Dendritic Research Articles

Wear and oxidation resistances of AlCrFeNiTi-based high entropy alloys

Based on the AlCoCrFeNi high entropy alloy, by replacing Co with Ti, new cobalt-free high entropy alloys AlCrFeNiTi and its derivative AlCrFeNiTiMn0.5 were designed and prepared by arc melting, and as-cast microstructures, wear and oxidation behaviors were investigated. The results show that two alloys are all composed of an ordered body centered cubic (BCC) and a disordered BCC phases, and typical dendritic structures including dendritic (DR), interdentritic (ID) and ‘eutectic phase’ areas were observed, implying the feature of near-eutectic high entropy alloys for AlCrFeNiTi and AlCrFeNiTiMn0.5. Adhesive wear mechanisms for these two alloys are indentified mainly as delamination wear, while oxidative wear is gradually dominated in AlCrFeNiTi. The wear resistance declines with addition of a half mole of Mn into AlCrFeNiTi due to rising wear coefficients of alloys. The oxidation kinetics of AlCrFeNiTi and AlCrFeNiTiMn0.5 alloys oxidized at 900 °C meet the exponential rate law, and the oxidation rate of AlCrFeNiTi is lower than that of AlCrFeNiTiMn0.5, suggesting a better oxidation resistance. Comparing with AlCrFeNiTi, a new Mn2O3 subscale was formed between the outer TiO2 scale and the underlying Al2O3 subscale on surfaces of AlCrFeNiTiMn0.5, which is consistent with theoretical analysis from the view of forming ability level by the first principles calculations.

Effects of annealing on the microstructural evolution and phase transition in an AlCrCuFeNi2 high-entropy alloy

An AlCrCuFeNi2 high entropy alloy (HEA) was prepared by arc melting, followed by annealing at different temperatures. The elemental distributions, phase formation, morphology and microstructural evolution of the HEA in both the as-cast state and annealed state were investigated. The results indicate that the HEA undergoes elements segregation, precipitation and spinodal decomposition. The as-cast alloy consists of Cr-Fe-Ni rich FCC dendritic (DR) phase and BCC interdendritic (ID) phase. Spherical Fe-Cr rich BCC precipitates were found to disperse in Al-Ni rich B2 (ordered BCC) matrix. After heat treatments, the distribution of elements clearly changes, along with changes of the constituent phase and morphology. After annealing at 600°C, the DR region remains Cr-Fe-Ni rich phase, while some spherical precipitates transform into the needle-like structure within the ID region. The L12 (ordered FCC) nanorod-shaped phase ((Ni,Cu)3Al) and plate-like Al-Ni rich phase form within the DR region when annealing up to 900°C. The L12 phase almost dissolves in the FCC matrix due to the order-disorder transition and an obvious coarsening of the Fe-Cr rich phase occurs after annealing at 1100°C. The segregation of Cu atoms at the interface between DR and ID regions is found at the as-cast state, while a uniform distribution of Cu atoms in DR and ID regions was observed after annealing at 1100°C.

Th17 immunity to tuberculosis requires CD40-CD40L interaction and is enhanced by CD40 engagement on Mycobacterium tuberculosis-infected dendritic cells

Abstract Mycobacterium tuberculosis (Mtb) impairs dendritic (DC) cell functions and induces suboptimal antigen-specific CD4 T cell immune responses that are poorly protective. Mucosal T-helper 17 (Th17) cells have emerged as important for protecting against tuberculosis (TB), but the mechanisms involved in generating antigen-specific Th17 responses during Mtb infection are not well defined. We previously reported that Mtb restricts Th17 responses and impairs CD40 expression on dendritic cells (DCs). We now demonstrate that generating IL-17 responses to Mtb requires CD40-dependent costimulation. CD40-deficient DCs were unable to induce antigen-specific IL-17 responses after Mtb infection despite the production of Th17-polarizing innate cytokines. Disrupting the interaction between CD40 on DCs and its ligand CD40L on antigen-specific CD4 T cells, genetically or via antibody blockade, significantly reduced generation of antigen specific IL-17 responses; cytokines were unable to compensate for this reduction. Importantly, engaging CD40 on DCs with a multimeric CD40 agonist (CD40LT) enhanced antigen-specific IL-17 generation in ex vivo DC-T cell co-culture assays. Further, intratracheal instillation of Mtb-infected DCs treated with CD40LT significantly augmented antigen-specific Th17 responses in vivo in the lungs and lung-draining lymph nodes of mice. Finally, boosting CD40-CD40L interactions promoted balanced Th1/Th17 responses in a setting of mucosal DC transfer, and conferred enhanced control of lung bacterial burdens following aerosol challenge with Mtb. Our results demonstrate that targeting the CD40-CD40L pathway represents a novel strategy to enhance early Th17 responses and improves adaptive immunity to TB.

Sphingosine-1-phosphate receptor-1 (S1P1) is expressed by lymphocytes, dendritic cells, and endothelium and modulated during inflammatory bowel disease.

The sphingosine-1-phosphate receptor-1 (S1P1) agonist ozanimod ameliorates ulcerative colitis, yet its mechanism of action is unknown. Here we examine the cell subsets that express S1P1 in intestine using S1P1-eGFP mice, the regulation of S1P1 expression in lymphocytes after administration of DSS, after colitis induced by transfer of CD4+CD45RBhi cells and by crossing a mouse with TNF-driven ileitis with S1P1-eGFP mice. We then assayed the expression of enzymes that regulate intestinal S1P levels, and the effect of FTY720 on lymphocyte behavior and S1P1 expression. We found that not only T and B cells express S1P1, but also dendritic (DC) and endothelial cells. Furthermore, chronic but not acute inflammatory signals increased S1P1 expression, while the enzymes that control tissue S1P levels in mice and humans with IBD were uniformly dysregulated, favoring synthesis over degradation. Finally, we observed that FTY720 reduced T cell velocity and induced S1P1 degradation and retention of naïve but not effector T cells. Our data demonstrate that chronic inflammation modulates S1P1 expression and tissue S1P levels and suggests that the anti-inflammatory properties of S1PR agonists might not be solely due to their lymphopenic effects, but also due to potential effects on DC migration and vascular barrier function.

Role of decidual CD14+ macrophages in the homeostasis of maternal–fetal interface and the differentiation capacity of the cells during pregnancy and parturition

ObjectiveDecidual macrophages (dMΦs) have been implicated in fetal tolerance, but little information is known regarding their differentiation capacity and interactions with T cells. The present study aimed to investigate the immunological characteristics of dMΦs at mid and term pregnancy. MethodsThe dMΦs were analyzed for their phenotypes and cytokine production by flow cytometry and ELISA, respectively. The transendothelial trafficking model was implemented to allow the dMΦs to differentiate. The differentiated cells from dMΦs were also measured for their phenotypes and cytokine production with same methods. The capacity of dMΦs or the differentiated cells from dMΦs to stimulate allogeneic T lymphocyte proliferation was evaluated by T lymphocyte stimulation assays. T cell differentiation was determined by flow cytometry. ResultsThe dMΦs in the mid-pregnancy (Mid-dMΦs) resembled the M2 phenotype. The differentiated cells from Mid-dMΦs had little stimulatory capacity on T cell proliferation and favored regulatory T cell differentiation. The dMΦs at term differentiated into dendritic (DC)-like cells, stimulating T cell activation, proliferation, and differentiation into IFN-γ-producing T cellsdecidual ConclusionsThe present study suggests that the differences in phenotypes and cytokine production between Mid- and Term-dMΦs relate to their different roles in the homeostasis of the maternal–fetal interface. Mid-dMΦs differentiate into DC-like cells with immunosuppressive properties, playing an important role in maintaining homeostasis required for a successful pregnancy. Term-dMΦs differentiate into DC-like cells with immunostimulatory properties, likely involved in the activation of labor. The different differentiation capacities of dMΦs in the varied pregnancy stages may be due to the placental microenvironment.

Tumor-derived alpha fetoprotein directly impacts human natural killer cell activity and viability

Alpha-fetoprotein (AFP) is an oncofetal antigen produced by hepatocellular carcinomas (HCC). Previous studies demonstrated that tumor-derived AFP (tAFP) is a glycoprotein that has an immunosuppressive role on natural killer (NK), T, B, and dendritic (DC) cells which may play a role in HCC pathogenesis. Defects in NK cells have been attributed to tAFP-mediated immunosuppression of DC. However, a direct tAFP effect on NK cells remains unexplored. Here we compared the ability of cord blood-derived AFP (nAFP) to that of tAFP to modulate human NK cell activity and longevity in vitro. Short-term exposure to tAFP and, especially, nAFP proteins induced a unique pro-inflammatory, IL-2 hyperresponsive phenotype in healthy donor NK cells as measured by CD69 upregulation, IL-1β, IL-6 and TNF secretion, and enhanced tumor cell killing. In contrast, extended co-culture with tAFP, but not nAFP, inhibited NK cell proliferation and viability. NK cell activation was directly mediated by the AFP protein itself, while their viability was affected by the low molecular mass cargo that co-purified with tAFP. Overall, these data show that nAFP and tAFP induce similar yet distinct changes in NK cell function and viability, respectively. Defining the impact of circulating AFP on NK cells may be crucial to understand the NK cell functional deficits described in HCC patients.

Phase Transformation and Aging Behavior of Al0.5CoCrFeNiSi0.2 High-Entropy Alloy

An Al0.5CoCrFeNiSi0.2 high-entropy alloy was prepared by vacuum arc melting. The alloy was aged from 700 to 1100 °C. The effects of aging on the phase transformation and mechanical performances were explored. The as-cast alloy showed a dendritic (DR) microstructure. The DR region was an Fe,Cr-rich FCC phase, while the interdendritic (ID) region was a spinodal structure composed of Fe,Cr-rich BCC (A2) and Ni,Al-rich BCC (B2) phases. At aging temperatures between 700 and 900 °C, the Fe,Cr-rich BCC (A2) phase in the ID region transformed into σ and Fe,Cr-rich FCC phases. Meanwhile, some Ni,Al-rich FCC phase particles precipitated from the DR region. During aging at 1100 °C, the DR microstructure disappeared, and a microstructure composed of Fe,Cr-rich FCC and Ni,Al-rich BCC (B2) phases both possessing a lamellar shape was developed. The alloy exhibited evident hardening and lower tensile strain when the aging temperature was lower than 1000 °C, which was mainly attributed to the generation of the σ phase in the ID region. However, a contrasting behavior was observed when the aging temperature was higher than 1000 °C, which was attributed to the redissolution of the σ phase and the microstructure coarsening.

Human Vδ2+ γδ T Cells Differentially Induce Maturation, Cytokine Production, and Alloreactive T Cell Stimulation by Dendritic Cells and B Cells

Human γδ T cells expressing the Vγ9Vδ2 T cell receptor can induce maturation of dendritic cells (DC) into antigen-presenting cells (APC) and B cells into antibody-secreting plasma cells. Since B cells are capable of presenting antigens to T cells, we investigated if Vγ9Vδ2 T cells can influence antigen-presentation by these cells. We report that Vγ9Vδ2 T cells induced expression of CD86, HLA-DR, and CD40 by B cells and stimulated the release of IL-4, IL-6, TNF-α, and IgG, IgA, and IgM. Vγ9Vδ2 T cells also augmented the ability of B cells to stimulate proliferation but not IFN-γ or IL-4 release by alloreactive T cells. In contrast, Vγ9Vδ2 T cells induced expression of CD86 and HLA-DR and the release of IFN-γ, IL-6, and TNF-α by DC and these DC stimulated proliferation and IFN-γ production by conventional T cells. Furthermore, CD86, TNF-α, IFN-γ, and cell contact were found to be important in DC activation by Vγ9Vδ2 T cells but not in the activation of B cells. These data suggest that Vγ9Vδ2 T cells can induce maturation of B cells and DC into APC, but while they prime DC to stimulate T helper 1 (TH1) responses, they drive maturation of B cells into APC that can stimulate different T cell responses. Thus, Vγ9Vδ2 T cells can control different arms of the immune system through selective activation of B cells and DC in vitro, which may have important applications in immunotherapy and for vaccine adjuvants.

Alpha fetoprotein directly induces a unique pro-inflammatory, IL-2 hyperresponsive phenotype in human natural killer cells

Alpha fetoprotein (AFP) is an oncofetal antigen commonly produced by hepatocellular carcinomas (HCC). Previous studies have shown that tumor-derived AFP (tAFP) has an immunosuppressive role on natural killer (NK), T, B, and dendritic (DC) cells which may play a role in HCC pathogenesis. Defects in NK cell frequency and function have partially been attributed to tAFP-mediated immunosuppression of DC function. However, a direct tAFP effect on NK cells remains unclear. Here we examine the ability of cord blood-derived AFP (nAFP) and tAFP to modulate human NK cell activity in vitro. We show that exposure to tAFP or, especially, nAFP proteins induces a unique pro-inflammatory NK cell activation profile as measured by CD69 upregulation, IL-1β and IL-6 secretion, and enhanced tumor cell killing. Interestingly, AFP-treated plus interleukin-2 (IL-2) stimulated cultures promote a degree of NK cell activation that is higher than that of NK cells activated with IL-2 alone by both phenotypic and functional measures, including elevated IFN-γ and GM-CSF secretion. To confirm that the observed effects are directly mediated by AFP protein, we confirmed that NK cells can readily bind to and take up nAFP and tAFP. The observed synergism between AFP and IL-2 may be mediated by the ability of AFP to modulate IL-2 receptor signaling, as shown by the ability of AFP to upregulate CD25, and downregulate CD122 and CD132 on NK cells. Overall, these data show that nAFP and tAFP induce a unique pro-inflammatory, IL-2 hyperresponive phenotype in NK cells. Defining the impact of circulating AFP on NK cells may be crucial to understand the NK cell functional deficits described in HCC patients, and for the development of an effective HCC-targeting immunotherapy.

Abstract 3664: High frequency of a circulating monocytic subpopulation of myeloid-derived suppressor cells predicts worst clinical outcome in untreated non-small lung cancer patients

Abstract Background: The immune system can either prevent and control the tumor growth or promote tumor escape. Myeloid-derived suppressor cells (MDSCs) represent a heterogeneous population of immature cells with immune suppressive properties and their overexpression seems to confer a worse prognosis in cancer patients. Objective: The aim of the study was to investigate the presence of new phenotypically immature subpopulations of MDSCs which could be associated with advanced/metastatic NSCLC patients' clinical outcome. Materials & Methods: Circulating subtypes of MDSCs (monocytic and granulocytic), as well as the CD4+ , CD8+ , dendritic (DC), monocytes and B cells were assessed in peripheral blood of of chemotherapy-naive patients with advanced/metastatic NSCLC (n=134) and healthy controls (n=28) were analyzed using flow cytometry. The phenotypic characterization of MDSCs subpopulations was determined in strictly immature myeloid cells. A correlation between the levels of MDSCs and other tested immune cells as well as their association with overall (OS) and progression- free survival (PFS) was evaluated; high expression of MDSCs was defined as the percentage of the cells above the 90% percentile of the controls. Results: Two monocytic [M-MDSC: CD14+CD15-CD11b+CD33+HLA-DR-Lin- and CD14+CD15+CD11b+CD33+HLA-DR-Lin-] and a granulocytic [G-MDSC: CD14-CD15+CD11b+CD33+HLA-DR-Lin-] subpopulation of MDSCs were significantly increased (p<0.001), while the frequencies of CD4+ cells, DC and monocytes were decreased in patients compared to healthy donors (p≤0.0001, p≤0.0004, p=0.015; respectively). The frequency of both M-MDSCs' subpopulations was inversely correlated to that of DCs (p≤0.04), while the G-MDSC subpopulation was inversely correlated to CD4+ T cells (p=0.006). Furthermore, the M-MDSCs expressed high levels of iNOS, whereas the G-MDSCs generated high levels of reactive oxygen species (ROS). Increased frequencies of M-MDSCs were associated with worse response to front-line treatment (p=0.02) and patients with normal levels of CD14+CD15+CD11b+CD33+HLA-DR-Lin- MDSCs had longer OS and PFS compared to those with high levels (p=0.008 and p=0.02, respectively). Multivariate analysis revealed that the increased number of CD14+CD15+CD11b+CD33+HLA-DR-Lin- MDSCs was independently associated with decreased PFS and OS. Conclusion: The data provide evidence that the increased frequency of two new M-MDSC subpopulations in patients with advanced/metastatic NSCLC is associated with an unfavorable clinical outcome. Citation Format: Eleni Kyriaki Vetsika, Filippos Koinis, Marianthi Gioulmpasani, Despoina Aggouraki, Anna Koutoulaki, Eirini Skalidaki, Dimitris Mavroudis, Vassilis Georgoulias, Athanasios Kotsakis. High frequency of a circulating monocytic subpopulation of myeloid-derived suppressor cells predicts worst clinical outcome in untreated non-small lung cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3664. doi:10.1158/1538-7445.AM2014-3664

Inflammasome activation is critical to the protective immune response during chemically induced squamous cell carcinoma.

Chronic inflammation affects most stages of tumorigenesis, including initiation, promotion, malignant differentiation, invasion and metastasis. Inflammasomes have been described as involved with persistent inflammation and are known to exert both pro and antitumour effects. We evaluated the influence of apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase (CASP)-1 in the antitumor immune response using a multistage model of squamous cell carcinoma (SCC) development. Absence of ASC and CASP-1 resulted in an earlier incidence and increased number of papilloma. Loss of inflammassome function in mice resulted in decreased presence of natural killer (NK), dendritic (DC), CD4+, CD8+ and CD45RB+ T cells in the tumor lesions as well as in lymph nodes (LN) compared with WT mice. Increased percentage of CD4+CD25+Foxp3+ T cells was associated with association with inflammasome loss of function. Moreover, significant differences were also found with neutrophils and macrophage infiltrating the lesions. Myeloperoxidase (MPO), but not elastase (ELA), activity oscillated among the groups during the SCC development. Levels of proinflammatory cytokines IL-1β, IL-18, Tumor Necrosis Factor (TNF)-α and Interferon (IFN)-γ were decreased in the tumor microenvironment in the absence of inflammasome proteins. These observations suggest a link between inflammasome function and SCC tumorigenesis, indicating an important role for inflammasome activation in the control of SCC development.

ManLAM-aptamer as an immunoadjuvant improves immune-protective effects of BCG (VAC7P.968)

Abstract Tuberculosis (TB) remains an important worldwide health threat. However vaccination with Mycobacterium bovis bacille Calmette-Guerin (BCG), the only available licensed vaccine of TB, provides limited protection against adult pulmonary tuberculosis. Mannosylated lipoarabinomannan (ManLAM) is a kind of glycolipid on the cell wall surface of BCG and has been implicated with remarkable immunosuppressive effect on the host immune cells. In the present study, we used Systematic Evolution of Ligands by EXponential enrichment (SELEX) to generate a single-stranded DNA aptamer that specifically bound to ManLAM of BCG with high affinity. We found that vaccination with BCG combined with BM2 recruited dendritic (DC) cells into peripheral lymph nodes and decreased the numbers of regulatory B cells and regulatory T cells. It could also significantly enhanced the Th1-type cellular response and induced conversion of IgG2a isoforms in mice. We demonstrated that vaccination with BCG plus BM2 significantly reduced the progression of Mycobacterial tuberculosis (M. tb) H37Rv infection, bacterial loads in lungs and spleens of mice via aerosol infection with M. tb H37Rv, and provided much better protective immune response against M. tb than vaccination with BCG alone. Our data provide a new strategy for TB control and the aptamer BM2 holds great promise for developing a new potential immunoadjuvant of BCG.

Role of Innate Immunity against Human Papillomavirus (HPV) Infections and Effect of Adjuvants in Promoting Specific Immune Response

During the early stages of human papillomavirus (HPV) infections, the innate immune system creates a pro-inflammatory microenvironment by recruiting innate immune cells to eliminate the infected cells, initiating an effective acquired immune response. However, HPV exhibits a wide range of strategies for evading immune-surveillance, generating an anti-inflammatory microenvironment. The administration of new adjuvants, such as TLR (Toll-like receptors) agonists and alpha-galactosylceramide, has been demonstrated to reverse the anti-inflammatory microenvironment by down-regulating a number of adhesion molecules and chemo-attractants and activating keratinocytes, dendritic (DC), Langerhans (LC), natural killer (NK) or natural killer T (NKT) cells; thus, promoting a strong specific cytotoxic T cell response. Therefore, these adjuvants show promise for the treatment of HPV generated lesions and may be useful to elucidate the unknown roles of immune cells in the natural history of HPV infection. This review focuses on HPV immune evasion mechanisms and on the proposed response of the innate immune system, suggesting a role for the surrounding pro-inflammatory microenvironment and the NK and NKT cells in the clearance of HPV infections.

AB0227 PSGL-1 deficiency develops systemic sclerosis in mice

BackgroundPrevious studies in our lab have established that PSGL-1 signaling in human monocyte-derived dendritic (DC) cells induces a tolerogenic program that enables them to promote the differentiation of naïve T...

Effect of Zr Addition on Microstructure and Properties of FeCrNiMnCoZr x and Al0.5FeCrNiMnCoZr x High Entropy Alloys

In this study, the effect of Zr addition on phase formation, microstructure, and hardness of FeCrNiMnCoZr x and Al0.5FeCrNiMnCoZr x were investigated. High entropy alloys (HEA) were synthesized using arc melting technique in argon (Ar) atmosphere (x = 0, 0.1, 0.2, 0.3). Ingots were homogenized for 24 h at 900 °C in Ar atmosphere. Phase formation, microstructure and hardness of HEAs were investigated using field emission scanning electron microscope, X-ray diffraction and Vickers microhardness tester. Electron micrographs of HEAs showed majorly dendritic(DR) and interdendritic(ID) phases. For both FeCrNiMnCoZr x and Al0.5FeCrNiMnCoZr x alloys, amount of ID phases is seen to increase with increased Zr content. Aluminium containing HEAs showed fine needle-shaped precipitates dispersed throughout the matrix phase. XRD results confirmed the presence of mixed FCC/BCC phases in FeCrNiMnCoZr x alloys and BCC as majority phase in Al0.5FeCrNiMnCoZr x alloys. As the Zr content increased, hardness of HEA increased.

IL-36γ/IL-1F9, an Innate T-bet Target in Myeloid Cells

By concerted action in dendritic (DC) and T cells, T-box expressed in T cells (T-bet, Tbx21) is pivotal for initiation and perpetuation of Th1 immunity. Identification of novel T-bet-regulated genes is crucial for further understanding the biology of this transcription factor. By combining siRNA technology with genome-wide mRNA expression analysis, we sought to identify new T-bet-regulated genes in predendritic KG1 cells activated by IL-18. One gene robustly dependent on T-bet was IL-36γ, a recently described novel IL-1 family member. Promoter analysis revealed a T-bet binding site that, along with a κB site, enables efficient IL-36γ induction. Using knock-out animals, IL-36γ reliance on T-bet was extended to murine DC. IL-36γ expression by human myeloid cells was confirmed using monocyte-derived DC and M1 macrophages. The latter model was employed to substantiate dependence of IL-36γ on endogenous T-bet in human primary cells. Ectopic expression of T-bet likewise mediated IL-36γ production in HaCaT keratinocytes that otherwise lack this transcription factor. Additional experiments furthermore revealed that mature IL-36γ has the capability to establish an inflammatory gene expression profile in human primary keratinocytes that displays enhanced mRNA levels for TNFα, CCL20, S100A7, inducible NOS, and IL-36γ itself. Data presented herein shed further light on involvement of T-bet in innate immunity and suggest that IL-36γ, besides IFNγ, may contribute to functions of this transcription factor in immunopathology.

Acknowledgments

An AlCrCuFeNi2 high entropy alloy (HEA) was prepared by arc melting, followed by annealing at different temperatures. The elemental distributions, phase formation, morphology and microstructural evolution of the HEA in both the as-cast state and annealed state were investigated. The results indicate that the HEA undergoes elements segregation, precipitation and spinodal decomposition. The as-cast alloy consists of Cr-Fe-Ni rich FCC dendritic (DR) phase and BCC interdendritic (ID) phase. Spherical Fe-Cr rich BCC precipitates were found to disperse in Al-Ni rich B2 (ordered BCC) matrix. After heat treatments, the distribution of elements clearly changes, along with changes of the constituent phase and morphology. After annealing at 600 °C, the DR region remains Cr-Fe-Ni rich phase, while some spherical precipitates transform into the needle-like structure within the ID region. The L12 (ordered FCC) nanorod-shaped phase ((Ni,Cu)3Al) and plate-like Al-Ni rich phase form within the DR region when annealing up to 900 °C. The L12 phase almost dissolves in the FCC matrix due to the order-disorder transition and an obvious coarsening of the Fe-Cr rich phase occurs after annealing at 1100 °C. The segregation of Cu atoms at the interface between DR and ID regions is found at the as-cast state, while a uniform distribution of Cu atoms in DR and ID regions was observed after annealing at 1100 °C.

Immune response to cancer and its regulation in regional lymph nodes

Regional draining lymph nodes (LNs) play a pivotal role in initiating immune responses. However, the presence of metastases may compromise their normal immunological function. Preclinical studies indicate that despite metastases, early tumor-draining LNs are still a rich source of sensitized T cells. Recently, we found that dendritic (DC)-tumor fusion hybrids were capable of stimulating therapeutic T-cell generation in the LN. However, this response is regulated by a tumor-specific suppression mechanism(s). Reversal of these dysfunctions would help the success of immunotherapy.

HIV-1-specific exosome targeted CD8+ T cell Vaccine. (45.24)

Abstract Severe immuno-suppression due to immuno-tolerance and CD4 T cells depletion is the distinguishing feature of HIV-1 infection. Although highly active antiviral therapy (HAART) is available to halt progression of AIDS, it has its own inherent defects, including incomplete protection with associated side effects, and economical concerns. In addition, most of the currently available vaccines fail to induce effective CD8 CTL against HIV-1 in absence of CD4 T cells. Previously, our laboratory showed the efficacy of novel exosome (EXO)-targeted CD4 T cell vaccine (aTexo) with uptake of ova-specific dendritic (DC)-cell-released exosomes in the absence of CD4 T cells using highly metastasizing tumor model. In the present study, we find that aTexo vaccine prepared from active CD8 T cells is capable to induce CD8 CTLs, and its effect is mediated through IL-2 secretion, acquired pMHC I complexes, and partially on CD80 and CD40L costimulators. We have constructed an adenovirus-gp120 vector, which will be used to harvest gp-120-specific exosomes following DC infection, and established various tumor cell lines by transfecting gp-120 gene. We propose to generate gp-120-specific aTexo vaccine and test its efficacy in the presence or absence of CD4 T cells. These studies may significantly impact the development of novel therapeutic vaccines against HIV-1 infection.

Mesenchymal Stem Cells Overexpressing IFN-β Inhibit Breast Cancer Growth and Metastases through Stat3 Signaling in a Syngeneic Tumor Model

We previously demonstrated that mesenchymal stem/stromal cells (MSC) are recruited to tumors and that IFN-β produced by MSC inhibited tumor growth in xenograft models. Because of a deficient immune system, murine xenograft models cannot fully recapitulate tumor and immune cell interactions during progression. Therefore we investigated the capacity of MSC to migrate to and engraft into primary breast tumor sites and subsequently explore mechanisms of tumor inhibition by MSC-delivered IFN-β in a syngeneic, immunocompetent murine model. Herein we report that 1) systemically administrated MSC migrate to established 4T1 breast cancer sites and localize among the tumor-stroma border and throughout the tumor mass; 2) high levels of IFN-β secreted by MSC are detectable in the tumor microenvironment but not in circulation; 3) intratumorally produced IFN-β inactivates constitutive phosphorylation of signal transducer activator transcription factor 3 (Stat3), Src, and Akt and down-regulates cMyc and MMP2 expression in 4T1 cells, and 4) in mice with established breast cancer IFN-β expressing MSC administered systemically resulted in inhibition of primary cancer growth and in dramatic reduction of pulmonary and hepatic metastases. 5) MSC-IFN-β treated, but not control mice, maintained normal levels of splenic mature dendritic (DC), CD8+ T cells and CD4+/Foxp3+ regulatory T-cells (Treg). Our findings suggest that MSC are capable of migrating to tumor sites in an immunocompetent environment, that IFN-β produced by MSC suppresses breast cancer growth through inhibition of Stat3 signaling, and dramatically reduces pulmonary and hepatic metastases. The online version of this article (doi:10.1007/s12307-010-0041-8) contains supplementary material, which is available to authorized users.