title: A noninfectious simian/human immunodeficiency virus DNA vaccine that protects macaques against AIDS.
abstract: 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.
authors.affiliation: Marion Merrell Dow Laboratory of Viral Pathogenesis, The University of Kansas Medical Center, 3901 Rainbow Blvd., Kansas City, KS 66160, USA.
authors.forename: Dinesh K
authors.initials: DK
authors.lastname: Singh
authors.forename: Zhenqian
authors.initials: Z
authors.lastname: Liu
authors.forename: Darlene
authors.initials: D
authors.lastname: Sheffer
authors.forename: Glenn A
authors.initials: GA
authors.lastname: Mackay
authors.forename: Marilyn
authors.initials: M
authors.lastname: Smith
authors.forename: Sukhbir
authors.initials: S
authors.lastname: Dhillon
authors.forename: Ramakrishna
authors.initials: R
authors.lastname: Hegde
authors.forename: Fenglan
authors.initials: F
authors.lastname: Jia
authors.forename: Istvan
authors.initials: I
authors.lastname: Adany
authors.forename: Opendra
authors.initials: O
authors.lastname: Narayan
mesh_terms: 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
chemical_list: 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
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