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The immunotoxic potential of drug candidates is assessed through the examination of results from a variety of studies and endpoints. While the functional assessment of CD8+ cytotoxic T-lymphocytes (CTL) is well-characterized in the clinic, the lack of a robust macaque CTL functional assay has been an important hurdle in evaluating and accurately quantifying cell-mediated CD8+ T-cell effector responses in the nonclinical setting. This paper describes the development of an assay to measure CTL activity in peripheral blood mononuclear cells (PBMC) isolated from Cynomolgus macaques. A human EGFR/CD3 Bispecific T-cell Engager (BiTE®) was used to mount a robust CD8+ T-cell response in the presence of target-expressing cells. Upon target engagement, degranulation of CD107a and production of interferon (IFN)-γ both reliably indicated a robust functional response in CD8+ T-cells. The BiTE®-mediated stimulation method proved to be favorable when compared to other methods of stimulation in the absence of target cells. These studies demonstrated acceptable longitudinal variability of the functional assay and sensitivity to dexamethasone-mediated immunosuppression. Taken together, the results indicated an assay leveraging CD3-bispecific antibodies and target-expressing cells can provide a robust approach to the in vitro or ex vivo assessment of CTL function in Cynomolgus macaques. Because the impairment of CTL activity by immunomodulators is recognized to be an important contributor to decreased antiviral defense and increased carcinogenicity risk, we believe that this novel assay to be a valuable addition to the immunotoxicology assessment of therapeutic drug candidates. [ABSTRACT FROM AUTHOR]

The requirements for the eradication of HIV in infected individuals are unknown. Intermittent administration of the immune activator interleukin-2 (IL-2) in combination with highly-active anti-retroviral therapy (HAART) has been suggested as an effective strategy to realize long-term control of HIV replication in vivo . However, potential latent virus reservoirs are considered to be a major impediment in achieving this goal. In this paper, a new mathematical model is designed and used to monitor the interactions between HIV, CD4+ T-cells, CD8+ T-cells, productively infected and latently infected CD4+ T-cells, and to evaluate therapeutic strategies during the first 3 years of HIV infection. The model shows that current anti-HIV therapies, including intermittent IL-2 and HAART, are insufficient in achieving eradication of HIV. However, it suggests that the HIV eradication may indeed be theoretically feasible if such therapy is administered continuously (without interruption) under some specified conditions. These conditions may realistically be achieved using an agent (such as a putative anti-HIV vaccine) that brings about a concomitant increase in the proliferation of HIV-specific CD4+ T- and CD8+ T-cells and the differentiation of CD8+ T-cells into anti-HIV cytotoxic T lymphocytes (CTLs). [ABSTRACT FROM AUTHOR]

Underdeveloped tree crops (⩽ 30 bone-dry tons ha−1) offer a main harvest of biomass-trees, which lack the size and the form for producing a log. However, about 1/3 of the available stems may yield at least one log, which could significantly increase the overall value of the harvest. Under those circumstances, it may make sense to sort log-trees from biomass-trees at an early stage, rather than having a harvester or a processor go through all of the treesonly to toss almost 70 out of 100 trees into the biomass pile. The Authors set up a controlled experiment to quantify the eventual benefit obtained by early pre-sorting, performed by the feller-buncher. Pre-sorting resulted in a 15% productivity loss for the feller-buncher, which was repaid by a 100% productivity increase for the processor. Considering the different hourly cost of the two machines, three Euros were saved for each additional Euro invested in pre-sorting. Pre-sorting made whole-tree harvesting (WTH) a significantly cheaper option than cut-to-length (CTL) , whereas the two systems would be almost equally expensive when no pre-sorting was applied. Pre-sorting would also facilitate multiple-tree delimbing, implemented through the introduction of a chainflail; that is likely to further reduce harvesting cost, returning financial viability to the harvesting of underdeveloped plantations. [ABSTRACT FROM AUTHOR]

The mechanization of roundwood harvesting in Sweden has historically cut costs rapidly. However, machinery and work methods have conceptually stayed the same since the introduction of the single-grip harvester. Current trends indicate that the next major wave of development will be based on automation, and that teleoperation is a step in this direction. This study aims to evaluate the economic potential of semi-automated tele-extraction compared to standard forwarding within the Nordic CTL two-machine harvesting system. The initial scenario examined autonomous terrain transportation with teleoperation during loading and unloading. A Discrete Event Simulation was implemented in the AnyLogic software. Input data included 1100 Swedish harvest sites, with in total 1.6 million m3 and extraction distances from 20 to 1500 m. Scenarios with different numbers of teleoperators for a fleet of ten forwarders were tested. The optimum number of operators was seven per ten forwarders, resulting in a potential extraction cost reduction of 7% compared to standard forwarding. Extraction distances further than 500 m enabled use of five or six teleoperators, resulting in potential cost reductions of up to 15%. The highest potential cost reductions were linked to increased time used for autonomous driving, increasing the teleoperator availability for other machines. The simulation model provides a framework for further evaluation of new scenarios of automation and teleoperation of forwarders. [ABSTRACT FROM AUTHOR]

Long-term machine-derived data sets comprising 140,000 trees were collected from four harvesters of equal age and similar working conditions, into two machine size classes, viz. two Ponsse Bears and two smaller Ponsse Beavers. Productivity functions for each size class were modelled using a nonlinear mixed effects approach. Based on these functions, unit costs and their sensitivity to utilization rates and cost of capital were assessed. Results showed that despite considerably higher capital costs (32%) on the Bear, a 50% higher mean productivity resulted in a unit cost only 17% higher than the Beaver in a disadvantageous scenario (high interest rates and low utilisation), and a 6% lower unit cost than the Beaver in an advantageous scenario (low interest and high utilisation), within the range of tree sizes observed. Between these extremes, only marginal differences in unit costs were observed. This demonstrates that the difference in ownership and operating costs between larger and smaller harvesters is largely negated by the difference in productivity rates. These results can provide useful insight into timber harvester investment decisions. Harvesters from two adjacent size classes can be used interchangeably at the same unit cost within a wide range of tree sizes despite productivity differences. It should be noted that increased repair costs and an eventual reduction in expected economic lifetime on a smaller harvester, or the negative effects of using a larger harvester in smaller trees, e.g. thinning operations, were not taken into account in this work. [ABSTRACT FROM AUTHOR]

Globally, almost 2 billion cubic meters of industrial roundwood are harvested yearly. Two of the most common methods of harvest and extraction are cut-to-length (CTL) and full-tree or tree-length (FT/TL). The aim of this study was to compile data on annual volumes of industrial roundwood harvested by the main methods in forestry countries. To quantify the effect of potential explanatory variables, the data were subjected to linear regression analysis, using shares of roundwood volumes harvested by fully mechanized CTL and/or FT/TL as response variables. Generally, high diesel price and Gross Domestic Product appear to favor CTL, while high shares of steep terrain (>20°) in forest land decrease the leve l of both mechanization and CTL, and low Social Security Rate (SSR) favor FT/TL. Two models were created for CTL, one with an R2 of 0.64 and another more complex with an R2 of 0.75. A separate model for mechanization (CTL and FT/TL together) showed an R2 0.57. The CTL models could potentially be used to predict shares of roundwood volumes harvested by CTL in countries not included in this study. Predictions for countries with large harvested volumes, e.g. China and India, are presented here, but they require validation, as does the model's applicability for countries with small harvested volumes. Countries with less than 10% of steep slope forests are almost exclusively mechanized according to the model. For FT/TL, the proposed model is probably not sufficiently robust for prediction, but it highlights SSR as one important explanatory variable. [ABSTRACT FROM AUTHOR]

Virus-specific CD8+T-cell responses play a major role in the control of HIV replication, and induction of HIV-specific T-cell responses is an important strategy for AIDS vaccine development. Optimization of the delivery system and immunogen would be the key for the development of an effective T cell-based AIDS vaccine. Heterologous prime-boost vaccine regimens using multiple viral vectors are a promising protocol for efficient induction of HIV-specific T-cell responses, and the development of a variety of potent viral vectors have been attempted. This review describes the current progress of the development of T cell-based AIDS vaccines using viral vectors, focusing on Sendai virus vectors, whose phase I clinical trials have been performed. [ABSTRACT FROM PUBLISHER]

The nuclear factor κ light chain enhancer of activated B cells (NFκB) is constitutively active in most cancers, controlling multiple cellular processes including proliferation, invasion and resistance to therapy. NFκB is primarily regulated through the association with inhibitory proteins that are known as inhibitors of NFκB (IκBs). Increased NFκB activity in tumor cells has been correlated with decrease stability of IκB proteins, in particular IκBα. In responso to a large number of stimuli, IκB proteins are degraded by the proteasome. Cytotoxic T lymphocytes (CTLs) recognize HLA-restricted antigenic peptides that are generated by proteasomal degradation in target cells. In the present study, we demonstrate the presence of naturally occurring IκBα-specific T cells in the peripheral blood of patients suffering from several unrelated tumor types, i.e., breast cancer, malignant melanoma and renal cell carcinoma, but not of healthy controls. Furthermore, we show that such IκBα-specific T cells are granzyme B-releasing, cytotoxic cells. Hence, the increased proteasomal degradation of IκBα in cancer induces IκBα-specific CTLs. [ABSTRACT FROM AUTHOR]

Recent development in antiretroviral treatment against HIV can help AIDS patients to fight against HIV. But the question that whether the disease is to be partially or totally eradicated from HIV infected individuals still remains unsolved. Usually, the most effective treatment for the disease is HAART which can only control the disease progression. But as the immune system becomes weak, the patients can not fight against other diseases. Immune cells are activated and proliferated by IL-2 after the identification of antigen. IL-2 production is impaired in HIV positive patients and intermitted administration of immune activator IL-2 together with HAART which is a more effective treatment to fight against the disease. Thus, its expediency is essential and is yet to be explored. In this article we anticipated a mathematical model of the effect of IL-2 together with RTIs therapy in HIV positive patients. Our analytical as well as numerical study shows that the optimal schedule of treatment for best result is to be obtained by systematic drug therapy. But at the last stage of treatment, the infection level raises again due to minimisation of drug dosage. Thus we study the perfect adherence of the drugs and found out if RTIs are taken with sufficient interval then for fixed interval of IL-2 therapy, certain amount of drug dosages may be able to sustain the immune system at pre-infection stage and the infected CD4+T cells are going towards extinction. [ABSTRACT FROM AUTHOR]

The influence of the number of apheresis-stimulation-infusion(s) cycles, and the time in culture before the infusion (one vs. two weeks), on the generation of tumor antigen-specific cytotoxic T-lymphocytes (CTL) was investigated in a phase I/II clinical adoptive immunotherapy trial. Two previously treated metastatic breast cancer patients with no evidence of disease, in complete remission (CR), were enrolled. Each apheretic peripheral blood mononuclear cell (PBMC) sample was stimulated twice with MUC-1 before infusion back into the patients. Killer T-cells responses against MUC-1-expressing MCF-7 (CTL), nonspecific natural killer (NK) and lymphokine-activated killer (LAK) target cell lines, as well as, cytokine production were measured before each infusion. Patients received 2 infusions per month for 4 months. There were no tumor recurrences or toxicity. CTL, NK and LAK cells, type 1 cytokine, gamma-interferon (G-INF), and CD4+ and CD8+ memory T-lymphocytes were initially generated, produced or induced, respectively, and then declined. The CTL, NK and LAK cells were only induced at the first infusion of the first month. Thus, maintaining PBMC in culture longer than the first infusion was of no benefit with regards to retaining functional killer T-cells. In conclusion, this study implies that one treatment is optimal. [ABSTRACT FROM AUTHOR]

The influence of tumor burden on the generation of tumor antigen-specific cytotoxic T-lymphocytes (CTL) was investigated in a phase I/II clinical adoptive immunotherapy trial. Four previously treated metastatic breast cancer patients, two with macroscopic disease and two with no evidence of disease, in complete remission (CR), were enrolled. Each apheretic peripheral blood mononuclear cell (PBMC) sample was stimulated twice with MUC-1 before infusion back into the patients. CTL responses against MCF-7 cell line and cytokine production were measured before infusion. Patients received two monthly CTL infusions and were monitored for toxicity, tumor response as well as tumor marker levels. The CTL generated from patients with high tumor burdens had less cytokine production and lower cytotoxicity of MCF-7 than the CTL of patients in CR. The differences between the two groups were observed after the two MUC1 in vitro stimulations of the cells obtained in first apheresis. This difference increased after the two MUC1 stimulations of the cells obtained in the second apheresis. The cytotoxicity function was sustained from the first infusion to the second apheresis only for the patients in CR. This suggests that tumor burden had an inverse effect on the function of the generated CTL. [ABSTRACT FROM AUTHOR]

Background Cells from AML patients can differentiate into the phenotype of DC when cultured with GM-CSF and IL-4. Such cytokine-treated AML-derived DC (AML-DC) can stimulate autologous T cells. In this study we examined whether an anti-CTLA-4 MAb (MDX-010) could enhance the generation of autologous anti-AML T cells. Methods MAb MDX-010 was added to AML PBMC cultures in the presence of GM-CSF and IL-4, a previously reported AML-DC culture method of generating anti-AML T cells. T-cell activation and proliferation were examined thereafter. Results Addition of MDX-010 to GM-CSF/IL-4-conditioned AML-DC cultures induced a mean seven-fold increase in the numbers of autologous T cells compared with cultures without MDX-010 ( P + IFN-? + CD69 + and 9.8±4.1% vs. 4±2.1% for CD8 + IFN-? + CD69 + with or without MDX-010; n =7; P <0.007, P <0.003, respectively). Discussion CTLA-4 blockade enhances the activity and numbers of AML-reactive T cells that can be stimulated by autologous AML-DC and may enhance the efficacy of adoptive immunotherapy of AML. [ABSTRACT FROM AUTHOR]

Background Recent reports have described a new strategy for differentiation and maturation of monocyte (Mo)-derived DCs within only 48 h of in vitro culture (fast-DC). Here we assess the efficacy of the fast-DC to process and present different Aspergillus fumigatus and CMV Ag preparations to autologous T cells, compared with DCs generated in standard 7-day cultures (standard-DC). Methods Adherent blood Mo were treated with GM-CSF and IL-4 (1 day for fast-DC, 5 days in the standard-DC) to generate immature DCs, and then were matured for either 1-2 days (fast-DC) or 2 days (standard-DC) with inflammatory cytokines. DCs were pulsed with A. fumigatus or CMV Ag preparation immediately prior to maturation, or infected after maturation with adeno-pp65. Mature DCs were then used to prime Ag-specific proliferative and cytotoxic T lymphocytes (CTL) responses. Results Fast-DC were CD14 - and expressed mature DC surface markers to the same degree as standard-DC, and maintained this phenotype after withdrawing cytokine from the cultures. Fast-DC and standard-DC were equally capable of inducing A. fumigatus and CMV-specific T-cell proliferation, as well as priming Ag-specific CTL activity. The Aspergillus - and CMV-specific CTL were of mixed CD3 + /CD4 + and CD3 + /CD8 + phenotype, and specifically killed autologous DC pulsed with A. fumigatus Ag and autologous CMV infected fibroblasts, respectively. Discussion Fast-DC are as effective as standard-DC in the generation of Ag-specific T-cell responses. Moreover, use of fast-DC not only reduces labor and supply cost, as well as workload and time, but also increases the number of DCs derived from adherent Mo, which may facilitate the use of DCs in clinical trials of cellular immunotherapy. [ABSTRACT FROM AUTHOR]

Virosomes are reconstituted viral envelopes which lack the genetic material but retain the cell entry and membrane fusion characteristics of the virus they are derived from. Thus, influenza virosomes are taken up by cells via receptor-mediated endocytosis, which directs the particles to the endosomal cell compartment. Subsequently, the virosomal membrane fuses with the endosomal membrane induced by the mildly acidic pH within the endosomes. This fusion process establishes continuity between the lumen of the virosome and the cell cytosol. Upon interaction of virosomes with antigen-presenting cells (APCs), protein antigens encapsulated within virosomes will be delivered to the cell cytosol, and thus, into the MHC class I presentation pathway. Indeed, virosome-mediated delivery of antigens in vivo results in efficient priming of a class I MHCrestricted cytotoxic T lymphocyte (CTL) response. [ABSTRACT FROM AUTHOR]

Forkhead box P3 (FOXP3)-specific cytotoxic CD8+ T cells are present among human peripheral blood mononuclear cells (PBMCs), especially in cancer patients. Such T lymphocytes are able not only to specifically recognize dendritic cells (DCs) that have been exposed to recombinant FOXP3 and regulatory T cells, but also to kill FOXP3+ malignant T cells. The natural occurrence of FOXP3-specific cytotoxic T lymphocytes among human PBMCs suggests a general role for these cells in the complex network of immune regulation. [ABSTRACT FROM AUTHOR]