Cancer remains the leading cause of death worldwide despite intense efforts in developing innovative treatments. Current approaches in cancer therapy are mainly directed to a selective targeting of cancer cells to avoid potential side effects associated with conventional therapy. In this respect, Natural killer (NK) cells have gained growing attention and are now being considered as promising therapeutic tools for cancer therapy owing to their intrinsic ability to rapidly recognize and kill cancer cells, while sparing normal healthy cells. NK cells play a key role in the first line of defense against transformed and virus-infected cells. NK cells sense their target through a whole array of receptors, both activating and inhibitory. Functional outcome of NK cell against target cells is determined by the balance of signals transmitted from diverse activating and inhibiting receptors. Despite significant progress made in the role of NK cells attack as a pivotal sentinel in tumor surveillance, the molecular has been that regulate NK cell responses remain unclear, which restricts the use of NK cells as a therapeutic measure. Accordingly, current efforts for NK cell-based cancer therapy have largely relied on the strategies that are based on the manipulation of inhibitory receptor function. However, if we better understand the mechanisms governing NK cell activation, including those mediated by diverse activating receptors, this knowledge can be applied to the development of optimal design for cancer immunotherapy by targeting NK cells.
Activation Analysis ; Cause of Death ; Immunotherapy ; Killer Cells, Natural ; Nitriles ; Pyrethrins ; Receptors, Immunologic

OBJECTIVETo investigate the effect of triggering receptor expressed on myeloid cells 2 (TREM-2) overexpression on airway inflammation and remodeling in mice with asthma.

METHODSA total of 40 BALB/c mice were randomly divided into normal control, asthma, empty vector, and TREM-2 overexpression groups (n=10 each). Ovalbumin (OVA) sensitization and challenge were performed to establish the model of asthma. The mice in the control group were given normal saline, and those in the empty vector and TREM-2 overexpression groups were transfected with adenovirus vector and TREM-2 adenovirus, respectively. RT-PCR and Western blot were used to measure the expression of TREM-2, MMP-2, MMP-9, ADAM33, and ADAM8. Bronchoalveolar lavage fluid (BALF) was collected to perform cell counting and classification. ELISA was used to measure the total serum level of IgE and the levels of cytokines in BALF.

RESULTSCompared with the control group, the asthma group showed significant reductions in the mRNA and protein expression of TREM-2 (P<0.05), a significantly increased level of Th2 cytokine (P<0.05), and significantly increased numbers of total cells and classified cells. Compared with the asthma group, the TREM-2 overexpression group showed a significantly reduced level of Th2 cytokine (P<0.05), a significantly reduced level of IgE (P<0.05), and significantly reduced numbers of total cells and classified cells (P<0.05), as well as significantly downregulated expression of the inflammatory factors and growth factors MMP-2, MMP-9, TGF-β1, ADAM8, and ADAM33 (P<0.05).

CONCLUSIONSTREM-2 overexpression significantly alleviates airway inflammation and airway remodeling in mice with asthma and may become a potential target for the prevention and treatment of childhood asthma.

Airway Remodeling ; Animals ; Asthma ; etiology ; immunology ; Cytokines ; analysis ; Female ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; RNA, Messenger ; analysis ; Receptors, Immunologic ; genetics ; physiology

OBJECTIVETo construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells.

METHODSThe designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method.

RESULTSThe proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation.

CONCLUSIONSThese new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.

Antigens, Surface ; analysis ; Base Sequence ; Biological Assay ; Cells, Cultured ; DNA ; analysis ; genetics ; Gene Expression Profiling ; methods ; Humans ; Lipopolysaccharide Receptors ; analysis ; Lymphocyte Antigen 96 ; Membrane Glycoproteins ; analysis ; Molecular Probe Techniques ; Molecular Sequence Data ; Monocytes ; metabolism ; RNA Probes ; analysis ; genetics ; Receptors, Cell Surface ; analysis ; Receptors, Immunologic ; analysis ; Ribonucleases ; Toll-Like Receptor 4 ; Toll-Like Receptors

UNLABELLEDThe purpose of this study was to evaluate the effects of cellular immunity activation on P58(+) cells expressing killer cell inhibitory receptor (KIR) and their regulatory function on cellular immunity, and provid theoretical data for preventing graft-vers-host disease (GVHD) in stem cell transplantation therapy. The mononuclear cells from human peripheral blood were incubated with IL-2, Con A and Lipostin (LP) for 72 hours. The KIR expressing cells, P58.1(+) and P58.2(+) cells, were analyzed by flow cytometry. The results showed that the percentages of CD3(+), CD4(+), CD8(+), CD16(+)CD56(+), P58.1(+) and P58.2(+) cells were greatly increased after treated with IL-2, Con A and LP, separately or in combination, and the percentages of above cells in combined treatment groups were higher than those of single stimulated groups, especially the percentage of cells in the IL-2 + LP group was significantly higher than those in IL-2 and LP singly treated groups.

IN CONCLUSIONIL-2, Con A and LP possess the ability to induce the expression of KIR and stimulate proliferation of P58.1(+) and P58.2(+) cells while to activate the celluar immunity response, the expression of P58 gene may be regulated by the activation of cellular immunity.

Adult ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD56 Antigen ; analysis ; CD8 Antigens ; analysis ; Cell Count ; Cell Division ; drug effects ; Concanavalin A ; pharmacology ; Flow Cytometry ; Humans ; Interleukin-2 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Receptors, IgG ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR ; Receptors, KIR2DL3

The study was aimed at the exploration of relationship between T cells expressing killer cell inhibitor receptors (KIR, CD158 and CD94) and graft-versus-host disease (GVHD) after hematopoietic stem cell transplantation. The expression rates of CD158a, CD158b and CD94 on T cells and NK cell were detected by flow cytometry and donor/recipient HLA-Cw was analyzed using PCR after peripheral blood stem cell transplantation (PBSCT) and umbilical cord blood transplantation (UCBT). After both PBSCT and UCBT, the rates of CD3(+)CD158a(+) and CD3(+)CD158b(+) T cells increased, especially the rate of CD8(+)CD158b(+) T cells. In both acute and chronic GVHD groups, the rate of CD3(+)CD158b(+) T cells increased, especially in acute GVHD. The CD94 mainly expressed on CD3(+)CD8(+) T cells. The percentage of the expression of CD94 on CD4(+) and CD8(+) cells after UCBT and PBSCT increased significantly. The expression of KIR in GVHD (early stage of transplantation) increased but the expression of KIR in chronic GVHD (advanced stage of transplantation) decreased. Five patients who HLA-Cw matched had no severe GVHD. In four patients who underwent allo-PBSCT and UCBT from related HLA-matched donors, only 2 patients had no aGVHD. Four patients underwent transplantation from unrelated HLA-matched donors had GVHD. These observations suggested that there is some relationship between GVHD and KIR expression on T cells. CD158b might be an inhibitory molecule of T cell activated at early stage after transplantation. Understanding the mechanism of GVHD with the expression of KIR on T cells, especially those binding the HLA-Cw might shed light on the establishment of the specific immunotolerance for the prevention of GVHD. To pay attention to HLA-Cw typing is very important to reduce GVHD and increase GVL effect in related or unrelated HLA-matched transplantation.
Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; Genotype ; Graft vs Host Disease ; etiology ; HLA-C Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Lectins, C-Type ; Receptors, Immunologic ; analysis ; Receptors, Interleukin-2 ; analysis ; Receptors, KIR ; Receptors, KIR2DL1 ; Receptors, KIR2DL3 ; T-Lymphocytes ; immunology

AIM: A quantitative ELISA kit for the detection of human secreted CD306/LAIR-2 was developed using monoclonal and polyclonal antibodies which were raised against a highly purified recombinant human secreted CD306/LAIR-2. METHODS: Anti-human secreted CD306/LAIR-2 monoclonal and polyclonal antibodies were raised by immunizing mouse or rabbit with recombinant human secreted CD306/LAIR-2. The monoclonal antibody was purified by protein G affinity, whereas the polyclonal antibody was purified by protein A affinity. The best match pair of antibodies were found and used to develop a double antibody sandwich ELISA kit for the detection of human secreted CD306/LAIR-2 in human samples. RESULTS: A human secreted CD306/LAIR-2 ELISA kit was formulated with highly purified recombinant human secreted CD306/LAIR-2, highly specific monoclonal and polyclonal antibodies. This kit realized the quantitative measurement of recombinant human CD306/LAIR-2 and natural CD306/LAIR-2 in human serum samples. CONCLUSIONS The developed human secreted CD306/LAIR-2 ELISA kit is a reliable quantitation immunoassay kit.
Animals ; Antibodies, Monoclonal ; immunology ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Rabbits ; Receptors, Immunologic ; analysis ; blood ; immunology ; Recombinant Proteins

BACKGROUNDTriggering receptors expressed on myeloid cells (TREM) proteins are a family of cell surface receptors expressed broadly by cells of the myeloid lineage. The aim of this study was to investigate the clinical significance of soluble TREM-1 (sTREM-1) in pleural effusions, and to determine the effects of pneumonia on pleural sTREM-1 concentrations.

METHODSPleural fluid was collected from 109 patients who presented to the respiratory institute (35 with malignant pleural effusion, 31 with tuberculous pleural effusion, 21 with bacterial pleural effusion, and 22 with transudate). The concentrations of sTREM-1, tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were determined in effusion and serum samples by enzyme linked immunosorbent assay (ELISA).

RESULTSThe concentrations of sTREM-1 in bacterial pleural effusion were significantly higher than those in malignant, tuberculous, and transudative groups (all P < 0.001). An sTREM-1 cutoff value of 768.1 ng/L had a sensitivity of 86% and a specificity of 93%. Pleural sTREM-1 levels were positively correlated with levels of TNF-alpha and IL-1beta. Patients with complicating bacterial pneumonia did not have elevated concentration of sTREM-1 in pleural effusion when compared with patients without pneumonia.

CONCLUSIONSDetermination of pleural sTREM-1 may improve the ability of clinicians to differentiate pleural effusion patients of bacterial origin from those with other etiologies. The occurrence of bacterial pneumonia did not affect pleural sTREM-1 concentrations.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cross-Sectional Studies ; Humans ; Interleukin-1beta ; analysis ; Membrane Glycoproteins ; analysis ; Middle Aged ; Pleural Effusion ; diagnosis ; metabolism ; Pneumonia ; metabolism ; Prospective Studies ; Receptors, Immunologic ; analysis ; Triggering Receptor Expressed on Myeloid Cells-1 ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo investigate the regulatory effect of IFN-gamma on recognition of target cells by human natural killer (NK) cells.

METHODSThe cytotoxic activity of human NK cell lines (NK92, NKL) was detected by MTT method. Expression of NK cell receptors (NKG2D, NKG2A/B, KIR2DL1 and KIR2DS1) and MICA on target cells (the ligand of NKG2D) was measured by RT-PCR.

RESULTSBoth NK92 and NKL cells exerted higher cytotoxicity to tumor cells with MICA expression, while tumors without MICA expression could resist NK cell lysis. IFN-gamma (> 1000 U/ml) inhibited NK lysis of tumor cells with MICA expression through down-regulating the expression of NKG2D, but up-regulating the expression of NKG2A/B and KIR2DL1.

CONCLUSIONIFN-gamma has a negative effect on activation and cytotoxicity of human NK cells by altering the balance between the expression of activating and inhibitory receptors on NK cells in favor of inhibition. This may serve to limit NK cell over-activation in vivo.

Cell Division ; drug effects ; Cytotoxicity, Immunologic ; drug effects ; Histocompatibility Antigens Class I ; analysis ; physiology ; Humans ; Interferon-gamma ; pharmacology ; Killer Cells, Natural ; immunology ; NK Cell Lectin-Like Receptor Subfamily C ; NK Cell Lectin-Like Receptor Subfamily K ; Receptors, Immunologic ; metabolism ; Receptors, KIR2DL1 ; Receptors, Natural Killer Cell ; Recombinant Proteins ; Tumor Cells, Cultured

OBJECTIVETo explore the change of RAGE-NF-κB signaling pathway during the course of hyperoxia-induced lung injury in newborn rats, and the effect of glucocorticoid on this pathway.

METHODSTwenty-four Sprague-Dawley neonatal rats were randomly divided into three groups (n=8 each) : sham control (control group), hyperoxia-induced acute lung injury (model group) and glucocorticoid-treated acute lung injury (glucocorticoid group). Rats were sacrificed at 13 days after birth. RAGE and NF-κB expression levels in lung tissues were detected by reverse transcription polymerase chain reaction, Western blot and immunohistochemistry analysis. The levels of tumor necrosis factor α (TNF-α) and sRAGE in bronchoalveolar lavage fluid (BALF) and serum were measured using ELISA. Lung damage was evaluated by histological examinations.

RESULTSRAGE and NF-κB mRNA and protein expression levels in lung tissues were significantly increased in the model and glucocorticoid groups compared with the control group (P<0.05). Serum RAGE concentrations were significantly increased but RAGE concentrations in BALF were significantly reduced in the model and glucocorticoid groups compared with the control group (P<0.05). RAGE and NF-κB expression at both mRNA and protein levels in lung tissues was significantly lower in the glucocorticoid group than in the model group (P<0.05). RAGE concentrations were significantly lower in serum (P<0.05), but were higher in BALF (P<0.05) in the glucocorticoid group than in the model group.

CONCLUSIONSRAGE-NF-κB pathway plays an important role in hyperoxia-induced lung injury in neonatal rats, and glucocorticoid administration may play a protective role against the lung injury by down-regulating RAGE-NF-κB signaling pathway.

Animals ; Animals, Newborn ; Glucocorticoids ; pharmacology ; Hyperoxia ; complications ; Lung Injury ; prevention & control ; NF-kappa B ; analysis ; genetics ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; analysis ; genetics ; physiology ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; analysis

Killer cell inhibitory receptors (KIR) are members of the immunoglobulin superfamily (Ig-SF) and transmembrane molecules type I expressed on human natural killer cells and some T lymphocytes. They are ligands for HLA class I antigens. According to the "missing-self" hypothesis, KIR deliver inhibitory signals that prevent target-cell lysis upon binding to the self MHC class I antigens. KIR regulates NK cell function concerned with the control graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect after allogeneic hematopoietic stem cell transplantation (HSCT). The KIR repertoire is substantially influenced by the polymorphic and polygeneic nature of the KIR gene family, and there exist the polymorphism of KIR ligands mainly HLA-C molecule. So it is difficult to achieve the beneficial effect of NK cells on the outcome of partly HLA-mismatched hematopoietic cell transplantation. This review aims to provide background and previous observations on the KIRs which are thought to influence the outcome of HSCT.
Animals ; Graft vs Host Disease ; etiology ; Graft vs Leukemia Effect ; HLA-C Antigens ; physiology ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, Immunologic ; analysis ; chemistry ; physiology ; Receptors, KIR ; T-Lymphocytes ; chemistry ; Transplantation, Homologous