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FEBS Letters Oct 2005CD14, CD68 and/or mouse F4/80 or human epidermal growth factor module-containing mucin-like receptor 1 (EMR1) are widely used as macrophage-specific markers. Since... (Comparative Study)
Comparative Study
CD14, CD68 and/or mouse F4/80 or human epidermal growth factor module-containing mucin-like receptor 1 (EMR1) are widely used as macrophage-specific markers. Since macrophages infiltrate several tissues during inflammatory processes, CD14, CD68 and EMR1-F4/80 have been employed to discriminate between tissue-containing macrophages, like adipose tissue (AT), and other cells. Using real-time PCR experiments, we show that isolated adipocytes from humans and mice AT express high levels of CD14 and CD68 mRNA, whereas EMR1-F4/80 is mainly present in the macrophage-containing stroma-vascular fraction. Furthermore, fibroblasts-like cells (adipoblasts), preadipocytes and adipocytes from the murine cell lines, 3T3-F442A and BFC-1, express CD14 and CD68 mRNA and protein as determined by fluorescence-activated cell sorter, but not F4/80 which, as expected, is strongly expressed in the macrophage cell line RAW264.7. These results reinforce the view that EMR1-F4/80 is the best macrophage marker to date and show that CD14 and CD68 are not macrophage-specific proteins.
Topics: Adipocytes; Animals; Antigens, CD; Antigens, Differentiation; Antigens, Differentiation, Myelomonocytic; Biomarkers; Calcium-Binding Proteins; Gene Expression; Humans; Lipopolysaccharide Receptors; Macrophages; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mucins; RNA, Messenger; Receptors, G-Protein-Coupled
PubMed: 16213494
DOI: 10.1016/j.febslet.2005.09.032 -
Oncotarget Apr 2016Simultaneous targeting of multiple tumor-associated antigens (TAAs) in cancer immunotherapy is presumed to enhance tumor cell selectivity and to reduce immune escape.The...
Simultaneous targeting of multiple tumor-associated antigens (TAAs) in cancer immunotherapy is presumed to enhance tumor cell selectivity and to reduce immune escape.The combination of B lymphoid marker CD19 and myeloid marker CD33 is exclusively present on biphenotypic B/myeloid leukemia cells. Triplebody 33-3-19 binds specifically to both of these TAAs and activates T cells as immune effectors. Thereby it induces specific lysis of established myeloid (MOLM13, THP-1) and B-lymphoid cell lines (BV173, SEM, Raji, ARH77) as well as of primary patient cells. EC50 values range from 3 pM to 2.4 nM. In accordance with our hypothesis, 33-3-19 is able to induce preferential lysis of double- rather than single-positive leukemia cells in a target cell mixture: CD19/CD33 double-positive BV173 cells were eliminated to a significantly greater extent than CD19 single-positive SEM cells (36.6% vs. 20.9% in 3 hours, p = 0.0048) in the presence of both cell lines. In contrast, equivalent elimination efficiencies were observed for both cell lines, when control triplebody 19-3-19 or a mixture of the bispecific single chain variable fragments 19-3 and 33-3 were used. This result highlights the potential of dual-targeting agents for efficient and selective immune-intervention in leukemia patients.
Topics: Antibody-Dependent Cell Cytotoxicity; Antigens, CD19; Antigens, Differentiation, Myelomonocytic; Antigens, Neoplasm; Antineoplastic Agents; Humans; Immunotherapy; Leukemia, Biphenotypic, Acute; Sialic Acid Binding Ig-like Lectin 3; Single-Chain Antibodies
PubMed: 26981773
DOI: 10.18632/oncotarget.8022 -
Clinical and Experimental Immunology Aug 1989A characteristic of active cytomegalovirus (CMV) infection is its suppressive effect on in vitro assays of immune function. The expression of CD11b by the Cd4+ and Cd8+...
Phenotypic study of CD4+ and CD8+ lymphocyte subsets in relation to cytomegalovirus carrier status and its correlate with pokeweed mitogen-induced B lymphocyte differentiation.
A characteristic of active cytomegalovirus (CMV) infection is its suppressive effect on in vitro assays of immune function. The expression of CD11b by the Cd4+ and Cd8+ lymphocytes allows the identification of subsets with distinct regulatory functions of pokeweed mitogen (PWM) induced B cell differentiation. In order to relate that result with our previous observation that CMV carriers have significantly increased numbers of CD4+, HNK1+ and CD8+, HNK1+ lymphocytes in their peripheral blood compared with non-carriers, we performed a three-colour flow cytometric analysis of the co-expression of Cd11b and HNK1 by CD4+ and CD8+ lymphocytes obtained from 27 CMV carriers and 42 non-carriers. The differences between CMV carriers and non-carriers were significant for the CD4+, HNK1+ lymphocytes (median [5th and 95th percentiles], 59 [18 and 123 versus 24/7 and 73 per mm3, respectively; P less than 0.001) and CD8+, HNK1+ lymphocytes (59 [18 259] versus 52 [23 and 139] per mm3; P less than 0.001), but not for the CD4+, CD11b+ lymphocytes (59 [18 and 135] versus 52 [17 and 104] per mm3) and the CD8+, CD11b+ lymphocytes (85 [34 and 293] versus 82 [21 and 248] per mm3). The CD4+, HNK1+ and CD8+, HNK1+ lymphocytes that were increased in CMV carriers compared with non-carriers included mostly CD11b-, but also CD11b+ lymphocytes. After sorting CD4+ and CD8+ lymphocytes for four CMV carriers into HNK1+ and HNK1- fractions, we analyzed their regulatory functions on PWM-driven B cell Helper function to PWM-driven B cell differentiation was exclusively associated with the CD4+, HNK1- lymphocytes; the CD4+, HNK1+ generally did not show helper or suppressor activity in this assay. Both CD8+, HNK1+ and CD8+, HNK1- lymphocytes showed suppressor activity. Thus, the NHK1 marker does not constitute a phenotypical correlate for suppressor cells of PWM-driven B-cell differentiation.
Topics: Adolescent; Adult; Antigens, Differentiation; Antigens, Differentiation, T-Lymphocyte; B-Lymphocytes; CD57 Antigens; CD8 Antigens; Carrier State; Cytomegalovirus Infections; Female; Humans; Lymphocyte Activation; Macrophage-1 Antigen; Male; Membrane Glycoproteins; Pokeweed Mitogens; T-Lymphocytes
PubMed: 2476271
DOI: No ID Found -
PloS One 2013CD6 has recently been identified and validated as risk gene for multiple sclerosis (MS), based on the association of a single nucleotide polymorphism (SNP), rs17824933,...
CD6 has recently been identified and validated as risk gene for multiple sclerosis (MS), based on the association of a single nucleotide polymorphism (SNP), rs17824933, located in intron 1. CD6 is a cell surface scavenger receptor involved in T-cell activation and proliferation, as well as in thymocyte differentiation. In this study, we performed a haptag SNP screen of the CD6 gene locus using a total of thirteen tagging SNPs, of which three were non-synonymous SNPs, and replicated the recently reported GWAS SNP rs650258 in a Spanish-Basque collection of 814 controls and 823 cases. Validation of the six most strongly associated SNPs was performed in an independent collection of 2265 MS patients and 2600 healthy controls. We identified association of haplotypes composed of two non-synonymous SNPs [rs11230563 (R225W) and rs2074225 (A257V)] in the 2(nd) SRCR domain with susceptibility to MS (P max(T) permutation = 1×10(-4)). The effect of these haplotypes on CD6 surface expression and cytokine secretion was also tested. The analysis showed significantly different CD6 expression patterns in the distinct cell subsets, i.e. - CD4(+) naïve cells, P = 0.0001; CD8(+) naïve cells, P<0.0001; CD4(+) and CD8(+) central memory cells, P = 0.01 and 0.05, respectively; and natural killer T (NKT) cells, P = 0.02; with the protective haplotype (RA) showing higher expression of CD6. However, no significant changes were observed in natural killer (NK) cells, effector memory and terminally differentiated effector memory T cells. Our findings reveal that this new MS-associated CD6 risk haplotype significantly modifies expression of CD6 on CD4(+) and CD8(+) T cells.
Topics: Adult; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Chromosome Mapping; Cluster Analysis; Cytokines; Female; Gene Order; Genetic Loci; Genetic Predisposition to Disease; Haplotypes; Humans; Linkage Disequilibrium; Lymphocyte Activation; Male; Multiple Sclerosis; Polymorphism, Single Nucleotide; Protein Interaction Domains and Motifs; Spain; White People; Young Adult
PubMed: 23638056
DOI: 10.1371/journal.pone.0062376 -
Cell Death & Disease Jan 2016GADD45 gene has been implicated in cell cycle arrest, cell survival or apoptosis in a cell type specific and context-dependent manner. Members of GADD45 gene family have...
GADD45 gene has been implicated in cell cycle arrest, cell survival or apoptosis in a cell type specific and context-dependent manner. Members of GADD45 gene family have been found differentially expressed in several podocyte injury models, but their roles in podocytes are unclear. Using an in vivo zebrafish model of inducible podocyte injury that we have previously established, we found that zebrafish orthologs of gadd45b were induced upon the induction of podocyte injury. Podocyte-specific overexpression of zebrafish gadd45b exacerbated edema, proteinuria and foot-process effacement, whereas knockdown of gadd45b by morpholino-oligos in zebrafish larvae ameliorated podocyte injury. We then explored the role of GADD45B induction in podocyte injury using in vitro podocyte culture. We confirmed that GADD45B was significantly upregulated during the early phase of podocyte injury in cultured human podocytes and that podocyte apoptosis induced by TGF-β and puromycin aminonucleoside (PAN) was aggravated by GADD45B overexpression but ameliorated by shRNA-mediated GADD45B knockdown. We also showed that ROS inhibitor NAC suppressed PAN-induced GADD45B expression and subsequent activation of p38 MAPK pathway in podocytes and that inhibition of GADD45B diminished PAN-induced p38 MAPK activation. Taken together, our findings demonstrated that GADD45B has an important role in podocyte injury and may be a therapeutic target for the management of podocyte injury in glomerular diseases.
Topics: Animals; Antigens, Differentiation; Apoptosis; Podocytes; Reactive Oxygen Species; Transfection; Zebrafish; Zebrafish Proteins
PubMed: 26794661
DOI: 10.1038/cddis.2015.300 -
Immunity Aug 2002B7-1 and B7-2 are generally thought to have comparable structures and affinities for their receptors, CD28 and CTLA-4, each of which is assumed to be bivalent. We show...
B7-1 and B7-2 are generally thought to have comparable structures and affinities for their receptors, CD28 and CTLA-4, each of which is assumed to be bivalent. We show instead (1) that B7-2 binds the two receptors more weakly than B7-1, (2) that, relative to its CTLA-4 binding affinity, B7-2 binds CD28 2- to 3-fold more effectively than B7-1, (3) that, unlike B7-1, B7-2 does not self-associate, and (4) that, in contrast to CTLA-4 homodimers, which are bivalent, CD28 homodimers are monovalent. Our results indicate that B7-1 markedly favors CTLA-4 over CD28 engagement, whereas B7-2 exhibits much less bias. We propose that the distinct structures and binding properties of B7-1 and B7-2 account for their overlapping but distinct effects on T cell responses.
Topics: Abatacept; Animals; Antigens, CD; Antigens, Differentiation; B7-1 Antigen; B7-2 Antigen; CD28 Antigens; CHO Cells; CTLA-4 Antigen; Cricetinae; Immunoconjugates; Kinetics; Membrane Glycoproteins; Protein Structure, Tertiary; Recombinant Fusion Proteins; Solutions
PubMed: 12196291
DOI: 10.1016/s1074-7613(02)00362-x -
Journal of Clinical Pathology Oct 1990The expression of myeloperoxidase (MPO) was studied in 100 cases of acute leukaemia (83 with acute myeloid leukaemia (AML) and 17 acute lymphoblastic leukaemia (ALL) by... (Comparative Study)
Comparative Study
The expression of myeloperoxidase (MPO) was studied in 100 cases of acute leukaemia (83 with acute myeloid leukaemia (AML) and 17 acute lymphoblastic leukaemia (ALL) by both a conventional cytochemical method and the immunocytochemical antiperoxidase (APAAP) technique using the monoclonal antibody MPO7. In each case the staining was evaluated by light microscopical examination (percentage of positive cells). Of the 83 cases of AML, 78 (93.9%) were positive for MPO7 compared with 70 (84.3%) by cytochemistry. Antibodies against the myeloid markers CD13 and CD33 were positive in 71 (85.5%) and 70 (84.3%) cases, respectively. Importantly, all cases of ALL were negative for both MPO7 and cytochemical MPO staining even when they were positive for CD13 and CD33. These results indicate that the anti-myeloperoxidase antibody MPO7 is the most sensitive and specific reagent for the diagnosis of AML and should therefore be included in routine immunophenotyping panels.
Topics: Antibodies, Monoclonal; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers, Tumor; CD13 Antigens; Humans; Immunoenzyme Techniques; Leukemia, Myeloid; Peroxidase; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Sensitivity and Specificity; Sialic Acid Binding Ig-like Lectin 3
PubMed: 1977771
DOI: 10.1136/jcp.43.10.847 -
Proceedings of the National Academy of... Dec 1991T-cell antigen receptor (TCR) heterodimers of both the alpha beta and gamma delta types are expressed at the surface of T cells only in association with a complex of...
T-cell antigen receptor (TCR) heterodimers of both the alpha beta and gamma delta types are expressed at the surface of T cells only in association with a complex of invariant chains called CD3. The requirement for individual CD3 components to achieve TCR surface expression was examined by cotransfection of a non-T-cell line with TCR alpha and beta, as well as CD3 delta, epsilon, gamma, and zeta, cDNAs. Both transient and stable transfectants expressing TCR and CD3 epitopes at the cell surface were generated. By transfection of TCR and CD3 components in different combinations, the TCR chains, as well as the CD3 epsilon and zeta chains, were each shown to be essential for reconstituting surface expression. On the other hand, CD3 delta and gamma chains could be used alternatively, providing evidence for two different types of TCR/CD3 complexes.
Topics: Antibodies, Monoclonal; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; CD3 Complex; Cell Membrane; Electrophoresis, Polyacrylamide Gel; Flow Cytometry; HeLa Cells; Humans; Macromolecular Substances; Models, Structural; Receptors, Antigen, T-Cell; Transfection
PubMed: 1835791
DOI: 10.1073/pnas.88.23.10619 -
Journal of Clinical Pathology Apr 1989Seventy six peripheral T cell lymphomas were examined immunohistologically to test their reactivity with a panel of monoclonal antibodies against 11 T cell associated...
Seventy six peripheral T cell lymphomas were examined immunohistologically to test their reactivity with a panel of monoclonal antibodies against 11 T cell associated antigens (CD1-8, CD27, UCHL1, and the T cell antigen receptor). Sixty two (82%) lymphomas showed aberrant phenotypes, and four main categories were distinguished as follows: (i) lack of one or several pan-T cell antigens (49, 64% of the cases); (ii) loss of both the CD4 and CD8 antigens (11, 15% of the cases); (iii) coexpression of the CD4 and CD8 antigens (13, 17% of the cases); and (iv) expression of the CD1 antigen (eight, 11% of the cases). No correlation was seen between the occurrence of aberrant phenotypes and the histological subtype. It is concluded that the demonstration of an aberrant phenotype is a valuable supplement to histological assessment in the diagnosis of peripheral T cell lymphomas. It is recommended that the panel of monoclonal antibodies against T cell differentiation antigens should be fairly large, as apparently any antigen may be lost in the process of malignant transformation.
Topics: Antibodies, Monoclonal; Antigens, CD1; Antigens, Differentiation; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; Biopsy; CD8 Antigens; Humans; Lymphoma; Phenotype; T-Lymphocytes
PubMed: 2469701
DOI: 10.1136/jcp.42.4.398 -
Frontiers in Immunology 2018Sepsis is an unmet clinical need constituting one of the most important causes of death worldwide, a fact aggravated by the appearance of multidrug resistant strains due...
Sepsis is an unmet clinical need constituting one of the most important causes of death worldwide, a fact aggravated by the appearance of multidrug resistant strains due to indiscriminate use of antibiotics. Host innate immune receptors involved in pathogen-associated molecular patterns (PAMPs) recognition represent a source of broad-spectrum therapies alternative or adjunctive to antibiotics. Among the few members of the ancient and highly conserved scavenger receptor cysteine-rich superfamily (SRCR-SF) sharing bacterial-binding properties there is CD6, a lymphocyte-specific surface receptor. Here, we analyze the bacterial-binding properties of three conserved short peptides (11-mer) mapping at extracellular SRCR domains of human CD6 (CD6.PD1, GTVEVRLEASW; CD6.PD2 GRVEMLEHGEW; and CD6.PD3, GQVEVHFRGVW). All peptides show high binding affinity for PAMPs from Gram-negative (lipopolysaccharide; from 3.5 to 3,000 nM) and Gram-positive (lipoteichoic acid; from 36 to 680 nM) bacteria. The CD6.PD3 peptide possesses broad bacterial-agglutination properties and improved survival of mice undergoing polymicrobial sepsis in a dose- and time-dependent manner. Accordingly, CD6.PD3 triggers a decrease in serum levels of both pro-inflammatory cytokines and bacterial load. Interestingly, CD6.PD3 shows additive survival effects on septic mice when combined with Imipenem/Cilastatin. These results illustrate the therapeutic potential of peptides retaining the bacterial-binding properties of native CD6.
Topics: Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Conserved Sequence; Gram-Negative Bacteria; Gram-Positive Bacteria; Humans; Immunity, Innate; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Models, Animal; Pathogen-Associated Molecular Pattern Molecules; Peptides; Protein Binding; Receptors, Scavenger; Sepsis; Teichoic Acids
PubMed: 29706953
DOI: 10.3389/fimmu.2018.00627