-chain Fv-Fc fusion proteins against West Nile virus. J Virol 79, 14606-14613. [CrossRef]

Halstead, S.B. 1998. Pathogenesis of dengue: challenges to molecular biology. Science 239, 476-481. [CrossRef]

Halstead, S.B. 2003. Neutralization and antibody-dependent enhancement of dengue viruses. Adv Virus Res 60, 421-467. [CrossRef]

Henchal, E.A., Putnak, J.R. 1990. Dengue viruses. Clin Microbiol Rev 3, 376-396. [CrossRef]

Huse, K., Böhme, H.J., Scholz, G.H. 2002. Purification of antibodies by affinity chromatography. Journal of Biochemical and Biophysical Methods 51(3), 217-231. [CrossRef]

Kehry, M.R., Castle, B.E. 1994. Regulation of CD40 ligand expression and use of recombinantCD40 ligand for studyingBcell growth and differentiation. SeminImmunol 6, 287-294. [CrossRef]

Kihira, Y., Aiba, S. 1992. Artificial immunoglobulin G-binding protein mimetic to staphylococcal protein A. Its production and application to purification of immunoglobulin G. J Chromatogr B: Biomed Sci Appl 597, 277-283.

Lai, C.Y., Tsai, W.Y., Lin, S.R., Kao, C.L., Hu, H.P., King, C.C., Wu, H.C., Chang, G.J., Wang, W.K. 2008. Antibodies to envelope glycoprotein of dengue virus during the natural course of infection are predominantly cross-reactive and recognize epitopes containing highly conserved residues at the fusion loop of domain II. J Virol 82(13), 6631- 6643. [CrossRef]

Leickt, L., Grubb, A., Ohlson, S. 1998. Screening for weak monoclonal antibodies in hybridoma technology. J Mol Recognit 11(1-6), 114- 116. [CrossRef]

Liang, W.Q., Fournier, M.J. 1997. Synthesis of functional eukaryotic ribosomal RNAs in trans: development of a novel in vivo rDNA system for dissecting ribosome biogenesis. Proc Natl Acad Sci USA 94, 2864-2868. [CrossRef]

Meulenbroek, A.J., Zeijlemaker, W.P. 1996. Human IgG subclasses: useful diagnostic markers for immunocompetence. Amsterdam CLB, 1-51.

Ong, S.H., Yip, J.T., Chen, Y.L., Liu, W., Harun, S., Lystiyaningsih, E., Heriyanto, B., Beckett, C.G., Mitchell, W.P., Hibberd, M.L., Suwandono, A., Vasudevan, S.G., Schreiber, M.J. 2008. Periodic re-emergence of endemic strains with strong epidemic potential-a proposed explanation for the 2004 Indonesian dengue epidemic. Infect Genet Evol 8(2), 191-204. [CrossRef]

Pauli, G., Gregersen, J-P., Ludwig, H. 1984. Plaque/ focus immunoassay: a simple method for detecting antiviral monoclonal or other antibodies and viral antigens in cell. J Immunol Methods 74, 337-344. [CrossRef]

Putnak, R., Fuller, J., Vander Zanden, L., Innis, B.L., Vaughn, D.W. 2003. Vaccination of rhesus macaques against dengue-2 virus with a plasmid DNA vaccine encoding the viral pre-membrane and envelope genes. Am J Trop Med Hyg 68(4), 469-476.

Rantam, F.A., Purwati, Soegijanto, S., Susilowati, H., Sudiana, K., Hendrianto, E., Soetijipto. 2013. Analysis of recombinant, multivalent dengue virus containing envelope (E) proteins from serotypes - 1, - 3, and - 4 and expressed in baculovirus. Trials Vaccinol DOI: 10.1016/j.trivac.2013.10.001. [CrossRef]

Rantam, F.A., Ferdiansyah, Nasronudin, Purwati. 2009. Stem Cell Exploration. Airlangga University Press, Surabaya, 70-75.

Rothman, A.L. 2010. Cellular immunology of sequential dengue virus infection and its role in disease pathogenesis. Curr Top Microbiol Immunol 338, 83-98. [CrossRef]

Sarathy, V.V., White, M., Li, L., Gorder, S.R., Pyles, R.B., Campbell, G.A., Milligan, G.N., Bourne, N., Barrett, A.D. 2015. A lethal murine infection model for dengue virus 3 in AG129 mice deficient in type I and II interferon receptors leads to systemic disease. J Virol 89(2), 1254-1266. [CrossRef]

Sui, J., Li, W., Roberts, A., Matthews, L.J., Murakami, A., Vogel, L., Wong, S.K., Subbarao, K., Farzan, M., Marasco, W.A. 2005. Evaluation of human monoclonal antibody 80R for immunoprophylaxis of severe acute respiratory syndrome by an animal study, epitope mapping, and analysis of spike variants. J Virol 79, 5900-5906. [CrossRef]

Sumarmo, 1987. Dengue haemorrhagic fever in Indonesia. Southeast Asian J Trop Med Public Health 18, 269-274. Tagliabue, A., Rappuoli, R. 2008. Vaccine adjuvants: the dream becomes real. Hum Vaccine 4, 347-349. [CrossRef]

Wu, S.J., Grouard-Vogel, G., Sun, W., Mascola, J.R., Brachtel, E., Putvatana, R., Louder, M.K., Filgueira, L., Marovich, M.A., Wong, H.K., Blauvelt, A., Murphy, G.S., Robb, M.L., Innes, B.L., Birx, D.L., Hayes, C.G,. Frankel, S.S. 2000. Human skin Langerhans cells are targets of dengue virus infection. Nat Med 6(7), 816-820. [CrossRef]

Yamanaka, A., Mulyatno, K.C., Susilowati, H., Hendrianto, E., Ginting, A.P., Sary, D.D., Rantam, F.A., Soegijanto, S., Konishi, E. 2011. Displacement of the predominant dengue virus from type 2 to type 1 with a subsequent genotype shift from IV to I in Surabaya, Indonesia 2008–2010. PLoS One 6, e27322. [CrossRef]

Yokoyama, T., Ayabe, S., Miyagi, H., Sugano, T., Otsu, A., Sato, H., Kageyama, S., Fujii, T., Shiraki, K. 2001. Varicella-zoster virus gH:gL contains a structure reactive with the anti-human gamma chain of IgG near the glycosylation site. J Gen Virol 82, 331-334. [CrossRef]

Yoshida, T., Yoshida, R., Ma, B.Y., Mikolajczak, S., Kelvin, D.J., Ochi. A. 2010. A novel mitogen fusion protein against CD40+ cells with potent vaccine adjuvant properties. Vaccine 28(21), 3688-3695. [CrossRef]
Acta Veterinaria Eurasia-Cover
  • ISSN: 2618-639X
  • Başlangıç: 1975
  • Yayıncı: İstanbul Üniversitesi-Cerrahpaşa
Sayıdaki Diğer Makaleler

Kerman Shahid Bahonar Üniversitesi Veteriner Hastanesi’ne Getirilen Köpeklerden Toplanan Uterus Örneklerinde Nested Trans-PCR ile C. burnetii Saptanması

Mohammad KHALILI, Mahdieh REZAEI, Farnoush Bakhshaee SHAHRBABAKI, Zeinab ABIRI

Kıvırcık Kuzularda PCR-RFLP Metodu Kullanılarak Calpastatin Geninin MspI ve NcoI Enzimleri ile Genetik Çeşitliliğinin Belirlenmesi

Kozet AVANUS

İran'ın Kuzeybatı Bölgesindeki Küçük Çaplı Yetiştiricilik Sisteminde Sığırlar ile Birlikte Yetiştirilen Manda (Bubalus Bubalis) Yavrularında Gastrointesinal Parazitlerin Prevalansı

Sima VALIPOUR, Mina MAGHSOUDLO, Mousa TAVASSOLI, Bahram DALIR-NAGHADEH

Bir Kedide Birbirlerine Bağlı Çeşitli Organ Sistemlerinin Bozulması Sonucunda Gelişen Perirenal Psöydokist Olgusu

Özge ERDOĞAN BAMAÇ, Utku BAKIREL, Sinem ÜLGEN SAKA

Detection of C. burnetii in Uterine Samples Collected from Referred Dogs to the Veterinary Hospital of Shahid Bahonar University of Kerman by Nested Trans-PCR

Mahdieh REZAEI, Mohammad KHALILI, Farnoush Bakhshaee SHAHRBABAKI, Zeinab ABIRI

Perirenal Pseudocyst in Consequence of Disorders of Several Interdependent Organ Systems in a Cat

Sinem ÜLGEN SAKA, ÖZGE ERDOĞAN BAMAÇ, UTKU BAKIREL

Calvarial Hyperostosis Syndrome in an American Pit Bull Terrier

DAMLA HAKTANIR, Ebru ERAVCI YALIN, YALÇIN DEVECİOĞLU, Alper DEMİRUTKU, AYDIN GÜREL

Tendürek Müflon Populasyonunun Genetik Çeşitliliği

Balal SADEGHI

Genetic Diversity of Tendürek Mouflon Population

Balal SADEGHI

Mezenkimal Kök Hücreler Üzerinde Dengue Virüsüne Karşı (DENV-1,2,3,4) Fab IgG Hibrit Antikorunun Nötralize Edici Potansiyeli

Eryk HENDRIANTO, Fedik A. RANTAM, Rahaju ERNAWATI, Helen SUSILOWATI, Rofiqul A’LA, Rizki Kriestya MAYASAR, Jola RAHMAHANI, Lia Nur AINI