A noninfectious simian/human immunodeficiency virus DNA vaccine that protects macaques against AIDS.
Simian/human immunodeficiency virus SHIV(KU2) replicates with extremely high titers in macaques. In order to determine whether the DNA of the viral genome could be used as a vaccine if the DNA were rendered noninfectious, we deleted the reverse transcriptase gene from SHIVKU2 and inserted this DNA (DeltartSHIVKU2) into a plasmid that was then used to test gene expression and immunogenicity. Transfection of Jurkat and human embryonic kidney epithelial (HEK 293) cells with the DNA resulted in production of all of the major viral proteins and their precursors and transient export of a large quantity of the Gag p27 into the supernatant fluid. As expected, no infectious virus was produced in these cultures. Four macaques were injected intradermally with 2 mg of the DNA at 0, 8, and 18 weeks. The animals developed neutralizing antibodies and low enzyme-linked immunospot assay (E-SPOT) titers against SHIVKU2. These four animals and two unvaccinated control animals were then challenged with heterologous SHIV89.6P administered into their rectums. The two control animals developed viral RNA titers exceeding 10(6) copies/ml of plasma, and these titers were accompanied by the loss of CD4+ T cells by 2 weeks after challenge. The two control animals died at weeks 8 and 16, respectively. All four of the immunized animals became infected with the challenge virus but developed lower titers of viral RNA in plasma than the control animals, and the titers decreased over time in three of the four macaques. The fourth animal remained viremic and died at week 47. Whereas the control animals failed to develop E-SPOT responses, all four of the immunized animals developed anamnestic E-SPOT responses after challenge. The animal that died developed the highest E-SPOT response and was the only one that produced neutralizing antibodies against the challenge virus. These results established that noninfectious DNA of pathogenic SHIV could be used as a vaccine to prevent AIDS, even though the immunological assays used did not predict the manner in which the challenge virus would replicate in the vaccinated animals.
Singh
Dinesh K
DK
Marion Merrell Dow Laboratory of Viral Pathogenesis, The University of Kansas Medical Center, 3901 Rainbow Blvd., Kansas City, KS 66160, USA.
Liu
Zhenqian
Z
Sheffer
Darlene
D
Mackay
Glenn A
GA
Smith
Marilyn
M
Dhillon
Sukhbir
S
Hegde
Ramakrishna
R
Jia
Fenglan
F
Adany
Istvan
I
Narayan
Opendra
O
D016915:AIDS Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D000163:Acquired Immunodeficiency Syndrome / Q000517:prevention & control*; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018791:CD4 Lymphocyte Count; D036022:Capsid Proteins / Q000378:metabolism; D002478:Cells, Cultured; D017353:Gene Deletion; D015683:Gene Products, gag / Q000378:metabolism; D006678:HIV / Q000276:immunology*; D015483:HIV Antibodies / Q000096:biosynthesis; D054303:HIV Reverse Transcriptase / Q000235:genetics; D006801:Humans; D007271:Injections, Intradermal; D008214:Lymphocytes / Q000276:immunology; D008254:Macaca nemestrina; D009500:Neutralization Tests; D012367:RNA, Viral / Q000097:blood; D018449:SAIDS Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D016097:Simian Acquired Immunodeficiency Syndrome / Q000517:prevention & control*; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D014162:Transfection; D014611:Vaccination; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000096:biosynthesis; D014766:Viremia; D054301:gag Gene Products, Human Immunodeficiency Virus
D016915:AIDS Vaccines; D000914:Antibodies, Viral; D036022:Capsid Proteins; D015683:Gene Products, gag; D015483:HIV Antibodies; D012367:RNA, Viral; D018449:SAIDS Vaccines; D019444:Vaccines, DNA; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C074737:p27 gag protein, Human immunodeficiency virus; D054303:HIV Reverse Transcriptase
J Biol Regul Homeost Agents. 2000 Jan-Mar;14(1):22-6
10763887
J Virol. 2003 Jun;77(11):6305-13
12743287
AIDS Res Hum Retroviruses. 2000 Sep 1;16(13):1247-57
10957722
Nat Med. 2000 Oct;6(10):1094-5
11017132
Virology. 2001 Jan 5;279(1):241-56
11145906
Science. 2001 Apr 6;292(5514):69-74
11393868
J Med Primatol. 2001 Aug;30(4):190-6
11555137
J Virol. 2001 Oct;75(20):9771-9
11559810
J Infect Dis. 2001 Oct 15;184(8):1007-14
11574915
Immunol Lett. 2001 Nov 1;79(1-2):57-61
11595290
Transfus Clin Biol. 2001 Aug;8(4):350-8
11642027
Dev Biol (Basel). 2000;104:93-100
11713830
Vaccine. 2002 May 6;20(15):1949-55
11983252
Vaccine. 2002 May 6;20(15):1956-60
11983253
Virology. 2002 Mar 15;294(2):270-81
12009868
J Virol. 2002 Jun;76(12):6138-46
12021347
Virology. 2002 Mar 30;295(1):133-46
12033772
Vaccine. 2002 Sep 10;20(27-28):3322-31
12213402
Nat Med. 2003 Jul;9(7):847-52
12835704
AIDS. 2004 May 21;18(8):1213-6
15166541
FASEB J. 2004 Sep;18(12):1459-61
15240562
J Immunol. 2004 Sep 15;173(6):4100-7
15356160
Nature. 1992 Mar 12;356(6365):152-4
1545867
Science. 1992 Dec 18;258(5090):1938-41
1470917
Virology. 1993 Aug;195(2):406-12
8337820
J Virol. 1996 May;70(5):3189-97
8627799
Nat Med. 1996 Mar;2(3):338-42
8612235
J Virol. 1996 Jun;70(6):3724-33
8648707
AIDS Res Hum Retroviruses. 1997 May 20;13(8):635-45
9168232
Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9378-83
9256490
J Med Primatol. 1997 Feb-Apr;26(1-2):51-8
9271189
J Virol. 1997 Oct;71(10):7711-8
9311855
Science. 1998 Apr 17;280(5362):427-31
9545219
AIDS Res Hum Retroviruses. 1998 Aug 10;14(12):1035-43
9718118
J Virol. 1998 Oct;72(10):7852-9
9733822
J Virol. 1998 Nov;72(11):9069-78
9765452
AIDS Res Hum Retroviruses. 1998 Oct;14 Suppl 3:S255-9
9814952
Virology. 2002 Sep 15;301(1):130-5
12359453
J Virol. 1998 Dec;72(12):10281-5
9811776
J Exp Med. 1998 Dec 21;188(12):2199-204
9858506
AIDS Res Hum Retroviruses. 1999 Mar 1;15(4):391-4
10082123
J Virol. 1999 May;73(5):4009-18
10196297
J Virol. 1999 Jun;73(6):4952-61
10233957
Virology. 1999 Aug 1;260(2):295-307
10417264
AIDS. 2002 Nov 8;16(16):2137-43
12409734
J Virol. 2002 Dec;76(23):12173-84
12414957
J Virol. 2003 Jan;77(2):989-98
12502815
Cell. 2002 Dec 13;111(6):853-66
12526811
Virology. 2003 Feb 15;306(2):334-46
12642106
Virology. 2000 Aug 15;274(1):149-64
10936096