The fixed cell images were obtained at ×63 magnification on a Zeiss Axioplan microscope with a 63×, 1.4 NA oil-immersion objective (Zeiss, Oberkochen, Germany), 2 taking 9 z stacks 300 nm apart, which completely encompassed the breadth of the cells, unless otherwise stated.
It will therefore be important to demonstrate the breadth of the T cell response generated by a vaccine, not only in terms of the number of epitopes targeted, but also the ability of epitope-specific responses to accommodate clade-specific viral diversity.
The LPP used here were the same as used in our 4-day T-helper IFN-γ-ELISPOT assays (see M&M section, the 4 LPP span the whole Influenza M1 protein) to measure Influenza-specific T-cell responses as well as to obtain some information about the breadth of the T-cell response [ 11, 19].
Standardized peptide reagents to define the magnitude and potential breadth of the T-cell response, especially to circulating strains of HIV-1, are needed.
Enhancing thymic function, together with suppression of immune activation and manipulation of Treg function will be essential to restoring the function and breadth of the T-cell compartment of the cellular immune system during HIV-1 infection.
In an analysis of the potential effect of LAMP targeting on the breadth of the T-cell repertoire in macaques, PBMCs from the immunized macaques were tested at week 20, after 3 DNA immunizations, for T-cell responses to 12 different pools consisting of ten peptides each, of the 15-aa Gag peptide set (Figure 7).
The breadth of the T-cell response to vaccination is dependent upon the size and specificity of the immunogen, and vaccine studies to date have induced T-cell responses with a wide variation in breadth.
We have shown previously that deleting the immune evasion genes from murine cytomegalovirus (MCMV) that target class I MHC presentation, has no impact on the size or breadth of the CD8+ T cell response elicited by infection, suggesting that the majority of MCMV-specific CD8+ T cells in vivo are not directly primed by infected professional APCs.
A glimpse of our lack of understanding of the breadth of T cell reactivity becomes apparent when analysing the MHC restriction of the epitopes identified to date.
