Macrophages and Natural Killer Cells: Regulation and Function

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Western blot analysis was carried out as previously described Arcuri et al. For detection, a chemiluminescence kit Amersham Biosciences Corp.

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The plates were washed again and streptavidin horseradish peroxidase Zymed, San Francisco, CA, USA was added to each well and incubated for 20 min at room temperature. After 20 min, the reaction was stopped by adding H 2 SO 4. MIF concentration was expressed as ng per ml of medium. The Fisher Least Significant Difference test was used as post-test to determine significant differences between groups. The first part of the study was undertaken to evaluate the effect of MIF on the cytolytic activity of purified uNK cells.

Uterine NK cells were isolated as previously described Vigano et al. Uterine NK cells were incubated in the presence of different concentrations of the recombinant human cytokine, and the NK-mediated cytolysis was tested in a 51 Cr release assay with K as the target cells. As presented in Table 1 , in agreement with a previous study Vigano et al.

Natural killer cells limit the clearance of senescent lung adenocarcinoma cells

Western blot analysis revealed the presence of a single band with the apparent molecular mass of 12 kDa, co-migrating with recombinant human MIF in both unstimulated Fig. Binding specificity was confirmed by probing identical blots with the anti-MIF antibody preabsorbed with the recombinant cytokine. The cytokine was detected in all the media examined, with an equivalent concentration in unstimulated and ILactivated uNK cells Table 2.

As shown in Fig.

The identity of the PCR product was confirmed by restriction analysis with RsaI, which demonstrated, in the bp fragment, the presence of a restriction site yielding products of the expected size not shown. To evaluate the effect of the endogenous MIF on uNK cells, purified cultures were incubated in the presence of increasing concentrations of anti-MIF antibody or corresponding amounts of goat IgG, and the NK cell-mediated cytolysis was tested in a 51 Cr release assay.

Natural Killer Cells: How Do They Kill Selectively?

In uNK cells treated with the anti-MIF antibody the cytolytic activity increased in a dose-dependent manner, while it was unaffected by the control antibody. Statistical significance was achieved for both the concentrations of anti-MIF antibody tested Table 3. During decidualization, progesterone acts on the oestrogen-primed human endometrium to transform stromal cells into decidua cells, initially around spiral arteries and subsequently throughout the endometrium.

The specific functions of these uNK cells are still debated. However, recent experimental data have shown no reversal of the inhibition of lysis in the presence of antibodies against trophoblast MHC class I antigens, suggesting that these molecules are probably less relevant for placental cell survival than previously supposed Avril et al.

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Targeting Innate Immunity

It is well known that CD56 bright NK cells have an intrinsic capacity for high-level production of type-1 and type-2 cytokines, whereas CD56 dim cells produce substantially less Cooper et al. Interestingly, uNK cells are also known to produce cytokines that normally are not produced by blood NK cells, such as leukaemia inhibitory factor Sharkey et al. First described as a T cell-derived protein, MIF has recently been shown to be produced by a variety of cell types, including monocytes, endothelial cells, keratinocytes and anterior pituitary cells, suggesting multifunctional physiological effects Nishihira A critical role for this factor in the mechanisms underlying the establishment of pregnancy has also been supposed.

MIF has been demonstrated to be expressed in the glandular epithelium and stroma of the cyclic endometrium as well as in the decidua and trophoblasts Arcuri et al.

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Serum concentrations of the cytokine are much higher in the first, second and third trimesters of pregnancy than in the non-pregnant status and, more importantly, these concentrations were found to be decreased in women with recurrent miscarriage. In particular, serum MIF levels were lower in abortion-prone women with a normal foetal chromosome karyotype than in those with an abnormal foetal chromosome karyotype, thus suggesting a role for the cytokine in the aetiology of abortion Yamada et al. Notwithstanding these observations, the precise significance of MIF in the context of gestation remains poorly defined.

The results reported herein clearly indicate that MIF is synthesized and released in significant concentrations by uNK cells. These findings are in agreement with recent data obtained by Koopman and colleagues comparing the gene expression profiles of uterine and peripheral NK cells using microarray analysis.


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Results from the present study also showed that a neutralizing anti-MIF antibody is able to significantly increase uNK cell cytolytic activity, thus supporting the concept that the cytokine exerts an immunomodulatory function on this lymphoid population. These results are supported by the significant, although slight, inhibition of the cytolytic activity observed with rhMIF. The limited effect of the exogenously administered cytokine suggests uNK MIF levels are probably sufficient to trigger a response.

A similar phenomenon has already been reported by Bacher and colleagues who found that, in T cells, although a neutralizing anti-MIF antibody inhibited cell proliferation and IL-2 production - indicating that MIF was a required participant to these effects — purified rMIF was not mitogenic by itself.


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  • Hum Reprod ; 24 : Eur J Immunol ; 30 : Vilches C, Parham P. KIR: Diverse, rapidly evolving receptors of innate and adaptive immunity.

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    Annu Rev Immunol ; 20 : J Exp Med ; : Macrophages and angiogenesis. J Leukoc Biol ; 55 : Macrophages and apoptotic cell clearance during pregnancy. Am J Reprod Immunol ; 51 : J Clin Endocrinol Metab ; 96 : Pre-eclampsia is associated with dendritic cell recruitment into the uterine decidua. J Pathol ; : Aberrant positioning of trophoblast and lymphocytes in the feto-maternal interface with pre-eclampsia. Virchows Arch ; : Serious foetal growth restriction is associated with reduced proportions of natural killer cells in decidua basalis.

    Persistence of decidual NK cells and KIR genotypes in healthy pregnant and preeclamptic women: a case-control study in the third trimester of gestation.

    Introduction

    Reprod Biol Endocrinol ; 9 : 8. Dendritic cells DCs play a major role in both inflammatory diseases and cancer. DCs within the tumor are inefficient in taking up, processing and presenting the tumor antigens effectively. Reprogramming DC metabolism and targeting the tumor antigens via surface receptors could improve anti-tumor immune response. We have recently reported the identification of novel, dominant ILC2-dependent circuits of immunosuppression in cancer patients. ILC2 may therefore represent attractive cell targets to reprogram the immunosuppressive tumor microenvironment.

    By decoding the transcriptional programs of human ILC2 we have identified candidate targets for ILC2 functional reprogramming. By therapeutically interfering with these circuits we are exploring the impact of these discoveries in preclinical mouse models, in view of Phase I clinical trials. Founded in , Sino Biological, Inc. We help our customers to obtain the best reagents and services to accelerate the pace of research and drug discovery to improve human health.

    These are informal, moderated discussions with brainstorming and interactive problem solving, allowing participants from diverse backgrounds to exchange ideas and experiences and develop future collaborations around a focused topic. View details on the topics and moderators here. Table 1: From Bench to Bedside to Industry. Michael V. In recent years there has been a dramatic increase in regulatory filings for allogenic cellular therapies across three regulatory agencies: the U.

    Increased clinical experience in East and West have identified promising cell therapy candidates that meet the demand for safe, immediate use, and more cost-viable cell therapies while also raising vigilance about cell therapies associated with severe adverse events. This presentation will review common features in pre-clinical package development that hinder entry into clinic and discuss common adverse events for recent cell therapy trials in three nations. Our approach can overcome the hurdles of cell therapy in the treatment of solid tumors by modulating the tumor microenvironment TME through macrophages with key characteristics: recruitment and access to the solid tumor TME, ability to survive in the hostile solid tumor milieu, maintenance of an anti-tumor phenotype in the presence of immunosuppressive factors, capacity to selectively destroy cancer cells, and activation of an adaptive immune response by presenting engulfed tumor material.

    The role of IL4R expression on survival outcomes and therapeutic benefit from the Phase 2b clinical trial of MDNA55 in one of the most aggressive tumor types will be presented. We have screened macrophage responses to over 4, compounds, including FDA-approved drugs, bioactive compounds and natural products, and identified compounds that can polarize macrophages from M1 to M2 or vice versa. We have validated selected compounds to drive tumor-associated macrophages to inflammatory phenotype in vivo to achieve anti-tumor effect alone or in combination with antibody therapeutics. These studies elucidate molecular basis underlying macrophage heterogeneity and provide a basis for modulating macrophage activities for disease intervention.

    This talk will address the complexity of the TME and how it contributes to failure of therapy, understanding of how targeting soluble TNF offers a solution to the resistance to therapy when part of combination therapy; and results of the Phase I trial of INB03 in patients with advanced cancer.

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