title: HIV-1 p24(gag) derived conserved element DNA vaccine increases the breadth of immune response in mice.
abstract: Viral diversity is considered a major impediment to the development of an effective HIV-1 vaccine. Despite this diversity, certain protein segments are nearly invariant across the known HIV-1 Group M sequences. We developed immunogens based on the highly conserved elements from the p24(gag) region according to two principles: the immunogen must (i) include strictly conserved elements of the virus that cannot mutate readily, and (ii) exclude both HIV regions capable of mutating without limiting virus viability, and also immunodominant epitopes located in variable regions. We engineered two HIV-1 p24(gag) DNA immunogens that express 7 highly Conserved Elements (CE) of 12-24 amino acids in length and differ by only 1 amino acid in each CE ('toggle site'), together covering >99% of the HIV-1 Group M sequences. Altering intracellular trafficking of the immunogens changed protein localization, stability, and also the nature of elicited immune responses. Immunization of C57BL/6 mice with p55(gag) DNA induced poor, CD4(+) mediated cellular responses, to only 2 of the 7 CE; in contrast, vaccination with p24CE DNA induced cross-clade reactive, robust T cell responses to 4 of the 7 CE. The responses were multifunctional and composed of both CD4(+) and CD8(+) T cells with mature cytotoxic phenotype. These findings provide a method to increase immune response to universally conserved Gag epitopes, using the p24CE immunogen. p24CE DNA vaccination induced humoral immune responses similar in magnitude to those induced by p55(gag), which recognize the virus encoded p24(gag) protein. The inclusion of DNA immunogens composed of conserved elements is a promising vaccine strategy to induce broader immunity by CD4(+) and CD8(+) T cells to additional regions of Gag compared to vaccination with p55(gag) DNA, achieving maximal cross-clade reactive cellular and humoral responses.
authors.affiliation: Human Retrovirus Pathogenesis Section, Center for Cancer Research, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
authors.forename: Viraj
authors.initials: V
authors.lastname: Kulkarni
authors.forename: Margherita
authors.initials: M
authors.lastname: Rosati
authors.forename: Antonio
authors.initials: A
authors.lastname: Valentin
authors.forename: Brunda
authors.initials: B
authors.lastname: Ganneru
authors.forename: Ashish K
authors.initials: AK
authors.lastname: Singh
authors.forename: Jian
authors.initials: J
authors.lastname: Yan
authors.forename: Morgane
authors.initials: M
authors.lastname: Rolland
authors.forename: Candido
authors.initials: C
authors.lastname: Alicea
authors.forename: Rachel Kelly
authors.initials: RK
authors.lastname: Beach
authors.forename: Gen-Mu
authors.initials: GM
authors.lastname: Zhang
authors.forename: Sylvie
authors.initials: S
authors.lastname: Le Gall
authors.forename: Kate E
authors.initials: KE
authors.lastname: Broderick
authors.forename: Niranjan Y
authors.initials: NY
authors.lastname: Sardesai
authors.forename: David
authors.initials: D
authors.lastname: Heckerman
authors.forename: Beatriz
authors.initials: B
authors.lastname: Mothe
authors.forename: Christian
authors.initials: C
authors.lastname: Brander
authors.forename: David B
authors.initials: DB
authors.lastname: Weiner
authors.forename: James I
authors.initials: JI
authors.lastname: Mullins
authors.forename: George N
authors.initials: GN
authors.lastname: Pavlakis
authors.forename: Barbara K
authors.initials: BK
authors.lastname: Felber
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D015658:HIV Infections / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D019444:Vaccines, DNA / Q000276:immunology*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000276:immunology*
chemical_list: D019444:Vaccines, DNA; D054301:gag Gene Products, Human Immunodeficiency Virus
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