Recombinant receptor-binding domain of SARS-CoV spike protein expressed in mammalian, insect and E. coli cells elicits potent neutralizing antibody and protective immunity.
Severe acute respiratory syndrome (SARS) is a newly emerging infectious disease. The potential recurrence of the disease from animal reservoirs highlights the significance of development of safe and efficient vaccines to prevent a future SARS epidemic. In this study, we expressed the recombinant receptor-binding domain (rRBD) in mammalian (293T) cells, insect (Sf9) cells, and E. coli, respectively, and compared their immunogenicity and protection against SARS-CoV infection in an established mouse model. Our results show that all rRBD proteins expressed in the above systems maintained intact conformation, being able to induce highly potent neutralizing antibody responses and complete protective immunity against SARS-CoV challenge in mice, albeit the rRBD expressed in 293T cells elicited stronger humoral immune responses with significantly higher neutralizing activity (P<0.05) than those expressed in Sf9 and E. coli cells. These results suggest that all three rRBDs are effective in eliciting immune responses and protection against SARS-CoV and any of the above expression systems can be used for production of rRBD-based SARS subunit vaccines. Preference will be given to rRBD expressed in mammalian cells for future evaluation of the vaccine efficacy in a non-human primate model of SARS because of its ability to refold into a native conformation more readily and to induce higher level of neutralizing antibody responses than those expressed in E. coli and insect cells.
Du
Lanying
L
Lindsley F. Kimball Research Institute, New York Blood Center, New York, NY 10065, USA.
Zhao
Guangyu
G
Chan
Chris C S
CC
Sun
Shihui
S
Chen
Min
M
Liu
Zhonghua
Z
Guo
Hongxiang
H
He
Yuxian
Y
Zhou
Yusen
Y
Zheng
Bo-Jian
BJ
Jiang
Shibo
S
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D007313:Insecta; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000276:immunology / Q000517:prevention & control*; D064370:Spike Glycoprotein, Coronavirus; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines
Vaccine. 2007 Jul 9;25(28):5220-31
17559989
Emerg Infect Dis. 2005 Aug;11(8):1312-4
16110580
Vaccine. 2006 Jun 29;24(26):5498-508
16725238
Cell Mol Life Sci. 2004 Nov;61(21):2738-43
15549175
Methods Mol Biol. 2009;497:211-21
19107420
Cell. 2003 Dec 12;115(6):652-3
14675530
Protein Expr Purif. 2006 Oct;49(2):211-8
16631378
Immunol Lett. 2008 Aug 15;119(1-2):71-7
18533276
Viral Immunol. 2006 Summer;19(3):518-24
16987069
Annu Rev Med. 2005;56:357-81
15660517
Biochem Biophys Res Commun. 2004 Nov 12;324(2):773-81
15474494
Nature. 2004 Apr 1;428(6982):561-4
15024391
J Immunol. 2008 Jan 15;180(2):948-56
18178835
Vaccine. 2006 Apr 24;24(17):3624-31
16497416
Vaccine. 2007 Aug 21;25(34):6334-40
17640780
Vaccine. 2008 Mar 20;26(13):1644-51
18289745
J Biotechnol. 2007 Jan 10;127(3):335-47
16959350
Biochem Biophys Res Commun. 2007 Jul 20;359(1):174-9
17533109
Vaccine. 2007 Jan 2;25(1):136-43
16919855
Clin Vaccine Immunol. 2007 Jul;14(7):894-901
17494640
Protein Sci. 2006 Jan;15(1):182-9
16322573
Science. 2003 Oct 10;302(5643):276-8
12958366
J Biol Chem. 2004 May 14;279(20):20836-49
14996844
Nat Rev Microbiol. 2009 Mar;7(3):226-36
19198616
J Biol Chem. 2004 Jan 30;279(5):3197-201
14670965
Nature. 2003 Nov 27;426(6965):450-4
14647384
J Antimicrob Chemother. 2005 Nov;56(5):919-22
16183666
Virology. 2006 Sep 15;353(1):6-16
16793110
Biochem Biophys Res Commun. 2003 Dec 26;312(4):1159-64
14651994
J Virol. 2004 Nov;78(22):12672-6
15507655
Vaccine. 2005 Mar 18;23(17-18):2273-9
15755610
J Biol Chem. 2004 Nov 19;279(47):49414-9
15345712
Vaccine. 2007 Apr 12;25(15):2832-8
17092615
Biochem Biophys Res Commun. 2004 Jun 18;319(1):283-8
15158473
Lancet. 2003 Oct 25;362(9393):1353-8
14585636
Lancet. 2004 Mar 20;363(9413):938-47
15043961
Intervirology. 2008;51(3):189-95
18724066
J Immunol. 2006 May 15;176(10):6085-92
16670317
Biochem Biophys Res Commun. 2004 Jan 30;314(1):235-41
14715271
Biochem Biophys Res Commun. 2009 Jul 10;384(4):486-90
19422787
J Virol. 2006 Apr;80(8):4079-87
16571824
Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14040-5
16169905
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6641-6
15096611
J Virol. 2005 Sep;79(18):11892-900
16140765
Pharmacol Rev. 2006 Sep;58(3):621-81
16968952
Science. 2003 May 30;300(5624):1394-9
12730500
Lancet. 2003 Dec 6;362(9399):1895-6
14667748
J Immunol. 2005 Apr 15;174(8):4908-15
15814718
Retrovirology. 2007 May 17;4:33
17509143
Vaccine. 2008 Apr 16;26(17):2164-74
18346823
J Virol. 2004 May;78(9):4552-60
15078936
Science. 2003 May 30;300(5624):1399-404
12730501
Science. 2005 Oct 28;310(5748):676-9
16195424
Vaccine. 2008 Nov 25;26(50):6338-43
18824060
Vaccine. 2006 Jun 5;24(23):4905-13
16621188
J Immunol Methods. 2005 Jan;296(1-2):37-44
15680149
Vaccine. 2009 Feb 5;27(6):857-63
19084043