bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epitope-Specific Antibody Response to IgE by Mimotope Immunization
Authors: Rudolf Michael P. , Vogel Monique , Kricek Franz , Ruf Christine , Zürcher Adrian W. , Reuschel Roland , Auer Manfred , Miescher Sylvia , Stadler Beda M. 
Published: December 2022

AbstractWe have previously described a mouse monoclonal anti-human IgE antibody (BSW17) capable of recognizing receptor-bound IgE without inducing mediator release from human basophils or mast cells. Moreover, immune complexes of IgE and BSW17 are not able to bind to the IgE receptor. An initial attempt to map the precise epitope recognized by this mAb by using Fcε-derived peptides of variable length was unsuccessful. However, by screening random peptide phage display libraries we isolated circular nona- and octapeptides specifically recognized by BSW17. These constrained peptides mimic at least a part of a conformational epitope and are thus called mimotopes. These mimotopes, either phage displayed or synthetically synthesized, did not react with any other anti-human IgE antibody tested, but efficiently inhibited the binding of human IgE to BSW17 only. The use of Rhodol-Green-labeled free cyclic peptide proved that these interactions were not carrier dependent. Immunization of rabbits with phage clones displaying the specific peptides on the surface induced an anti-human IgE response specific for the epitope of BSW17. Therefore, we conclude that such mimotopes or mimotope-derived peptides might be used for vaccination to induce in vivo a beneficial anti-IgE response as a novel immunotherapy.
Several Common HLA-DR Types Share Largely Overlapping Peptide Binding Repertoires
Authors: Southwood Scott , Sidney John , Kondo Akihiro , del Guercio Marie-France , Appella Ettore , Hoffman Stephen , Kubo Ralph T. , Chesnut Robert W. , Grey Howard M. , Sette Alessandro 
Published: December 2022

AbstractThe peptide binding specificities of HLA-DRB1*0401, DRB1*0101, and DRB1*0701 have been analyzed by the use of large collections of synthetic peptides corresponding to naturally occurring sequences. The results demonstrated that nearly all peptides binding to these DR molecules bear a motif characterized by a large aromatic or hydrophobic residue in position 1 (Y, F, W, L, I, V, M) and a small, noncharged residue in position 6 (S, T, C, A, P, V, I, L, M). In addition, allele-specific secondary effects and secondary anchors were defined, and these parameters were utilized to derive allele-specific motifs and algorithms. By the combined use of such algorithms, peptides capable of degenerate DRB1*0101, DRB1*0401, and DRB1*0701 binding were identified. Additional experiments utilizing a panel of quantitative assays specific for nine additional common DR molecules identified a large set of DR molecules, which includes at least the DRB1*0101, DRB1*0401, DRB1*0701, DRB5*0101, DRB1*1501, DRB1*0901, and DRB1*1302 allelic products, characterized by overlapping peptide-binding repertoires. These results have implications for understanding the molecular interactions involved in peptide-DR binding, as well as the genetic and structural basis of MHC polymorphism. These results also have potential practical implications for the development of epitope-based prophylactic and therapeutic vaccines.
Revealing the In Vivo Behavior of CD4+ T Cells Specific for an Antigen Expressed inEscherichia coli
Authors: Chen Zong-ming , Jenkins Marc K. 
Published: December 2022

AbstractThe clonal expansion and anatomic location of microbe-specific CD4+ Th cells was studied by tracking the fate of adoptively transferred DO11.10 TCR transgenic T cells specific for OVA peptide 323–339/I-Ad in BALB/c mice infected s.c. with Escherichia coli expressing a MalE-OVA fusion protein. After infection, the DO11.10 T cells accumulated in the T cell-rich paracortical regions of the draining lymph nodes, proliferated there for several days, and then moved into the B cell-rich follicles before they slowly disappeared from the lymph nodes. These changes occurred despite the fact that viable organisms were never found in the lymph nodes. The DO11.10 T cells also accumulated in the s.c. infection site, but about 1 day later than in the draining lymph nodes. Injection of purified MalE-OVA fusion protein alone induced a transient accumulation of DO11.10 T cells in the paracortical regions, but these T cells never entered follicles and the mice did not produce anti-OVA antibodies. The DO11.10 T cells that survived in animals injected with MalE-OVA alone were hyporesponsive to in vitro Ag restimulation and did not produce IL-2 and IFN-γ, whereas DO11.10 T cells from mice infected with MalE-OVA-expressing bacteria produced both lymphokines. These results suggest that Ag-specific T cells are first activated in secondary lymphoid organs following primary bacterial infection and then migrate to the infection site. Furthermore, productive activation of the T cells during the primary response is dependent on bacterial components other than the Ag itself.
Ribavirin Inhibits Viral-Induced Macrophage Production of TNF, IL-1, the Procoagulant fgl2 Prothrombinase and Preserves Th1 Cytokine Production But Inhibits Th2 Cytokine Response
Authors: Ning Qin , Brown Deron , Parodo Jean , Cattral Mark , Gorczynski Reginald , Cole Edward , Fung Laisum , Ding Jin Wen , Liu Ming Feng , Rotstein Ori , Phillips M. James , Levy Gary 
Published: December 2022

AbstractRibavirin, a synthetic guanosine analogue, possesses a broad spectrum of activity against DNA and RNA viruses. It has been previously shown to attenuate the course of fulminant hepatitis in mice produced by murine hepatitis virus strain 3. We therefore studied the effects of ribavirin on murine hepatitis virus strain 3 replication, macrophage production of proinflammatory mediators including TNF, IL-1, and the procoagulant activity (PCA), fgl2 prothrombinase; and Th1/Th2 cytokine production. Although ribavirin had inhibitory effects on viral replication (&lt;1 log), even at high concentrations complete eradication of the virus was not seen. In contrast, at physiologic concentrations (up to 500 μg/ml), ribavirin markedly reduced viral-induced parameters of macrophage activation. With ribavirin treatment, the concentrations of PCA, TNF-α and IL-1β all decreased to basal concentrations: PCA from 941 ± 80 to 34 ± 11 mU/106 cells; TNF-α from 10.73 ± 2.15 to 2.74 ± 0.93 ng/ml; and IL-1β from 155.91 ± 22.62 to 5.74 ± 0.70 pg/ml. The inhibitory effects of ribavirin were at the level of gene transcription as evidenced by Northern analysis. Both in vitro and in vivo, ribavirin inhibited the production of IL-4 by Th2 cells, whereas it did not diminish the production of IFN-γ in Th1 cells. In contrast, ribavirin had no inhibitory effect on TNF-α and IL-1β production in LPS-stimulated macrophages. These results suggest that the beneficial effects of ribavirin are mediated by inhibition of induction of macrophage proinflammatory cytokines and Th2 cytokines while preserving Th1 cytokines.
Cutting Edge: Ca2+-Dependent Exocytosis in Mast Cells Is Stimulated by the Ca2+ Sensor, Synaptotagmin I
Authors: Baram Dana , Linial Michal , Mekori Yoseph A. , Sagi-Eisenberg Ronit 
Published: December 2022

Abstract
Mast cells secrete a variety of biologically active substances that mediate inflammatory responses. Synaptotagmin(s) (Syts) are a gene family of proteins that are implicated in the control of Ca2+-dependent exocytosis. In the present study, we investigated the possible occurrence and functional involvement of Syt in the control of mast cell exocytosis. Here, we demonstrate that both connective tissue type and mucosal-like mast cells express Syt-immunoreactive proteins, and that these proteins are localized almost exclusively to their secretory granules. Furthermore, expression of Syt I, the neuronal Ca2+ sensor, in rat basophilic leukemia cells (RBL-2H3), a tumor analogue of mucosal mast cells, resulted in prominent potentiation and acceleration of Ca2+-dependent exocytosis. Therefore, these findings implicate Syt as a Ca2+ sensor that mediates regulated secretion in mast cells to calcium ionophore.
Monoclonal Antibodies to Mitochondrial E2 Components Define Autoepitopes in Primary Biliary Cirrhosis
Authors: Migliaccio Christopher , Nishio Akiyoshi , Van de Water Judy , Ansari Aftab A. , Leung Patrick S. C. , Nakanuma Yasuni , Coppel Ross L. , Gershwin M. Eric 
Published: December 2022

AbstractPrimary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the presence of antimitochondrial Abs (AMA). The autoantigens recognized by AMA are the E2 components of the pyruvate dehydrogenase complex (PDC-E2), the branched chain 2-oxoacid dehydrogenase complex E (BCOADC-E2), and the 2-oxoglutarate dehydrogenase complex E (OGDC-E2). Previous studies using murine monoclonal and human combinatorial Abs to PDC-E2 have demonstrated an intense linear staining pattern in the apical region of biliary epithelial cells (BEC) in PBC but not control liver. We therefore examined whether mAbs to the other mitochondrial autoantigens BCOADC-E2 and OGDC-E2 demonstrated disease-specific patterns of reactivity. Using an expressed recombinant “trihybrid” protein containing the lipoyl domains of PDC-E2, OGDC-E2, and BCOADC-E2, we immunized BALB/c mice to produce 35 mAbs specific for one or more of the above mitochondrial autoantigens. Seven of these mAbs uniquely stained the apical region of BEC in PBC. Of these seven, one was reactive to PDC-E2, two recognized BCOADC-E2, three were reactive to OGDC-E2, and one recognized all three Ags. Our current data demonstrate that, similar to our previous studies regarding PDC-E2, mAbs to BCOADC-E2 and OGDC-E2, or a molecule that cross-reacts with the inner lipoyl domain of all three enzymes, also show a uniquely intense staining pattern in the apical region of BEC in patients with PBC when compared with diseased controls. The abundance of such disease-specific determinants in the target cells of PBC raises interesting possibilities regarding the role of these autoantigens in the pathogenesis of this disease.
Acceleration oflprLymphoproliferative and Autoimmune Disease by Transgenic Protein Kinase CK2α
Authors: Rifkin Ian R. , Channavajhala Padma L. , Kiefer Heather L.B. , Carmack Adrienne J. , Landesman-Bollag Esther , Beaudette Britte C. , Jersky Brian , Salant David J. , Ju Shyr-Te , Marshak-Rothstein Ann , Seldin David C. 
Published: December 2022

AbstractMRL-lpr/lpr mice have a Fas receptor mutation that leads to abnormalities of apoptosis, lymphoproliferation, and a lupus-like autoimmune disease associated with the production of autoantibodies. Other than Fas pathway defects, little is known about molecular abnormalities that predispose to autoimmunity. Protein kinase CK2 (also termed casein kinase II), a serine-threonine protein kinase whose targets include many critical regulators of cellular growth, is highly expressed in a lymphoproliferative disease of cattle and in many human cancers. Overexpression of the CK2α catalytic subunit in lymphocytes of transgenic mice leads to T cell lymphoma. We hypothesized that CK2 dysregulation and Fas mutation might cooperatively augment lymphocyte proliferation and transformation. We find that in MRL-lpr/lpr mice bearing the CK2α transgene, the lymphoproliferative process is dramatically exacerbated, as these mice develop massive splenomegaly and lymphadenopathy by 12 wk of age in association with increased autoantibody production and accelerated renal disease. The lymphoid organs are filled with the unusual B220+CD4−CD8− T cells typically seen in MRL-lpr/lpr mice, not the B220−CD4+CD8+ or B220−CD4−CD8+ T cells typically seen in CK2α transgenic lymphomas. The T cells do not fulfill the criteria for transformation, as they are polyclonal and not transplantable or immortal in cell culture. Thus, although the lpr lymphoproliferative and autoimmune syndrome is potentiated by the presence of the CK2α transgene, this combination of apoptotic and proliferative abnormalities appears to be insufficient to transform lymphoid cells.
CD40 Engagement Triggers Switching to IgA1 and IgA2 in Human B Cells Through Induction of Endogenous TGF-β: Evidence for TGF-β But Not IL-10-Dependent Direct Sμ→Sα and Sequential Sμ→Sγ, Sγ→Sα DNA Recombination
Authors: Zan Hong , Cerutti Andrea , Dramitinos Patricia , Schaffer András , Casali Paolo 
Published: December 2022

AbstractIgA are major effectors of antimicrobial defense in the respiratory and digestive tracts. We have analyzed the requirements for and the modalities of switching to IgA using our recently identified monoclonal model of human germinal center differentiation, CL-01 B cells. CL-01 cells bear surface IgM (sIgM) and sIgD and switch to all seven downstream isotypes in response to physiologic stimuli. In these cells, CD40 engagement by CD40 ligand induces production of endogenous TGF-β and IL-10, expression of germline Iα1-Cα1 and Iα2-Cα2 transcripts, mature VHDJH-Cα1 and VHDJH-Cα2 transcripts, and IgA secretion. These events are associated with not only direct Sμ→Sα, but also sequential Sμ→Sγ, Sγ→Sα DNA recombination, and are ablated by neutralizing anti-TGF-β but not IL-10 Ab, and indicating that TGF-β, not IL-10, is a crucial mediator of the transcriptional activation and recombination of human Cα1 and Cα2 genes. Our findings in CL-01 cells were reproduced in freshly isolated naive sIgM+ sIgD+ B lymphocytes. Thus, engagement of CD40, in the absence of other (known) stimuli, is sufficient to effectively induce switching to IgA in human B cells. This is effected by direct and sequential DNA recombination events, which are both dependent upon endogenous TGF-β secreted by the CD40L-induced B cells.
Glucocorticoids Inhibit Bioactive IL-12p70 Production by In Vitro-Generated Human Dendritic Cells Without Affecting Their T Cell Stimulatory Potential
Authors: Vieira Pedro L. , Kaliński Paweł , Wierenga Eddy A. , Kapsenberg Martien L. , de Jong Esther C. 
Published: December 2022

AbstractGlucocorticoids (GC) are known to affect the immune response at several stages. However, little is known about how GC influence the initiation of the specific immune response at the level of dendritic cells (DC), the highly professional APC for T cells. Therefore, we studied whether GC modulate the cytokine production and T cell stimulatory function of DC. In LPS-stimulated DC, GC strongly reduced the secretion of the Th1-skewing factor IL-12p70 and, to a lesser extent, the production of the proinflammatory cytokines IL-6 and TNF-α. Regarding the T cell stimulatory function of DC, GC did not influence the cell surface expression of HLA-DR or the costimulatory molecules CD40 and CD80 and did not influence the ability of DC to take up Ag. Consequently, GC pretreatment of DC indeed did not affect their ability to stimulate CD4+ Th cell proliferation in response to superantigen. However, as a result of their defective production of bioactive IL-12, GC-pretreated DC have a reduced ability to promote the production of IFN-γ in CD4+ Th lymphocytes, as shown by the observation that IFN-γ production could be restored by exogenous IL-12. In contrast, GC treatment of DC enhanced the secretion of the antiinflammatory cytokine IL-10 and the type 2 cytokine IL-5 by the T cells. It is concluded that, in addition to their role as potent inhibitors of inflammation via the direct suppression of cytokine production in T cells, GC may further inhibit T cell-mediated inflammation indirectly via the suppression of IL-12 production by DC.
HLA-B27 Subtype Polymorphism and CTL Epitope Choice: Studies with EBV Peptides Link Immunogenicity with Stability of the B27:Peptide Complex
Authors: Brooks Jill M. , Colbert Robert A. , Mear John P. , Leese Alison M. , Rickinson Alan B. 
Published: December 2022

AbstractHLA-B27-restricted CTL responses to EBV are principally directed against two of the EBV nuclear Ags, EBNAs 3B and 3C. We have previously described a target epitope derived from EBNA 3C (residues 258–266, sequence RRIYDLIEL) that is immunodominant in the context of at least three different B27 subtypes, including B*2705 and B*2702. In this study, we show that this peptide binds well to B*2705 and B*2702 in a cell surface binding assay, and that the two B27:peptide complexes are relatively stable, with t1/2 of 20 and 37 h, respectively. We now identify another B27-restricted epitope derived from EBNA 3B (residues 243–253, sequence RRARSLSAERY), which again accords well with the B*2705/B*2702 consensus motifs, having an arginine residue at position 2 and a tyrosine residue at the carboxyl terminus. In this case, five of five B*2702-positive donors respond to the epitope, whereas there was no response in any B*2705-positive donor studied. This peptide binds at least as well to B*2705 as to its restriction element B*2702; however, the two class I:peptide complexes show marked differences in stability, with t1/2 of 9 and 42 h, respectively. Thus, the stability of B27:peptide complexes can vary markedly between different B27 subtypes in ways that may not be apparent from cell surface binding assays and cannot be predicted from currently known peptide consensus motifs, yet which may critically influence CTL epitope choice.
B7.1 Is a Quantitatively Stronger Costimulus Than B7.2 in the Activation of Naive CD8+ TCR-Transgenic T Cells
Authors: Fields Patrick E. , Finch Rosalynde J. , Gray Gary S. , Zollner Richard , Thomas Jenifer L. , Sturmhoefel Knut , Lee Kwan , Wolf Stanley , Gajewski Thomas F. , Fitch Frank W. 
Published: December 2022

AbstractUsing a TCR transgenic mouse bred onto a recombinase-activating gene-2-deficient background, we have examined the influence of B7.1 and B7.2 on activation of naive, CD8+ T cells in vitro. We found that B7.1 was a more potent costimulus than B7.2 for induction of proliferation and IL-2 production by naive CD8+ T cells. This difference appeared to be quantitative in nature, as determined using transfectants expressing various defined levels of B7.1 or B7.2, or using purified B7.1 or B7.2 fusion proteins. In contrast to the quantitative differences seen in stimulation of naive T cells, B7.1 and B7.2 were comparable in their ability to costimulate responses in T cells previously primed in vitro. In addition, primed, but not naive, T cells were capable of proliferating and producing IL-2 in response to a TCR stimulus alone, apparently in the absence of B7 costimulation. Lastly, we found that B7.1 and B7.2 were equivalently capable of driving differentiation of naive CD8+ T cells into an IL-4-producing phenotype when exogenous IL-4 was added to the primary culture or to an IFN-γ-producing phenotype in the presence of IL-12. These results indicate that signals generated by B7.1 and B7.2 are qualitatively similar, but that B7.1 is quantitatively stronger than B7.2. Further, our results indicate that the activation state of the responding T cell may influence the efficiency with which the T cell can respond to a costimulatory signal provided by either B7.1 or B7.2.
Normal Human CD4+ Memory T Cells Display Broad Heterogeneity in Their Activation Threshold for Cytokine Synthesis
Authors: Waldrop Shar L. , Davis Kenneth A. , Maino Vernon C. , Picker Louis J. 
Published: December 2022

AbstractCD4+ memory T cells coordinate immune responses against viruses and other pathogens via the Ag-induced secretion of potent effector cytokines. The efficacy of these responses depends on both the overall number of pathogen-specific memory T cells and the particular array of cytokines that these cells are programmed to secrete. Here, we provide evidence that heterogeneity in Ag triggering thresholds constitutes an additional critical determinant of memory T cell function. Using a novel assay that allows single-cell detection of Ag-specific T cell cytokine production, we demonstrate that CMV-specific CD4+ memory cells from human peripheral blood display pronounced differences in their costimulatory requirements for Ag-induced triggering of IFN-γ and IL-2 secretion, ranging from cells that trigger with little costimulation (e.g., resting APC alone) to cells requiring potent costimulation through multiple pathways (resting APC plus multiple costimulatory mAbs, or activated APC). These differences in costimulatory requirements are independent of clonal differences in TCR signaling intensity, consistent with an intrinsic activation-threshold heterogeneity that is “downstream” from the TCR. Thus, “effective” frequencies of Ag-specific CD4+ memory T cells appear to depend on the activation status of available APC, a dependence that would allow the immune system to rapidly adjust the number of functional Ag-specific memory T cells in a particular effector site according to local conditions.
Oral Tolerance to Low Dose β2-Glycoprotein I: Immunomodulation of Experimental Antiphospholipid Syndrome
Authors: Blank Miri , George Jacob , Barak Vivian , Tincani Angela , Koike Takao , Shoenfeld Yehuda 
Published: December 2022

AbstractOral tolerance was induced in BALB/c mice by feeding low dose β2-glycoprotein I (β2GPI). The β2GPI-fed mice did not develop serologic and clinical markers of experimental antiphospholipid syndrome (APS) upon immunization with the autoantigen. The treated group was characterized by low titers of serum anti-β2GPI and anticardiolipin Abs in the serum, lack of fetal resorptions, low incidence of thrombocytopenia, and normal aPTT (activated partial thromboplastin time) values. β2GPI given orally before priming with β2GPI resulted in complete prevention of experimental APS development; β2GPI given at an early stage of the disease reduced clinical manifestations. However, administration of β2GPI 70 days postimmunization had a less significant effect on disease expression. Tolerized mice exhibited a diminished T lymphocyte proliferation response to β2GPI in comparison with β2GPI-immunized mice fed with OVA. When nontolerant β2GPI-primed T lymphocytes were mixed with T lymphocytes derived from tolerized mice, a significant inhibition of proliferation upon exposure to β2GPI was observed. The induction of suppression was β2GPI specific and driven, as well as TGF-β mediated. The β2GPI-specific response of T lymphocytes from the β2GPI-fed mice was reversed by anti-TGF-β Abs. The tolerance was adoptively transferred by CD8+ T cells from the tolerized mice into naive mice. Those CD8+ cells were MHC class I restricted, found to secrete TGF-β, and had no cytolytic activity. Oral administration of β2GPI suppressed priming of CTLs in the recipient mice. In sum, β2GPI-induced oral tolerance has an immunomodulatory effect in experimental APS, demonstrating the importance of β2GPI in the pathogenesis of the disease.
Mutational Analysis of CD28-Mediated Costimulation of Jun-N-Terminal Kinase and IL-2 Production
Authors: Barz Corinna , Nagel Thomas , Truitt Kenneth E. , Imboden John B. 
Published: December 2022

Abstract
The accessory molecule CD28 delivers a costimulus that acts in concert with TCR signals to promote T cell activation. Activation of Jun-N-terminal kinases (JNK) requires simultaneous stimulation of the TCR and CD28 and, therefore, likely plays an important role in signal integration during costimulation. We investigated the effects of mutations in the 41-amino acid cytoplasmic domain of murine CD28 on its ability to deliver costimuli for JNK activation and IL-2 production when expressed in Jurkat T cells. Our results indicate that the costimulus for JNK activation requires the membrane-proximal 24 amino acids of the CD28 cytoplasmic domain and is not mediated by the tyrosine-based recruitment of signaling molecules, including phosphatidylinositol 3-kinase. Deletion of the carboxyl-terminal 17 amino acids does not affect the ability of CD28 to augment JNK activation but impairs its ability to enhance TCR-mediated production of IL-2, demonstrating that optimal costimulation of IL-2 production requires CD28 signals in addition to the activation of JNK.