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Faculty Paper

Gary Splitter
University of Wisconsin-Madison

"Discordant Brucella melitensis Antigens Yield Cognate CD8+ T cells in vivo"

Brucella spp. are intracellular bacteria that cause the most frequent zoonosis in the world. Though recent work has advanced the field of Brucella vaccine development, there remains no safe human vaccine. In order to produce a safe and effective human vaccine, the immune response to Brucella spp. requires greater understanding. Induction of Brucella specific CD8(+) T cells is considered an important aspect of the host response; however, the CD8(+) T cell response is not clearly defined. Discovering the epitope containing antigens recognized by Brucella specific CD8(+) T cells and correlating them with microarray data will aid in determining proteins critical for vaccine development that cover a kinetic continuum during infection. Developing tools to take advantage of the BALB/c mouse model of Brucella melitensis infection will help to clarify the correlates of immunity and improve the efficacy of this model. Two H-2(d) CD8(+) T cell epitopes have been characterized, and a group of immunogenic proteins have provoked IFN-gamma production by CD8(+) T cells. RYCINSASL and NGSSSMATV induced cognate CD8(+) T cells after peptide immunization that showed specific killing in vivo. Importantly, we found by microarray analysis that the genes encoding these epitopes are differentially expressed following macrophage infection, further emphasizing that these discordant genes may play an important role in the pathogenesis of Brucella melitensis infection.

Details : Durward M, Harms J, Magnani D, Eskra L. Infect Immun. 2009 Nov 2.

 

Paper of Interest

John Glass
J. Craig Venter Institute | Rockville, Maryland

"Creating bacterial strains from genomes that have been cloned and engineered in yeast"

We recently reported the chemical synthesis, assembly, and cloning of a bacterial genome in yeast. To produce a synthetic cell, the genome must be transferred from yeast to a receptive cytoplasm. Here we describe methods to accomplish this. We cloned a Mycoplasma mycoides genome as a yeast centromeric plasmid and then transplanted it into Mycoplasma capricolum to produce a viable M. mycoides cell. While in yeast, the genome was altered by using yeast genetic systems and then transplanted to produce a new strain of M. mycoides. These methods allow the construction of strains that could not be produced with genetic tools available for this bacterium.

Details : Lartigue C, Vashee S, Algire MA, Chuang RY, Benders GA, MaL, Noskov VN, Denisova EA, Gibson DG, Assad-Garcia N, Alperovich N, Thomas DW, Merryman C, Hutchison CA 3rd, Venter JC, Glass JI. Science. 2009 Sep 25;325(5948):1693-6.