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研究生：	鄭亦淩 Cheng, Yi-Ling
論文名稱：	探討新冠病毒棘蛋白受體結合區與登革病毒蛋白間的抗原相似性 Study on the Antigenic Similarity between SARS-CoV-2 S1-RBD and Dengue Virus Proteins
指導教授：	葉才明 Yeh, Trai-Ming
學位類別：	碩士 Master
系所名稱：	醫學院 - 醫學檢驗生物技術學系 Department of Medical Laboratory Science and Biotechnology
論文出版年：	2022
畢業學年度：	110
語文別：	英文
論文頁數：	66
中文關鍵詞：	新冠病毒、登革病毒、快篩試驗、抗原相似性、抗體依賴性增強
外文關鍵詞：	COVID-19, dengue virus, SARS-CoV-2, diagnosis, antibody-dependent enhancement
相關次數：	點閱：143 下載：3
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嚴重急性呼吸道症候群冠狀病毒2型，以下簡稱為新冠病毒，自西元2019年12月造成全球性大流行。多項研究指出新冠病毒感染可能會在登革病毒血清檢測中造成假陽性反應，反之亦然；這代表新冠病毒與登革病毒感染人體後，可能產生具有交叉反應的抗體。目前針對這些交叉反應抗體在新冠病毒或登革病毒感染中是否具有保護力或反而會增加疾病嚴重程度尚不明瞭。在本篇研究中，我們發現登革病患血清中對新冠病毒棘蛋白受體結合區(SARS-CoV-2 S1-RBD)具有交叉反應的抗體顯著高於健康對照組。接著，我們從SARS-CoV-2 S1-RBD重組蛋白免疫的兔血清中純化出抗SARS-CoV-2 S1-RBD免疫球蛋白G (anti-SARS-CoV-2 S1-RBD IgG)，結果顯示，anti-SARS-CoV-2 S1-RBD IgG可以與登革病毒膜蛋白與非結構蛋白1產生交叉反應；我們也利用噬菌體表達隨機胜肽庫系統進一步找出anti-SARS-CoV-2 S1-RBD IgG辨認到登革病毒膜蛋白與非結構蛋白1的潛在抗原決定區。在細胞實驗結果中，我們發現anti-SARS-CoV-2 S1-RBD IgG能夠有效抑制登革病毒感染與登革病毒非結構蛋白1所造成的血管內皮細胞滲漏。而在登革感染小鼠模式的結果顯示，給予anti-SARS-CoV-2 S1-RBD IgG能夠有效減緩登革病毒感染所造成的小鼠出血時間延長，並降低血清中登革病毒非結構蛋白1的濃度。最後，我們也在某些新冠病患的個體中發現具有與登革病毒蛋白交叉反應的抗體存在，且新冠病患血清組相較於健康對照組的血清能夠有效抑制登革病毒感染。綜上所述，本研究證實SARS-CoV-2 S1-RBD能透過與登革病毒蛋白的抗原相似性產生對登革病毒具有交叉反應的抗體，並減緩登革病毒感染所造成的致病機轉。

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread globally since December 2019. Several studies have reported that SARS-CoV-2 infections may produce false-positive reactions in dengue virus (DENV) serology tests and vice versa. However, it remains unclear whether SARS-CoV-2 and DENV cross-reactive antibodies provide cross-protection against each other disease or promote disease severity. In this study, we confirmed that antibodies against the SARS-CoV-2 spike protein and its receptor-binding domain (S1-RBD) were significantly increased in dengue patients compared to normal controls. In addition, anti-SARS-CoV-2 S1-RBD IgG purified from anti-S1-RBD hyperimmune rabbit serum could cross-react with both DENV envelope protein (E) and nonstructural protein 1 (NS1). The potential epitopes of DENV E and NS1 recognized by these antibodies were identified by a phage-displayed random peptide library. In addition, DENV infection and DENV NS1-induced endothelial hyperpermeability in vitro were inhibited in the presence of anti-SARS-CoV-2 S1-RBD IgG. Passive transfer of anti-SARS-CoV-2 S1-RBD IgG into mice also reduced the prolonged bleeding time and decreased the serum NS1 levels in DENV-infected mice. Finally, DENV cross-reactive antibodies were found in some individuals after SARS-CoV-2 infection, and COVID-19 patients’ sera could inhibit DENV infection compared to normal controls in vitro. Thus, our results suggest that the antigenic cross-reactivity between the SARS-CoV-2 RBD and DENV can induce the production of anti-SARS-CoV-2 S1-RBD antibodies that cross-react with DENV, which may hinder dengue pathogenesis.

中文摘要 I
Abstract II
Table of Contents III
List of figures VII
Abbreviations Index VIII
Introduction 1
1 SARS-CoV-2 1
1.1 Clinical symptoms of SARS-CoV-2 infection 1
1.2 Genome structure of SARS-CoV-2 1
1.3 SARS-CoV-2 spike protein 2
1.4 Development of SARS-CoV-2 vaccines 2
2 Dengue virus (DENV) 4
2.1 Epidemiology of DENV infection 4
2.2 Clinical symptoms of DENV infection 4
2.3 Genome structure of DENV 4
2.4 DENV envelope (E) protein 5
2.5 DENV precursor membrane (prM) protein 5
2.6 DENV nonstructural protein 1 (NS1) 6
2.7 DENV pathogenesis	6
2.8 Diagnostic methods of DENV 7
3 The cross-reacting antibodies between SARS-CoV-2 and DENV 7
Objective and Specific Aims 9
Material and Methods 11
1 Materials 11
1.1 Cell lines 11
1.2 Antibodies 11
1.3 Recombinant proteins and synthetic peptides	12
1.4 Primers 13
1.5 Plasmids 13
1.6 Reagents 13
1.7 Beads 15
1.8 Medium 15
1.9 Antibiotics 15
1.10 Kits 16
1.11 Consumables and Devices 16
1.12 Instruments 17
2 Methods	18
2.1 Gibson assembly 18
2.2 Expression and Purification of SARS-CoV-2 S1-RBD Recombinant Protein	18
2.3 Reverse transcription PCR (RT-PCR) 18
2.4 ACE2 activity test 19
2.5 Immunization and antibody purification 19
2.6 Enzyme-Linked Immunosorbent Assay (ELISA) 20
2.7 Western Blotting 21
2.8 SARS-CoV-2 Pseudovirus Neutralization Test 21
2.9 Viral stocks	22
2.10 Immunofluorescence Assay	22
2.11 Epitope Mapping using a Phage-Display Random Peptide Library	23
2.12 Fluorescent Focus Assay (FFA) and Focus Reduction Neutralization Test (FRNT) 23
2.13 Antibody-Dependent Enhancement (ADE) 24
2.14 Transwell Permeability Assay 24
2.15 Mouse Model of DENV Infection 25
2.16 Bleeding Time	25
2.17 NS1 Quantitative ELISA 26
2.18 Human serum collection 26
2.19 Statistical Analysis 27
Results 28
1 Cloning and functional test of SARS-CoV-2 S1-RBD 28
1.1 Construction of SARS-CoV-2 S1 RBD 28
1.2 Expression and purification of SARS-CoV-2 S1-RBD protein 28
1.3 SARS-CoV-2 S1-RBD protein binds to ACE2 in the human Caco-2 cell line 29
1.4 SARS-CoV-2 S1-RBD protein enhances ACE2 activity 29
2 Anti-SARS-CoV-2 S1-RBD IgG purification and its cross-reactivity to DENV proteins 30
2.1 Antibodies Against the SARS-CoV-2 Spike Protein in Archived Dengue Patient Sera 30
2.2 Anti-SARS-CoV-2 S1-RBD IgG Purification and Characterization 30
2.3 Anti-SARS-CoV-2 S1-RBD IgG Inhibits SARS-CoV-2 S1-RBD Binding to Recombinant ACE2 Protein in a Competitive ELISA and SARS-CoV-2 Pseudovirus Infection 31
2.4 Anti-SARS-CoV-2 S1-RBD IgG Cross-Reacts with DENV Proteins 32
3 Identification of the cross-reacting sequences/epitopes in different DENV proteins recognized by anti-SARS-CoV-2 S1-RBD IgG 32
3.1 Identification of Sequences/Epitopes Recognized by Anti-SARS-CoV-2 S1-RBD IgG using a Phage-Displayed 12-mer Random Peptide Library Kit 32
3.2 The Peptide TQFEKASVNTTR Competitively Inhibits Anti-SARS-CoV-2 S1-RBD IgG Binding to the SARS-CoV-2 S1-RBD and DENV E Protein 33
4 Influence of anti-SARS-CoV-2 S1-RBD IgG on DENV infection 34
4.1 Anti-SARS-CoV-2 S1-RBD IgG Inhibits DENV Infection and NS1-Induced Endothelial Hyperpermeability without Causing ADE In Vitro 34
4.2 Anti-SARS-CoV-2 S1-RBD IgG Protects Mice from DENV-Induced Prolonged Bleeding Time and Decreases NS1 Levels in Mouse Sera 35
4.3 Possible epitope of DENV NS1 recognized by anti-SARS-CoV-2 S1-RBD IgG 35
4.4 COVID-19 patients’ sera inhibit DENV infection in vitro 36
Discussion 37
1 The impact of antibody cross-reactivity between SARS-CoV-2 and DENV on the diagnosis of both diseases 37
2 The impact of antibody cross-reactivity between SARS-CoV-2 and DENV on the severity of both diseases 38
3 Anti-SARS-CoV-2 S1-RBD IgG cross-reactivity with DENV E 39
4 Anti-SARS-CoV-2 S1-RBD IgG cross-reactivity with DENV NS1 40
Conclusion 41
Reference	42
Figures 47
Appendix 63
                                    

Alen, M. M., K. Dallmeier, J. Balzarini, J. Neyts and D. Schols (2012). "Crucial role of the N-glycans on the viral E-envelope glycoprotein in DC-SIGN-mediated dengue virus infection." Antiviral Res 96(3): 280-287.
Avirutnan, P., N. Punyadee, S. Noisakran, C. Komoltri, S. Thiemmeca, K. Auethavornanan, A. Jairungsri, R. Kanlaya, N. Tangthawornchaikul and C. Puttikhunt (2006). "Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement." J. Infect. Dis. 193(8): 1078-1088.
Beatty, P. R., H. Puerta-Guardo, S. S. Killingbeck, D. R. Glasner, K. Hopkins and E. Harris (2015). "Dengue virus NS1 triggers endothelial permeability and vascular leak that is prevented by NS1 vaccination." Sci. Transl. Med. 7(304): 304ra141-304ra141.
Bokhari, S., F. Mahmood and S. Bokhari (2020). "Case Report: Diagnosis of COVID-19 versus Tropical Diseases in Pakistan." Am J Trop Med Hyg 103(1): 77-78.
Cao, X. (2020). "COVID-19: immunopathology and its implications for therapy." Nat Rev Immunol 20(5): 269-270.
Chen, H. R., Y. C. Chuang, Y. S. Lin, H. S. Liu, C. C. Liu, G. C. Perng and T. M. Yeh (2016). "Dengue Virus Nonstructural Protein 1 Induces Vascular Leakage through Macrophage Migration Inhibitory Factor and Autophagy." PLoS Negl Trop Dis 10(7): e0004828.
Costin, J. M., E. Zaitseva, K. M. Kahle, C. O. Nicholson, D. K. Rowe, A. S. Graham, L. E. Bazzone, G. Hogancamp, M. F. Sierra, R. H. Fong, S.-T. Yang, L. Lin, J. E. Robinson, B. J. Doranz, L. V. Chernomordik, S. F. Michael, J. S. Schieffelin and S. Isern (2013). "Mechanistic Study of Broadly Neutralizing Human Monoclonal Antibodies against Dengue Virus That Target the Fusion Loop." J. Virol. 87(1): 52-66.
Crill, W. D. and J. T. Roehrig (2001). "Monoclonal antibodies that bind to domain III of dengue virus E glycoprotein are the most efficient blockers of virus adsorption to Vero cells." J. Virol. 75(16): 7769-7773.
El-Qushayri, A. E., A. M. A. Kamel, A. Reda and S. Ghozy (2022). "Does dengue and COVID-19 co-infection have worse outcomes? A systematic review of current evidence." Rev Med Virol: e2339.
Halstead, S. B. (1989). "Antibody, macrophages, dengue virus infection, shock, and hemorrhage: a pathogenetic cascade." Rev Infect Dis 11 Suppl 4: S830-839.
Henchal, E. A. and J. R. Putnak (1990). "The dengue viruses." Clin Microbiol Rev 3(41): 376-396.
Hoffmann, M., H. Kleine-Weber, S. Schroeder, N. Krüger, T. Herrler, S. Erichsen, T. S. Schiergens, G. Herrler, N. H. Wu, A. Nitsche, M. A. Müller, C. Drosten and S. Pöhlmann (2020). "SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor." Cell 181(2): 271-280.e278.
Huang, Y., C. Yang, X. F. Xu, W. Xu and S. W. Liu (2020). "Structural and functional properties of SARS-CoV-2 spike protein: potential antivirus drug development for COVID-19." Acta Pharmacol Sin 41(9): 1141-1149.
Khairunisa, S. Q., I. H. Amarullah, S. Churrotin, A. L. Fitria, M. Amin, M. I. Lusida and S. Soegijanto (2021). "Potential Misdiagnosis between COVID-19 and Dengue Infection Using Rapid Serological Test." Infect Dis Rep 13(2): 540-551.
Krammer, F. (2020). "SARS-CoV-2 vaccines in development." Nature 586(7830): 516-527.
Kuhn, R. J., W. Zhang, M. G. Rossmann, S. V. Pletnev, J. Corver, E. Lenches, C. T. Jones, S. Mukhopadhyay, P. R. Chipman and E. G. Strauss (2002). "Structure of dengue virus: implications for flavivirus organization, maturation, and fusion." Cell 108(5): 717-725.
Lai, Y. C., Y. C. Chuang, C. C. Liu, T. S. Ho, Y. S. Lin, R. Anderson and T. M. Yeh (2017). "Antibodies Against Modified NS1 Wing Domain Peptide Protect Against Dengue Virus Infection." Sci Rep 7(1): 6975.
Lee, W. S., A. K. Wheatley, S. J. Kent and B. J. DeKosky (2020). "Antibody-dependent enhancement and SARS-CoV-2 vaccines and therapies." Nat Microbiol 5(10): 1185-1191.
Lei, H. Y., T. M. Yeh, H. S. Liu, Y. S. Lin, S. H. Chen and C. C. Liu (2001). "Immunopathogenesis of dengue virus infection." J Biomed Sci 8(51): 377-388.
Li, L., S. M. Lok, I. M. Yu, Y. Zhang, R. J. Kuhn, J. Chen and M. G. Rossmann (2008). "The flavivirus precursor membrane-envelope protein complex: structure and maturation." Science 319(5871): 1830-1834.
Libraty, D. H., P. R. Young, D. Pickering, T. P. Endy, S. Kalayanarooj, S. Green, D. W. Vaughn, A. Nisalak, F. A. Ennis and A. L. Rothman (2002). "High Circulating Levels of the Dengue Virus Nonstructural Protein NS1 Early in Dengue Illness Correlate with the Development of Dengue Hemorrhagic Fever." J. Infect. Dis. 186(8): 1165-1168.
Lu, J. and P. D. Sun (2020). "High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity." J Biol Chem. 295(52):18579-18588
Luo, Y.-Y., J.-J. Feng, J.-M. Zhou, Z.-Z. Yu, D.-Y. Fang, H.-J. Yan, G.-C. Zeng and L.-F. Jiang (2013). "Identification of a novel infection-enhancing epitope on dengue prM using a dengue cross-reacting monoclonal antibody." BMC Microbiol. 13(1): 194.
Lustig, Y., S. Keler, R. Kolodny, N. Ben-Tal, D. Atias-Varon, E. Shlush, M. Gerlic, A. Munitz, R. Doolman, K. Asraf, L. I. Shlush and A. Vivante (2020). "Potential antigenic cross-reactivity between SARS-CoV-2 and Dengue viruses." Clin Infect Dis. 73(7): 2444-2449.
Martina, B. E., P. Koraka and A. D. Osterhaus (2009). "Dengue virus pathogenesis: an integrated view." Clin Microbiol Rev 22(4): 564-581.
Messina, J. P., O. J. Brady, N. Golding, M. U. G. Kraemer, G. R. W. Wint, S. E. Ray, D. M. Pigott, F. M. Shearer, K. Johnson, L. Earl, L. B. Marczak, S. Shirude, N. Davis Weaver, M. Gilbert, R. Velayudhan, P. Jones, T. Jaenisch, T. W. Scott, R. C. Reiner, Jr. and S. I. Hay (2019). "The current and future global distribution and population at risk of dengue." Nat Microbiol 4(9): 1508-1515.
Modhiran, N., D. Watterson, D. A. Muller, A. K. Panetta, D. P. Sester, L. Liu, D. A. Hume, K. J. Stacey and P. R. Young (2015). "Dengue virus NS1 protein activates cells via Toll-like receptor 4 and disrupts endothelial cell monolayer integrity." Sci Transl Med 7(304): 304ra142.
Modis, Y., S. Ogata, D. Clements and S. C. Harrison (2003). "A ligand-binding pocket in the dengue virus envelope glycoprotein." PNAS 100(12): 6986-6991.
Nath, H., A. Mallick, S. Roy, S. Sukla, K. Basu, A. De and S. Biswas (2021). "Archived dengue serum samples produced false-positive results in SARS-CoV-2 lateral flow-based rapid antibody tests." J Med Microbiol 70(6): 001369.
Nath, H., A. Mallick, S. Roy, S. Sukla and S. Biswas (2021). "Computational modelling supports that dengue virus envelope antibodies can bind to SARS-CoV-2 receptor binding sites: Is pre-exposure to dengue virus protective against COVID-19 severity?" Comput Struct Biotechnol J 19: 459-466.
Nicolete, V. C., P. T. Rodrigues, I. C. Johansen, R. M. Corder, J. Tonini, M. A. Cardoso, J. G. de Jesus, I. M. Claro, N. R. Faria, E. C. Sabino, M. C. Castro and M. U. Ferreira (2021). "Interacting Epidemics in Amazonian Brazil: Prior Dengue Infection Associated With Increased Coronavirus Disease 2019 (COVID-19) Risk in a Population-Based Cohort Study." Clin Infect Dis 73(11): 2045-2054.
Nunthavichitra, S., S. Prapaso, V. Luvira, S. Muangnoicharoen, P. Leaungwutiwong and W. Piyaphanee (2020). "Case Report: COVID-19 Presenting as Acute Undifferentiated Febrile Illness-A Tropical World Threat." Am J Trop Med Hyg 103(1): 83-85.
Op De Beeck, A., R. Molenkamp, M. Caron, A. Ben Younes, P. Bredenbeek and J. Dubuisson (2003). "Role of the transmembrane domains of prM and E proteins in the formation of yellow fever virus envelope." J Virol 77(2): 813-820.
Pokidysheva, E., Y. Zhang, A. J. Battisti, C. M. Bator-Kelly, P. R. Chipman, C. Xiao, G. G. Gregorio, W. A. Hendrickson, R. J. Kuhn and M. G. Rossmann (2006). "Cryo-EM reconstruction of dengue virus in complex with the carbohydrate recognition domain of DC-SIGN." Cell 124(3): 485-493.
Prapty, C., R. Rahmat, Y. Araf, S. K. Shounak, A. A. Noor, T. I. Rahaman, M. J. Hosen, C. Zheng and M. G. Hossain (2022). "SARS-CoV-2 and dengue virus co-infection: Epidemiology, pathogenesis, diagnosis, treatment, and management." Rev Med Virol: e2340.
Premkumar, L., B. Segovia-Chumbez, R. Jadi, D. R. Martinez, R. Raut, A. Markmann, C. Cornaby, L. Bartelt, S. Weiss, Y. Park, C. E. Edwards, E. Weimer, E. M. Scherer, N. Rouphael, S. Edupuganti, D. Weiskopf, L. V. Tse, Y. J. Hou, D. Margolis, A. Sette, M. H. Collins, J. Schmitz, R. S. Baric and A. M. de Silva (2020). "The receptor binding domain of the viral spike protein is an immunodominant and highly specific target of antibodies in SARS-CoV-2 patients." Sci Immunol 5(48).
Rana, M. S., M. Usman, M. M. Alam, A. Ikram, M. Salman and M. Umair (2022). "Impact of previous SARS-CoV-2 infection on the rate of mortality in dengue. A preliminary report from Pakistan." J Infect. 84(5): 722-746.
Ravikumar, N., M. S. Randhawa, K. Nallasamy, S. K. Angurana, M. Kumar, G. K. Mohi, R. K. Ratho and M. Jayashree (2021). "Impact of Recent SARS-CoV-2 Infection on the Course and Severity of Dengue in Children: A Prospective Observational Study from North India." Am J Trop Med Hyg 105(3): 751-755.
Rothman, A. L. (2004). "Dengue: defining protective versus pathologic immunity." J Clin Invest 113(7): 946-951.
Santoso, M. S., S. Masyeni, S. Haryanto, B. Yohan, M. L. Hibberd and R. T. Sasmono (2021). "Assessment of dengue and COVID-19 antibody rapid diagnostic tests cross-reactivity in Indonesia." Virol J 18(1): 54.
Shang, J., G. Ye, K. Shi, Y. Wan, C. Luo, H. Aihara, Q. Geng, A. Auerbach and F. Li (2020). "Structural basis of receptor recognition by SARS-CoV-2." Nature 581(7807): 221-224.
Thomas, S. J. (2015). "NS1: A corner piece in the dengue pathogenesis puzzle?" Sci Transl Med 7(304): 304fs337.
Ulrich, H., M. M. Pillat and A. Tarnok (2020). "Dengue Fever, COVID-19 (SARS-CoV-2), and Antibody-Dependent Enhancement (ADE): A Perspective." Cytometry.
Vabret, N., G. J. Britton, C. Gruber, S. Hegde, J. Kim, M. Kuksin, R. Levantovsky, L. Malle, A. Moreira, M. D. Park, L. Pia, E. Risson, M. Saffern, B. Salome, M. Esai Selvan, M. P. Spindler, J. Tan, V. van der Heide, J. K. Gregory, K. Alexandropoulos, N. Bhardwaj, B. D. Brown, B. Greenbaum, Z. H. Gumus, D. Homann, A. Horowitz, A. O. Kamphorst, M. A. Curotto de Lafaille, S. Mehandru, M. Merad, R. M. Samstein and P. Sinai Immunology Review (2020). "Immunology of COVID-19: Current State of the Science." Immunity 52(6): 910-941
Vanroye, F., D. V. D. Bossche, I. Brosius, B. Tack, M. V. Esbroeck and J. Jacobs (2021). "COVID-19 Antibody Detecting Rapid Diagnostic Tests Show High Cross-Reactivity When Challenged with Pre-Pandemic Malaria, Schistosomiasis and Dengue Samples." Diagnostics 11(7), 1163.
Walls, A. C., Y. J. Park, M. A. Tortorici, A. Wall, A. T. McGuire and D. Veesler (2020). "Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein." Cell 181(2): 281-292.e286.
Wan, S. W., P. W. Chen, C. Y. Chen, Y. C. Lai, Y. T. Chu, C. Y. Hung, H. Lee, H. F. Wu, Y. C. Chuang, J. Lin, C. P. Chang, S. Wang, C. C. Liu, T. S. Ho, C. F. Lin, C. K. Lee, B. A. Wu-Hsieh, R. Anderson, T. M. Yeh and Y. S. Lin (2017). "Therapeutic Effects of Monoclonal Antibody against Dengue Virus NS1 in a STAT1 Knockout Mouse Model of Dengue Infection." J Immunol 199(8): 2834-2844.
World Health, O. (2009). Dengue guidelines for diagnosis, treatment, prevention and control : new edition. Geneva, World Health Organization.
Yan, G., C. K. Lee, L. T. M. Lam, B. Yan, Y. X. Chua, A. Y. N. Lim, K. F. Phang, G. S. Kew, H. Teng, C. H. Ngai, L. Lin, R. M. Foo, S. Pada, L. C. Ng and P. A. Tambyah (2020). "Covert COVID-19 and false-positive dengue serology in Singapore." Lancet Infect Dis 20(5): 536.
Yang, Z.-y., W.-p. Kong, Y. Huang, A. Roberts, B. R. Murphy, K. Subbarao and G. J. Nabel (2004). "A DNA vaccine induces SARS coronavirus neutralization and protective immunity in mice." Nature 428(6982): 561-564.

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