Cassette-based presentation of SIV epitopes with recombinant gas vesicles from halophilic archaea.
In earlier studies we demonstrated recombinant gas vesicles from Halobacterium sp. NRC-1, expressing a model six amino acid insert, or native vesicles displaying chemically coupled TNP, each were immunogenic, and antigenic. Long-lived responses displaying immunologic memory were elicited without exogenous adjuvant. Here we report the generation and expression of cassettes containing SIV derived DNA. The results indicate a cassette-based display/delivery system derived from recombinant halobacterial gas vesicle genes is highly feasible. Data specifically support four conclusions: (i) Recombinants carrying up to 705 bp of SIV DNA inserted into the gvpC gene form functional gas vesicles; (ii) SIV peptides contained as part of the expressed recombinant, surface exposed GvpC protein are recognized by antibody elicited in monkeys exposed to native SIV in vivo; (iii) in the absence of adjuvant, mice immunized with the recombinant gas vesicle (r-GV) preparations mount a solid, titratable antibody response to the test SIV insert that is long lived and exhibits immunologic memory; (iv) recombinant organelles, created through the generation of cassettes encoding epitopes inserted into the gvpC DNA, can be used to construct a multiepitope display (MED) library, a potentially cost effective vehicle to express and deliver peptides of SIV, HIV or other pathogens.
Stuart
Elizabeth S
ES
Department of Microbiology, University of Massachusetts, Amherst, MA 01003, USA. esstuart@microbio.umass.edu
Morshed
Fazeela
F
Sremac
Marinko
M
DasSarma
Shiladitya
S
D000818:Animals; D022162:Cytoplasmic Vesicles / Q000235:genetics*; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D006217:Halobacterium / Q000235:genetics / Q000502:physiology*; D007106:Immune Sera / Q000276:immunology; D007156:Immunologic Memory; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology
D000939:Epitopes; D007106:Immune Sera; D011506:Proteins; D011994:Recombinant Proteins; D014765:Viral Vaccines; C038524:gas vesicle protein