Location rather than CD62L phenotype is critical in the early establishment of influenza-specific T cell memory

The rapid recall of influenza virus-specific CD8+ T cell effector function is protective, although our understanding of T cell memory remains incomplete. Recent debate has focused particularly on the CD62L lymph node homing receptor. The present analysis shows that although functional memory can be established from both CD62Lhi and CD62Llo CD8+ T cell subsets soon after initial encounter between naive precursors and antigen, the optimal precursors are CD8+CD44hiCD25lo immune lymphocytes isolated from draining lymph nodes on day 3.5 after influenza virus infection. Analysis of primed T cells at different times after challenge indicates that the capacity to transfer memory is diminished at the peak of the primary cytotoxic T lymphocyte response, challenging speculations that the transition to memory first requires full differentiation to effector status. It seems that location rather than CD62Lhi/lo phenotype may be the more profitable focus for further dissection of the early establishment of T cell memory.

In vitro osteoblast-like cell proliferation on nano-hydroxyapatite coatings with different morphologies on a titanium-niobium shape memory alloy

The morphology of nanomaterials significantly affects their physical, chemical, and biological properties. In the present study, nano-hydroxyapatite coatings with different morphologies were produced on the surface of a titanium-niobium shape memory alloy via a hydrothermal process. The effect of the nano-hydroxyapatite coatings on the in vitro proliferation of SaOS-2 osteoblast-like cells was investigated. Factors including crystallinity, surface micro-roughness, and surface energy of the nano-hydroxyapatite coatings were discussed. Results show that in vitro proliferation of the osteoblast-like cells was significantly enhanced on the nano-hydroxyapatite-coated titanium-niobium alloy compared to the titanium-niobium alloy without coating. The cell numbers on the nano-hydroxyapatite-coated titanium-niobium alloy changed consistently with the surface energy of the hydroxyapatite coatings. This study suggests that surface energy as a characteristic parameter influencing the in vitro proliferation of osteoblast-like cells was predominant over the crystallinity and surface micro-roughness of the nano-hydroxyapatite coatings.

Shape memory alloy scaffolds for bone tissue engineering

Titanium alloy scaffolds for bone tissue engineering are receiving increasing attention because their porous structure and mechanical properties can be adjusted to match those of bone. In particular, there is an enormous potential to increase the life of such implant material if the porous structure can be imparted with shape memory properties. In the present study, TiNi scaffolds with a porous structure and high porosities up to 75% were fabricated by powder metallurgy. The porous structure was characterized by scanning electron microscope. The mechanical properties, the shape memory and superelastic effects were investigated by differential scanning calorimetry, nanoindentation and compressive tests. Results indicate that the porous TiNi scaffolds display an open-cell porous structure which provides new bone tissue ingrowth ability. The mechanical properties of the TiNi scaffolds can be tailored to match those of natural bone. Furthermore, the TiNi scaffolds show good shape memory and superelastic effects.

Titanium–nickel shape memory alloy foams for bone tissue engineering

Titanium–nickel (TiNi) shape memory alloy (SMA) foams with an open-cell porous structure were fabricated by space-holder sintering process and characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis. The mechanical properties and shape memory properties of the TiNi foam samples were investigated using compressive test. Results indicate that the plateau stresses and elastic moduli of the foams under compression decrease with the increase of their porosities. The plateau stresses and elastic moduli are measured to be from 1.9 to 38.3 MPa and from 30 to 860 MPa for the TiNi foam samples with porosities ranged from 71% to 87%, respectively. The mechanical properties of the TiNi alloy foams can be tailored to match those of bone. The TiNi alloy foams exhibit shape memory effect (SME), and it is found that the recoverable strain due to SME decreases with the increase of foam porosity.

Tracking phenotypically and functionally distinct T cell subsets via T cell repertoire diversity

Antigen-specific T cell receptors (TCRs) recognise complexes of immunogenic peptides (p) and major histocompatibility complex (MHC) glycoproteins. Responding T cell populations show profiles of preferred usage (or bias) toward one or few TCRβ chains. Such skewing is also observed, though less commonly, in TCRα chain usage. The extent and character of clonal diversity within individual, antigen-specific T cell sets can be established by sequence analysis of the TCRVβ and/or TCRVα CDR3 loops. The present review provides examples of such TCR repertoires in prominent responses to acute and persistent viruses. The determining role of structural constraints and antigen dose is discussed, as is the way that functionally and phenotypically distinct populations can be defined at the clonal level. In addition, clonal dissection of “high” versus “low” avidity, or “central” versus “effector” memory sets provides insights into how these antigen specific T cell responses are generated and maintained. As TCR diversity potentially influences both the protective capacity of CD8+ T cells and the subversion of immune control that leads to viral escape, analysing the spectrum of TCR selection and maintenance has implications for improving the functional efficacy of T cell responsiveness and effector function.

An in vivo cytotoxicity threshold for influenza A virus-specific effector and memory CD8+ T cells

Influenza A virus infection of C57BL/6 (B6) mice is characterized by prominent CD8 T cell responses to H2Db complexed with peptides from the viral nucleoprotein (NP366, ASNENMETM) and acid polymerase (PA224, SSLENFRAYV). An in vivo cytotoxicity assay that depends on the adoptive transfer of peptide-pulsed, syngeneic targets was used in this study to quantitate the cytotoxic potential of DbNP366- and DbPA224-specific acute and memory CD8 T cells following primary or secondary virus challenge. Both T cell populations displayed equivalent levels of in vivo effector function when comparable numbers were transferred into naive B6 hosts. Cytotoxic activity following primary infection clearly correlated with the frequency of tetramer-stained CD8 T cells. This relationship looked, however, to be less direct following secondary exposure, partly because the numbers of CD8DbNP366 T cells were greatly in excess. However, calculating the in vivo E:T ratios indicated that in vivo lysis, like many other biological functions, is threshold dependent. Furthermore, the capacity to eliminate peptide-pulsed targets was independent of the differentiation state (i.e., primary or secondary effectors) and was comparable for the two T cell specificities that were analyzed. These experiments provide insights that may be of value for adoptive immunotherapy, where careful consideration of both the activation state and the number of effector cells is required.

Protection and compensation in the influenza virus-specific CD8+ T cell response.

Influenza virus-specific CD8+ T cells generally recognize peptides derived from conserved, internal proteins that are not subject to antibody-mediated selection pressure. Prior exposure to any one influenza A virus (H1N1) can prime for a secondary CD8+ T cell response to a serologically different influenza A virus (H3N2). The protection afforded by this recall of established CD8+ T cell memory, although limited, is not negligible. Key characteristics of primary and secondary influenza-specific host responses are probed here with recombinant viruses expressing modified nucleoprotein (NP) and acid polymerase (PA) genes. Point mutations were introduced into the epitopes derived from the NP and PA such that they no longer bound the presenting H2Db MHC class I glycoprotein, and reassortant H1N1 and H3N2 viruses were made by reverse genetics. Conventional (C57BL/6J, H2b, and Ig+/+) and Ig-/- (muMT) mice were more susceptible to challenge with the single NP [HKx31 influenza A virus (HK)-NP] and PA (HK-PA) mutants, but unlike the Ig-/- mice, Ig+/+ mice were surprisingly resistant to the HK-NP/-PA double mutant. This virus was found to promote an enhanced IgG response resulting, perhaps, from the delayed elimination of antigen-presenting cells. Antigen persistence also could explain the increase in size of the minor KbPB1703 CD8+ T cell population in mice infected with the mutant viruses. The extent of such compensation was always partial, giving the impression that any virus-specific CD8+ T cell response operates within constrained limits. It seems that the relationship between protective humoral and cellular immunity is neither simple nor readily predicted.

Epitope-specific CD4+, but not CD8+, T-cell responses induced by recombinant influenza A viruses protect against Mycobacterium tuberculosis infection

Tuberculosis remains a global health problem, in part due to failure of the currently available vaccine, BCG, to protect adults against pulmonary forms of the disease. We explored the impact of pulmonary delivery of recombinant influenza A viruses (rIAVs) on the induction of Mycobacterium tuberculosis (M. tuberculosis)-specific CD4(+) and CD8(+) T-cell responses and the resultant protection against M. tuberculosis infection in C57BL/6 mice. Intranasal infection with rIAVs expressing a CD4(+) T-cell epitope from the Ag85B protein (PR8.p25) or CD8(+) T-cell epitope from the TB10.4 protein (PR8.TB10.4) generated strong T-cell responses to the M. tuberculosis-specific epitopes in the lung that persisted long after the rIAVs were cleared. Infection with PR8.p25 conferred protection against subsequent M. tuberculosis challenge in the lung, and this was associated with increased levels of poly-functional CD4(+) T cells at the time of challenge. By contrast, infection with PR8.TB10.4 did not induce protection despite the presence of IFN-γ-producing M. tuberculosis-specific CD8(+) T cells in the lung at the time of challenge and during infection. Therefore, the induction of pulmonary M. tuberculosis epitope-specific CD4(+), but not CD8(+) T cells, is essential for protection against acute M. tuberculosis infection in the lung.

Anodization parameters influencing the morphology and electrical properties of TiO2 nanotubes for living cell interfacing and investigations

Nanotube structures have attracted tremendous attention in recent years in many applications. Among such nanotube structures, titania nanotubes (TiO2) have received paramount attention in the medical domain due to their unique properties, represented by high corrosion resistance, good mechanical properties, high specific surface area, as well as great cell proliferation, adhesion and mineralization. Although lot of research has been reported in developing optimized titanium nanotube structures for different medical applications, however there is a lack of unified literature source that could provide information about the key parameters and experimental conditions required to develop such optimized structure. This paper addresses this gap, by focussing on the fabrication of TiO2 nanotubes through anodization process on both pure titanium and titanium alloys substrates to exploit the biocompatibility and electrical conductivity aspects, critical factors for many medical applications from implants to in-vivo and in-vitro living cell studies. It is shown that the morphology of TiO2 directly impacts the biocompatibility aspects of the titanium in terms of cell proliferation, adhesion and mineralization. Similarly, TiO2 nanotube wall thickness of 30-40nm has shown to exhibit improved electrical behaviour, a critical factor in brain mapping and behaviour investigations if such nanotubes are employed as micro-nano-electrodes.