1,062 results on '"CD2 Antigens"'
Search Results
2. Complement receptor 1 is the human erythrocyte receptor for Plasmodium vivax erythrocyte binding protein.
- Author
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Lee SK, Crosnier C, Valenzuela-Leon PC, Dizon BLP, Atkinson JP, Mu J, Wright GJ, Calvo E, Gunalan K, and Miller LH
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- Humans, Receptors, Cell Surface, Erythrocytes, Reticulocytes, CD2 Antigens, Cell Adhesion Molecules, Plasmodium vivax, Malaria, Falciparum
- Abstract
The discovery that Africans were resistant to infection by Plasmodium vivax ( P. vivax ) led to the conclusion that P. vivax invasion relied on the P. vivax Duffy Binding Protein (PvDBP) interacting with the Duffy Antigen Receptor for Chemokines (DARC) expressed on erythrocytes. However, the recent reporting of P. vivax infections in DARC-negative Africans suggests that the parasite might use an alternate invasion pathway to infect DARC-negative reticulocytes. To identify the parasite ligands and erythrocyte receptors that enable P. vivax invasion of both DARC-positive and -negative erythrocytes, we expressed region II containing the Duffy Binding-Like (DBL) domain of P. vivax erythrocyte binding protein (PvEBP-RII) and verified that the DBL domain binds to both DARC-positive and -negative erythrocytes. Furthermore, an AVidity-based EXtracelluar Interaction Screening (AVEXIS) was used to identify the receptor for PvEBP among over 750 human cell surface receptor proteins, and this approach identified only Complement Receptor 1 (CR1, CD35, or C3b/C4b receptor) as a PvEBP receptor. CR1 is a well-known receptor for P. falciparum Reticulocyte binding protein Homology 4 (PfRh4) and is present on the surfaces of both reticulocytes and normocytes, but its expression decreases as erythrocytes age. Indeed, PvEBP-RII bound to a subpopulation of both reticulocytes and normocytes, and this binding was blocked by the addition of soluble CR1 recombinant protein, indicating that CR1 is the receptor of PvEBP. In addition, we found that the Long Homology Repeat A (LHR-A) subdomain of CR1 is the only subdomain responsible for mediating the interaction with PvEBP-RII., Competing Interests: Competing interests statement:The authors declare no competing interest.
- Published
- 2024
- Full Text
- View/download PDF
3. The coreceptor CD2 uses plasma membrane microdomains to transduce signals in T cells
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Kaizuka, Yoshihisa, Douglass, Adam D, Vardhana, Santosh, Dustin, Michael L, and Vale, Ronald D
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Biochemistry and Cell Biology ,Biological Sciences ,1.1 Normal biological development and functioning ,Underpinning research ,Inflammatory and immune system ,Animals ,Antigen-Presenting Cells ,Antigens ,CD ,CD2 Antigens ,CD58 Antigens ,Cell Adhesion ,Cell Membrane ,Drug Synergism ,Humans ,Intercellular Adhesion Molecule-1 ,Membrane Microdomains ,Receptors ,Antigen ,T-Cell ,Signal Transduction ,T-Lymphocytes ,Antigens ,CD58 ,Antigens ,CD2 ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
The interaction between a T cell and an antigen-presenting cell (APC) can trigger a signaling response that leads to T cell activation. Prior studies have shown that ligation of the T cell receptor (TCR) triggers a signaling cascade that proceeds through the coalescence of TCR and various signaling molecules (e.g., the kinase Lck and adaptor protein LAT [linker for T cell activation]) into microdomains on the plasma membrane. In this study, we investigated another ligand-receptor interaction (CD58-CD2) that facilities T cell activation using a model system consisting of Jurkat T cells interacting with a planar lipid bilayer that mimics an APC. We show that the binding of CD58 to CD2, in the absence of TCR activation, also induces signaling through the actin-dependent coalescence of signaling molecules (including TCR-zeta chain, Lck, and LAT) into microdomains. When simultaneously activated, TCR and CD2 initially colocalize in small microdomains but then partition into separate zones; this spatial segregation may enable the two receptors to enhance signaling synergistically. Our results show that two structurally distinct receptors both induce a rapid spatial reorganization of molecules in the plasma membrane, suggesting a model for how local increases in the concentration of signaling molecules can trigger T cell signaling.
- Published
- 2009
4. Findings in Immunologic Receptors Reported from National Institutes of Health (NIH) [CRISPR Activation Screen to Optimize Chimeric Antigen Receptor (CAR) T Cell Immunophenotype].
- Abstract
A recent study conducted by researchers at the National Institutes of Health (NIH) focused on immunologic receptors and their role in adoptive cellular therapies (ACT). The study found that CAR T-cells, which are genetically engineered immune cells, can become functionally impaired due to exhaustion when persistently activated and exposed to the tumor microenvironment. The researchers used CRISPR activation screening to identify genes that enhance T cell phenotypes associated with durable function. They found that the activation of certain genes, such as IL23 and KLF4, resulted in a more potent and durable CAR T-cell immunophenotype. This research has implications for the development of more effective adoptive cellular therapies. [Extracted from the article]
- Published
- 2023
5. Research from Shandong University Yields New Findings on B-Cell Lymphoma (The Characterization and Prognostic Value of Tumor Immune Microenvironment in Diffuse Large B-Cell Lymphoma).
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PROGNOSIS ,DIFFUSE large B-cell lymphomas ,TUMOR microenvironment ,LYMPHOMAS ,LYMPHATIC diseases - Abstract
A study conducted by researchers at Shandong University in China has investigated the tumor immune microenvironment (TME) in diffuse large B-cell lymphoma (DLBCL) and its correlation with patient prognosis. The study analyzed transcriptomic data and clinical information from DLBCL patients and identified 22 distinct immune cell types within the TME. A novel TME immune score based on the proportion of these immune cells was established and found to be significantly associated with worse prognosis. The study also identified CD2 as a prognostic biomarker in DLBCL patients and revealed its association with different T-cell activation states. These findings may contribute to the development of personalized management strategies for DLBCL patients. [Extracted from the article]
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- 2023
6. The SH3 domain of p56lck binds to proline-rich sequences in the cytoplasmic domain of CD2.
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Bell, GM, Fargnoli, J, Bolen, JB, Kish, L, and Imboden, JB
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2.1 Biological and endogenous factors ,Aetiology ,Amino Acid Sequence ,Animals ,Base Sequence ,CD2 Antigens ,Cells ,Cultured ,Interleukin-2 ,Lymphocyte Specific Protein Tyrosine Kinase p56(lck) ,Molecular Sequence Data ,Mutagenesis ,Peptide Fragments ,Proline ,Protein Binding ,Protein Conformation ,Rats ,Receptors ,Cell Surface ,Recombinant Proteins ,Sequence Deletion ,Signal Transduction ,Spodoptera ,Structure-Activity Relationship ,T-Lymphocytes ,src Homology Domains ,src-Family Kinases ,Medical and Health Sciences ,Immunology - Abstract
CD2, a cell surface glycoprotein expressed on T cells and natural killer cells, can couple to signaling pathways that result in T cell proliferation. An Src-like protein tyrosine kinase, p56lck, coprecipitates with CD2, and perturbation of CD2 by monoclonal antibodies results in an increase in the activity of p56lck, suggesting that an interaction with p56lck contributes to CD2-mediated signaling. Herein, we investigate the mechanism by which CD2 associates with p56lck. We demonstrate that CD2 and p56lck associate when coexpressed in nonlymphoid cells, that this association requires the cytoplasmic domain of CD2, and that the SH3 domain of p56lck mediates its interactions with CD2. Using truncation mutants of CD2, we identify two regions in the cytoplasmic domain of CD2 involved in binding p56lck. Each region contains a proline-rich sequence that, in the form of a synthetic peptide, directly binds p56lck. Thus, proline-rich sequences in the cytoplasmic domain of CD2 allow this transmembrane receptor to bind to the SH3 domain of p56lck.
- Published
- 1996
7. Single-cell measurements of two-dimensional binding affinity across cell contacts
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Simon J. Davis, Victoria Junghans, Peter Jönsson, Manto Chouliara, Ana Filipa L.O.M. Santos, and Tommy Dam
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Lipid Bilayers ,Cell ,Population ,CD2 Antigens ,Biophysics ,Cell Communication ,Ligands ,Jurkat cells ,Jurkat Cells ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,education ,Lipid bilayer ,Receptor ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,Chemistry ,CD48 ,Ligand (biochemistry) ,Affinities ,Rats ,medicine.anatomical_structure ,030217 neurology & neurosurgery ,Protein Binding - Abstract
The two-dimensional (2D) affinity between protein molecules across contacting cells is a key parameter regulating and initiating several cellular processes. However, measuring 2D affinity can be challenging and experimental data are limited. In addition, the obtained 2D affinities are typically averaged over the cell population. We here present a method to measure 2D affinity on single cells binding to polyhistidine-tagged fluorescent ligands anchored to a supported lipid bilayer (SLB). By decreasing the density of ligands in the SLB using imidazole a new steady-state accumulation in the contact is obtained, and from this change, both the 2D affinity and the number of receptors on the cell can be determined. The method was validated on an SLB containing rat CD2 binding to the rat CD48 mutant T92A expressed on Jurkat T cells. The addition of imidazole did not influence the average 2D affinity (1/Kd), and the spread in affinities within the cell population was low, Kd = 4.9 ± 0.9 molecules/μm2 (mean ± SD), despite an order of magnitude spread in ligand accumulation due to differences in receptor density. It was also found that cell contact size increased both with ligand density and with the number of receptors per cell, but that the contact size stayed approximately constant when lowering the ligand density, above a density of around 10 rCD2 molecules/μm2, after the contact first had formed, indicative of a heterogeneous process. In summary, this method not only allows for single-cell affinities to be measured, but it can also reduce measurement and analysis time and improve measurement accuracy. Due to the low spread in 2D Kd within the cell population, the analysis can further be restricted to the cells showing the strongest binding, paving the way for using this method to study weak binding events.
- Published
- 2021
8. CD28 interaction with B7 costimulates primary allogeneic proliferative responses and cytotoxicity mediated by small, resting T lymphocytes.
- Author
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Azuma, M, Cayabyab, M, Buck, D, Phillips, JH, and Lanier, LL
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Clinical Research ,Immunization ,Vaccine Related ,5.2 Cellular and gene therapies ,Aetiology ,Development of treatments and therapeutic interventions ,2.1 Biological and endogenous factors ,Inflammatory and immune system ,Animals ,Antibodies ,Monoclonal ,Antigens ,CD ,Antigens ,Differentiation ,T-Lymphocyte ,Antigens ,Surface ,B-Lymphocytes ,B7-1 Antigen ,CD2 Antigens ,CD28 Antigens ,CD3 Complex ,Cytotoxicity ,Immunologic ,Flow Cytometry ,Lymphocyte Activation ,Mice ,Mice ,Inbred BALB C ,Receptors ,Antigen ,T-Cell ,Receptors ,Immunologic ,T-Lymphocytes ,T-Lymphocytes ,Cytotoxic ,Transfection ,Tumor Cells ,Cultured ,Medical and Health Sciences ,Immunology - Abstract
Engagement of the CD3/T cell antigen receptor complex on small, resting T cells is insufficient to trigger cell-mediated cytotoxicity or to induce a proliferative response. In the present study, we have used genetic transfection to demonstrate that interaction of the B7-BB1 B cell activation antigen with the CD28 T cell differentiation antigen costimulates cell-mediated cytotoxicity and proliferation initiated by either anti-CD2 or anti-CD3 monoclonal antibody (mAb). Moreover, a B7-negative Burkitt's lymphoma cell line that fails to stimulate an allogeneic mixed lymphocyte response is rendered a potent stimulator after transfection with B7. The mixed leukocyte reaction proliferative response against the B7 transfectant is inhibited by either anti-CD28 or B7 mAb. We also demonstrate that freshly isolated small, resting human T cells can mediate anti-CD3 or anti-CD2 mAb-redirected cytotoxicity against a murine Fc receptor-bearing mastocytoma transfected with human B7. These preexisting cytotoxic T lymphocytes in peripheral blood are present in both the CD4 and CD8 subsets, but are preferentially within the CD45RO+ "memory" population. While small, resting T cells apparently require costimulation by CD28/B7 interactions, this requirement is lost after T cell activation. Anti-CD3 initiates a cytotoxic response mediated by in vitro cultured T cell clones in the absence of B7 ligand. The existence of functional cytolytic T cells in the small, resting T cell population may be advantageous in facilitating rapid responses to immune challenge.
- Published
- 1992
9. The OX-44 molecule couples to signaling pathways and is associated with CD2 on rat T lymphocytes and a natural killer cell line.
- Author
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Bell, GM, Seaman, WE, Niemi, EC, and Imboden, JB
- Subjects
Biotechnology ,Cancer ,Hematology ,Vaccine Related ,Immunization ,1.1 Normal biological development and functioning ,Underpinning research ,Animals ,Antibodies ,Monoclonal ,Antigens ,CD ,Antigens ,Differentiation ,T-Lymphocyte ,Antigens ,Surface ,CD2 Antigens ,Cell Line ,Cytotoxicity ,Immunologic ,Killer Cells ,Natural ,Lymphocyte Activation ,Mice ,Mice ,Inbred BALB C ,Rats ,Rats ,Inbred F344 ,Receptors ,Antigen ,T-Cell ,Receptors ,Immunologic ,Signal Transduction ,T-Lymphocytes ,Tetraspanin 25 ,Medical and Health Sciences ,Immunology - Abstract
The MRC OX-44 molecule, which is expressed on all peripheral leukocytes, identifies the subset of thymocytes capable of proliferating in response to alloantigens and lectins (Paterson, D.J., J.R. Green, W.A. Jefferies, M. Puklavec, and A.F. Williams. 1987. J. Exp. Med. 165:1). When we isolated monoclonal antibodies (mAbs) on the basis of their ability to activate the phosphatidylinositol signaling pathway in RNK-16 cells (a rat leukemia line with natural killer activity), three of the resulting mAbs recognized the OX-44 molecule. Addition of these mAbs to RNK-16 elicits protein tyrosine phosphorylation, generates inositol phosphates, and increases the concentration of cytoplasmic free calcium. These responses require the addition of intact mAb and are not observed with F(ab')2 fragments. One of these mAbs (7D2) is mitogenic for freshly isolated rat splenic T cells and synergizes with a mAb to the T cell antigen receptor in this activation. A 50-60-kD glycoprotein coprecipitates with the OX-44 molecule from RNK-16 cells and rat splenic T cells. Peptide mapping and reprecipitation studies indicate that the coprecipitating molecule is CD2. Thus, the OX-44 molecule can couple to multiple signaling pathways and associates with CD2 on both RNK-16 and rat T cells.
- Published
- 1992
10. Multidimensional single-cell analysis identifies a role for CD2-CD58 interactions in clinical antitumor T cell responses
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Gabrielle Romain, Paolo Strati, Ali Rezvan, Mohsen Fathi, Irfan N. Bandey, Jay R T. Adolacion, Darren Heeke, Ivan Liadi, Mario L. Marques-Piubelli, Luisa M. Solis, Ankit Mahendra, Francisco Vega, Laurence J.N. Cooper, Harjeet Singh, Mike Mattie, Adrian Bot, Sattva S. Neelapu, and Navin Varadarajan
- Subjects
T-Lymphocytes ,Antigens, CD19 ,CD2 Antigens ,Receptors, Antigen, T-Cell ,Humans ,Lymphoma, Large B-Cell, Diffuse ,General Medicine ,Single-Cell Analysis ,CD58 Antigens ,Immunotherapy, Adoptive - Abstract
The in vivo persistence of adoptively transferred T cells is predictive of antitumor response. Identifying functional properties of infused T cells that lead to in vivo persistence and tumor eradication has remained elusive. We profiled CD19-specific chimeric antigen receptor (CAR) T cells as the infusion products used to treat large B cell lymphomas using high-throughput single-cell technologies based on time-lapse imaging microscopy in nanowell grids (TIMING), which integrates killing, cytokine secretion, and transcriptional profiling. Our results show that the directional migration of CD19-specific CAR T cells is correlated with multifunctionality. We showed that CD2 on T cells is associated with directional migration and that the interaction between CD2 on T cells and CD58 on lymphoma cells accelerates killing and serial killing. Consistent with this, we observed that elevated CD58 expression on pretreatment tumor samples in patients with relapsed or refractory large B cell lymphomas treated with CD19-specific CAR T cell therapy was associated with complete clinical response and survival. These results highlight the importance of studying dynamic T cell-tumor cell interactions in identifying optimal antitumor responses.
- Published
- 2022
11. Rosetting T cells in Hodgkin lymphoma are activated by immunological synapse components HLA class II and CD58
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Lydia Visser, Johanna Veldman, Natasja Muller, Anke van den Berg, Arjan Diepstra, Magdalena Huberts-Kregel, Bouke G. Hepkema, Stem Cell Aging Leukemia and Lymphoma (SALL), and Translational Immunology Groningen (TRIGR)
- Subjects
CD4-Positive T-Lymphocytes ,0301 basic medicine ,EXPRESSION ,Rosette Formation ,Immunological Synapses ,CD58 ,Immunology ,CD2 Antigens ,Receptors, Antigen, T-Cell ,LINES ,ADHESION ,Lymphocyte Activation ,LYMPHOCYTES ,Biochemistry ,Peripheral blood mononuclear cell ,Immunological synapse ,Gene Knockout Techniques ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,CYTOMETRY ,Cell Line, Tumor ,Protein Interaction Mapping ,REVEALS ,Cell Adhesion ,medicine ,Humans ,REED-STERNBERG CELLS ,RECEPTOR ,Cell adhesion molecule ,Chemistry ,T-cell receptor ,Histocompatibility Antigens Class II ,AMPLIFICATION ,Cell Biology ,Hematology ,CD58 Antigens ,medicine.disease ,Hodgkin Disease ,Coculture Techniques ,Cell biology ,030104 developmental biology ,Reed–Sternberg cell ,Cell culture ,030220 oncology & carcinogenesis ,NODES ,CRISPR-Cas Systems - Abstract
A unique feature of Hodgkin lymphoma (HL) is the presence of CD4+ T cells that surround, protect, and promote survival of tumor cells. The adhesion molecules involved in this so-called T-cell rosetting are important components of the immunological synapse (IS). However, it is unknown whether this synapse is fully assembled and leads to T-cell activation by enabling interaction between the T-cell receptor (TCR) and human leukocyte antigen class II (HLA-II). We established a novel rosetting model by coculturing HLA-II–matched peripheral blood mononuclear cells with HL cell lines and showed IS formation with activation of rosetting T cells. HLA-II downregulation by class II transactivator knockout did not affect the extent of rosetting, but almost completely abrogated T-cell activation. Intriguingly, the level of CD58 expression correlated with the extent of rosette formation, and CD58 knockout or CD2 blockade reduced both rosette formation and T-cell activation. The extension of our findings to primary HL tissue by immunohistochemistry and proximity ligation assays showed interaction of CD2 with CD58 and of TCR-associated CD4 with HLA-II. In conclusion, T-cell rosetting in HL is established by formation of the IS, and activation of rosetting T cells critically depends on the interaction of both TCR-HLA-II and CD2-CD58.
- Published
- 2020
12. Modulation of co‐stimulatory signal from CD2–CD58 proteins by a grafted peptide
- Author
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Pravin Parajuli, Achyut Dahal, Rushikesh Sable, Leeza Shrestha, Seetharama D. Jois, Veena Taneja, and Ted J. Gauthier
- Subjects
T-Lymphocytes ,Trypsin inhibitor ,CD2 Antigens ,Peptide ,Binding, Competitive ,Peptides, Cyclic ,01 natural sciences ,Biochemistry ,Article ,Cell Line ,Protein–protein interaction ,Drug Discovery ,Cell Adhesion ,Humans ,Amino Acid Sequence ,Protein Interaction Maps ,Surface plasmon resonance ,Cell adhesion ,Pharmacology ,Alanine ,chemistry.chemical_classification ,Binding Sites ,Protein Stability ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,Alanine scanning ,CD58 Antigens ,0104 chemical sciences ,Molecular Docking Simulation ,010404 medicinal & biomolecular chemistry ,Enzyme ,Drug Design ,Biophysics ,Molecular Medicine ,Trypsin Inhibitors ,Protein Binding - Abstract
Peptides were designed to inhibit the protein-protein interaction of CD2 and CD58 to modulate the immune response. This work involved the design and synthesis of eight different peptides by replacing each amino acid residue in peptide 6 with alanine as well as grafting the peptide to the sunflower trypsin-inhibitor framework. From the alanine scanning studies, mutation at position 2 of the peptide was shown to result in increased potency to inhibit cell adhesion interactions. The most potent peptide from the alanine scanning was further studied for its detailed three-dimensional structure and binding to CD58 protein using surface plasmon resonance and flow cytometry. This peptide was used to graft to the sunflower trypsin inhibitor to improve the stability of the peptide. The grafted peptide, SFTI-a1, was further studied for its potency as well as its thermal, chemical, and enzymatic stability. The grafted peptide exhibited improved activity compared to our previously grafted peptide and was stable against thermal and enzymatic degradation.
- Published
- 2020
13. Human Oral Epithelial Cells Suppress T Cell Function via Prostaglandin E2 Secretion
- Author
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Jose L. Sanchez-Trincado, Hector F. Pelaez-Prestel, Esther M. Lafuente, and Pedro A. Reche
- Subjects
CD4-Positive T-Lymphocytes ,Transcription, Genetic ,Immunology ,CD40 Ligand ,T cells ,CD2 Antigens ,chemical and pharmacologic phenomena ,CD8-Positive T-Lymphocytes ,immunomodulation ,Lymphocyte Activation ,T-Lymphocytes, Regulatory ,Dinoprostone ,Cell Line ,Interferon-gamma ,Immune Tolerance ,Immunology and Allergy ,Humans ,dendritic cells ,CD40 Antigens ,Original Research ,Immunity ,hemic and immune systems ,Epithelial Cells ,RC581-607 ,CD58 Antigens ,Interleukin-12 ,oral epithelial cells ,PGE2 ,viral infection ,Immunologic diseases. Allergy - Abstract
The oral mucosa is constantly exposed to a plethora of stimuli including food antigens, commensal microbiota and pathogens, requiring distinct immune responses. We previously reported that human oral epithelial cells (OECs) suppress immune responses to bacteria, using H413 and TR146 OEC lines and primary OECs in co-culture with dendritic cells (DCs) and T cells (OEC-conditioned cells). OECs reduced DCs expression of CD80/CD86 and IL-12/TNFα release and impaired T cell activation. Here, we further evaluated the immunosuppression by these OECs and investigated the underlying mechanisms. OEC-conditioned DCs did not induce CD4 T cell polarization towards Treg, judging by the absence of FoxP3 expression. OECs also repressed T-bet/IFNγ expression in CD4 and CD8 T cells activated by DCs or anti-CD3/CD28 antibodies. This inhibition depended on OEC:T cell ratio and IFNγ repression occurred at the transcriptional level. Time-lapse experiments showed that OECs inhibited early steps of T cell activation, consistent with OECs inability to suppress T cells stimulated with PMA/ionomycin. Blocking CD40/CD40L, CD58/CD2 and PD-L1/PD-1 interactions with specific antibodies did not disrupt T cell suppression by OECs. However, preventing prostaglandin E2 (PGE2) synthesis or blocking PGE2 binding to the cognate EP2/EP4 receptors, restored IFNγ and TNFα production in OEC-conditioned T cells. Finally, treating OECs with poly(I:C), which simulates viral infections, limited T cell suppression. Overall, these results point to an inherent ability of OECs to suppress immune responses, which can nonetheless be eluded when OECs are under direct assault.
- Published
- 2022
14. CD2 is a surface marker for mouse and rat spermatogonial stem cells
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Guiying Chen, Takashi Shinohara, Hiroko Morimoto, and Mito Kanatsu-Shinohara
- Subjects
Male ,endocrine system ,Population ,CD2 Antigens ,Spermatogonial stem cells ,Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Surface marker ,Animals ,Spermatogenesis ,skin and connective tissue diseases ,education ,Gene ,030304 developmental biology ,Transplantation ,0303 health sciences ,education.field_of_study ,030219 obstetrics & reproductive medicine ,Adult Germline Stem Cells ,integumentary system ,Transfection ,Flow Cytometry ,CD2 ,Spermatogonia ,Rats ,Cell biology ,Apoptosis ,Original Article ,Animal Science and Zoology ,Stem cell - Abstract
The spermatogonial stem cell (SSC) population in testis is small, and the lack of SSC markers has severely handicapped research on these cells. During our attempt to identify genes involved in SSC aging, we found that CD2 is expressed in cultured SSCs. Flow cytometric analysis and spermatogonial transplantation experiments showed that CD2 is expressed in SSCs from mature adult mouse testes. Cultured SSCs transfected with short hairpin RNAs (shRNAs) against CD2 proliferated poorly and showed an increased frequency of apoptosis. Moreover, functional analysis of transfected cells revealed impairment of SSC activity. Fluorescence activated cell sorting and spermatogonial transplantation experiments showed that CD2 is expressed not only in mouse but also in rat SSCs. The results indicate that CD2 is a novel SSC surface marker conserved between mouse and rat SSCs.
- Published
- 2020
15. Researchers Submit Patent Application, "Methods For Activation And Expansion Of Tumor Infiltrating Lymphocytes", for Approval (USPTO 20230108584).
- Subjects
TUMOR-infiltrating immune cells ,PATENT applications ,PATENT offices ,MONONUCLEAR leukocytes ,CD28 antigen - Abstract
The method of claim 7, wherein the CD3 agonist is an anti-CD3 antibody, optionally a humanized anti-CD3 antibody, or a soluble monospecific complex comprising two anti-CD3 antibodies linked together. "Surprisingly, the streamlined methods provided herein, in some embodiments, offer a 30-50% increase in fold TIL (e.g., edited TIL) expansion over current TIL expansion protocols, while also supporting proliferation of effector T cells and enrichment of a central memory T cell phenotype, even in the absence of IL-2. The method of any one of the preceding claims, wherein the agonist of the T cell costimulatory molecule is selected from: a CD28 agonist, a CD137 agonist, a CD2 agonist, and combinations thereof. [Extracted from the article]
- Published
- 2023
16. CD58 Immunobiology at a Glance
- Author
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Yalu Zhang, Qiaofei Liu, Quan Liao, and Sen Yang
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Graft Rejection ,0301 basic medicine ,Immunological Synapses ,Lymphoma ,T-Lymphocytes ,CD58 ,Immunology ,CD2 Antigens ,Review ,Autoimmune Diseases ,Immunological synapse ,Immunomodulation ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Antigen ,Antigens, Neoplasm ,Neoplasms ,Cell Adhesion ,Tumor Microenvironment ,Humans ,Protein Isoforms ,Immunology and Allergy ,Intestinal Mucosa ,Cell adhesion ,Receptor ,immune evasion ,Tumor microenvironment ,Leukemia ,T cell activation ,Chemistry ,lymphocyte functional antigen-3 ,Endothelial Cells ,RC581-607 ,CD58 Antigens ,CD2 ,Cell biology ,Killer Cells, Natural ,Transplantation ,030104 developmental biology ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,Cytomegalovirus Infections ,Cytokines ,Tumor Escape ,LFA-3 ,Immunologic diseases. Allergy ,Signal Transduction - Abstract
The glycoprotein CD58, also known as lymphocyte-function antigen 3 (LFA-3), is a costimulatory receptor distributed on a broad range of human tissue cells. Its natural ligand CD2 is primarily expressed on the surface of T/NK cells. The CD2-CD58 interaction is an important component of the immunological synapse (IS) that induces activation and proliferation of T/NK cells and triggers a series of intracellular signaling in T/NK cells and target cells, respectively, in addition to promoting cell adhesion and recognition. Furthermore, a soluble form of CD58 (sCD58) is also present in cellular supernatant in vitro and in local tissues in vivo. The sCD58 is involved in T/NK cell-mediated immune responses as an immunosuppressive factor by affecting CD2-CD58 interaction. Altered accumulation of sCD58 may lead to immunosuppression of T/NK cells in the tumor microenvironment, allowing sCD58 as a novel immunotherapeutic target. Recently, the crucial roles of costimulatory molecule CD58 in immunomodulation seem to be reattracting the interests of investigators. In particular, the CD2-CD58 interaction is involved in the regulation of antiviral responses, inflammatory responses in autoimmune diseases, immune rejection of transplantation, and immune evasion of tumor cells. In this review, we provide a comprehensive summary of CD58 immunobiology.
- Published
- 2021
17. Polymorphic estrogen receptor binding site causes CD2-dependent sex bias in the susceptibility to autoimmune diseases
- Author
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Yibo He, Kutty Selva Nandakumar, Mike Aoun, Roman A. Zubarev, Pierre Sabatier, Rikard Holmdahl, Gonzalo Fernandez Lahore, Michael Förster, Martina Johannesson, and Erik Lönnblom
- Subjects
Male ,Science ,T-Lymphocytes ,T cell ,T cells ,CD2 Antigens ,General Physics and Astronomy ,Autoimmunity ,Locus (genetics) ,Immunogenetics ,Biology ,Lymphocyte Activation ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Autoimmune Diseases ,Arthritis, Rheumatoid ,Mice ,Immune system ,Downregulation and upregulation ,medicine ,Animals ,Humans ,Genetic Predisposition to Disease ,Rheumatoid arthritis ,Sex Characteristics ,Binding Sites ,Polymorphism, Genetic ,Multidisciplinary ,Estradiol ,Estrogen receptor binding ,General Chemistry ,Sexual dimorphism ,Disease Models, Animal ,medicine.anatomical_structure ,Gene Expression Regulation ,Immunology ,Female - Abstract
Complex autoimmune diseases are sexually dimorphic. An interplay between predisposing genetics and sex-related factors probably controls the sex discrepancy in the immune response, but the underlying mechanisms are unclear. Here we positionally identify a polymorphic estrogen receptor binding site that regulates Cd2 expression, leading to female-specific differences in T cell-dependent mouse models of autoimmunity. Female mice with reduced Cd2 expression have impaired autoreactive T cell responses. T cells lacking Cd2 costimulation upregulate inhibitory Lag-3. These findings help explain sexual dimorphism in human autoimmunity, as we find that CD2 polymorphisms are associated with rheumatoid arthritis and 17-β-estradiol-regulation of CD2 is conserved in human T cells. Hormonal regulation of CD2 might have implications for CD2-targeted therapy, as anti-Cd2 treatment more potently affects T cells in female mice. These results demonstrate the relevance of sex-genotype interactions, providing strong evidence for CD2 as a sex-sensitive predisposing factor in autoimmunity., The Cia21 locus on chromosome 3 has been associated with rheumatoid arthritis severity in females. Here the authors show this locus houses a non-coding polymorphic estrogen receptor binding site and how it regulates neighbouring gene expression of CD2, implicating CD2 signalling in the sexual dimorphism of a variety of T cell-dependent autoimmune diseases.
- Published
- 2021
18. Keratinocytes costimulate naive human T cells via CD2: a potential target to prevent the development of proinflammatory Th1 cells in the skin
- Author
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Beate Niesler, Christian Orlik, Jutta Schröder-Braunstein, Knut Schäkel, Jüri Habicht, Yvonne Samstag, Daniel Deibel, Emre Balta, Guido H. Wabnitz, Johanna Küblbeck, and Sabina Ganskih
- Subjects
0301 basic medicine ,Chemokine ,Autoimmunity ,Lymphocyte Activation ,nonprofessional antigen-presenting cells ,0302 clinical medicine ,Immunology and Allergy ,STAT1 ,Phosphorylation ,human T cells ,Skin ,biology ,Chemistry ,Cell Differentiation ,Intercellular Adhesion Molecule-1 ,Lymphocyte Function-Associated Antigen-1 ,Up-Regulation ,STAT1 Transcription Factor ,Infectious Diseases ,costimulation ,Cytokines ,Protein Binding ,LFA-1 ,keratinocytes ,Receptors, CCR7 ,CD58 ,Immunology ,CD2 Antigens ,Article ,Proinflammatory cytokine ,Interferon-gamma ,03 medical and health sciences ,Immune system ,Th1 cells ,Psoriasis ,medicine ,Humans ,Secretion ,Th17 cells ,Inflammation ,Epidermis (botany) ,Dendritic Cells ,CD58 Antigens ,CD2 ,medicine.disease ,inflammatory skin diseases ,030104 developmental biology ,biology.protein ,Cancer research ,Leukocyte Common Antigens ,Epidermis ,030215 immunology - Abstract
The interplay between keratinocytes and immune cells, especially T cells, plays an important role in the pathogenesis of chronic inflammatory skin diseases. During psoriasis, keratinocytes attract T cells by releasing chemokines, while skin-infiltrating self-reactive T cells secrete proinflammatory cytokines, e.g., IFNγ and IL-17A, that cause epidermal hyperplasia. Similarly, in chronic graft-versus-host disease, allogenic IFNγ-producing Th1/Tc1 and IL-17-producing Th17/Tc17 cells are recruited by keratinocyte-derived chemokines and accumulate in the skin. However, whether keratinocytes act as nonprofessional antigen-presenting cells to directly activate naive human T cells in the epidermis remains unknown. Here, we demonstrate that under proinflammatory conditions, primary human keratinocytes indeed activate naive human T cells. This activation required cell contact and costimulatory signaling via CD58/CD2 and CD54/LFA-1. Naive T cells costimulated by keratinocytes selectively differentiated into Th1 and Th17 cells. In particular, keratinocyte-initiated Th1 differentiation was dependent on costimulation through CD58/CD2. The latter molecule initiated STAT1 signaling and IFNγ production in T cells. Costimulation of T cells by keratinocytes resulting in Th1 and Th17 differentiation represents a new explanation for the local enrichment of Th1 and Th17 cells in the skin of patients with a chronic inflammatory skin disease. Consequently, local interference with T cell–keratinocyte interactions may represent a novel strategy for the treatment of Th1 and Th17 cell-driven skin diseases.
- Published
- 2019
19. Structure-based identification of inhibitors disrupting the CD2–CD58 interactions
- Author
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Adèle D. Laurent, Laurence Leherte, Daniel P. Vercauteren, Neha Tripathi, Modélisation Et Spectroscopie (ModES), Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), and Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
- Subjects
T-Lymphocytes ,CD58 ,[SDV]Life Sciences [q-bio] ,CD2 Antigens ,Drug Evaluation, Preclinical ,Computational biology ,Molecular Dynamics Simulation ,Molecular dynamics ,Ligands ,01 natural sciences ,Drug design ,Docking ,Structure-Activity Relationship ,Cell surface receptor ,0103 physical sciences ,Drug Discovery ,Humans ,[CHIM]Chemical Sciences ,Amino Acid Sequence ,Organic Chemicals ,Physical and Theoretical Chemistry ,ADME ,Virtual screening ,Binding Sites ,010304 chemical physics ,Chemistry ,[CHIM.ORGA]Chemical Sciences/Organic chemistry ,CD58 Antigens ,CD2–CD58 interactions ,In vitro ,3. Good health ,0104 chemical sciences ,Computer Science Applications ,Molecular Docking Simulation ,010404 medicinal & biomolecular chemistry ,Docking (molecular) ,Pharmacophore ,Databases, Chemical ,Protein Binding - Abstract
International audience; The immune system has very intricate mechanisms of fighting against the invading infections which are accomplished by a sequential event of molecular interactions in the body. One of the crucial phenomena in this process is the recognition of T-cells by the antigen-presenting cells (APCs), which is initiated by the rapid interaction between both cell surface receptors, i.e., CD2 located on T-cells and CD58 located on APCs. Under various pathological conditions, which involve undesired immune response, inhibiting the CD2-CD58 interactions becomes a therapeutically relevant opportunity. Herein we present an extensive work to identify novel inhibiting agents of the CD2-CD58 interactions. Classical molecular dynamics (MD) simulations of the CD2-CD58 complex highlighted a series of crucial CD58 residues responsible for the interactions with CD2. Based on such results, a pharmacophore map, complementary to the CD2-binding site of CD58, was created and employed for virtual screening of ~ 300,000 available compounds. On the ~ 6000 compounds filtered from pharmacophore mapping, ADME screening leads to ~ 350 molecules. Molecular docking was then performed on these molecules, and fifteen compounds emerged with significant binding energy (< - 50 kcal/mol) for CD58. Finally, short MD simulations were performed in triplicate on each complex (i) to provide a microscopic view of the ligand binding and (ii) to rule out possibly weak binders of CD58 from the identified hits. At last, we suggest eight compounds for in vitro testing that were identified as promising hits to bind CD58 with a high binding affinity
- Published
- 2021
20. Siplizumab Induces NK Cell Fratricide Through Antibody-Dependent Cell-Mediated Cytotoxicity
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Christian Binder, Felix Sellberg, Filip Cvetkovski, Stefan Berg, Erik Berglund, and David Berglund
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lcsh:Immunologic diseases. Allergy ,0301 basic medicine ,medicine.drug_class ,Immunology ,CD2 Antigens ,antibody-dependent cell-mediated cytotoxicity ,Antibodies, Monoclonal, Humanized ,Lymphocyte Activation ,Monoclonal antibody ,Lymphocyte Depletion ,siplizumab ,NK alloreactivity ,Jurkat Cells ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Immunology and Allergy ,NK cell ,Cytotoxicity ,Siplizumab ,Original Research ,Antibody-dependent cell-mediated cytotoxicity ,biology ,Chemistry ,Receptors, IgG ,Antibody-Dependent Cell Cytotoxicity ,Immunology in the medical area ,Autologous lymphocyte ,CD2 ,Mixed lymphocyte reaction ,Actin cytoskeleton ,Molecular biology ,Killer Cells, Natural ,030104 developmental biology ,spontaneous cytotoxicity ,Immunologi inom det medicinska området ,biology.protein ,Antibody ,lcsh:RC581-607 ,030215 immunology ,medicine.drug - Abstract
The glycoprotein CD2 is expressed on T and NK cells and contributes to cell-cell conjugation, agonistic signaling and actin cytoskeleton rearrangement. CD2 has previously been shown to have an important function in natural NK cell cytotoxicity but to be expendable in antibody-mediated cytotoxicity. Siplizumab is a monoclonal anti-CD2 IgG1 antibody that is currently undergoing clinical trials in the field of transplantation. This study investigated the effect of CD2 binding and Fc γ receptor binding by siplizumab (Fc-active) and Fc-silent anti-CD2 monoclonal antibodies in allogeneic mixed lymphocyte reaction and autologous lymphocyte culture. Further, induction of NK cell fratricide and inhibition of natural cytotoxicity as well as antibody-dependent cytotoxicity by these agents were assessed. Blockade of CD2 via monoclonal antibodies in the absence of Fc γ receptor binding inhibited NK cell activation in allogeneic mixed lymphocyte reaction. In contrast, siplizumab increased NK cell activation in both mixed lymphocyte reaction and autologous lymphocyte culture due to FcγRIIIA binding. However, experiments using purified NK cells did not show an inhibitory effect of CD2 blockade on natural cytotoxicity or antibody-dependent cytotoxicity. Lastly, it was shown that siplizumab induces NK cell fratricide. Concluding, siplizumab is a promising biopharmaceutical drug candidate for depletion of T and NK cells with minimal off-target effects.
- Published
- 2021
21. CD2 Is a Novel Immune-Related Prognostic Biomarker of Invasive Breast Carcinoma That Modulates the Tumor Microenvironment
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Yanzhu Chen, Lin Zhang, Feng Liu, and Zhishang Meng
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0301 basic medicine ,Adult ,ImmuneScore ,Candidate gene ,medicine.medical_treatment ,Immunology ,CD2 Antigens ,Breast Neoplasms ,Biology ,Transcriptome ,03 medical and health sciences ,0302 clinical medicine ,breast invasive carcinoma ,medicine ,Biomarkers, Tumor ,Tumor Microenvironment ,Immunology and Allergy ,Humans ,Protein Interaction Maps ,RNA-Seq ,tumor immunology ,KEGG ,Lymph node ,Aged ,Neoplasm Staging ,Original Research ,Aged, 80 and over ,Tumor microenvironment ,Cancer ,Immunotherapy ,RC581-607 ,Middle Aged ,medicine.disease ,Prognosis ,CD2 ,Survival Analysis ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer research ,Biomarker (medicine) ,Female ,Immunologic diseases. Allergy ,Algorithms - Abstract
Female breast cancer (BCa) is the most commonly occurring cancer worldwide. The tumor microenvironment (TME) plays an essential role in tumor invasion, angiogenesis, unlimited proliferation, and even immune escape, but we know little about the TME of BCa. In this study, we aimed to find a TME-related biomarker for BCa, especially for invasive breast carcinoma (BRCA), that could predict prognosis and immunotherapy efficacy. Based on RNA-seq transcriptome data and the clinical characteristics of 1222 samples (113 normal and 1109 tumor samples) from The Cancer Genome Atlas (TCGA) database, we used the ESTIMATE algorithm to calculate the ImmuneScore and StromalScore and then identified differentially expressed genes (DEGs) between the high and low ImmuneScore groups and the high and low StromalScore groups. Thereafter, a protein–protein interaction (PPI) network analysis and univariate Cox regression analyses of overall survival were used to identify potential key genes. Five candidate genes were identified, comprising CD2, CCL19, CD52, CD3E, and ITK. Thereafter, we focused on CD2, analyzing CD2 expression and its association with survival. CD2 expression was associated with tumor size (T stage) to some extent, but not with overall TNM stage, lymph node status (N stage), or distant metastasis (M stage). High CD2 expression was associated with longer survival. METABRIC data were used to validate the survival result (n = 276). Gene set enrichment analysis (GSEA) showed that the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways that were significantly associated with high CD2 expression were mainly immune-related pathways. Furthermore, CD2 expression was correlated with 16 types of tumor-infiltrating immune cells (TICs). Hence, CD2 might be a novel biomarker in terms of molecular typing, and it may serve as a complementary approach to TNM staging to improve clinical outcome prediction for BCa patients.
- Published
- 2021
22. A case report on concurrent occurrence of systemic mastocytosis and myeloid sarcoma presenting with extensive skin involvements and the results of genetic study
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Xinye Wang, Daobin Zhou, Xianyong Jiang, Lu Zhang, Hao Cai, and Xuan Wang
- Subjects
Male ,Pathology ,Dasatinib ,0302 clinical medicine ,myeloid sarcoma ,030212 general & internal medicine ,Systemic mastocytosis ,Sarcoma, Myeloid ,Skin ,Antibiotics, Antineoplastic ,medicine.diagnostic_test ,biology ,Cytarabine ,Kit ,General Medicine ,Mast cell ,DNA-Binding Proteins ,Proto-Oncogene Proteins c-kit ,medicine.anatomical_structure ,Treatment Outcome ,Bone marrow suppression ,030220 oncology & carcinogenesis ,Drug Therapy, Combination ,Sarcoma ,medicine.drug ,Research Article ,Adult ,medicine.medical_specialty ,Antimetabolites, Antineoplastic ,CD2 Antigens ,Histamine Antagonists ,Platelet Transfusion ,systemic mastocytosis ,03 medical and health sciences ,Mastocytosis, Systemic ,medicine ,Myeloid sarcoma ,Humans ,Clinical Case Report ,Protein Kinase Inhibitors ,CD117 ,business.industry ,Daunorubicin ,Interleukin-2 Receptor alpha Subunit ,medicine.disease ,Skin biopsy ,Mutation ,biology.protein ,Leukotriene Antagonists ,Lymph Nodes ,business ,Arid1a ,Transcription Factors - Abstract
Introduction: Systemic mastocytosis is a rare disease due to mast cell accumulation in various extracutaneous sites. Systemic mastocytosis with an associated clonal hematologic non-MC lineage disease is the second most common subtype of systemic mastocytosis. The most common mutation associated with both systemic mastocytosis and myeloid sarcoma is mutation in Kit. Here, we identified the novel KIT D816V and ARID1A G1254S mutations co-occurring in systemic mastocytosis with myeloid sarcoma. Patient Concerns: A 33-year old male patient presented multiple skin lesions for 10 years. Symptoms accelerated in 2017 with decreased body weight. Physical examination revealed enlarged lymph nodes in his neck, axilla and inguinal region; conjunctival hemorrhage; gingival hyperplasia. Skin biopsy showed mast cell infiltration. Flow cytometry detected CD2, CD25 and CD117 positive cells in lymph nodes. Codon 816 KIT mutation D816V and codon 1245 ARID1A mutation G1254S were found in peripheral blood. MPO, CD117, CD68 positive cells in lymph nodes indicated co-existing myeloid sarcoma. Diagnosis: Systemic mastocytosis with an associated clonal hematologic non-MC lineage disease of myeloid sarcoma Interventions: Cytarabine and daunorubicin for myeloid sarcoma and dasatinib for systemic mastocytosis were initiated. Anti-histamine and anti-leukotrienes therapy were used to prevent NSAIDs-induced shock. Platelets were infused to treat bone marrow suppression. Outcomes: Patient was discharged after recovered from bone marrow suppression. Dasatinib continued on outpatient. Conclusion: This is the first case of patient with systemic mastocytosis and myeloid sarcoma simultaneously presenting extensive skin involvements. Mutations of Kit and Arid1a emphasis the importance to notice possibility of various tumors occurring in patients with multiple mutations. In addition, cysteine-leukotrienes-receptor antagonists should always be used to prevent anaphylactic shock due to mast cell activation.
- Published
- 2020
23. Siplizumab, an Anti-CD2 Monoclonal Antibody, Induces a Unique Set of Immune Modulatory Effects Compared to Alemtuzumab and Rabbit Anti-Thymocyte Globulin In Vitro
- Author
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Binder, Christian, Sellberg, Felix, Cvetkovski, Filip, Berglund, Erik, and Berglund, David
- Subjects
Immunology ,CD2 Antigens ,costimulation blockade ,Antibodies, Monoclonal, Humanized ,Lymphocyte Activation ,siplizumab ,Immunomodulation ,T cell biology ,Antineoplastic Agents, Immunological ,T-Lymphocyte Subsets ,Cell Line, Tumor ,Animals ,Humans ,Alemtuzumab ,Cells, Cultured ,Original Research ,Antilymphocyte Serum ,immune modulation ,Receptors, IgG ,Antibody-Dependent Cell Cytotoxicity ,Immunology in the medical area ,Forkhead Transcription Factors ,Complement System Proteins ,Immunologi inom det medicinska området ,immunotherapy ,Rabbits - Abstract
Antibodies are commonly used in organ transplant induction therapy and to treat autoimmune disorders. The effects of some biologics on the human immune system remain incompletely characterized and a deeper understanding of their mechanisms of action may provide useful insights for their clinical application. The goal of this study was to contrast the mechanistic properties of siplizumab with Alemtuzumab and rabbit Anti-Thymocyte Globulin (rATG). Mechanistic assay systems investigating antibody-dependent cell-mediated cytotoxicity, antibody-dependent cell phagocytosis and complement-dependent cytotoxicity were used to characterize siplizumab. Further, functional effects of siplizumab, Alemuzumab, and rATG were investigated in allogeneic mixed lymphocyte reaction. Changes in T cell activation, T cell proliferation and frequency of naive T cells, memory T cells and regulatory T cells induced by siplizumab, Alemtuzumab and rATG in allogeneic mixed lymphocyte reaction were assessed via flow cytometry. Siplizumab depleted T cells, decreased T cell activation, inhibited T cell proliferation and enriched naive and bona fide regulatory T cells. Neither Alemtuzumab nor rATG induced the same combination of functional effects. The results presented in this study should be used for further in vitro and in vivo investigations that guide the clinical use of immune modulatory biologics.
- Published
- 2020
24. A novel CD2 staining–based flow cytometric assay for assessment of natural killer cell cytotoxicity
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Lei Lei, Rui Teng, Jinsong Hu, Michael O'Dwyer, Ramone A. Williamson, Peigen Gao, Ang Li, Nan Lv, Dan Zhang, Ping Chen, and Yanmeng Wang
- Subjects
Cytotoxicity, Immunologic ,0301 basic medicine ,Microbiology (medical) ,HL60 ,Clinical Biochemistry ,Population ,Cell ,CD2 Antigens ,Apoptosis ,Flow cytometry ,Natural killer cell ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,medicine ,Humans ,Immunology and Allergy ,education ,Cytotoxicity ,Research Articles ,Fluorescent Dyes ,education.field_of_study ,Staining and Labeling ,biology ,medicine.diagnostic_test ,Chemistry ,flow cytometry ,Biochemistry (medical) ,Public Health, Environmental and Occupational Health ,Reproducibility of Results ,Hematology ,natural killer cell ,CD2 ,Molecular biology ,Killer Cells, Natural ,Medical Laboratory Technology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,cytotoxicity ,Antibody ,Research Article ,K562 cells - Abstract
Background Assessing cytotoxicity is fundamental to studying natural killer (NK) cell function. Various radioactive and non‐radioactive cytotoxicity assays measuring target cell death have been developed. Among these methods, the most commonly used 51Chromium‐release assay (CRA) and flow cytometry–based cytotoxicity assays (FCCs) are the major representatives. Nonetheless, several drawbacks, including dye leakage and the potential effects of prior labeling on cells, curb the broad applicability of the FCCs. Methods Here, we report a rapid FCC for quantifying target cell death after co‐incubation with NK cells. In this assay, after 4 hours of NK cell‐target cell co‐incubation, fluorochrome‐conjugated CD2 antibody was used to identify NK cells, and SYTOX Green and Annexin V‐FITC were further used to detect target cell death in CD2‐negative population. In parallel, both CRA and FCC assay using CFSE/ 7‐AAD were performed to validate the reproducibility and replicability. Results We observed that CD2 is exclusively positive on NK cells other than the most common hematological target tumor cells, such as K562, HL60, MOLM13, Raji, NCI‐H929, rpmi8226, MM.1S, and KMS11. Assessment of target cell death using the CD2‐based FCC shows a significantly higher percent specific lysis of the target cells compared to the standard CRA and the FCC assay using CFSE and 7‐AAD. Conclusions We demonstrated that this CD2‐based FCC is a fast, simple, and reliable method for evaluating NK cell cytotoxicity., A CD2 staining‐based flow cytometric assay for measuring natural killer cell cytotoxicity. After incubation of NK cells with target cells at different E:T ratios for 4 hours, fluorochrome‐conjugated CD2 antibody was used to identify NK cells, nucleic acid dye SYTOX Green and early apoptotic marker Annexin V were further used to detect target cell death in CD2‐negative population. Compared to 51Chromium release assay and other traditional flow cytometric assays, this CD2 based flow cytometric cytotoxicity assay is highly sensitive and reliable, since there is no any interference before or during the incubation by labeling cells.
- Published
- 2020
25. Anti-CD2 Antibody-Coated Nanoparticles Containing IL-2 Induce NK Cells That Protect Lupus Mice
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David A. Horwitz, Aijing Liu, Sean Bickerton, Giuseppe Castaldo, Giuseppe Matarese, Tarek M. Fahmy, Antonio La Cava, Horwitz, D. A., Liu, A., Bickerton, S., Castaldo, G., Matarese, G., Fahmy, T. M., and La Cava, A.
- Subjects
0301 basic medicine ,lcsh:Immunologic diseases. Allergy ,medicine.medical_treatment ,Immunology ,Population ,CD2 Antigens ,NK cells ,medicine.disease_cause ,Autoimmunity ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,systemic lupus erythematosus ,Transforming Growth Factor beta ,medicine ,cytokine ,Immunology and Allergy ,Animals ,Lupus Erythematosus, Systemic ,NK cell ,education ,Original Research ,education.field_of_study ,Systemic lupus erythematosus ,biology ,Chemistry ,nanoparticle ,autoimmunity ,immune regulation ,Immunotherapy ,medicine.disease ,Molecular biology ,cytokines ,Killer Cells, Natural ,Mice, Inbred C57BL ,030104 developmental biology ,Mice, Inbred DBA ,biology.protein ,Interleukin-2 ,Nanoparticles ,immunotherapy ,Antibody ,lcsh:RC581-607 ,CD8 ,030215 immunology ,Transforming growth factor - Abstract
We recently reported that the treatment with nanoparticles (NPs) loaded with tolerogenic cytokines suppressed the manifestations of lupus-like disease induced by the transfer of donor CD4+ T cells from DBA/2 mice into (C57BL/6 × DBA/2)F1 (BDF1) mice. Although the protective effects were ascribed to the induction of adaptive CD4+ and CD8+ T regulatory cells, the results suggested that another population of immune cells could be involved. Here we report that NK cells critically contribute to the protection from lupus-like disease conferred by NPs to BDF1 mice, and that this effect is TGF-β-dependent.
- Published
- 2020
26. Phosphoproteomics of CD2 signaling reveals AMPK-dependent regulation of lytic granule polarization in cytotoxic T cells
- Author
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Gabriele Turacchio, Gioia Boncompagni, Michael Volkmar, Mario Milco D'Elios, Oreste Acuto, Raphael Heilig, Nicoletta Resta, Vanessa Zurli, Giuseppe Campoccia, Cosima T. Baldari, Tommaso Montecchi, Anna Kabanova, Rienk Offringa, Roman Fischer, Isabel Poschke, and Giuliana Wimmer
- Subjects
Proteomics ,CD2 Antigens ,AMP-Activated Protein Kinases ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Cytotoxic T cell ,Humans ,Phosphorylation ,Protein kinase A ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Chemistry ,Secretory Vesicles ,Autophagy ,Phosphoproteomics ,AMPK ,Cell Biology ,Phosphoproteins ,Cell biology ,Cell killing ,030220 oncology & carcinogenesis ,CD8 ,Signal Transduction ,T-Lymphocytes, Cytotoxic - Abstract
Understanding the costimulatory signaling that enhances the activity of cytotoxic T cells (CTLs) could identify potential targets for immunotherapy. Here, we report that CD2 costimulation plays a critical role in target cell killing by freshly isolated human CD8+ T cells, which represent a challenging but valuable model to gain insight into CTL biology. We found that CD2 stimulation critically enhanced signaling by the T cell receptor in the formation of functional immune synapses by promoting the polarization of lytic granules toward the microtubule-organizing center (MTOC). To gain insight into the underlying mechanism, we explored the CD2 signaling network by phosphoproteomics, which revealed 616 CD2-regulated phosphorylation events in 373 proteins implicated in the regulation of vesicular trafficking, cytoskeletal organization, autophagy, and metabolism. Signaling by the master metabolic regulator AMP-activated protein kinase (AMPK) was a critical node in the CD2 network, which promoted granule polarization toward the MTOC in CD8+ T cells. Granule trafficking was driven by active AMPK enriched on adjacent lysosomes, revealing previously uncharacterized signaling cross-talk between vesicular compartments in CD8+ T cells. Our results thus establish CD2 signaling as key for mediating cytotoxic killing and granule polarization in freshly isolated CD8+ T cells and strengthen the rationale to choose CD2 and AMPK as therapeutic targets to enhance CTL activity.
- Published
- 2020
27. Expansion and CD2/CD3/CD28 stimulation enhance Th2 cytokine secretion of human invariant NKT cells with retained anti-tumor cytotoxicity
- Author
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Geoffrey Neale, Kelly A. Andrews, Xiaodian Sun, Daniel E. Geraghty, Asha Pillai, Shalini Pereira, Paige Tedrick, Anouk A.J. Hamers, Kim E. Nichols, and Katherine Verbist
- Subjects
0301 basic medicine ,Cancer Research ,CD3 Complex ,Cell Transplantation ,medicine.medical_treatment ,Immunology ,CD2 Antigens ,Apoptosis ,Blood Donors ,Lymphocyte Activation ,Jurkat cells ,Article ,Antibodies ,03 medical and health sciences ,Jurkat Cells ,0302 clinical medicine ,Th2 Cells ,Cancer immunotherapy ,Co-stimulation ,CD28 Antigens ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Humans ,Genetics (clinical) ,Cells, Cultured ,Cell Proliferation ,Transplantation ,Chemistry ,Gene Expression Profiling ,CD28 ,Cell Biology ,Natural killer T cell ,Granzyme B ,030104 developmental biology ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Cytokines ,Natural Killer T-Cells ,Immunotherapy ,K562 Cells - Abstract
Background aims Key obstacles in human iNKT cell translational research and immunotherapy include the lack of robust protocols for dependable expansion of human iNKT cells and the paucity of data on phenotypes in post-expanded cells. Methods We delineate expansion methods using interleukin (IL)-2, IL-7 and allogeneic feeder cells and anti-CD2/CD3/CD28 stimulation by which to dependably augment Th2 polarization and direct cytotoxicity of human peripheral blood CD3+Vα24+Vβ11+ iNKT cells. Results Gene and protein expression profiling demonstrated augmented Th2 cytokine secretion (IL-4, IL-5, IL-13) in expanded iNKT cells stimulated with anti-CD2/CD3/CD28 antibodies. Cytotoxic effector molecules including granzyme B were increased in expanded iNKT cells after CD2/CD3/CD28 stimulation. Direct cytotoxicity assays using unstimulated expanded iNKT cell effectors revealed α-galactosyl ceramide (α-GalCer)-dependent killing of the T-ALL cell line Jurkat. Moreover, CD2/CD3/CD28 stimulation of expanded iNKT cells augmented their (α-GalCer-independent) killing of Jurkat cells. Co-culture of expanded iNKT cells with stimulated responder cells confirmed contact-dependent inhibition of activated CD4+ and CD8+ responder T cells. Discussion These data establish a robust protocol to expand and novel pathways to enhance Th2 cytokine secretion and direct cytotoxicity in human iNKT cells, findings with direct implications for autoimmunity, vaccine augmentation and anti-infective immunity, cancer immunotherapy and transplantation.
- Published
- 2020
28. CD2 Regulates Pathogenesis of Asthma Induced by House Dust Mice Extract
- Author
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Kellie Mullany, Ryan M. Griffin, Rupali Das, Hariharan Subramanian, Ananth K. Kammala, Kanedra Thaxton, Reynold A. Panettieri, and Tanwir Hashem
- Subjects
0301 basic medicine ,Pathogenesis ,Mice ,0302 clinical medicine ,Immunology and Allergy ,Medicine ,Lung ,Original Research ,Mice, Knockout ,Mice, Inbred BALB C ,Interleukin-13 ,Pyroglyphidae ,Interleukin ,respiratory system ,house dust mite extract ,microRNAs ,medicine.anatomical_structure ,Th2-allergic response ,interleukin (IL)-13 ,Cytokines ,Female ,costimulatory molecules ,medicine.symptom ,Bronchoalveolar Lavage Fluid ,lcsh:Immunologic diseases. Allergy ,Immunology ,CD2 Antigens ,Inflammation ,03 medical and health sciences ,Immune system ,Th2 Cells ,Respiratory Hypersensitivity ,Animals ,Interleukin 4 ,Asthma ,business.industry ,Mucin ,asthma ,medicine.disease ,CD2 ,respiratory tract diseases ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Gene Expression Regulation ,lcsh:RC581-607 ,business ,030215 immunology - Abstract
Characteristic of allergic asthma, CD4+Th2 lymphocytes secrete Th2 cytokines, interleukin (IL)-4, IL-13, and IL-5 that mediate the inflammatory immune response. Surface expression of CD2 and its ligand, CD58, is increased on the monocytes and eosinophils of asthma patients, which correlate with elevated serum IgE levels, suggesting that CD2 may contribute to allergic airway inflammation. Using a murine model of asthma, we observed that house dust mice extract (HDME)-exposed Balb/c mice have increased airway hyperresponsiveness (AHR), lung inflammation, goblet cell hyperplasia, and elevated levels of Th2 cytokines in the lungs, as well as increased serum IgE levels as compared to the control mice. In contrast, with the exception of serum IgE levels, all the other parameters were significantly reduced in HDME-treated Cd2 -/- mice. Interestingly, Il13 but not Il4 or Il5 gene expression in the lungs was dramatically decreased in HDME-exposed Cd2 -/- mice. Of note, the gene expression of IL-13 downstream targets (Muc5b and Muc5ac) and high affinity IL-13Rα2 were upregulated in the lungs of HDME-exposed Balb/c mice but were significantly reduced in HDME-exposed Cd2 -/- mice. Consistently, gene expression of microRNAs regulating mucin production, inflammation, airway smooth muscle cell proliferation and IL-13 transcripts were increased in the lungs of HDME-exposed Cd2 -/- mice. Given the established role of IL-13 in promoting goblet cell hyperplasia, lung inflammation and AHR in allergic asthma, our studies reveal a unique role for CD2 in the regulation of Th2-associated allergic asthma.
- Published
- 2020
29. CD2 Immunobiology
- Author
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Binder, Christian, Cvetkovski, Filip, Sellberg, Felix, Berg, Stefan, Visbal, Horacio Paternina, Sachs, David H., Berglund, Erik, and Berglund, David
- Subjects
LFA-3 (lymphocyte functional antigen-3) ,Immunological Synapses ,T cell activation ,T-Lymphocytes ,Immunology ,CD2 Antigens ,Immunology in the medical area ,Alefacept ,Review ,costimulation blockade ,Lymphocyte Activation ,CD2 ,siplizumab ,costimulation ,Immunologi inom det medicinska området ,Immunologi ,Animals ,Humans ,CD58 - Abstract
The glycoprotein CD2 is a costimulatory receptor expressed mainly on T and NK cells that binds to LFA3, a cell surface protein expressed on e.g., antigen-presenting cells. CD2 has an important role in the formation and organization of the immunological synapse that is formed between T cells and antigen-presenting cells upon cell-cell conjugation and associated intracellular signaling. CD2 expression is upregulated on memory T cells as well as activated T cells and plays an important role in activation of memory T cells despite the coexistence of several other costimulatory pathways. Anti-CD2 monoclonal antibodies have been shown to induce immune modulatory effects in vitro and clinical studies have proven the safety and efficacy of CD2-targeting biologics. Investigators have highlighted that the lack of attention to the CD2/LFA3 costimulatory pathway is a missed opportunity. Overall, CD2 is an attractive target for monoclonal antibodies intended for treatment of pathologies characterized by undesired T cell activation and offers an avenue to more selectively target memory T cells while favoring immune regulation.
- Published
- 2020
30. Proximity ligation assay to study protein–protein interactions of proteins on two different cells
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Seetharama D. Jois, Sitanshu S. Singh, Nithya Jambunathan, Rushikesh Sable, Konstantin G. Kousoulas, and Sandeep Pallerla
- Subjects
0301 basic medicine ,Models, Molecular ,assays ,CD58 ,bioanalysis ,Cell ,CD2 Antigens ,Proximity ligation assay ,Jurkat cells ,General Biochemistry, Genetics and Molecular Biology ,Protein–protein interaction ,03 medical and health sciences ,Jurkat Cells ,0302 clinical medicine ,Immune system ,medicine ,Humans ,Cells, Cultured ,biology ,Chemistry ,Histocompatibility Antigens Class II ,Membrane Proteins ,Proteins ,flourescence ,CD58 Antigens ,Flow Cytometry ,Small molecule ,Synoviocytes ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,030220 oncology & carcinogenesis ,biology.protein ,Antibody ,Reports ,Biotechnology ,Protein Binding - Abstract
Protein–protein interactions (PPI) by homo-, hetero- or oligo-merization in the cellular environment regulate cellular processes. PPI can be inhibited by antibodies, small molecules or peptides, and this inhibition has therapeutic value. A recently developed method, the proximity ligation assay (PLA), provides detection of PPI in the cellular environment. However, most applications using this assay are for proteins expressed in the same cell. We employ PLA for the first time to study PPI of cell surface proteins on two different cells. Inhibition of PPI using a peptide inhibitor is also quantified using this assay; PLA is used to detect PPI of CD2 and CD58 between Jurkat cells (T cells) and human fibroblast-like synoviocyte-rheumatoid arthritis cells that are important in the immune response in the autoimmune disease rheumatoid arthritis. This assay provides direct evidence of inhibition of PPI of two proteins on different cell surfaces., METHOD SUMMARY The proximity ligation assay (PLA) technique has been used to study protein–protein interactions (PPI) within a particular cell or to study the PPI of cell surface proteins that are expressed on the same cell. Here, for the first time, we report a PLA to be used on the surface of different cells. We have used this technique to visualize the interactions between proteins present on Jurkat (a nonadherent cell line) and HFLS-RA (an adherent cell line) cells in the same experiment. The assay was performed using the Duolink®II assay kit (Sigma Aldrich or Olink Bioscience, Uppsala, Sweden).
- Published
- 2018
31. Hydrodynamic trapping measures the interaction between membrane-associated molecules
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Ana Filipa L.O.M. Santos, Jana Hladilkova, Peter Jönsson, Victoria Junghans, Simon J. Davis, and Mikael Lund
- Subjects
0301 basic medicine ,Streptavidin ,Flexibility (anatomy) ,Glycosylation ,Lipid Bilayers ,CD2 Antigens ,lcsh:Medicine ,02 engineering and technology ,010402 general chemistry ,Hydrodynamic trapping ,01 natural sciences ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Membrane associated ,medicine ,Molecule ,Humans ,Lipid bilayer ,lcsh:Science ,030304 developmental biology ,0303 health sciences ,Multidisciplinary ,Membranes ,Chemistry ,lcsh:R ,Membrane Proteins ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,030104 developmental biology ,medicine.anatomical_structure ,Membrane protein ,CD4 Antigens ,Biophysics ,Hydrodynamics ,Leukocyte Common Antigens ,lcsh:Q ,0210 nano-technology - Abstract
How membrane proteins distribute and behave on the surface of cells is determined by the molecules’ interaction potential. However, measuring this potential, and how it varies with protein-to-protein distance, has been challenging. We here present how a method we call hydrodynamic trapping can achieve this. Our method uses the focused liquid flow from a micropipette to locally accumulate molecules protruding from a lipid membrane. The interaction potential, as well as information about the dimensions of the studied molecule, are obtained by relating the degree of accumulation to the strength of the trap. We have used this to study four representative proteins, with different height-to-width ratios and protein properties; from the globular streptavidin, to the rod-like immune cell proteins CD2, CD4 and CD45. The obtained data illustrates how protein shape, glycosylation and flexibility influence the behaviour of membrane proteins as well as underline the general applicability of the method.
- Published
- 2018
32. Pharmacokinetic and pharmacodynamic study of a clinically effective anti‐CD2 monoclonal antibody
- Author
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David H. Sachs, Felix Sellberg, Jane Fontenot, Megan Sykes, J Hope, David Berglund, Erik Berglund, Christian Binder, and Adam Griesemer
- Subjects
Male ,0301 basic medicine ,Pan troglodytes ,medicine.drug_class ,Biopsy ,Immunology ,CD2 Antigens ,Pharmacology ,Antibodies, Monoclonal, Humanized ,Monoclonal antibody ,Lymphocyte Depletion ,Immunophenotyping ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Lymphocytes ,Siplizumab ,Lymph node ,biology ,business.industry ,Antibodies, Monoclonal ,General Medicine ,Rats ,Killer Cells, Natural ,Transplantation ,030104 developmental biology ,medicine.anatomical_structure ,Immunoglobulin G ,Monoclonal ,biology.protein ,Cytokines ,Female ,Lymph ,Antibody ,business ,Biomarkers ,030215 immunology ,medicine.drug - Abstract
The humanized IgG1κ monoclonal antibody siplizumab and its rat parent monoclonal IgG2b antibody BTI-322 are directed against the CD2 antigen. Siplizumab is species-specific, reacting with human and chimpanzee cells but not with cells from any other species, including other non-human primates. Because siplizumab treatment has recently shown great potential in clinical transplantation, we now present the results of our previous pharmacokinetic, pharmacodynamic and safety studies of both antibodies. Fourteen chimpanzees received 1-3 doses of 0.143 to 5.0 mg/kg iv The effects were followed with flow cytometry on peripheral lymphocytes and staining of lymph nodes. Side effects were recorded. Serum antibody concentrations were followed. Across the doses, a rapid, transient depletion of CD2, CD3, CD4 and CD8 lymphocytes and NK cells was observed for both antibodies. Immune reconstitution was more rapid for BTI-322 compared to siplizumab. Paracortical lymph node T cell depletion was moderate, estimated at 45% with doses of >0.6 mg/kg. Restoration of lymph node architecture was seen after two weeks to two months for all animals. All four subjects receiving BTI-322 experienced AEs on the first dosing day, while the eight subjects dosed with siplizumab experienced few mild, transient AEs. Infusion with siplizumab and BTI-322 resulted in rapid depletion of CD2+ cells in circulation and tissue. Siplizumab had a longer t1/2 and fewer AEs compared to BTI-322.
- Published
- 2019
33. Subpopulations of swine γδ T cells defined by TCRγ and WC1 gene expression
- Author
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Colin P. Farrell, John C. Schwartz, Lisa-Maria Prawits, Cynthia L. Baldwin, John A. Hammond, Alexandria Gillespie, Janice C. Telfer, Sabine E. Hammer, Angela Schlerka, and Lauren Le Page
- Subjects
Membrane Glycoproteins ,Swine ,T-Lymphocytes ,Receptor expression ,Immunology ,T-cell receptor ,Haplotype ,CD2 Antigens ,Receptors, Antigen, T-Cell, gamma-delta ,Ruminants ,Biology ,Ligand (biochemistry) ,Molecular biology ,Genes, T-Cell Receptor ,Gene expression ,Homologous chromosome ,Animals ,Cattle ,Receptor ,Gene ,Developmental Biology - Abstract
γδ T cells constitute a major portion of lymphocytes in the blood of both ruminants and swine. Subpopulations of swine γδ T cells have been distinguished by CD2 and CD8α expression. However, it was not clear if they have distinct expression profiles of their T-cell receptor (TCR) or WC1 genes. Identifying receptor expression will contribute to understanding the functional differences between these subpopulations and their contributions to immune protection. Here, we annotated three genomic assemblies of the swine TCRγ gene locus finding four gene cassettes containing C, J and V genes, although some haplotypes carried a null TRGC gene (TRGC4). Genes in the TRGC1 cassette were homologs of bovine TRGC5 cassette while the others were not homologous to bovine genes. Here we evaluated three principal populations of γδ T cells (CD2+/SWC5-, CD2-/SWC5+, and CD2-/SWC5-). Both CD2− subpopulations transcribed WC1 co-receptor genes, albeit with different patterns of gene expression but CD2+ cells did not. All subpopulations transcribed TCR genes from all four cassettes, although there were differences in expression levels. Finally, the CD2+ and CD2− γδ T-cell populations differed in their representation in various organs and tissues, presumably at least partially reflective of different ligand specificities for their receptors.
- Published
- 2021
34. HPTE-Induced Embryonic Thymocyte Death and Alteration of Differentiation Is Not Rescued by ERα or GPER Inhibition but Is Exacerbated by Concurrent TCR Signaling
- Author
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Aaditya Patel, Eddie Avellaneda, Victoria Sanchez, Jacqueline Marquez, Atalie Lim, Cristina Zambrano, Sara Moeller, Priscilla Escalante Cobb, Christine Broussard, and Kanya Godde
- Subjects
Male ,estrogen receptor alpha (ERα) ,Diethylstilbestrol ,Estrogen receptor ,Endocrine Disruptors ,Receptors, G-Protein-Coupled ,Mice ,embryonic thymocytes ,Biology (General) ,reproductive and urinary physiology ,Cells, Cultured ,Spectroscopy ,Thymocytes ,Cell Death ,Estrogen receptor binding ,Chemistry ,Cell Differentiation ,General Medicine ,Computer Science Applications ,Cell biology ,Thymocyte ,Receptors, Estrogen ,Female ,GPER ,geographic locations ,Signal Transduction ,medicine.drug ,animal structures ,QH301-705.5 ,medicine.drug_class ,CD2 Antigens ,Receptors, Antigen, T-Cell ,Article ,Catalysis ,Inorganic Chemistry ,Phenols ,parasitic diseases ,medicine ,Animals ,Physical and Theoretical Chemistry ,QD1-999 ,Molecular Biology ,organic chemicals ,T cell development ,Organic Chemistry ,T-cell receptor ,Estrogen Receptor alpha ,endocrine disrupting chemicals ,thymocyte differentiation ,Mice, Inbred C57BL ,HPTE ,G protein-coupled estrogen receptor (GPER) ,Estrogen ,estrogenic endocrine disruptors ,Estrogen receptor alpha - Abstract
Organochlorine pesticides, such as DDT, methoxychlor, and their metabolites, have been characterized as endocrine disrupting chemicals (EDCs), suggesting that their modes of action involve interaction with or abrogation of endogenous endocrine function. This study examined whether embryonic thymocyte death and alteration of differentiation induced by the primary metabolite of methoxychlor, HPTE, rely upon estrogen receptor binding and concurrent T cell receptor signaling. Estrogen receptor inhibition of ERα or GPER did not rescue embryonic thymocyte death induced by HPTE or the model estrogen diethylstilbestrol (DES). Moreover, adverse effects induced by HPTE or DES were worsened by concurrent TCR and CD2 differentiation signaling, compared with EDC exposure post-signaling. Together, these data suggest that HPTE- and DES-induced adverse effects on embryonic thymocytes do not rely solely on ER alpha or GPER but may require both. These results also provide evidence of a potential collaborative signaling mechanism between TCR and estrogen receptors to mediate adverse effects on embryonic thymocytes, as well as highlight a window of sensitivity that modulates EDC exposure severity.
- Published
- 2021
35. Targeted insertion of an anti-CD2 monoclonal antibody transgene into the GGTA1 locus in pigs using FokI-dCas9
- Author
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Andrew M. Lew, Nella Fisicaro, Wayne J. Hawthorne, Peter J. Cowan, Mark B. Nottle, Jamie L. Brady, Evelyn J Salvaris, Ivan Vassiliev, Stephen M. McIlfatrick, and Philip J. O'Connell
- Subjects
0301 basic medicine ,Male ,Nuclear Transfer Techniques ,Swine ,Xenotransplantation ,medicine.medical_treatment ,Transgene ,Science ,CD2 Antigens ,Biology ,Article ,Animals, Genetically Modified ,03 medical and health sciences ,Pregnancy ,CRISPR-Associated Protein 9 ,medicine ,Gene silencing ,CRISPR ,Animals ,Humans ,Gene Knock-In Techniques ,Transgenes ,Deoxyribonucleases, Type II Site-Specific ,Gene ,Multidisciplinary ,Cas9 ,Gene targeting ,Antibodies, Monoclonal ,Reproducibility of Results ,Fibroblasts ,Galactosyltransferases ,Molecular biology ,030104 developmental biology ,Gene Targeting ,biology.protein ,Medicine ,Female ,Antibody - Abstract
Xenotransplantation from pigs has been advocated as a solution to the perennial shortage of donated human organs and tissues. CRISPR/Cas9 has facilitated the silencing of genes in donor pigs that contribute to xenograft rejection. However, the generation of modified pigs using second-generation nucleases with much lower off-target mutation rates than Cas9, such as FokI-dCas9, has not been reported. Furthermore, there have been no reports on the use of CRISPR to knock protective transgenes into detrimental porcine genes. In this study, we used FokI-dCas9 with two guide RNAs to integrate a 7.1 kilobase pair transgene into exon 9 of the GGTA1 gene in porcine fetal fibroblasts. The modified cells lacked expression of the αGal xenoantigen, and secreted an anti-CD2 monoclonal antibody encoded by the transgene. PCR and sequencing revealed precise integration of the transgene into one allele of GGTA1, and a small deletion in the second allele. The cells were used for somatic cell nuclear transfer to generate healthy male knock-in piglets, which did not express αGal and which contained anti-CD2 in their serum. We have therefore developed a versatile high-fidelity system for knocking transgenes into the pig genome for xenotransplantation purposes.
- Published
- 2017
36. Leishmania donovani resistant to Ambisome or Miltefosine exacerbates CD58 expression on NK cells and promotes trans-membrane migration in association with CD2
- Author
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Fauzia Jamal, Sanjiva Bimal, Shubhankar K. Singh, Vikash Kumar, Krishna Pandey, Manish Kumar, Shyam Narayan, Pradeep Das, Pushkar Shivam, Sarita Kumari, A. K. Gupta, and Vidya Nand Ravi Das
- Subjects
0301 basic medicine ,Cellular immunity ,Phosphorylcholine ,CD58 ,Immunology ,CD2 Antigens ,Drug Resistance ,Leishmania donovani ,Stimulation ,Biology ,Lymphocyte Activation ,Biochemistry ,Peripheral blood mononuclear cell ,03 medical and health sciences ,Immunity ,Amphotericin B ,medicine ,Humans ,Immunology and Allergy ,Molecular Biology ,Miltefosine ,Hematology ,CD58 Antigens ,biology.organism_classification ,medicine.disease ,Molecular biology ,CD56 Antigen ,Killer Cells, Natural ,030104 developmental biology ,Visceral leishmaniasis ,Leukocytes, Mononuclear ,Leishmaniasis, Visceral ,medicine.drug - Abstract
Visceral leishmaniasis (VL) is a disease that is associated with compromised immunity and drug un-responsiveness as well as with the emergence of drug resistance in Leishmania donovani (Ld). Ld down-modulates cellular immunity by manipulating signaling agents, including a higher expression of the adhesion molecule CD58. The expression of CD58 and CD2 on natural killer (NK) cells facilitates intercellular adhesion and signaling. The influence of drug-resistant Ld on the expression of CD58 and CD2 was addressed in this study. The mean florescence intensity (MFI) of CD58 but not of CD2 was twofold higher on CD56+ cells during VL, but was down-regulated after treatment. In addition, MFI of CD58 on CD56+ cells was further exacerbated in VL subjects who had relapsed after Ambisome or Miltefosine treatment. The same pattern of CD58 expression was also obtained upon stimulation of healthy peripheral blood mononuclear cells with Miltefosine- or Ambisome-resistant Ld. The ratio of CD56+CD58+IFN-γ+/CD56+CD58+IL-10+ cells was reduced by 6.98-fold after stimulation with Ld. Further, an antagonist to CD58 or its counter-receptor CD2 down-regulated CD56+ NK cell recruitment across a polycarbonate trans-membrane at Ld infection sites. This study reports that factors associated with drug resistance in Ld probably promote higher expression of CD58 on CD56+ cells and their migration to the infection site in association with CD2.
- Published
- 2017
37. Probiotic bacteria prevent Salmonella – induced suppression of lymphoproliferation in mice by an immunomodulatory mechanism
- Author
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R. Doug Wagner and Shemedia J. Johnson
- Subjects
0301 basic medicine ,Male ,Lymphocyte ,medicine.medical_treatment ,T-Lymphocytes ,lcsh:QR1-502 ,Apoptosis ,Probiotic ,lcsh:Microbiology ,Mice ,Peyer's Patches ,0302 clinical medicine ,Salmonella ,Mesenteric lymph nodes ,Lymphocytes ,B-Lymphocytes ,Mice, Inbred BALB C ,biology ,NF-kappa B ,Salmonella enterica ,medicine.anatomical_structure ,Cytokine ,Cytokines ,Female ,Signal Transduction ,Research Article ,Microbiology (medical) ,CD2 Antigens ,PTPRC ,Microbiology ,Immunomodulation ,03 medical and health sciences ,Immune system ,Antigen ,medicine ,Animals ,RNA, Messenger ,Cell Proliferation ,Immunosuppression Therapy ,Salmonella Infections, Animal ,Innate immune system ,Probiotics ,biology.organism_classification ,Disease Models, Animal ,030104 developmental biology ,Toll-Like Receptor 6 ,Gene Expression Regulation ,Signal transduction genes ,biology.protein ,Leukocyte Common Antigens ,Lymph Nodes ,Gene expression ,Spleen ,030215 immunology - Abstract
Background Salmonella enterica infections often exhibit a form of immune evasion. We previously observed that probiotic bacteria could prevent inhibition of lymphoproliferation and apoptosis responses of T cells associated with S. enterica infections in orally challenged mice. Results In this study, changes in expression of genes related to lymphocyte activation in mucosa-associated lymphoid tissues (MALT) of mice orally infected with S. enterica with and without treatment with probiotic bacteria were evaluated. Probiotic bacteria increased expression of mRNA for clusters of differentiation antigen 2 (Cd2), protein tyrosine phosphatase receptor type C (Ptprc), and Toll-like receptor 6 (Tlr6) genes related to T and B cell activation in mouse intestinal tissue. The probiotic bacteria were also associated with reduced mRNA expression of a group of genes (RelB, Myd88, Iκκa, Jun, Irak2) related to nuclear factor of kappa light chains enhancer in B cells (NF-κB) signal transduction pathway-regulated cytokine responses. Probiotic bacteria were also associated with reduced mRNA expression of apoptotic genes (Casp2, Casp12, Dad1, Akt1, Bad) that suggest high avidity lymphocyte sparing. Reduced CD2 immunostaining in mesenteric lymph nodes (MLN) was suggestive of reduced lymphocyte activation in probiotic-treated mice. Reduced immunostaining of TLR6 in MALT of probiotic-treated, S. enterica-infected mice suggests that diminished innate immune sensitivity to S. enterica antigens is associated with preventing lymphocyte deletion. Conclusions The results of this study are consistent with prevention of S. enterica-induced deletion of lymphocytes by the influence of probiotic bacteria in mucosal lymphoid tissues of mice.
- Published
- 2017
38. A Novel Role for Triglyceride Metabolism in Foxp3 Expression
- Author
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Zhanru Yu, Stephen P. Cobbold, Annemieke Ten Bokum, Benedikt M. Kessler, Herman Waldmann, and Duncan Howie
- Subjects
0301 basic medicine ,lcsh:Immunologic diseases. Allergy ,Immunology ,CD2 Antigens ,T-Lymphocytes, Regulatory ,Cell Line ,regulatory T (Treg) cell ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Lipid droplet ,Immunology and Allergy ,Animals ,Humans ,Diacylglycerol O-Acyltransferase ,Protein kinase C ,Protein Kinase C ,Triglycerides ,Diacylglycerol kinase ,Original Research ,chemistry.chemical_classification ,tolerance ,Chemistry ,Fatty Acids ,FOXP3 ,Lipid metabolism ,Forkhead Transcription Factors ,Metabolism ,Lipid Droplets ,differentiation ,lipotoxicity ,metabolomics ,Cell biology ,030104 developmental biology ,Enzyme ,Lipotoxicity ,CD52 Antigen ,Metabolome ,Female ,lipids (amino acids, peptides, and proteins) ,lcsh:RC581-607 ,metabolism ,030215 immunology - Abstract
Lipid metabolism plays a key role in many cellular processes. We show here that regulatory T cells have enhanced lipid storage within subcellular lipid droplets (LD). They also express elevated amounts of both isoforms of diacylglycerol acyl transferase (DGAT1 & 2), enzymes required for the terminal step of triacylglycerol synthesis. In regulatory T-cells (Tregs), the conversion of diacylglycerols to triacylglycerols serves two additional purposes other than lipid storage. First, we demonstrate that it protects T cells from the toxic effects of saturated long chain fatty acids. Second, we show that Triglyceride formation is essential for limiting activation of protein kinase C via free diacyl glycerol moieties. Inhibition of DGAT1 resulted in elevated active PKC and nuclear NFKB, as well as impaired Foxp3 induction in response to TGFβ. Thus, Tregs utilize a positive feedback mechanism to promote sustained expression of Foxp3 associated with control of LD formation.
- Published
- 2019
39. Bioinformatics analysis of genes associated with the patchy-type alopecia areata: CD2 may be a new therapeutic target
- Author
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Shiqing Feng, Weixiao Liu, Guangzhi Ning, Jiaxiao Shi, Peng Peng, Peng Mi, and Cong Xing
- Subjects
differentially expressed genes ,Alopecia Areata ,patchy-type alopecia areata ,CD2 Antigens ,lcsh:Medicine ,Computational biology ,030204 cardiovascular system & hematology ,General Biochemistry, Genetics and Molecular Biology ,Pathogenesis ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Gene expression ,protein‑protein interaction network ,Medicine ,Humans ,KEGG ,Gene ,Autoimmune disease ,integumentary system ,business.industry ,function enrichment ,Gene Expression Profiling ,lcsh:R ,Computational Biology ,Human Genetics ,Genetic Therapy ,Alopecia areata ,medicine.disease ,cd2 ,Hair loss ,030220 oncology & carcinogenesis ,business - Abstract
Background: Alopecia areata (AA) is mainly a T cell-medicated autoimmune disease with non-scarring hair loss and limited treatment options. Of these, the patchy-type alopecia areata (AAP) is the most common and relatively easy to treat due to smaller areas of the scalp affected. To understand the pathogenesis of AAP and explore the therapeutic target, we focus on the molecular signatures by comparing AAP and normal subjects. Methods: The gene expression profile (GSE68801) was obtained from Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified between AAP patients and normal controls using the GEO2R. Then the Gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and Protein-Protein interaction (PPI) network analysis were performed for DEGs. Results: A total of 185 DEGs were identified, including 45 up-regulated genes and 140 down-regulated genes. The up-regulated DEGs were related to the immune response and chemokine signaling pathway. Meanwhile, down-regulated DEGs were enriched in keratin filament and intermediate filament. Subsequently, the top 10 hub genes were picked out in the PPI network, among them, CD2 showed the highest connectivity degree and central roles. Conclusion: Our data suggest that the CD2 may be a new therapeutic target for AAP. Further study is needed to explore the value of CD2 in the treatment of alopecia areata.
- Published
- 2019
40. A dynamic CD2-rich compartment at the outer edge of the immunological synapse boosts and integrates signals
- Author
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Mikhail A Kutuzov, H Rada, Matteo Morotti, Ahmed Ashour Ahmed, A Kvalvaag, S Yusuf, Nina Wietek, Jehan Afrose, David Depoil, Salvatore Valvo, Enas Abu-Shah, Michael Meyer-Hermann, Matthias Friedrich, Thomas Starkey, Viveka Mayya, Lee Lyw., Michael L. Dustin, Elizabeth R. Mann, Philippos Demetriou, Kseniya Korobchevskaya, Anastasios Siokis, and Sarah McCuaig
- Subjects
0301 basic medicine ,Immunological Synapses ,CD58 ,T cell ,Immunology ,Programmed Cell Death 1 Receptor ,CD2 Antigens ,Receptors, Antigen, T-Cell ,Biology ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,Article ,Immunological synapse ,03 medical and health sciences ,0302 clinical medicine ,Lymphocytes, Tumor-Infiltrating ,Neoplasms ,medicine ,Cell Adhesion ,Immune Tolerance ,Immunology and Allergy ,Humans ,Cell adhesion ,Cells, Cultured ,T-cell receptor ,CD28 ,Receptor Cross-Talk ,CD58 Antigens ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Signal transduction ,Single-Cell Analysis ,CD8 ,030215 immunology ,Protein Binding ,Signal Transduction - Abstract
The CD2–CD58 recognition system promotes adhesion and signaling and counters exhaustion in human T cells. We found that CD2 localized to the outer edge of the mature immunological synapse, with cellular or artificial APC, in a pattern we refer to as a ‘CD2 corolla’. The corolla captured engaged CD28, ICOS, CD226 and SLAM-F1 co-stimulators. The corolla amplified active phosphorylated Src-family kinases (pSFK), LAT and PLC-γ over T cell receptor (TCR) alone. CD2–CD58 interactions in the corolla boosted signaling by 77% as compared with central CD2–CD58 interactions. Engaged PD-1 invaded the CD2 corolla and buffered CD2-mediated amplification of TCR signaling. CD2 numbers and motifs in its cytoplasmic tail controlled corolla formation. CD8+ tumor-infiltrating lymphocytes displayed low expression of CD2 in the majority of people with colorectal, endometrial or ovarian cancer. CD2 downregulation may attenuate antitumor T cell responses, with implications for checkpoint immunotherapies. The adhesion receptor CD2 plays an important role in the full activation of T cells. Dustin and colleagues show that CD2 occupies a region in the periphery of the immunological synapse where it amplifies cognate antigen signals, whereas the presence of PD-1 disrupts this effect.
- Published
- 2019
41. Safety and pharmacodynamics of anti-CD2 monoclonal antibody treatment in cynomolgus macaques - an experimental study
- Author
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Felix Sellberg, Megan Sykes, David Berglund, Keith A. Reimann, Robin Fröbom, David H. Sachs, Paula Alonso-Guallart, Nathaly Llore, Christian Binder, Makenzie Danton, Erik Berglund, Adam Griesemer, Alina Iuga, Hiroshi Sakai, and Dilrukshi Ekanayake-Alper
- Subjects
Male ,medicine.drug_class ,Lymphocyte ,T-Lymphocytes ,CD2 Antigens ,030230 surgery ,Monoclonal antibody ,Major histocompatibility complex ,Lymphocyte Depletion ,Article ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Medicine ,Animals ,Lymph node ,Transplantation ,biology ,business.industry ,Antibodies, Monoclonal ,medicine.disease ,Transplant rejection ,medicine.anatomical_structure ,Immunology ,biology.protein ,Macaca ,030211 gastroenterology & hepatology ,Antibody ,business ,CD8 - Abstract
BACKGROUND: Anti-CD2 treatment provides targeted immunomodulatory properties that have demonstrated clinical usefulness to condition the immune system and to treat transplant rejection. The treatment is species-specific due to structural CD2 antigen differences between non-human primates and humans. Herein, we report the safety profile and efficacy of two modifications of the same anti-CD2 monoclonal antibody in cynomolgus macaques. METHODS: Twelve subjects received one i.v. anti-CD2 (of rat or rhesus type) dose each, range 1–4 mg/kg, and were followed for 1–7 days. Treatment effects were evaluated with flow cytometry on peripheral blood and histopathological evaluation of secondary lymphoid organs. In vitro inhibitory activity on primary MHC disparate MLRs was determined. RESULTS: Upon anti-CD2 treatment, CD4(+), CD8(+) memory subsets were substantially depleted. Naïve T-cells and Tregs were relatively spared, and exhibited lower CD2 expression than memory T-cells. Early immune reconstitution was noted for naïve cells, while memory counts had not recovered after one week. Both antibodies displayed a concentration-dependent MLR inhibition. Lymph node examination revealed no significant lymphocyte depletion. None of the animals experienced any significant study drug-related adverse events. CONCLUSIONS: This study outlines the safety and pharmacodynamic profile of primate-specific anti-CD2 treatment, relevant for translation of anti-CD2-based animal models into clinical trials.
- Published
- 2019
42. Functional Phenotypic Diversity of Regulatory T Cells Remaining in Inflamed Skin
- Author
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Ryoyo Ikebuchi, Maika Fujimoto, Yasutaka Nakanishi, Hiromi Okuyama, Taiki Moriya, Yutaka Kusumoto, and Michio Tomura
- Subjects
0301 basic medicine ,medicine.medical_treatment ,Dermatitis, Contact ,T-Lymphocytes, Regulatory ,regulatory T cells ,Mice ,0302 clinical medicine ,Cell Movement ,T-Lymphocyte Subsets ,Neuropilin 1 ,Immunology and Allergy ,Cluster Analysis ,CTLA-4 Antigen ,IL-2 receptor ,contact hypersensitivity response ,Cells, Cultured ,Original Research ,integumentary system ,hemic and immune systems ,Forkhead Transcription Factors ,Biodiversity ,Phenotype ,CD52 Antigen ,Organ Specificity ,Single-Cell Analysis ,lcsh:Immunologic diseases. Allergy ,skin ,Immunology ,CD2 Antigens ,chemical and pharmacologic phenomena ,Biology ,diversity ,03 medical and health sciences ,Downregulation and upregulation ,single cells ,medicine ,Animals ,Humans ,Inflammation ,Immunology in the medical area ,Immunotherapy ,Blockade ,Granzyme B ,Mice, Inbred C57BL ,030104 developmental biology ,Immunologi inom det medicinska området ,lcsh:RC581-607 ,Function (biology) ,030215 immunology - Abstract
Regulatory T cells (Tregs) migrate between lymphoid and peripheral tissues for maintaining immune homeostasis. Tissue-specific function and functional heterogeneity of Tregs have been suggested, however, correlation between them and inter-tissue movement remain unknown. We used a contact hypersensitivity model of mice expressing a photoconvertible protein for tracking migratory cells. After marking cells in skin, we purified Tregs exhibiting a different migration pattern [Tregs recruiting to or remaining in the skin and emigrating from the skin to draining lymph nodes (dLNs) within half a day] and examined single-cell gene and protein expression profiles. Correlation and unsupervised clustering analyses revealed that Tregs in both skin and dLNs comprised two subpopulations, one highly expressing Nrp1 with variable CD25, Granzyme B, and/or CTLA-4 expression and another with 3 subsets strongly expressing CD25, Granzyme B, or CTLA-4 together with CD39. Characteristic subsets of Tregs remaining in the skin displayed higher CD25 and CD39 expression and lower Granzyme B and CTLA-4 expression compared with Tregs migrating to the skin. In addition, CCR5 expression in Tregs in skin was positively and negatively correlated with CD39 and Nrp-1 expression, respectively. To assess the predictive value of these data for immunotherapy, we blocked CCR5 signaling and found modest downregulation of CD39 and modest upregulation of Nrp1 expression in skin Tregs. Our data reveal a high functional diversity of Tregs in skin that is strongly related to trafficking behavior, particularly skin retention. Modulation of tissue-specific trafficking and function is a promising clinical strategy against autoimmune, infectious, and neoplastic diseases. Significance Statement Regulatory T cells (Tregs) are essential for maintaining immune homeostasis. To reveal tissue-specific immunoinhibitory functions and inter-tissue movement correlation based on Treg functional heterogeneity, we examined single-cell gene and protein expression profiles of Tregs recruited to, remaining in, or emigrating from the contact hypersensitivity-induced inflamed skin. Tregs in skin were composed of several subpopulations; one with high Nrp1 expression and another with 3 subsets strongly expressing CD25, Granzyme B, or CTLA-4 together with CD39. Tregs remaining in skin displayed highCD25, CD39, and CCR5 expression, and CCR5 signaling blockade downregulated CD39. A high Treg functional diversity in skin is strongly related to trafficking behavior. Tissue-specific trafficking and functional modulation are a promising clinical strategy against autoimmune, infectious, and neoplastic diseases.
- Published
- 2019
43. Constrained Cyclic Peptides as Immunomodulatory Inhibitors of the CD2:CD58 Protein–Protein Interaction
- Author
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Thomas Durek, Seetharama D. Jois, David J. Craik, Ted J. Gauthier, Sandeep Pallerla, Veena Taneja, and Rushikesh Sable
- Subjects
Models, Molecular ,0301 basic medicine ,CD2 Antigens ,Peptide ,Biology ,Peptides, Cyclic ,Biochemistry ,Article ,Mass Spectrometry ,Theta defensin ,Epitope ,Protein–protein interaction ,Mice ,03 medical and health sciences ,Immune system ,Adjuvants, Immunologic ,Cell Line, Tumor ,Cell Adhesion ,Animals ,Humans ,Amino Acid Sequence ,Cell adhesion ,Nuclear Magnetic Resonance, Biomolecular ,Peptide sequence ,chemistry.chemical_classification ,Circular Dichroism ,General Medicine ,Surface Plasmon Resonance ,CD58 Antigens ,Cyclic peptide ,030104 developmental biology ,chemistry ,Molecular Medicine ,Protein Binding - Abstract
The interaction between the cell-cell adhesion proteins CD2 and CD58 plays a crucial role in lymphocyte recruitment to inflammatory sites, and inhibitors of this interaction have potential as immunomodulatory drugs in autoimmune diseases. Peptides from the CD2 adhesion domain were designed to inhibit CD2:CD58 interactions. To improve the stability of the peptides, β-sheet epitopes from the CD2 region implicated in CD58 recognition were grafted into the cyclic peptide frameworks of sunflower trypsin inhibitor and rhesus theta defensin. The designed multicyclic peptides were evaluated for their ability to modulate cell-cell interactions in three different cell adhesion assays, with one candidate, SFTI-a, showing potent activity in the nanomolar range (IC50: 51 nM). This peptide also suppresses the immune responses in T cells obtained from mice that exhibit the autoimmune disease rheumatoid arthritis. SFTI-a was resistant to thermal denaturation, as judged by circular dichroism spectroscopy and mass spectrometry, and had a half-life of ∼24 h in human serum. Binding of this peptide to CD58 was predicted by molecular docking studies and experimentally confirmed by surface plasmon resonance experiments. Our results suggest that cyclic peptides from natural sources are promising scaffolds for modulating protein-protein interactions that are typically difficult to target with small-molecule compounds.
- Published
- 2016
44. Critical Role of CD2 Co-stimulation in Adaptive Natural Killer Cell Responses Revealed in NKG2C-Deficient Humans
- Author
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Johannes Landskron, Vivien Béziat, Marie Schaffer, Monika Enqvist, John Trowsdale, Vincent Yi Sheng Oei, Jyothi Jayaraman, Hans-Gustaf Ljunggren, Eivind Heggernes Ask, Quirin Hammer, Stella Larsson, Kjetil Taskén, Lisa L. Liu, Karl-Johan Malmberg, Ebba Sohlberg, James A. Traherne, Chiara Romagnani, Jodie P. Goodridge, Traherne, James [0000-0002-6003-8559], Trowsdale, John [0000-0002-0150-5698], and Apollo - University of Cambridge Repository
- Subjects
killer cells: natural ,0301 basic medicine ,CD2 Antigens ,Cytomegalovirus ,Adaptive Immunity ,CD8-Positive T-Lymphocytes ,Biology ,GPI-Linked Proteins ,Lymphocyte Activation ,Article ,General Biochemistry, Genetics and Molecular Biology ,CD49b ,Natural killer cell ,Interferon-gamma ,03 medical and health sciences ,Interleukin 21 ,0302 clinical medicine ,medicine ,Humans ,Extracellular Signal-Regulated MAP Kinases ,lcsh:QH301-705.5 ,Cells, Cultured ,Ribosomal Protein S6 ,Lymphokine-activated killer cell ,Janus kinase 3 ,Receptors, IgG ,cells: cultured ,receptors: IgG ,Natural killer T cell ,3. Good health ,Cell biology ,Killer Cells, Natural ,030104 developmental biology ,medicine.anatomical_structure ,lcsh:Biology (General) ,Immunology ,Interleukin 12 ,antigens: CD2 ,NK Cell Lectin-Like Receptor Subfamily C ,030215 immunology - Abstract
Infection by human cytomegalovirus (HCMV) leads to NKG2C-driven expansion of adaptive natural killer (NK) cells, contributing to host defense. However, approximately 4% of all humans carry a homozygous deletion of the gene that encodes NKG2C ($\textit{NKG2C}^{-/-}$). Assessment of NK cell repertoires in 60 $\textit{NKG2C}^{-/-}$ donors revealed a broad range of NK cell populations displaying characteristic footprints of adaptive NK cells, including a terminally differentiated phenotype, functional reprogramming, and epigenetic remodeling of the interferon (IFN)-$\gamma$ promoter. We found that both NKG2C$^{-}$ and NKG2C$^{+}$ adaptive NK cells expressed high levels of CD2, which synergistically enhanced ERK and S6RP phosphorylation following CD16 ligation. Notably, CD2 co-stimulation was critical for the ability of adaptive NK cells to respond to antibody-coated target cells. These results reveal an unexpected redundancy in the human NK cell response to HCMV and suggest that CD2 provides "signal 2" in antibody-driven adaptive NK cell responses.
- Published
- 2016
45. Use of Alefacept for Preconditioning in Multiply Transfused Pediatric Patients with Nonmalignant Diseases
- Author
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Leslie S. Kean, Muna Qayed, Ann E. Haight, Kuang-Yueh Chiang, John T. Horan, and Elizabeth Stenger
- Subjects
Male ,Transplantation Conditioning ,medicine.medical_treatment ,Lymphocyte ,Graft vs Host Disease ,Pilot Projects ,Hematopoietic stem cell transplantation ,0302 clinical medicine ,T-Lymphocyte Subsets ,Child ,Bone Marrow Transplantation ,0303 health sciences ,Graft Survival ,Anemia, Aplastic ,Hematology ,3. Good health ,Killer Cells, Natural ,Haematopoiesis ,medicine.anatomical_structure ,Female ,Nonmalignant diseases ,Cord Blood Stem Cell Transplantation ,Stem cell ,Unrelated Donors ,medicine.drug ,Recombinant Fusion Proteins ,CD2 Antigens ,Blood Component Transfusion ,Alefacept ,Rejection ,Article ,Dyskeratosis Congenita ,Lymphocyte Depletion ,Immunophenotyping ,03 medical and health sciences ,Immune system ,medicine ,Humans ,030304 developmental biology ,Transplantation ,business.industry ,Historically Controlled Study ,Infant ,Calcineurin ,Fanconi Anemia ,Immunology ,business ,Immunologic Memory ,CD8 ,Conditioning ,030215 immunology - Abstract
Transfusion-related alloimmunization is a potent barrier to the engraftment of allogeneic hematopoietic stem cells in patients with nonmalignant diseases (NMDs). Memory T cells, which drive alloimmunization, are relatively resistant to commonly used conditioning agents. Alefacept, a recombinant leukocyte function antigen-3/IgG1 fusion protein, targets CD2 and selectively depletes memory versus naive T cells. Three multiply transfused pediatric patients with NMD received a short course of high-dose i.v. alefacept (.25 mg/kg/dose on days -40 and -9 and .5 mg/kg/dose on days -33, -26, -19, and -12) before undergoing unrelated allogeneic transplant in the setting of reduced-intensity pretransplant conditioning and calcineurin inhibitor-based post-transplant graft-versus-host disease (GVHD) prophylaxis. Alefacept infusions were well tolerated in all patients. Peripheral blood flow cytometry was performed at baseline and during and after alefacept treatment. As expected, after the 5 weekly alefacept doses, each patient demonstrated selective loss of CD2(hi)/CCR7(-)/CD45RA(-) effector memory (Tem) and CD2(hi)/CCR7(+)/CD45RA(-) central memory (Tcm) CD4(+) and CD8(+) T cells with relative preservation of the CD2(lo) Tem and Tcm subpopulations. In addition, depletion of CD2(+) natural killer (NK) cells also occurred. Neutrophil recovery was rapid, and all 3 patients had 100% sorted (CD3/CD33) peripheral blood donor chimerism by day +100. Immune reconstitution (by absolute neutrophil, monocyte, and lymphocyte counts) was comparable with a cohort of historical control patients. All 3 patients developed GVHD but are all now off immune suppression and2 years post-transplant with stable full-donor engraftment. These results suggest that alefacept at higher dosing can deplete both memory T cells and NK cells and that incorporating CD2-targeted depletion into a reduced-intensity transplant regimen is feasible and safe in heavily transfused patients.
- Published
- 2015
46. Ptpn22 and Cd2 Variations Are Associated with Altered Protein Expression and Susceptibility to Type 1 Diabetes in Nonobese Diabetic Mice
- Author
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Fraser, Heather I, Howlett, Sarah, Clark, Jan, Rainbow, Daniel B, Stanford, Stephanie M, Wu, Dennis J, Hsieh, Yi-Wen, Maine, Christian J, Christensen, Mikkel, Kuchroo, Vijay, Sherman, Linda A, Podolin, Patricia L, Todd, John A, Steward, Charles A, Peterson, Laurence B, Bottini, Nunzio, Wicker, Linda S, and Apollo - University of Cambridge Repository
- Subjects
B-Lymphocytes ,T-Lymphocytes ,Molecular Sequence Data ,CD2 Antigens ,Mice, Transgenic ,Protein Tyrosine Phosphatase, Non-Receptor Type 22 ,Chromosomes, Mammalian ,Polymorphism, Single Nucleotide ,Mice ,Diabetes Mellitus, Type 1 ,Gene Expression Regulation ,Genetic Loci ,Mice, Inbred NOD ,Immunogenetics ,Animals ,Humans ,Genetic Predisposition to Disease ,Alleles ,Signal Transduction - Abstract
By congenic strain mapping using autoimmune NOD.C57BL/6J congenic mice, we demonstrated previously that the type 1 diabetes (T1D) protection associated with the insulin-dependent diabetes (Idd)10 locus on chromosome 3, originally identified by linkage analysis, was in fact due to three closely linked Idd loci: Idd10, Idd18.1, and Idd18.3. In this study, we define two additional Idd loci--Idd18.2 and Idd18.4--within the boundaries of this cluster of disease-associated genes. Idd18.2 is 1.31 Mb and contains 18 genes, including Ptpn22, which encodes a phosphatase that negatively regulates T and B cell signaling. The human ortholog of Ptpn22, PTPN22, is associated with numerous autoimmune diseases, including T1D. We, therefore, assessed Ptpn22 as a candidate for Idd18.2; resequencing of the NOD Ptpn22 allele revealed 183 single nucleotide polymorphisms with the C57BL/6J (B6) allele--6 exonic and 177 intronic. Functional studies showed higher expression of full-length Ptpn22 RNA and protein, and decreased TCR signaling in congenic strains with B6-derived Idd18.2 susceptibility alleles. The 953-kb Idd18.4 locus contains eight genes, including the candidate Cd2. The CD2 pathway is associated with the human autoimmune disease, multiple sclerosis, and mice with NOD-derived susceptibility alleles at Idd18.4 have lower CD2 expression on B cells. Furthermore, we observed that susceptibility alleles at Idd18.2 can mask the protection provided by Idd10/Cd101 or Idd18.1/Vav3 and Idd18.3. In summary, we describe two new T1D loci, Idd18.2 and Idd18.4, candidate genes within each region, and demonstrate the complex nature of genetic interactions underlying the development of T1D in the NOD mouse model.
- Published
- 2015
47. Development of a RACE-based RNA-Seq approach to characterize the T-cell receptor repertoire of porcine γδ T cells
- Author
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John A. Hammond, Melanie Leopold, Lisa-Maria Prawits, Armin Saalmüller, Sarina Ravens, Kerstin H. Mair, John C. Schwartz, Sabine E. Hammer, and Wilhelm Gerner
- Subjects
0301 basic medicine ,Swine ,T-Lymphocytes ,T cell ,Immunology ,Population ,CD2 Antigens ,RNA-Seq ,Adaptive Immunity ,Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Complementary DNA ,Endopeptidases ,medicine ,Animals ,Humans ,education ,Cells, Cultured ,education.field_of_study ,Sequence Analysis, RNA ,Gene Expression Profiling ,T-cell receptor ,Genetic Variation ,High-Throughput Nucleotide Sequencing ,RNA ,Receptors, Antigen, T-Cell, gamma-delta ,Acquired immune system ,Phenotype ,Molecular biology ,Immunity, Innate ,030104 developmental biology ,medicine.anatomical_structure ,030215 immunology ,Developmental Biology - Abstract
Recent data suggest that porcine γδ T cells exhibit a similar degree of functional plasticity as human and murine γδ T cells. Due to the high frequency of TCR-γδ+ cells in blood and secondary lymphatic organs, the pig is an attractive model to study these cells, especially their combined features of the innate and the adaptive immune system. Using a 5' RACE-like approach, we translated a human/murine NGS library preparation strategy to capture full-length V-(D)-J TRG and TRD clonotypes in swine. After oligo(dT) primed conversion of input RNA, the cDNA population was enriched for full-length V(D)J TCR transcripts with porcine-specific primers including Illumina adaptor sequences as overhangs for Illumina MiSeq analysis. After quality control and processing by FastQC and ea-utils, porcine TRG and TRD sequences were mapped against the human IMGT reference directory. Porcine blood-derived CD2+ and CD2‾ TCR-γδ+ cells exhibited two distinct clonotypes Vγ11JγP1 (74.6%) and Vγ10JγP1 (57.7%), respectively. Despite the high TCR-δ diversity among CD2+ cells (39 clonotypes), both subsets shared the same abundant Vδ1DδxJδ4 clonotype at approximately identically frequencies (CD2+: 31.2%; CD2‾: 37.0%). The flexible nature of this approach will facilitate the assessment of organ-specific phenotypes of γδ T cell subsets alongside with their respective TCR diversity at single cell resolution.
- Published
- 2020
48. Costimulatory Function of Cd58/Cd2 Interaction in Adaptive Humoral Immunity in a Zebrafish Model
- Author
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Tong Shao, Wei Shi, Jia-yu Zheng, Xiao-xiao Xu, Ai-fu Lin, Li-xin Xiang, and Jian-zhong Shao
- Subjects
0301 basic medicine ,lcsh:Immunologic diseases. Allergy ,Models, Molecular ,Cellular immunity ,CD58 ,ved/biology.organism_classification_rank.species ,Immunology ,CD2 Antigens ,adaptive humoral immunity ,Antigen-Presenting Cells ,Adaptive Immunity ,Lymphocyte Activation ,cd58 ,03 medical and health sciences ,Structure-Activity Relationship ,0302 clinical medicine ,Immune system ,T-Lymphocyte Subsets ,Immunology and Allergy ,Animals ,Cd4+ T cells ,Amino Acid Sequence ,Cloning, Molecular ,RNA, Small Interfering ,Model organism ,Zebrafish ,Original Research ,Gene knockdown ,B-Lymphocytes ,biology ,Base Sequence ,ved/biology ,Sequence Analysis, DNA ,costimulatory signals ,biology.organism_classification ,Acquired immune system ,CD58 Antigens ,cd2 ,Cell biology ,Immunity, Humoral ,Protein Transport ,030104 developmental biology ,Humoral immunity ,Antibody Formation ,RNA Interference ,lcsh:RC581-607 ,030215 immunology ,Protein Binding - Abstract
CD58 and CD2 have long been known as a pair of reciprocal adhesion molecules involved in the immune modulations of CD8+ T and NK-mediated cellular immunity in humans and several other mammals. However, the functional roles of CD58 and CD2 in CD4+ T-mediated adaptive humoral immunity remain poorly defined. Moreover, the current functional observations of CD58 and CD2 were mainly acquired from in vitro assays, and in vivo investigation is greatly limited due to the absence of a Cd58 homology in murine models. In this study, we identified cd58 and cd2 homologs from the model species zebrafish (Danio rerio). These two molecules share conserved structural features to their mammalian counterparts. Functionally, cd58 and cd2 were significantly upregulated on antigen-presenting cells and Cd4+ T cells upon antigen stimulation. Blockade or knockdown of Cd58 and Cd2 dramatically impaired the activation of antigen-specific Cd4+ T and mIgM+ B cells, followed by the inhibition of antibody production and host defense against bacterial infections. These results indicate that CD58/CD2 interaction was required for the full activation of CD4+ T-mediated adaptive humoral immunity. The interaction of Cd58 with Cd2 was confirmed by co-immunoprecipitation and functional competitive assays by introducing a soluble Cd2 protein. This study highlights a new costimulatory mechanism underlying the regulatory network of adaptive immunity and makes zebrafish an attractive model organism for the investigation of CD58/CD2-mediated immunology and disorders. It also provides a cross-species understanding of the evolutionary history of costimulatory signals from fish to mammals as a whole.
- Published
- 2018
49. Transcription factors early growth response gene (Egr) 2 and 3 control inflammatory responses of tolerant T cells
- Author
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Omodho, Becky, Miao, Tizong, Symonds, Alistair L.J., Singh, Randeep, Li, Suling, and Wang, Ping
- Subjects
Antigens, Differentiation, T-Lymphocyte ,CD2 Antigens ,T cells ,Lymphocyte Activation ,Autoantigens ,Enterotoxins ,Mice ,Antigens, CD ,T-Lymphocyte Subsets ,Immune Tolerance ,Animals ,Humans ,Lectins, C-Type ,Gene Knock-In Techniques ,Early Growth Response Protein 3 ,Early Growth Response Protein 2 ,Original Research ,Bone Marrow Transplantation ,Cell Proliferation ,Inflammation ,Mice, Knockout ,Transplantation Chimera ,tolerance ,Mice, Inbred C57BL ,Disease Models, Animal ,Hyaluronan Receptors ,Gene Expression Regulation ,Models, Animal ,Interleukin-2 ,Egr2 ,Tolerance ,Signal Transduction - Abstract
Introduction: Impaired proliferation and production of IL2 are the hallmarks of experimental T cell tolerance. However, in most autoimmune diseases, autoreactive T cells do not display hyper proliferation, but inflammatory phenotypes. Methods: We have now demonstrated that the transcription factors Egr2 and 3 are important for the control of inflammatory cytokine production by tolerant T cells, but not for tolerance induction. Results: In the absence of Egr2 and 3, T cell tolerance, as measured by impaired proliferation and production of IL2, can still be induced, but tolerant T cells produced high levels of inflammatory cytokines. Egr2 and 3 regulate expression of differentiation repressors and directly inhibit T-bet function in T cells. Indeed, decreased expression of differentiation repressors, such as Id3 and Tcf1, and increased expression of inflammatory transcription factors, such as RORgt and Bhlhe40 were found in Egr2/3 deficient T cells under tolerogenic conditions. In addition, T-bet was co-expressed with Egr2 in tolerant T cells and Egr2/3 defects leads to production of high levels of IFNg in tolerant T cells. Conclusions: Our findings demonstrated that despite impaired proliferation and IL2 production, tolerant T cells can display inflammatory responses in response to antigen stimulation and this is controlled at least partly by Egr2 and 3. Arthritis Research UK and Medical Research Council UK
- Published
- 2018
50. Investigating cyclic peptides inhibiting CD2-CD58 interactions through molecular dynamics and molecular docking methods
- Author
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Laurence Leherte, Adèle D. Laurent, Axel Petit, Daniel P. Vercauteren, Denis Jacquemin, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), and Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)
- Subjects
0301 basic medicine ,Stereochemistry ,Protein Conformation ,CD2 Antigens ,Crystal structure ,Molecular dynamics ,Molecular Dynamics Simulation ,Ligands ,Peptides, Cyclic ,03 medical and health sciences ,Drug Discovery ,Side chain ,[CHIM]Chemical Sciences ,Amino Acid Sequence ,Physical and Theoretical Chemistry ,Binding site ,chemistry.chemical_classification ,Binding Sites ,Hydrogen bond ,Chemistry ,ligands ,cyclic peptides ,Hydrogen Bonding ,Ligand (biochemistry) ,CD58 Antigens ,Cyclic peptide ,Computer Science Applications ,inhibitor ,Molecular Docking Simulation ,030104 developmental biology ,Docking (molecular) ,Molecular docking ,Thermodynamics ,CD58 ,Protein Binding - Abstract
The CD2–CD58 protein–protein interaction is known to favor the recognition of antigen presenting cells by T cells. The structural, energetics, and dynamical properties of three known cyclic CD58 ligands, named P6, P7, and RTD-c, are studied through molecular dynamics (MD) simulations and molecular docking calculations. The ligands are built so as to mimic the C and F β-strands of protein CD2, connected via turn inducers. The MD analyses focus on the location of the ligands with respect to the experimental binding site and on the direct and water-mediated hydrogen bonds (H bonds) they form with CD58. Ligand P6, with a sequence close to the experimental β-strands of CD2, presents characteristics that explain its higher experimental affinity, e.g., the lower mobility and flexibility at the CD58 surface, and the larger number and occurrence frequency of ligand-CD58 H bonds. For the two other ligands, the structural modifications lead to changes in the binding pattern with CD58 and its dynamics. In parallel, a large set of molecular docking calculations, carried out with various search spaces and docking algorithms, are compared to provide a consensus view of the preferred ligand binding modes. The analysis of the ligand side chain locations yields results that are consistent with the CD2–CD58 crystal structure and suggests various binding modes of the experimentally identified hot spot of the ligands, i.e., Tyr86. P6 is shown to form a number of contacts that are also present in the experimental CD2–CD58 structure.
- Published
- 2018
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