Identification of HLA-A3-restricted cytotoxic T lymphocyte epitopes from carcinoembryonic antigen and HER-2/neu by primary in vitro immunization with peptide-pulsed dendritic cells

The human carcinoembryonic antigen (CEA) and HER-2/neu are potential target antigens for CTL specific immunotherapy for common malignancies such as breast, lung, colon, and gastric carcinomas. Several CTL epitopes restricted by HLA-A2, the most common human histocompatibility molecule, have been previously reported. However, to develop CTL-based immunotherapies for the general population, it is necessary to identify epitopes restricted by other common histocompatibility alleles. Here, we describe two HLA-A3-restricted CTL epitopes from the CEA and HER-2/neu antigens. HLA-A3 binding synthetic peptides from CEA and HER-2/neu were tested for immunogenicity by in vitro primary CTL induction protocol using peripheral blood mononuclear cells from normal healthy volunteers. One peptide from CEA (CEA[9(61)]: HLFGYSWYK) and one peptide from HER-2/neu (HER2[9(754)]: VLRENTSPK) were shown to induce CTL that was capable of killing a tumor cell line expressing HLA-A3 and the corresponding tumor-associated antigen. Additional MHC binding studies with the most common HLA molecules belonging to the HLA-A3 superfamily (HLA-A*1101, -A*3101, -A*3301, and -A*6801), demonstrated that CEA[9(61)] binds five of five A3 supertype molecules with high affinity, and the HER2[9(754)] epitope was able to bind to four of the same five alleles. These results indicate that these two new CTL epitopes should be immunogenic in individuals expressing either HLA-A3, or other members of the HLA-A3 superfamily.     

Notch signalling mediates segmentation of the Drosophila leg

The legs of Drosophila are divided into segments along the proximodistal axis by flexible structures called joints. The separation between segments is already visible in the imaginal disc as folds of the epithelium, and cells at segment boundaries have different morphology during pupal development. We find that Notch is locally activated in distal cells of each segment, as demonstrated by the restricted expression of the Enhancer of split mbeta gene, and is required for the formation of normal joints. The genes fringe, Delta, Serrate and Suppressor of Hairless, also participate in Notch function during leg development, and their expression is localised within the leg segments with respect to segment boundaries. The failure to form joints when Notch signalling is compromised leads to shortened legs, suggesting that the correct specification of segment boundaries is critical for normal leg growth. The requirement for Notch during leg development resembles that seen during somite formation in vertebrates and at the dorsal ventral boundary of the wing, suggesting that the creation of boundaries of gene expression through Notch activation plays a conserved role in co-ordinating growth and patterning.     

Specific T cell recognition of peptides derived from prostate-specific antigen in patients with prostate cancer

Active, population-based surveillance for invasive group A streptococcal (GAS) disease was conducted in laboratories in metropolitan Atlanta from 1 January 1994 through 30 June 1995. Clinical and laboratory records were reviewed and isolates characterized. One hundred and eighty-three cases of invasive GAS disease were identified (annual incidence, 5.2 cases/100,000). The incidence was highest among blacks (9.7/100,000 per year; relative risk (RR), 1.92; confidence interval (CI), 1.69-2.19; P < .0001) and the elderly, particularly nursing home residents (RR, 13.66; CI, 7.07-26.40; P < .0001). The mean age of patients was 41.3 years (range, 0-95 years). Skin and soft-tissue infections were most common. Mortality was 14.4%; risk of death was significantly higher for patients with streptococcal toxic shock syndrome (STSS) (RR, 9.73; CI, 3.34-29; P = .0008) and individuals infected with M-type 1 (RR, 7.40; CI, 1.5-16; P = .0084). Fourteen percent of invasive GAS infections were STSS and 3% were necrotizing fasciitis. Invasive GAS disease was associated with varicella infection in children (RR, 12.19; CI, 5.58-26.62; P < .0001). M (or emm) types included M1 (16%), M12 (12%), and M3 (11%). Continued study of GAS disease is essential to further define risk factors and risk of secondary cases and to develop effective prevention strategies.     

Role of preimmunization virus sequences in cellular immunity in HIV-infected patients during HIV type 1 MN recombinant gp160 immunization

The effect of patient preimmunization virus sequences on CTL responses during gp160 immunization were studied. Ten HLA-A2+, HIV+ asymptomatic patients with CD4+ T cells >500/mm3 were given two courses of HIV-1 MN rgp160 vaccine over a 2-year period. Envelope epitope-specific CTL responses, using PBMCs, were measured against peptide-coated autologous B lymphoblastoid cell lines. Optimum CTL epitopes were determined by HLA-A2-binding affinity of 9- to 10-mer peptides containing the HLA-A2.1-binding motif. Ten of the high- or intermediate-binding peptides were conserved among >50% of reported clade B HIV strains. These peptide-specific CTL activities and the patient virus sequences in peptide-coding regions were monitored. Six patients showed envelope peptide-specific CTL responses, which correlated with the presence of whole envelope antigen-specific CTL responses. Five of these patients, who showed responses to epitopes in the gp41 region (aa 814-824), had preimmunization virus similar to the vaccine sequence in this region. Three patients who did not show these epitope-specific responses had initially different sequences in the HIV gene encoding that region. The epitope-specific CTL responses appear to reflect recall responses, as only patients infected with virus containing the vaccine sequence developed them and they could be recalled with a second set of vaccine injections. This appears to be reminiscent of the concept of T cell "original antigenic sin." This vaccine was also immunogenic as measured by gp160-specific lymphocyte-proliferative responses. However, increased immune responses did not impact the HIV load or CTL epitope sequences during therapy.     

Effect of Substrate Type and Temperature on the Tribological Properties of MoS_x Coatings

SPUTTERING deposited MoSx coatings are often usedas a solid lubricant in space applications,where a lowcoefficient of friction and long lifetime of coating areneeded.But the lubricating properties and endurancelives of sputtered MoSx coatings are strongly dependenton the sputtering parameters and the substrateconditions.Spalvin[1]showed that three groups can be dividedabout the effect of temperature on the nucleation ofMoSx coatings:(1)amorphous(-195°C);(2)crystalline-amorphous(-195°C …     

Structural and Tribological Properties of Nanostructured Supersonic Cold Sprayed Ni-20 wt.% Sn Coatings

80-μm-thick nanostructured coatings consisting of a Ni solid solution, Ni₃Sn, Ni₃Sn₂, and metastable NiSn intermetallic phases were deposited via supersonic cold spraying onto inconel 718 alloy substrates. These coatings have complex nanostructured metallurgical phases as revealed by transition electron microscopy, scanning electron microscopy, and x-ray diffraction techniques. Their mechanical properties were determined by nanoindentation measurements. Furthermore, the wear behavior of these nanostructured sprayed coatings was compared to the one of the industrial bulk or sprayed coated benchmark materials. It was found that the nanostructured coatings exhibit higher wear resistance than the industrial benchmarks, thanks to an appropriate balance of hard intermetallic phases and soft Ni matrix, as well as to their nanostructuring. Their frictional characteristics under reciprocating sliding are mainly determined by the formation of an oxide-based tribo-layer, which was analyzed by x-ray photoelectron spectroscopy. The role of intermetallic phases in these coatings on the friction and wear is also discussed.     

Synthesis of polymeric and hybrid nanoparticles for electroplating applications

Monodisperse polymeric particles with diameters in the range of 60-1400 nm were prepared by (emulsifier-free) emulsion polymerization and incorporated into electrolytic zinc coatings aiming to improve the corrosion resistance of electrogalvanized steel. Various types of polymeric nanoparticles were thus synthesized in order to assess the effect of the emulsifier, initiator and comonomer type on the particle morphology, stability and codeposition behavior. The polymerization experiments were carried out in laboratory-scale glass reactors and the most promising recipes were successfully scaled-up in a fully automated pilot-scale reactor. Replicates of some representative experiments, which were run both in lab and pilot-scale reactors, indicated excellent reproducibility of the polymerization process. Uniform, polymer-containing zinc coatings were produced by electrolytic codeposition of the nanoparticles from an acid zinc plating bath using a rotating disk electrode (RDE). Hybrid polystyrene/silica nanoparticles with increased silica content were also prepared via emulsifier-free emulsion polymerization, in the presence of an ultrafine aqueous silica sol, to be used in electrocoating applications. The effect of key process parameters, such as initial monomers molar ratio and pH on the size, morphology and silica content of the produced hybrid nanoparticles was investigated.     

New Insights Leading to Improved Process Control

Composite plating may nowadays be considered as a well-established industrial technology for the synthesis of some types of composite coatings like Ni-SiC in the automotive industry. Potentialities of that composite plating process are numerous but new developments are fastidious and expensive. This paper highlights some recent progresses made in determining the forces bringing and keeping particles in the neighbourhood of electrodes and in detecting the trajectory of particles near electrodes. The last possibility opens the way to the development of an electrochemical sensor suitable for on-line process control in composite plating.     

Hydrogen production from acid and enzymatic oat straw hydrolysates in an anaerobic sequencing batch reactor: Performance and microbial population analysis

Feasibility of hydrogen production from acid and enzymatic oat straw hydrolysates was evaluated in an anaerobic sequencing batch reactor at 35 °C and constant substrate concentration (5 g chemical oxygen demand/L). In a first experiment, hydrogen production was replaced by methane production. Selective pressures applied in a second experiment successfully prevented methane production. During this experiment, initial feeding with glucose/xylose, as model substrates, promoted biomass granulation. Also, the highest hydrogen molar yield (HMY, 2 mol H₂/mol sugar consumed) and hydrogen production rate (HPR, 278 mL H₂/L-h) were obtained with these model substrates. Gradual substitution of glucose/xylose by acid hydrolysate led to disaggregation of granules and lower HPR and HMY. When the model substrates were completely substituted by enzymatic hydrolysate, the HMY and HPR were 0.81 mol H₂/mol sugar consumed and 29.6 mL H₂/L-h, respectively. Molecular analysis revealed a low bacterial diversity in the stages with high hydrogen production and vice versa. Furthermore, Clostridium pasteurianum was identified as the most abundant species in stages with a high hydrogen production. Despite that feasibility of hydrogen production from hydrolysates was demonstrated, lower performance from hydrolysates than from model substrates was obtained.