Search

We created and tested multi-epitope DNA or protein vaccines with TLR4 ligand emulsion adjuvant (gluco glucopyranosyl lipid adjuvant in a stable emulsion [GLA-SE]) for their ability to protect against Toxoplasma gondii in HLA transgenic mice. Our constructs each included 5 of our best down-selected CD8+ T cell–eliciting epitopes, a universal CD4+ helper T lymphocyte epitope (PADRE), and a secretory signal, all arranged for optimal MHC-I presentation. Their capacity to elicit immune and protective responses was studied using immunization of HLA-A*11:01 transgenic mice. These multi-epitope vaccines increased memory CD8+ T cells that produced IFN-γ and protected mice against parasite burden when challenged with T. gondii. Endocytosis of emulsion-trapped protein and cross presentation of the antigens must account for the immunogenicity of our adjuvanted protein. Thus, our work creates an adjuvanted platform assembly of peptides resulting in cross presentation of CD8+ T cell–eliciting epitopes in a vaccine that prevents toxoplasmosis.

We created and tested multi-epitope DNA or protein vaccines with TLR4 ligand emulsion adjuvant (gluco glucopyranosyl lipid adjuvant in a stable emulsion [GLA-SE]) for their ability to protect against Toxoplasma gondii in HLA transgenic mice. Our constructs each included 5 of our best down-selected CD8(+) T cell-eliciting epitopes, a universal CD4(+) helper T lymphocyte epitope (PADRE), and a secretory signal, all arranged for optimal MHC-I presentation. Their capacity to elicit immune and protective responses was studied using immunization of HLA-A*11:01 transgenic mice. These multi-epitope vaccines increased memory CD8(+) T cells that produced IFN-γ and protected mice against parasite burden when challenged with T. gondii. Endocytosis of emulsion-trapped protein and cross presentation of the antigens must account for the immunogenicity of our adjuvanted protein. Thus, our work creates an adjuvanted platform assembly of peptides resulting in cross presentation of CD8(+) T cell-eliciting epitopes in a vaccine that prevents toxoplasmosis.

The complement system is a critical component of innate immunity that requires regulation to avoid inappropriate activation. This regulation is provided by many proteins, including complement factor H (CFH), a critical regulator of the alternative pathway of complement activation. Given its regulatory function, mutations in CFH have been implicated in diseases such as age-related macular degeneration and membranoproliferative glomerulonephritis, and central nervous system diseases such as Alzheimer's disease, Parkinson's disease, and a demyelinating murine model, experimental autoimmune encephalomyelitis (EAE). There have been few investigations on the transcriptional regulation of CFH in the brain and CNS. Our studies show that CFH mRNA is present in several CNS cell types. The murine CFH (mCFH) promoter was cloned and examined through truncation constructs and we show that specific regions throughout the promoter contain enhancers and repressors that are positively regulated by inflammatory cytokines in astrocytes. Database mining of these regions indicated transcription factor binding sites conserved between different species, which led to the investigation of specific transcription factor binding interactions in a 241 base pair (bp) region at -416 bp to -175 bp that showed the strongest activity. Through supershift analysis, it was determined that c-Jun and c-Fos interact with the CFH promoter in astrocytes in this region. These results suggest a relationship between cell cycle and complement regulation, and how these transcription factors and CFH affect disease will be a valuable area of investigation.

The complement cascade of the immune system is an important mediator of the inflammatory response to infection; however it is crucial that this pathway is tightly regulated to prevent uncontrolled activation, which can lead to damage to host tissues. The complement system has many regulators that control activation; both membrane-bound and soluble factors. This review will focus on what is currently known about the transcriptional regulation of the soluble complement regulatory genes C1-inhibitor, complement factor I, complement factor H and C4-binding protein. The absence or mutation of these regulators is all associated with specific disease, and yet their contribution to disease is often poorly understood. It is through full understanding of these genes that we can comprehend the diseases with which they are implicated, and thus prove why knowledge of the transcriptional regulation of these genes is valuable.

The complement system is a critical component of innate immunity that requires regulation to avoid inappropriate activation. This regulation is provided by many proteins including complement Factor H (CFH), a secreted protein of the alternative pathway. Despite its regulatory function, gene and promoter polymorphisms of CFH have been implicated in patients with age-related macular degeneration and atypical hemolytic uremic syndrome, indicating that a better understanding of this gene’s regulation is valuable. Presently, little work has been done to address the role of CFH transcriptional control in the central nervous system (CNS). We have cloned the murine CFH promoter and have made truncation constructs to examine CFH gene regulation in cells of the CNS. Our studies show that CFH mRNA is present in several CNS cell types, and that specific regions throughout the promoter contain enhancers and repressors that are positively and negatively regulated by inflammatory cytokines. Database mining of these regions indicates that there are potential transcription factor binding sites conserved between different species, which led us to investigate specific transcription factor binding interactions in these regions. Through these studies we hope to elucidate the transcriptional regulation of CFH in the CNS.

The complement system is an important component of innate immunity that requires proper regulation to avoid inappropriate activation. This regulation is provided by many proteins including complement Factor H (CFH), a secreted protein of the alternative pathway. Despite its regulatory function, gene and promoter polymorphisms of CFH have been implicated in disease of many patients with age-related macular degeneration and atypical hemolytic uremic syndrome, indicating that a better understanding of this gene's regulation is valuable. Presently, little work has been done to address the role of CFH in the central nervous system (CNS). We have cloned the murine CFH promoter and have made truncation constructs to examine CFH gene regulation in cells of the CNS. Our studies show that CFH mRNA is present in CNS cell types, and that specific regions throughout the promoter contain possible enhancers and repressors. Database mining of these regions indicates that there are potential transcription factor binding sites conserved between many different species, which led us to investigate specific transcription factor binding interactions in these regions. Through these studies we hope to elucidate the transcriptional regulation of CFH in the CNS. This work is supported by the National Multiple Sclerosis Society.