Search

The diversity of T cells in the human liver may reflect the composition of TILs in CRLM. Our ex vivo characterization of CRLM vs. adjacent liver tissue detected CD103+CD39+CD8+ TRM cells predominantly in CRLM, which prompted further assessments. These TRM cells responded to cognate antigens in vitro. As functional activities of autologous TILs are central to the implementation of personalized cancer treatments, we applied a patient-derived xenograft (PDX) model to monitor TILs’ capacity to control CRLM-derived tumors in vivo. We established PDX mice with CRLMs from two patients, and in vitro expansion of their respective TILs resulted in opposing CD4+ vs. CD8+ TIL ratios. These CRLMs also displayed mutated KRAS, which enabled trametinib-mediated inhibition of MEK. Regardless of the TIL subset ratio, persistent or transient control of CRLM-derived tumors of limited size by the transferred TILs was observed only after trametinib treatment. Of note, a portion of transferred TILs was observed as CD103+CD8+ TRM cells that strictly accumulated within the autologous CRLM-derived tumor rather than in the spleen or blood. Thus, the predominance of CD103+CD39+CD8+ TRM cells in CRLM relative to the adjacent liver and the propensity of CD103+CD8+ TRM cells to repopulate the autologous tumor may identify these TILs as strategic targets for therapies against advanced CRC.

本文採用教育詮釋學方法，進行E. Schütz普通教育學的探究。首先探討其思想淵源；其次闡明其主要內涵；接著評價其優劣得失；最後提出其重要啟示，以提供我國作為建立教育理論和解決教育問題的參考。Schütz普通教育學的思想淵源來自F. Nietzsche的批判哲學、E. Husserl的現象學、M. Heidegger的現象學、E.Fink的教育學和M. Foucault的後現代主義。主要內涵包括教育理論、陶冶理論、研究方法論、學術理論和教育學未來的課題。Schütz的普通教育學具有提出教育理論、陶冶理論、研究方法論、學術理論和教育學未來課題的新方向等優點，但是其普通教育學也存在著一些問題。儘管如此，Schütz的普通教育學仍然相當重要，可以補充傳統教學理論的不足、改善教育實踐偏頗的缺失、開啟研究方法論的新視野、闡明教育學學術的獨特性和提供教育學未來發展方向等重要啟示，因此相當值得我們加以重視。 This article used the method of educational hermeneutics to investigate E.Schütz’s general pedagogy. First, to discuss its origin of thought. Second, to explain its main contents. Third, to evaluate its advantages and disadvantages. Fourth, to address missions of the future pedagogy, in order to offer our country as a reference to establish educational theory and to improve of educational practice. The origin of thought about Schütz’s general pedagogy comes from F. Nietzsche’s critical philosophy, E. Husserl’s phenomenology, M. Heidegger’s phenomenology, E. Fink’s pedagogy and M. Foucault’s postmodernism. Its main contents include educational theory, Bildung theory, research methodology, academic theory and future missions of pedagogy. Schütz’s general pedagogy has some advantages, it proposes new directions of educational theory, Bildung theory, research methodology, academic theory and pedagogical mission. Certainly Schütz’s general pedagogy has some problems. Even though, Schütz’s general pedagogy is still very important. It can compensate the shortages of traditional instruction theory, improve the biases of educational practice, open the new horizon of research methodology, explain the unique academic character of pedagogy and offer the future development direction of pedagogy. Therefore, it deserves us to pay attention to Schütz’s general pedagogy.

The diversity of T cells in the human liver may reflect the composition of TILs in CRLM. Our ex vivo characterization of CRLM vs. adjacent liver tissue detected CD103+CD39+CD8+ T

Simple Summary Colorectal cancer (CRC) remains the third most common cancer. Associations between intratumoral T cells, also known as tumor infiltrating lymphocytes (TILs), and the CRC patients’ responses to treatment have been described. Traditionally, TILs and antigen presenting cells (APCs) are studied separately on preserved CRC biopsies, disregarding the adjacent colonic tissue that would also be exposed to the administrated chemotherapy or radiotherapy. Thus, combined data sets on the subset composite and functional capacity of APCs and T cells within the same tumor, as well as colonic tissue, remain infrequent. Our phenotypic and functional comparison of T cell and APC subsets in tumor vs. colon from patients with CRC may give further insights into their propensity to maintain CRC treatment-induced immune responses locally in tumor and off-target colonic tissue. Abstract Although mouse models of CRC treatments have demonstrated robust immune activation, it remains unclear to what extent CRC patients’ APCs and TILs interact to fuel or quench treatment-induced immune responses. Our ex vivo characterization of tumor and adjacent colon cell suspensions suggest that contrasting environments in these tissues promoted inversed expression of T cell co-stimulatory CD80, and co-inhibitory programmed death (PD)-ligand1 (PD-L1) on intratumoral vs. colonic APCs. While putative tumor-specific CD103+CD39+CD8+ TILs expressed lower CD69 (early activation marker) and higher PD-1 (extended activation/exhaustion marker) than colonic counterparts, the latter had instead higher CD69 and lower PD-1 levels. Functional comparisons showed that intratumoral APCs were inferior to colonic APCs regarding protein uptake and upregulation of CD80 and PD-L1 after protein degradation. Our attempt to model CRC treatment-induced T cell activation in vitro showed less interferon (IFN)-γ production by TILs than colonic T cells. In this model, we also measured APCs’ CD80 and PD-L1 expression in response to activated co-residing T cells. These markers were comparable in the two tissues, despite higher IFN- γ exposure for colonic APCs. Thus, APCs within distinct intratumoral and colonic milieus showed different activation and functional status, but were similarly responsive to signals from induced T cell activation.

Simple Summary Colorectal cancer (CRC) remains the third most common malignancy. Tumor-infiltrating lymphocytes (TILs) have emerged as correlates to CRC patient outcome after treatment. The pro- or anti-tumor responses of TILs are usually assessed in cell suspensions of fresh tumors that were surgically removed a few hours earlier. We propose a platform for concurrent enumeration and in vitro functional evaluation of TILs in cryopreserved tumor biopsies, offering the benefit of postponing tumor processing and analyses of TILs in cell suspensions until clinical post-treatment responses are established. Our platform is practical considering the inconsistent time when patient samples become available for research purposes and can be readily utilized by other laboratories. With a fresh portion of tumor biopsies as benchmark, we validated the recovery of viable TILs capable of interferon (IFN)-γ responses in the cryopreserved portion of same biopsies. Ultimately, this platform could provide sufficient information on TILs, to also predict patient outcome after CRC treatments. Abstract TILs comprise functionally distinct conventional and unconventional T cell subsets and their role in responses to CRC treatments is poorly understood. We explored recovery of viable TILs from cryopreserved tumor biopsies of (chemo)-radiated patients with rectal cancer to establish a platform for retrospective TIL analyses of frozen tumors from pre-selected study cohorts. Frequencies of TIL subsets and their capacity to mount IFN-γ responses in cell suspensions of fresh vs. cryopreserved portions of the same tumor biopsies were determined for platform validation. The percentages and proportions of CD4+ TILs and CD8+ cytotoxic T lymphocytes (CTLs) among total TILs were not affected by cryopreservation. While recovery of unconventional γδ T cells and mucosal-associated invariant T cells (MAIT cells) was stable after cryopreservation, the regulatory T cells (Tregs) were reduced, but in sufficient yields for quantification. IFN-γ production by in vitro-stimulated CD4+ TILs, CTLs, γδ T cells, and MAIT cells were proportionally similar in fresh and cryopreserved tumor portions, albeit the latter displayed lower levels. Thus, the proposed platform intended for TIL analyses on cryopreserved tumor biobank biopsies holds promises for studies linking the quantity and quality of TIL subsets with specific clinical outcome after CRC treatment.

mRNA vaccines are rapidly emerging as a powerful platform for infectious diseases because they are well tolerated, immunogenic, and scalable and are built on precise but adaptable antigen design. We show that two immunizations of modified non-replicating mRNA encoding influenza H10 hemagglutinin (HA) and encapsulated in lipid nanoparticles (LNP) induce protective HA inhibition titers and H10-specific CD4+ T cell responses after intramuscular or intradermal delivery in rhesus macaques. Administration of LNP/mRNA induced rapid and local infiltration of neutrophils, monocytes, and dendritic cells (DCs) to the site of administration and the draining lymph nodes (LNs). While these cells efficiently internalized LNP, mainly monocytes and DCs translated the mRNA and upregulated key co-stimulatory receptors (CD80 and CD86). This coincided with upregulation of type I IFN-inducible genes, including MX1 and CXCL10. The innate immune activation was transient and resulted in priming of H10-specific CD4+ T cells exclusively in the vaccine-draining LNs. Collectively, this demonstrates that mRNA-based vaccines induce type-I IFN-polarized innate immunity and, when combined with antigen production by antigen-presenting cells, lead to generation of potent vaccine-specific responses., mRNA vaccines have been proven to be suitable and efficient against pandemic pathogens. However, the immune processes after mRNA vaccination that lead to robust responses remain elusive. Now in Molecular Therapy, Liang et al. (2017) define the target cells and immune responses at the vaccination sites and in the lymph nodes that result in vaccine immunity.

The ALVAC-HIV clade B/AE and equivalent SIV-based/gp120 + Alum vaccines successfully decreased the risk of virus acquisition in humans and macaques. Here, we tested the efficacy of HIV clade B/C ALVAC/gp120 vaccine candidates + MF59 or different doses of Aluminum hydroxide (Alum) against SHIV-Cs of varying neutralization sensitivity in macaques. Low doses of Alum induced higher mucosal V2-specific IgA that increased the risk of Tier 2 SHIV-C acquisition. High Alum dosage, in contrast, elicited serum IgG to V2 that correlated with a decreased risk of Tier 1 SHIV-C acquisition. MF59 induced negligible mucosal antibodies to V2 and an inflammatory profile with blood C-reactive Protein (CRP) levels correlating with neutralizing antibody titers. MF59 decreased the risk of Tier 1 SHIV-C acquisition. The relationship between vaccine efficacy and the neutralization profile of the challenge virus appear to be linked to the different immunological spaces created by MF59 and Alum via CXCL10 and IL-1β, respectively., Author summary The ALVAC-based clade C /gp120 combination is currently being tested for efficacy and licensures in South Africa in the HVTN702 HIV vaccine trial using the MF59 adjuvant rather than Alum with the intent of inducing higher neutralizing antibodies and T cell responses. Here, we present studies in 71 macaques that evaluated the relative efficacy of identical ALVAC-HIV B/C HIV vaccines with Alum or MF59 following challenge exposure to SHIV-clade C viral stocks that differed in their neutralization profiles. Our results demonstrated that use of the proprietary Novartis Alum at lower doses affects the proportion of mucosal V2-specific IgG and IgA, and that IgAs and IgGs were respectively associated with an increased or decreased risk of virus acquisition. In contrast, animals immunized with the MF59 regimen had no V2 mucosal responses, and a high level of serum neutralizing antibodies (Tier 1) to easy to neutralize viruses that were associated with a decreased risk of Tier 1 SHIV-C acquisition. In vitro studies suggest the hypothesis that the low doses of proprietary Novartis Alum used in our studies may be the underlying reason for the decreased vaccine efficacy, via lower inflammasome activation and IL-1β production.

BACKGROUND: The live attenuated BPZE1 vaccine candidate induces protection against B. pertussis and prevents nasal colonization in animal models. Here we report on the responses in humans receiving a single intranasal administration of BPZE1. METHODS: We performed multiple assays to dissect the immune responses induced in humans (n = 12) receiving BPZE1, with particular emphasis on the magnitude and characteristics of the antibody responses. Such responses were benchmarked to adolescents (n = 12) receiving the complete vaccination program of the currently used acellular pertussis vaccine (aPV). Using immunoproteomics analysis, potentially novel immunogenic B. pertussis antigens were identified. RESULTS: All BPZE1 vaccinees showed robust B. pertussis–specific antibody responses with regard to significant increase in 1 or more of the following parameters: IgG, IgA, and memory B cells to B. pertussis antigens. BPZE1–specific T cells showed a Th1 phenotype, and the IgG exclusively consisted of IgG1 and IgG3. In contrast, all aPV vaccines showed a Th2-biased response. Immunoproteomics profiling revealed that BPZE1 elicited broader and different antibody specificities to B. pertussis antigens as compared with the aPV that primarily induced antibodies to the vaccine antigens. Moreover, BPZE1 was superior at inducing opsonizing antibodies that stimulated ROS production in neutrophils and enhanced bactericidal function, which was in line with the finding that antibodies against adenylate cyclase toxin were only elicited by BPZE1. CONCLUSION: The breadth of the antibodies, the Th1-type cellular response, and killing mechanisms elicited by BPZE1 may hold prospects of improving vaccine efficacy and protection against B. pertussis transmission. TRIAL REGISTRATION: ClinicalTrials.gov NCT02453048, NCT00870350. FUNDING: ILiAD Biotechnologies, Swedish Research Council (Vetenskapsrådet), Swedish Heart-Lung Foundation.

Background Herpes zoster ophthalmicus occurs primarily in elderly or immunocompromised individuals after reactivation of varicella zoster virus (VZV). Recurrences of zoster ophthalmicus are uncommon because the reactivation efficiently boosts anti-VZV immunity. A 28-year-old female presented to our clinic with a history of multiple recurrences of zoster ophthalmicus. Methods Whole-exome sequencing (WES), analyses of VZV T-cell immunity, and pathogen recognition receptor function in primary antigen-presenting cells (APCs) and fibroblasts were performed. Results Normal VZV-specific T-cell immunity and antibody response were detected. Whole-exome sequencing identified a heterozygous nonsynonymous variant (c.2324C > T) in the Toll-like receptor 3 (TLR3) gene resulting in formation of a premature stop-codon. This alteration could potentially undermine TLR3 signaling in a dominant-negative fashion. Therefore, we investigated TLR3 signaling responses in APCs and fibroblasts from the patient. The APCs responded efficiently to stimulation with TLR3 ligands, whereas the responses from the fibroblasts were compromised. Conclusions We report a novel TLR3 variant associated with recurrent zoster ophthalmicus. Toll-like receptor 3 responses that were unaffected in APCs but diminished in fibroblasts are in line with previous reports linking TLR3 deficiency with herpes simplex virus encephalitis. Mechanisms involving compromised viral sensing in infected cells may thus be central to the described immunodeficiency.

e14510 Background: CD4 is highly and uniformly expressed T-cell lymphomas (TCL) including PTCL and CTCL, suggesting its potential as a surface target for CAR-T therapy. However, there is a significant risk of potential antigen masking by CAR introduced into tumor cells, which thereby leads to escape of tumor cells from recognition by CAR-T cells. For example, Ruella M et al ( Nat Med, 2018) reported that contaminating malignant B cells transduced with anti-CD19 CAR (called CAR-B cells) in the manufacturing process led to the CD19 antigen masking by CAR molecules. Hence, CAR-B tumor cells could not be recognized by CAR-T cells, which resulted in relapse of CAR expressing B lymphoma cells. A similar antigen masking effect might occur for the anti-CD4 CAR-T product. Methods: To identify single-chain variable fragment (scFv) without antigen-masking effect, large panels of fully human monoclonal antibodies were converted to format of CAR modality with the scFv-4-1BB-CD3z structure and introduced into CD4+ and CD8+ T cells. After lentiviral transduction, residual CD4+ T cells were quantified and the CARs with complete elimination of CD4+ T cells were selected for further validation. The selected CAR constructs were then introduced into CD4+ TCL cells HH (CAR-HH cells) to mimic the potential risk of introducing CAR into contaminating malignant T cells in the manufacturing process. Thereafter, CAR-HH cells were subject to in vitro killing assay by LB1901. Last, identified CAR constructs were further tested for in vitro and in vivo anti-tumor efficacy and off-target binding and killing. Results: Introduction of LB1901 CAR into CD4+ and CD8+ T cells led to complete elimination of CD4+ T cells, suggesting no masking effect of CAR on CD4 antigen. Furthermore, the introduction of CAR into CD4+ HH cells did not protect HH cells from being recognized and eliminated by LB1901, further confirming that the CAR modality of LB1901 does not mask CD4 antigen. An in vivo anti-tumor efficacy study showed that LB1901 exhibited dose-dependent anti-tumor activity without significant adverse effect. As low as 0.3 million CAR+ cells completely suppressed tumor growth, suggesting the potent anti-tumor activity by LB1901. Immunohistochemical analysis of normal tissues with LB1901 scFV binder showed no off-target binding. Furthermore, no killing toward CD4- cell lines and primary cells derived from vital organs or antigen-independent cytokine release was observed in vitro. Conclusions: Altogether, the in vitro and in vivo studies showed that LB1901 did not “mask” the CD4 antigen but exhibited potent anti-tumor activity without off-target effects. A phase 1 study of LB1901 CAR-T in patients with relapsed or refractory PTCL or CTCL is ongoing in the US to assess the safety and tolerability of LB1901 CAR-T (NCT04712864).

本文採用教育詮釋學方法，進行Winkler普通教育學的探究。Winkler是「反思教育學」的代表人物，對於德國閱讀教育的推動、古文中學的改革、反思教育學的形成、社會教育理論的建立和社會教育工作的實施，具有重要貢獻，被列為德國當代著名的教育學家。其普通教育學的主要內涵包括教育概念的釐清、陶冶概念的分析、理論與實踐的關係、教育學說的批判和普通教育學的課題。 Winkler的普通教育學是一種反思教育學，能夠釐清教育概念的真正意義，瞭解陶冶概念的歷史演變，闡明教育理論與實踐的關係，批判錯誤的教育學說，明白普通教育學的課題。但是其普通教育學也存在著教育觀點否定兒童本性的價值、教育理論忽略跨學科的論述和教育活動結構仍然有待說明等問題。儘管如此，Winkler的觀點依然可以作為我國建立教育理論和解決教育問題的參考，對於我 國教育學術而言，具有促進普通教育學成為核心學科、提供教師作為實施教學的指引和解決後現代社會中的教育問題等啟示，相當值得我們加以重視！ This article adopted method of educational hermeneutics to investigate M.Winkler’s general pedagogy. Winkler is not only the delegate of reflective pedagogy, but also a famous educator in modern Germany. He made a great contribution to the promotion of reading education, reform of high schools, formation of reflective pedagogy, establishment of social pedagogy theories and implementation of social work. The main contents of Winkler’s general pedagogy include clarification of the concept of education, analysis of the concept of cultivation, relation between theories and practice, critique of educational doctrines and missions of general pedagogy. Winkler’s general pedagogy is a reflective pedagogy. It has the following advantages: first, clarifying the concept of general pedagogy; second, understanding historical development of the concept of cultivation; third, explaining the relation between educational theories and practice; fourth, criticizing mistakes of educational doctrines; fifth, understanding the missions of general pedagogy. Nevertheless, it also contains the following problems: first, it denies the natural value of children; second, it neglects the discourse across disciplines; third, it needs more explanation about the structure of educational activities. Winkler’s general pedagogy can be considered as a reference for the establishment of educational theories and the solutions to educational problems. It can help us to promote general pedagogy as a core subject, and provide teachers with a guide to instructional implementation and a solution for educational problems in the postmodern society. Winkler’s general pedagogy deserves more academic attention.

The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. While methods for obtaining and characterizing murine muscle cell suspensions have been reported, protocols for nonhuman primates (NHPs) have not been well defined. NHPs comprise important in vivo models for studies of immune cell function due to their high degree of resemblance with humans. In this study, we developed and systematically compared methods to collect vaccine-injected muscle tissue to be processed into single cell suspensions for flow cytometric characterization of immune cells. We found that muscle tissue processed by mechanical disruption alone resulted in significantly lower immune cell yields compared to enzymatic digestion using Liberase. Dendritic cell subsets, monocytes, macrophages, neutrophils, B cells, T cells and NK cells were readily detected in the muscle by the classical human markers. The methods for obtaining skeletal muscle cell suspension established here offer opportunities to increase the understanding of immune responses in the muscle, and provides a basis for defining immediate post-injection vaccine responses in primates.

Myeloid-derived suppressor cells (MDSCs) are major regulators of T cell responses in several pathological conditions. Whether MDSCs increase and influence T cell responses in temporary inflammation, such as after vaccine administration, is unknown. Using the rhesus macaque model, which is critical for late-stage vaccine testing, we demonstrate that monocytic (M)-MDSCs and polymorphonuclear (PMN)-MDSCs can be detected using several of the markers used in humans. However, whereas rhesus M-MDSCs lacked expression of CD33, PMN-MDSCs were identified as CD33+ low-density neutrophils. Importantly, both M-MDSCs and PMN-MDSCs showed suppression of T cell proliferation in vitro. The frequency of circulating MDSCs rapidly and transiently increased 24 h after vaccine administration. M-MDSCs infiltrated the vaccine injection site, but not vaccine-draining lymph nodes. This was accompanied by upregulation of genes relevant to MDSCs such as arginase-1, IDO1, PDL1, and IL-10 at the injection site. MDSCs may therefore play a role in locally maintaining immune balance during vaccine-induced inflammation.

The innate immune mechanisms by which adjuvants enhance the potency and protection of vaccine-induced adaptive immunity are largely unknown. We introduce a model to delineate the steps of how adjuvant-driven innate immune activation leads to priming of vaccine responses using rhesus macaques. Fluorescently labeled HIV-1 envelope glycoprotein (Env) was administered together with the conventional aluminum salt (alum) adjuvant. This was compared to Env given with alum with preabsorbed Toll-like receptor 7 (TLR7) ligand (alum-TLR7) or the emulsion MF59 because they show superiority over alum for qualitatively and quantitatively improved vaccine responses. All adjuvants induced rapid and robust immune cell infiltration to the injection site in the muscle. This resulted in substantial uptake of Env by neutrophils, monocytes, and myeloid and plasmacytoid dendritic cells (DCs) and migration exclusively to the vaccine-draining lymph nodes (LNs). Although less proficient than monocytes and DCs, neutrophils were capable of presenting Env to memory CD4+ T cells. MF59 and alum-TLR7 showed more pronounced cell activation and overall higher numbers of Env+ cells compared to alum. This resulted in priming of higher numbers of Env-specific CD4+ T cells in the vaccine-draining LNs, which directly correlated with increased T follicular helper cell differentiation and germinal center formation. Thus, strong innate immune activation promoting efficient vaccine antigen delivery to infiltrating antigen-presenting cells in draining LNs is an important mechanism by which superior adjuvants enhance vaccine responses.

Advances in HIV-1 vaccine clinical trials and preclinical research indicate that the virus envelope glycoproteins (Env) are likely to be an essential component of a prophylactic vaccine. Efficient Ag uptake and presentation by dendritic cells (DCs) is important for strong CD4+ Th cell responses and the development of effective humoral immune responses. In this study, we examined the capacity of distinct primary human DC subsets to internalize and present recombinant Env to CD4+ T cells. Consistent with their specific receptor expression, skin DCs bound and internalized Env via C-type lectin receptors, whereas blood DC subsets, including CD1c+ myeloid DCs, CD123+ plasmacytoid DCs (PDCs), and CD141+ DCs exhibited a restricted repertoire of C-type lectin receptors and relied on CD4 for uptake of Env. Despite a generally poor capacity for Ag uptake compared with myeloid DCs, the high expression of CD4 on PDCs allowed them to bind and internalize Env very efficiently. CD4-mediated uptake delivered Env to EEA1+ endosomes that progressed to Lamp1+ and MHC class II+ lysosomes where internalized Env was degraded rapidly. Finally, all three blood DC subsets were able to internalize an Env-CMV pp65 fusion protein via CD4 and stimulate pp65-specific CD4+ T cells. Thus, in the in vitro systems described in this paper, CD4-mediated uptake of Env is a functional pathway leading to Ag presentation, and this may therefore be a mechanism used by blood DCs, including PDCs, for generating immune responses to Env-based vaccines.

To study whether in-vivo recruitment of dendritic cells in response to antigen administration in the skin is altered during HIV-1 infection.Skin punch biopsies were collected from HIV-1-positive as well as seronegative individuals at 48 h after intradermal injection of inactivated antigens of mumps virus, Candida albicans, or purified protein derivate (PPD) from Mycobacterium tuberculosis.Cryosections were analyzed by in-situ staining and computerized imaging.Control skin biopsies showed that there was no difference in the number of skin-resident dendritic cells between seronegative and HIV-1-positive individuals. Antigen injection resulted in substantial infiltration of dendritic cells compared to the frequencies found in donor-matched control skin. In HIV-1-positive individuals, CD123(+)/CD303(+) plasmacytoid dendritic cells and CD11c myeloid dendritic cells, including the CD141(+) cross-presenting subset, were recruited at lower levels compared to healthy controls in response to PPD and mumps but not C. albicans. The level of dendritic cell recruitment correlated with the frequencies of T cells infiltrating the respective antigen sites. Ki67(+) cycling T cells at the injection sites were much more frequent in response to each of the antigens in the HIV-1-positive individuals, including those with AIDS, compared to healthy controls.Multiple dendritic cell subsets infiltrate the dermis in response to antigen exposure. There was no obvious depletion or deficiency in mobilization of dendritic cells in response to antigen skin tests during chronic HIV-1 infection. Instead, the levels of antigen-specific memory T cells that accumulate at the antigen site may determine the level of dendritic cell infiltration.

Neutrophils are critical cells of the innate immune system and rapidly respond to tissue injury and infection. Increasing evidence also indicates that neutrophils have versatile functions in contributing to adaptive immunity by internalizing and transporting antigen and influencing antigen-specific responses. Here, we demonstrate that freshly isolated human neutrophils can function as antigen-presenting cells (APCs) to memory CD4+ T cells. Neutrophils pulsed with the cognate antigens cytomegalovirus pp65 or influenza hemagglutinin were able to present the antigens to autologous antigen-specific CD4+ T cells in a major histocompatibility complex class II (MHC-II; HLA-DR)-dependent manner. Although myeloid dendritic cells and monocytes showed superior presenting ability, neutrophils consistently displayed antigen presentation capability. Upregulation of HLA-DR on neutrophils required the presence of the antigen-specific or activated T cells whereas exposure to innate stimuli such as Toll-like receptor ligands was not sufficient. Neutrophils sorted from vaccine-draining lymph nodes from rhesus macaques also showed expression of HLA-DR and were capable of presenting vaccine antigen to autologous antigen-specific memory CD4+ T cells ex vivo. Altogether, the data demonstrate that neutrophils can adapt a function as APCs and, in combination with their abundance in the immune system, may have a significant role in regulating antigen-specific T-cell responses.

本研究採用教育詮釋學的方法，探討Benner改革教育學的思想淵源、主要內涵、優劣得失，以及重要啟示。Benner深受Rousseau教育學思想、Kant批判哲學、Herbart普通教育學、Humboldt語言哲學、Hegel辯證哲學、Schleiermacher教育理論、Fink存在現象學、Derbolav實踐學和Kuhn科學哲學的影響，注重改革教育學與常規教育學的關係、教育學與政治學的關係、改革教育學歷史撰寫的形式、現代教育學在德國的發展、西方占領區和西德的國家學校改革及學校實驗等問題的探究，具有擴大改革教育學的探討範圍、轉變改革教育學的撰寫方式、澄清改革教育學的核心關係、充實改革教育學的實質內涵，以及指出教育改革運動的優劣得失等優點。儘管Benner改革教育學存在著一些問題，但是，仍然可以做為我國建構教育改革理論和進行教育改革活動的參考，在學術研究和教育實務上，值得我們加以重視。 This study explored the thinking background and major contents of Benner’s reform pedagogy and analyzed its advantages, disadvantages, and significant implications through the methodology of educational hermeneutics. Dietrich Benner was deeply influenced by the thinking of various ideologists including Rousseau’s pedagogy, Kant’s criticism, Herbart’s general pedagogy, Humboldt’s linguistic philosophy, Hegel’s dialectics, Schleiermacher’s educational theories, Fink’s existential phenomenology, Derbolav’s praxeology, and Kuhn’s philosophy of science. His research focused on the correlation between the reform pedagogy and the traditional pedagogy, and between pedagogy and political science, as well as the style of history writing for the reform pedagogy, the development of modern pedagogy in Germany, the reform and educational experiment in national schools in Allied-occupied and West Germany, etc. His research has contributed to the expanded study of reform pedagogy, facilitated changes in its writing style, clarified its core relations, enriched its contents, and highlighted the advantages and shortcomings of this initiative. Benner’s reform pedagogy is not without its defeces, but it offers worthwhile information in terms of both academic research and educational practice, when we endeavor to construct theories or design activities for educational reform in Taiwan.

Plasmacytoid dendritic cells (pDCs) are rarely present in normal skin but have been shown to infiltrate lesions of infections or autoimmune disorders. Here, we report that several DC subsets including CD123(+) BDCA-2/CD303(+) pDCs accumulate in the dermis in indurations induced by the tuberculin skin test (TST), used to screen immune sensitization by Mycobacterium tuberculosis. Although the purified protein derivate (PPD) used in the TST did not itself induce pDC recruitment or IFN-α production, the positive skin reactions showed high expression of the IFN-α-inducible protein MxA. In contrast, the local immune response to PPD was associated with substantial cell death and high expression of the cationic antimicrobial peptide LL37, which together can provide a means for pDC activation and IFN-α production. In vitro, pDCs showed low uptake of PPD compared with CD11c(+) and BDCA-3/CD141(+) myeloid DC subsets. Furthermore, supernatants from pDCs activated with LL37-DNA complexes reduced the high PPD uptake in myeloid DCs, as well as decreased their capacity to activate T-cell proliferation. Infiltrating pDCs in the TST reaction site may thus have a regulatory effect upon the antigen processing and presentation functions of surrounding potent myeloid DC subsets to limit potentially detrimental and excessive immune stimulation.

A recombinant vaccine containing Aventis Pasteur's canarypox vector (ALVAC)-HIV and gp120 alum decreased the risk of HIV acquisition in the RV144 vaccine trial. The substitution of alum with the more immunogenic MF59 adjuvant is under consideration for the next efficacy human trial. We found here that an ALVAC-simian immunodeficiency virus (SIV) and gp120 alum (ALVAC-SIV + gp120) equivalent vaccine, but not an ALVAC-SIV + gp120 MF59 vaccine, was efficacious in delaying the onset of SIVmac251 in rhesus macaques, despite the higher immunogenicity of the latter adjuvant. Vaccine efficacy was associated with alum-induced, but not with MF59-induced, envelope (Env)-dependent mucosal innate lymphoid cells (ILCs) that produce interleukin (IL)-17, as well as with mucosal IgG to the gp120 variable region 2 (V2) and the expression of 12 genes, ten of which are part of the RAS pathway. The association between RAS activation and vaccine efficacy was also observed in an independent efficacious SIV-vaccine approach. Whether RAS activation, mucosal ILCs and antibodies to V2 are also important hallmarks of HIV-vaccine efficacy in humans will require further studies.

Designing strategies for targeting antigens to dendritic cells is a major goal in vaccinology. Here, PLGA (poly lactic-co-glycolic acid) microspheres and with several surface modifications that affect to their uptake by human blood primary dendritic cells and monocytes have been evaluated. Higher uptake was found by all the cell types when cationic microspheres (PLGA modified with polyethylene imine) were used. These cationic particles were in vivo evaluated in mice. In addition, MPLA(1) or poly(I:C)(2) and α-GalCer(3) were also encapsulated to address their adjuvant effect. All the microspheres were able to produce humoral immune responses, albeit they were higher for cationic microspheres. Moreover, surface charge seemed to have a role on biasing the immune response; cationic microspheres induced higher IFN-γ levels, indicative of Th1 activation, while unmodified ones mainly triggered IL4 and IL17A release, showing Th2 activation. Thus, we have shown here the potential and versatility of these MS, which may be tailored to needs.

Use of Thermal Interface Materials (TIM) is a common thermal solution approach in handheld devices to reduce junction temperature and control device skin temperature. This work summarizes the thermal benefits of using a TIM for enhancing the user experience and increasing System on Chip (SoC) performance. On the other hand, TIM induces a load on the package which in turn can impose stress on the package solder joints. This paper explains the impact of a variety of parameters such as TIM material and thickness and system boundary conditions on thermal performance of the SoC/system and the load distribution on solder joints. The complexity of mechanical load distribution is discussed through extensive data collection and simulation in phone and tablet form factors. Design guidelines for selection of appropriate TIM are proposed to improve the thermal performance without compromising the reliability of the SoC package.Copyright © 2015 by ASME

Nonlive vaccine platforms that induce potent cellular immune responses in mucosal tissue would have broad application for vaccines against infectious diseases and tumors. Induction of cellular immunity could be optimized by targeted activation of multiple innate and costimulatory signaling pathways, such as CD40 or TLRs. In this study, we evaluated immune activation and elicitation of T cell responses in nonhuman primates after immunization with peptide Ags adjuvanted with an agonistic anti-CD40Ab, with or without the TLR3 ligand poly IC:LC. We found that i.v. administration of the anti-CD40Ab induced rapid and transient innate activation characterized by IL-12 production and upregulated costimulatory and lymph node homing molecules on dendritic cells. Using fluorescently labeled Abs for in vivo tracking, we found that the anti-CD40Ab bound to all leukocytes, except T cells, and disseminated to multiple organs. CD4+ and CD8+ T cell responses were significantly enhanced when the anti-CD40Ab was coadministered with poly IC:LC compared with either adjuvant given alone and were almost exclusively compartmentalized to the lung. Notably, Ag-specific T cells in the bronchoalveolar lavage were sustained at ∼5–10%. These data indicate that systemic administration of anti-CD40Ab may be particularly advantageous for vaccines and/or therapies that require T cell immunity in the lung.

In this paper, an efficient multi-to-single mode converter based on a tapered multimode fiber is presented. This novel tapered multimode fiber has a multimode input end and a single mode output end, which is fabricated by a reproducible chemical etching process. The core diameter of the thicker end of the fiber is around 150 μm (a typical multimode fiber case) while the core diameter of the thinner end of the fiber is around 10 μm (a typical single mode fiber case). By launching the light into the tapered multimode fiber from the thicker end, the multimode light field distribution at the thicker input end becomes a single mode light field distribution at the thinner output end. Thus, a unique multi-to-single mode conversion is achieved. The major advantages of this mode converter include: (1) high conversion efficiency (>95% based on theoretical analysis), (2) compact size and (3) low cost, which make it very useful for a variety of applications, such as: (1) efficiently coupling the light from a laser, in particular a semiconductor diode laser, into a single mode fiber; (2) highly sensitive evanescent wave based fiber optic sensors and (3) effective mode conversion and connection between multimode and single mode fibers. Experimental results have confirmed this theoretical prediction (i.e., a multi-to-single mode conversion can be realized via a special tapered multimode fiber).

A growing body of evidence suggests that pathological lesions express tissue-specific molecular targets or biomarkers within the tissue. Interleukin-4 receptor (IL-4R) is overexpressed in many types of cancer cells, including lung cancer. Here we investigated the properties of IL-4R-binding peptide-1 (IL4RPep-1), a CRKRLDRNC peptide, and its ability to target the delivery of liposomes to lung tumor. IL4RPep-1 preferentially bound to H226 lung tumor cells which express higher levers of IL-4R compared to H460 lung tumor cells which express less IL-4R. Mutational analysis revealed that C1, R2, and R4 residues of IL4RPep-1 were the key binding determinants. IL4RPep-1-labeled liposomes containing doxorubicin were more efficiently internalized in H226 cells and effectively delivered doxorubicin into the cells compared to unlabeled liposomes. In vivo fluorescence imaging of nude mice subcutaneously xenotransplanted with H226 tumor cells indicated that IL4RPep-1-labeled liposomes accumulate more efficiently in the tumor and inhibit tumor growth more effectively compared to unlabeled liposomes. Interestingly, expression of IL-4R was high in vascular endothelial cells of tumor, while little was detected in vascular endothelial cells of control organs including the liver. IL-4R expression in cultured human vascular endothelial cells was also up-regulated when activated by a pro-inflammatory cytokine tumor necrosis factor-α. Moreover, the up-regulation of IL-4R expression was observed in primary human lung cancer tissues. These results indicate that IL-4R-targeting nanocarriers may be a useful strategy to enhance drug delivery through the recognition of IL-4R in both tumor cells and tumor endothelial cells.

SummaryAnalysis of hematopoietic stem cell function in nonhuman primates provides insights that are relevant for human biology and therapeutic strategies. In this study, we applied quantitative genetic barcoding to track the clonal output of transplanted autologous rhesus macaque hematopoietic stem and progenitor cells over a time period of up to 9.5 months. We found that unilineage short-term progenitors reconstituted myeloid and lymphoid lineages at 1 month but were supplanted over time by multilineage clones, initially myeloid restricted, then myeloid-B clones, and then stable myeloid-B-T multilineage, long-term repopulating clones. Surprisingly, reconstitution of the natural killer (NK) cell lineage, and particularly the major CD16+/CD56− peripheral blood NK compartment, showed limited clonal overlap with T, B, or myeloid lineages, and therefore appears to be ontologically distinct. Thus, in addition to providing insights into clonal behavior over time, our analysis suggests an unexpected paradigm for the relationship between NK cells and other hematopoietic lineages in primates.

Vaccines constructed from rare-serotype recombinant adenovirus vectors (rAd) such as rAd serotype 28 (rAd28) and rAd35 are currently being explored as alternatives to rAd5-based vaccines because they circumvent the problems with pre-existing immunity that complicate the effectiveness of rAd5 vaccines. However, previous work has demonstrated that the immunogenicity of rAd28 and rAd35 is substantially lower than rAd5. Here we show that rAd28 and rAd35 increase apoptosis of antigen presenting cells (APCs), such as monocytes, relative to rAd5 and mock infected controls. APCs undergoing apoptosis showed an increased loss of vector-insert expression. Loss of vector-insert expression correlated with activation of NK cells, which resulted in apoptosis of co-cultured monocytes. Finally, we show that activation of NK cells is dependent on IFNα which is produced by exposure to rAd28 or rAd35, but not to rAd5. Taken together, these data demonstrate that IFNα-induced activation of NK cells leads to increased monocyte apoptosis and subsequent vector-insert loss. This may be a possible mechanism that results in reduced immunogenicity of rAd28 and rAd35-based vectors.

A recombinant vaccine containing Aventis Pasteur’s canarypox vector (ALVAC)–HIV and gp120 alum decreased the risk of HIV acquisition in the RV144 vaccine trial. The substitution of alum with the more immunogenic MF59 adjuvant is under consideration for the next efficacy human trial. We found here that an ALVAC–simian immunodeficiency virus (SIV) and gp120 alum (ALVAC–SIV + gp120) equivalent vaccine, but not an ALVAC–SIV + gp120 MF59 vaccine, was efficacious in delaying the onset of SIVmac251 in rhesus macaques, despite the higher immunogenicity of the latter adjuvant. Vaccine efficacy was associated with alum-induced, but not with MF59-induced, envelope (Env)-dependent mucosal innate lymphoid cells (ILCs) that produce interleukin (IL)-17, as well as with mucosal IgG to the gp120 variable region 2 (V2) and the expression of 12 genes, ten of which are part of the RAS pathway. The association between RAS activation and vaccine efficacy was also observed in an independent efficacious SIV-vaccine approach. Whether RAS activation, mucosal ILCs and antibodies to V2 are also important hallmarks of HIV-vaccine efficacy in humans will require further studies.

Background Most vaccines are delivered into the muscle although it contains very few potent antigen presenting cells, such as dendritic cells (DCs) that are critical for driving adaptive immune responses. Understanding the early mechanisms that dictate vaccine responses and why some adjuvants like the oil-in-water emulsion MF59 are shown to be more potent than alum is important for the design of new vaccines. Here, we investigated the recruitment of immune cells to the vaccine injection site and uptake of a clinically relevant HIV-1 envelope glycoprotein (Env) and MF59 in a non-human primate (NHP) model.

OA25.01 Background: The RV144 HIV vaccine trial resulted in limited, but significant protection, from HIV acquisition. Serum antibodies directed to the Env variable regions 1 and 2 (V1/V2) inversely correlated with the risk of HIV-1 infection and sieve analysis demonstrated immunologic pressure on two regions of the V2. Prior macaque studies demonstrated that a similar vaccine for SIV (ALVAC-SIV) induced protection from SIVmac251 acquisition in a low dose neonatal challenge model, but not in adult high dose challenge models. Methods: We challenged ALVAC-SIV/gp120/alum vaccinated adult macaques intrarectally with repeated low doses of SIVmac251 in a study powered to allow benchmarking against the results of RV144. An additional arm of the study evaluated these vaccines together with the oil-in-water emulsion MF59 adjuvant. Results: We found that alum protected macaques from SIVmac251 acquisition while MF59 did not despite its ability to elicit higher systemic T-cell and antibodies responses. MF59 altered homing of antibody producing cells and increased the frequency of CXCR3+ plasmablasts in blood that positively correlated with anti-envelope IgA serum levels and phagocytosis. Alum, in contrast, increased the frequency of plasmablasts expressing the mucosal integrin a4b7 that positively correlated with IgA responses to cyclic V2 in rectal mucosa. In the alum group mucosal IgG to cyclic V2 correlated with lower risk of SIVmac251 acquisition. However, mucosal IgG to linear and cyclic V2 correlated with an increased risk of SIVmac251 acquisition in the MF59 group. The two adjuvants modulated distinct signaling pathways and RAS, a signal transducer that facilitates cross talk among B-cells, T-cells and antigen presenting cells, was demonstrated to be a biomarker of vaccine efficacy in the alum group. Conclusions: These data highlight the importance of the quality of the mucosal antibodies to V2 in protection and suggest that activation of RAS may constitute a novel approach to improve vaccine efficacy against HIV.

The uplink performance of a 4G LTE system, which includes an eNodeB modem and higher layer network elements, is presented in this paper. The physical layer link performance of this pre-commercial prototype, along with comparisons to simulations, is presented. The performance characterization in the lab is done for the shared channel as well as the control channel. Further, we present a demonstration of adaptive modulation and coding capabilities in the LTE uplink.

Recently most manufacturing operations in FAB are done in automated manner. Following the predefined process routing information, every lot is automatically dispatched, transported, and processed by manufacturing execution system (MES). However all kinds of wafer sorting operation cannot be predefined in the route because unplanned wafer sorting is required according to the process result, condition of process equipment, or other reason. This paper describes concerns to automate sorter operation and proposes the framework to cover both planned and unplanned sorter operation in the same automated manner with event driven base.

A number of rhenium and manganese carbonyl complexes have been prepared from the reactions of the ligands f₄fars, f₆fars and .f₄fos with Re₂ (CO)₁₀, Mn₂(CO)₁₀ and ReMn(CO)₁₀. They can be classified into two types. The first type has the general formula (L-L)M₂(CO)₈ in which the ligand L-L bridges the two metal carbonyl moieties. The second type of complexes, i- f₄farsM₂(CO)₈, are isomers of the first type and have the structure (CO)₄M-(CH₃)₂As-M(CO)₄-(CH₃)₂AsC = CCF₂CF₂ which is the result of a ligand rearrangement reaction. The new complexes (L-L)M₂(CO)₈ are the first examples of 1igand bridged derivatives containing metal carbonyl fragments otherwise held together only by metal-metal bonds. They readily react with iodine with cleavage of the M-M bond to yield (L-L)[M(CO)₄I] ₂. The kinetics of these fast cleavage reactions were studied by the stopped-flow technique. The enthalpies of activation for f₄farsRe₂(CO)₈, f₆farsRe₂(CO)₈, and f₄fars(CO)₄,ReMn(CO)₄ are 6.9 ± 0.3, 9.3 ± 0.9, 14.2 + 0.9 kcal mole⁻¹ respectively while the approximate entropy values are -13.8±1.0, -11.1 ± 3.0 and 1.4 ± 3.2 e.u. respectively. No activation parameters were obtainable for the reaction of f₄farsMn₂(CO)₈, with iodine because it has a half-life time of less than 2 milliseconds at 25°C. Prolonged heating of f₄farsMn₂(CO)₈, (M = Re, Mn) results in the thermal rearrangement of these complexes to give their isomers i-(L-L)Mn₂(CO)₈,. The kinetics of these thermal rearrangement reactions were studied by ¹H n.m.r. spectroscopy. The enthalpies of activation for f₄farsMn₂(CO)₈, f₄farsRe₂(CO)₈, and f₄fars(CO)₄,ReMn(CO)₄ are 36.8 ± 2.0, 46.8 ± 0.5 and 39.2 ± 5.2 respectively while the entropy values are 15.0 ± 4.7, 23.2 x 1.1 and 20.9 ± 2.8 e.u. respectively. The mechanisms of these reactions and the iodine cleavage reactions are discussed and possible intermediates are suggested in the light of the kinetic and related chemical and spectroscopic studies. The iodine complexes f₄fars[M(CO)₄I]₂ (M = Re, Mn) are unstable and, on heating, give metal carbonyl iodides and the chelate complexes f4farsM(CO)₃I. The latter are related to f₄farsM(CO)₃Cl (M = Re, Mn) which were obtained as by-products in the reaction of the ligands with the parent carbonyls. As an appendix to this thesis, reactions of some fluorinated acetylenes with tetrakis(triphenylphosphine)-platinum(O) and the reaction of bis (trifluoromethyl)diazomethane with bis(trimethylsilyl)acetylene are described.