Two-dimensional (2D) NMR spectroscopy has become a routine part of molecular structure determination in liquids. In most modern experiments there is some correlation of transition frequencies between the two dimensions, and the mechanism responsible for the correlation dictates the information obtainable from the 2D spectrum. Two distinct mechanisms for magnetization transfer are considered : scalar coupling and cross-relaxation. The former gives rise to information about the proximity of spins through the electronic framework of the molecule and the latter provides information on spatial proximity and molecular motion. Two-dimensional experiments of both types are described, concentrating in particular on TOCSY and ROESY. It is shown how polarization transfer can be tailored and controlled, and how the unwanted coherent transfer due to J coupling can he eliminated in the ROESY experiment. Throughout the discussion, the multiple-pulse sequence used for each experiment is the central focal point, and its design and analysis are the main considerations.

Evidence obtained from both animal models and humans suggests that T cells specific for HSV-1 and HSV-2 glycoprotein D (gD) contribute to protective immunity against herpes infection. However, knowledge of gD-specific human T cell responses is limited to CD4+ T cell epitopes, with no CD8+ T cell epitopes identified to date. In this study, we screened the HSV-1 gD amino acid sequence for HLA-A*0201-restricted epitopes using several predictive computational algorithms and identified 10 high probability CD8+ T cell epitopes. Synthetic peptides corresponding to four of these epitopes, each nine to 10 amino acids in length, exhibited high-affinity binding in vitro to purified human HLA-A*0201 molecules. Three of these four peptide epitopes, gD53–61, gD70–78, and gD278–286, significantly stabilized HLA-A*0201 molecules on T2 cell lines and are highly conserved among and between HSV-1 and HSV-2 strains. Consistent with this, in 33 sequentially studied HLA-A*0201-positive, HSV-1-seropositive, and/or HSV-2-seropositive healthy individuals, the most frequent and robust CD8+ T cell responses, assessed by IFN-γ ELISPOT, CD107a/b cytotoxic degranulation, and tetramer assays, were directed mainly against gD53–61, gD70–78, and gD278–286 epitopes. In addition, CD8+ T cell lines generated by gD53–61, gD70–78, and gD278–286 peptides recognized infected target cells expressing native gD. Lastly, CD8+ T cell responses specific to gD53–61, gD70–78, and gD278–286 epitopes were induced in HLA-A*0201 transgenic mice following ocular or genital infection with either HSV-1 or HSV-2. The functional gD CD8+ T cell epitopes described herein are potentially important components of clinical immunotherapeutic and immunoprophylactic herpes vaccines.

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Vaccination with a mixture self-adjuvanting lipopeptides containing novel HSV-1 immunodominant gD T-cell epitopes protected mice from ocular herpes infection and disease. The strength of protective immunity induced by these lipopeptides together with their safety provide a molecularly defined vaccine formulation that could combat ocular herpes infection and disease in humans.

Refocused insensitive nucleus enhancement by polarization transfer (RINEPT) from protons (1H) to a J-coupled phosphorus (31P) has been incorporated into three-dimensional (3D) chemical-shift-imaging (CSI) sequence on a clinical imager. The technique is demonstrated on a phantom and in in vivo human brain. The polarization-transfer efficiency (approximately 1.2) is lower than the theoretical maximum of gamma1H/gamma31P approximately 2.4 resulting from 1H-1H homonuclear J couplings of similar magnitude competing with the 1H --> 31P transfer. Nevertheless, compared with direct 31P Ernst-angle excitation, signal gains of up to x1.8 were obtained mainly as a result of T1 differences between 31P and the 1H. Spectral interpretation is simplified by editing out all non-proton-coupled 31P signals. The duration, approximately 50 min, and power deposition, approximately 1 W x kg(-1), make the application suitable for human studies.