bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural Basis of the gp120 Superantigen-Binding Site on Human Immunoglobulins
Authors: Karray Saoussen , Juompan Laure , Maroun Rachid C. , Isenberg David , Silverman Gregg J. , Zouali Moncef 
Published: December 2022

AbstractB cell superantigens (SAg) interact with normal human nonimmune Igs (Igs), independently of the light chain isotype, and activate a large proportion of the B cell repertoire. Recently, the major envelope protein of HIV-1, gp120, was found to exhibit SAg-like properties for B cells with potential pathologic consequences for the infected host. This unconventional mode of interaction contrasts with its binding to immunization-induced Abs, which requires the tertiary structure of the heavy and light chain variable regions. In this report, we have examined the structural basis of the interaction between human Igs and gp120. We found that gp120 binding is restricted to Igs from the VH3 gene family and that the two VH genes 3-23 and 3-30, known to be overutilized during all stages of B cell development, frequently impart gp120 binding. We also provide evidence that the viral gp120 SAg can interact with only a subset of the human VH3+ Igs that can convey binding to the prototypic bacterial B cell SAg protein A from Staphylococcus aureus. Finally, we have identified amino acid positions present primarily in the first and third framework regions of the Ig heavy chain variable region, outside the conventional hypervariable loops, which correlate with gp120 binding. In a three-dimensional sequence-homology model, these residues partially overlap with the predicted SAg protein A binding site for VH3+ Igs.
Leishmania mexicanaCysteine Proteinase-Deficient Mutants Have Attenuated Virulence for Mice and Potentiate a Th1 Response
Authors: Alexander James , Coombs Graham H. , Mottram Jeremy C. 
Published: December 2022

AbstractLeishmania mexicana mutants lacking cysteine proteinase genes cpa (Δcpa), cpb (Δcpb), or both cpa and cpb (Δcpa/cpb) have been generated by targeted gene disruption. Δcpa mutants produce a disease phenotype in BALB/c mice close to that of wild-type L. mexicana, but Δcpb mutants are much less infective, producing very slowly growing small lesions, and Δcpa/cpb double mutants do not induce lesion growth. Immunologic analysis of Ab isotype during infection and splenocyte IFN-γ, IL-2, and IL-4 production following stimulation with Leishmania Ag or Con A indicates that there was a significant shift from a predominantly Th2-associated immune response in mice infected with wild-type L. mexicana to a Th1-associated response in mice inoculated with Δcpb or Δcpa/cpb. Significantly, Δcpa altered the balance of the immunologic response to a lesser extent than did the other mutants. Similar disease outcomes and switches in the Th1/Th2 balance were also observed when other L. mexicana-susceptible mouse strains were infected with the mutants. BALB/c and C57BL/6 mice vaccinated with Δcpa/cpb and CBA/Ca mice vaccinated with Δcpb or Δcpa/cpb were subsequently more resistant, to varying degrees, than were untreated mice to infection with wild-type parasites, as measured by development of lesions and parasite burden. These data implicate leishmanial cysteine proteinases not only as parasite virulence factors but also in modulation of the immune response and provide strong encouragement that cysteine proteinase-deficient L. mexicana mutants are candidate attenuated live vaccines.
Matrix Metalloproteinases Generate Angiostatin: Effects on Neovascularization
Authors: Cornelius Lynn A. , Nehring Leslie C. , Harding Elizabeth , Bolanowski Mark , Welgus Howard G. , Kobayashi Dale K. , Pierce Richard A. , Shapiro Steven D. 
Published: December 2022

Abstract
Angiostatin, a cleavage product of plasminogen, has been shown to inhibit endothelial cell proliferation and metastatic tumor cell growth. Recently, the production of angiostatin has been correlated with tumor-associated macrophage production of elastolytic metalloproteinases in a murine model of Lewis lung cell carcinoma. In this report we demonstrate that purified murine and human matrix metalloproteinases generate biologically functional angiostatin from plasminogen. Macrophage elastase (MMP-12 or MME) proved to be the most efficient angiostatin-producing MMP. MME was followed by gelatinases and then the stomelysins in catalytic efficiency; interstitial collagenases had little capacity to generate angiostatin. Both recombinant angiostatin and angiostatin generated from recombinant MME-treated plasminogen inhibited human microvascular endothelial cell proliferation and differentiation in vitro. Finally, employing macrophages isolated from MME-deficient mice and their wild-type littermates, we demonstrate that MME is required for the generation of angiostatin that inhibits the proliferation of human microvascular endothelial cells.
The First Subcomponent of Complement, C1q, Triggers the Production of IL-8, IL-6, and Monocyte Chemoattractant Peptide-1 by Human Umbilical Vein Endothelial Cells
Authors: van den Berg Rocco H. , Faber-Krol Maria C. , Sim Robert B. , Daha Mohamed R. 
Published: December 2022

AbstractWe and others have demonstrated previously the occurrence of cC1qR/CaR, a receptor for the collagen-like stalks of complement component C1q, on endothelial cells. In the present study we investigated whether binding of C1q to endothelial cells resulted in enhancement of cytokine or chemokine production. HUVEC produced 82 ± 91 pg/ml of IL-8, 79 ± 113 pg/ml of IL-6, and 503 ± 221 pg/ml of monocyte chemoattractant peptide-1 (MCP-1) under basal conditions. Incubation with C1q resulted in a time- and dose-dependent up-regulation of IL-8 (1012 ± 43 pg/ml), IL-6 (392 ± 20 pg/ml), and MCP-1 (2450 ± 101 pg/ml). This production is dependent on de novo protein synthesis, as demonstrated by the detection of specific mRNA after C1q stimulation, and inhibition of peptide production in the presence of cycloheximide. The production of all factors was inhibited (69 ± 7%) by the collagenous fragments of C1q, while the C1q globular heads only induced 13 ± 11% inhibition. When HUVEC were incubated with C1q in the presence of aggregated IgM, enhanced production of IL-8 (2500 ± 422 pg/ml), IL-6 (997 ± 21 pg/ml), and MCP-1 (5343 ± 302 pg/ml) was found. Furthermore, F(ab′)2 anti-calreticulin partially inhibited the production of IL-8, confirming at least the involvement of cC1qR/CaR. These experiments suggest that in an inflammatory response C1q not only is able to activate the complement pathway, but when presented in a proper fashion also might induce the production of factors that contribute to acute phase responses and recruitment of inflammatory cells.
Early Development and Spreading of Autoantibodies to Epitopes of IA-2 and Their Association with Progression to Type 1 Diabetes
Authors: Naserke Heike E. , Ziegler Anette-G. , Lampasona Vito , Bonifacio Ezio 
Published: December 2022

AbstractAutoimmunity precedes clinical type 1 diabetes, and indicators of maturing autoimmune responses may be useful markers for disease prediction. To study this, epitope maturation of autoantibodies to the related protein tyrosine phosphatase (PTP)-like autoantigens IA-2 and IA-2β was examined in sequential samples from birth in a cohort of 21 offspring developing multiple islet autoantibodies and a similar cohort of 48 relatives of patients with type 1 diabetes recruited at an older age. Initial reactivity in offspring was heterogeneous against the IA-2 juxtamembrane region (10/21) and PTP domains (13/21), and both specificity and extent of initial IA-2/IA-2β autoantibodies were associated with HLA class II genotype. Intra-IA-2 epitope spreading and/or intermolecular spreading to IA-2β epitopes were observed in seven offspring. In contrast, in older relatives, IA-2/IA-2β Ab reactivity was stable and spreading rare. Development of diabetes in children was associated with the presence of Abs to the IA-2 juxtamembrane region (risk by age 5 yr, 52% vs 0% in those with PTP domain Abs only; p &lt; 0.02), and 5 of 26 relatives who developed diabetes had IA-2 Abs only against the juxtamembrane region. The findings show that autoantibody reactivity to IA-2/IA-2β is dynamic in the young, show that the juxtamembrane region of IA-2 is an early IA-2 autoantibody target, and suggest that these Abs are a risk factor for development of type 1 diabetes in infancy.
Cutting Edge: Detection of a High Frequency of Virus-Specific CD4+ T Cells During Acute Infection with Lymphocytic Choriomeningitis Virus
Authors: Varga Steven M. , Welsh Raymond M. 
Published: December 2022

AbstractLymphocytic choriomeningitis virus (LCMV), like many viruses, induces a profound activation and expansion of CD8+ T cells. In contrast, CD4+ T cells do not increase in total number during the acute infection. We show here that mice infected with LCMV have a low but detectable frequency (&lt;1/300) of CD4+ T cells, as detected by IL-2 production in limiting dilution assays, to each of two class II peptides during the peak of the acute LCMV response and into long-term memory. However, during the peak of the acute CD4+ T cell response, &gt;20% of the CD4+ T cells secreted IFN-γ after stimulation with PMA and ionomycin, and &gt;10% of the CD4+ T cells secreted IFN-γ after stimulation with the LCMV peptides. Thus, these new sensitive assays reveal a heretofore unappreciated, yet profound Ag-specific CD4+ T cell response during viral infections.
Urocanic Acid Enhances IL-10 Production in Activated CD4+ T Cells
Authors: Holáň Vladimír , Kuffová Lucia , Zajícová Alena , Krulová Magdaléna , Filipec Martin , Holler Petr , Jančárek Alexander 
Published: December 2022

AbstractThe immunosuppressive effects of UV radiation have been well documented. This suppression has been attributed to the action of the cis form of urocanic acid (UCA), a photoproduct of trans-UCA, a natural constituent of the skin. Here, we show that mouse spleen cells preincubated with cis-UCA have a diminished proliferative response to allogeneic cells in MLC and to stimulation with anti-CD3 mAb. Cells preincubated with cis-UCA also had a decreased ability to serve as APC and to stimulate the proliferation of allogeneic lymphocytes in MLC. Simultaneously, the production of IL-2 and IFN-γ by cells preincubated with cis-UCA was decreased. However, IL-10 gene expression and IL-10 protein secretion by spleen cells stimulated in the presence of cis-UCA were significantly enhanced. The principal cell population displaying the cis-UCA-induced elevated production of IL-10 was CD4+ T cells, which were shown to be a direct target of cis-UCA action. This was also supported by the observation that production of IL-10 by stimulated splenic non-T cells or by macrophages was not altered by cis-UCA. The enhanced production of IL-10 by activated CD4+ T cells may represent a novel pathway of UVB radiation-induced, cis-UCA-mediated immunosuppression. We suggest that the elevated production of IL-10 by activated CD4+ T cells may account for the suppressor T cell phenomena described in UV-irradiated recipients.
Chemical Chaperones Enhance Superantigen and Conventional Antigen Presentation by HLA-DM-Deficient as well as HLA-DM-Sufficient Antigen-Presenting Cells and Enhance IgG2a Production In Vivo
Authors: Ghumman Birinder , Bertram Edward M. , Watts Tania H. 
Published: December 2022

AbstractChemical chaperones, first defined in studies of mutant cystic fibrosis transmembrane conductance regulator proteins, are small molecules that act as stabilizers of proteins in their native state and have the ability in some cases to rescue protein-folding mutants within cells. HLA-DM is an MHC II-specific molecular chaperone that facilitates peptide loading onto MHC II proteins and also stabilizes empty MHC II molecules prior to their acquisition of antigenic peptides. APC that lack HLA-DM exhibit quantitative defects in protein Ag as well as superantigen presentation. Here we show that both the superantigen and protein presentation defect in MHC II-transfected, HLA-DM-deficient T2 cells can be partially overcome by treating the APC with the chemical chaperones glycerol, DMSO, or trimethylamine oxide. These chemical chaperones also enhance superantigen and conventional Ag presentation by wild-type APC. In vivo, glycerol was found to act as an adjuvant and resulted in enhanced IgG2a production to trinitrophenyl-keyhole limpet hemocyanin (TNP-KLH). In vitro, the enhancement of Ag presentation by chemical chaperones was found to take place at the level of the APC and took several hours to develop. Subcellular fractionation experiments show that HLA-DM enhances presentation of peptides by dense endosome fractions whereas chemical chaperones enhance presentation by light membrane fractions (early endosome or plasma membrane). The mechanism by which these chemical chaperones augment Ag presentation is not defined, but flow cytometric analysis suggests that the enhancement may be due to a subtle effect on the stability of several different proteins at the cell surface.
Selective Ability of Mouse CD1 to Present Glycolipids: α-Galactosylceramide Specifically Stimulates Vα14+ NK T Lymphocytes
Authors: Burdin Nicolas , Brossay Laurent , Koezuka Yasuhiko , Smiley Stephen T. , Grusby Michael J. , Gui Ming , Taniguchi Masaru , Hayakawa Kyoko , Kronenberg Mitchell 
Published: December 2022

AbstractMouse CD1 (mCD1) glycoproteins are known to present peptides, while human CD1 molecules present glycolipids. In mice, mCD1-autoreactive NK T cells play critical roles in various immune responses, through the secretion of high amounts of cytokines. This study was initiated to determine whether glycolipids are involved in the autorecognition of mCD1 by NK T cells. α-Galactosylceramide (α-GalCer) was the only glycolipid tested capable of eliciting an mCD1-restricted response by splenic T cells. Moreover, splenic T cells derived from mCD1-deficient mice were not stimulated by α-GalCer, suggesting that the responsive T cells are selected by mCD1. Using cytoflow techniques, we confirmed that, in response to α-GalCer, IFN-γ-secreting cells displayed an NK T cell phenotype. The predominance of IFN-γ vs IL-4, however, is determined by the type of mCD1+ APC, suggesting the potential for APC regulation of cytokine production by NK T cells. Among a panel of 10 mCD1-autoreactive T cell hybridomas, only the ones that express the typical Vα14Jα281 TCR rearrangement of NK T cells responded to α-GalCer. Fixation or treatment of mCD1+ APCs with an inhibitor of endosomal acidification and the use of mCD1 mutants unable to traffic through endosome still allowed α-GalCer to stimulate NK T cells. Thus, endosomal trafficking and Ag processing are not required for glycolipid recognition. In summary, α-GalCer might be the autologous ligand, or a mimic of a glycolipid ligand, involved in the mCD1-mediated stimulation of NK T cells.
Fine Specificity of the Myelin-Reactive T Cell Repertoire: Implications for TCR Antagonism in Autoimmunity
Authors: Anderton Stephen M. , Manickasingham Shivanthi P. , Burkhart Christoph , Luckcuck Tracy A. , Holland Sam J. , Lamont Alan G. , Wraith David C. 
Published: December 2022

AbstractThe use of altered peptide ligands (APL) to modulate T cell responses has been suggested as a means of treating T cell-mediated autoimmune disorders. We have assessed the therapeutic potential of TCR antagonist peptides in autoimmunity using murine experimental autoimmune encephalomyelitis (EAE) as a model. The Tg4 transgenic mouse expresses an MHC class II-restricted TCR specific for the immunodominant encephalitogenic epitope of myelin basic protein, Ac1–9 (acetylated N-terminal nonamer). We have used T cell lines derived from Tg4 mice to define the TCR contact residues within Ac1–9. APL with appropriate substitutions at the primary TCR contact residue were effective antagonists of Tg4 T cells. These antagonist APL, however, were found to induce EAE in susceptible, nontransgenic strains of mice. Underlying this, the Ac1–9-specific T cell repertoire of normal mice, rather than reflecting the Tg4 phenotype, showed considerable diversity in fine specificity and was able to respond to the Tg4 antagonist APL. Defining antagonist APL in vitro using T cell clones, therefore, was not a reliable approach for the identification of APL with EAE-suppressing potential in vivo. Our findings highlight the complexities of the autoreactive T cell repertoire and have major implications for the use of APL in autoimmune diseases.
Mechanisms of Hyaluronan-Induced Up-Regulation of ICAM-1 and VCAM-1 Expression by Murine Kidney Tubular Epithelial Cells: Hyaluronan Triggers Cell Adhesion Molecule Expression Through a Mechanism Involving Activation of Nuclear Factor-κB and Activating Protein-1
Authors: Oertli Beat , Beck-Schimmer Beatrice , Fan Xiaohong , Wüthrich Rudolf P. 
Published: December 2022

AbstractThe matrix constituent hyaluronan (HA) markedly accumulates in inflammatory lesions. To gain insight into the biologic significance of this phenomenon we tested the hypothesis that HA could regulate cell adhesion molecule expression in epithelial cells. Using a clonal line of mouse cortical tubular (MCT) cells we found that fragmented intermediate m.w., but not high m.w., HA markedly increased ICAM-1 and VCAM-1 steady state mRNA and cell surface expression. Up-regulation of ICAM-1 and VCAM-1 mRNA by HA was preceded by a marked increase in NF-κB and activating protein-1 DNA binding activity in MCT cells. Transcript levels for the NF-κB inhibitor IκBα and for the activating protein-1 constituents c-jun and c-fos also increased in response to HA stimulation of tubular cells. Inhibition of NF-κB with the serine protease inhibitor N-tosyl-l-phenylalanine chloromethyl ketone blocked the HA-mediated expression of ICAM-1 and VCAM-1 in MCT cells. In conclusion, HA displays proinflammatory effects by directly stimulating the expression of the cell adhesion molecules ICAM-1 and VCAM-1 in mouse kidney epithelial cells. HA could thereby play an important role in leukocyte adhesion in inflammatory renal diseases.
Differential Usage of VH Gene Segments Is Mediated bycisElements
Authors: Yu Calvin C. K. , Larijani Mani , Miljanic Ivana N. , Wu Gillian E. 
Published: December 2022

AbstractIg diversity is generated in large part by the combinatorial joining of the Ig gene segments, VH, D, and JH, that together encode the variable domain of Ig. The final Ig repertoire, however, not only reflects the diversity generated through V(D)J recombinatorial joining, but it is also the product of a number of developmental restraints and selections. To avoid such restrictions and assess the recombination potential of individual Ig gene segments, we constructed Ig heavy (H) chain microlocus plasmids, each of which contain germline coding, recombination signal, and flanking sequences of a VH, D, and JH gene segment. These plasmids allow us to assess the recombination potential of the segments in the context of their natural flanking DNA sequences, but in the absence of any higher order chromatin structure or cellular selection. We found that the frequency and extent of deletions and additions at the recombination breakpoints are similar to those observed at rearranged Ig H chain loci in intact animals. The relative frequencies of the types of rearrangements—VD-J, V-DJ, VinvD-J (invD = inverted D), and VDJ—however, differ strongly. Moreover, V81x, the most used VH gene segment in intact mice, also is overused in this plasmid assay, 15 to 30 times that of another VH segment. This result indicates that the overuse of V81x in the early B cell repertoire can be a consequence of its DNA sequence and not of cellular activities.
Molecular Mapping with Functional Antibodies Localizes Critical Sites on the Human IL Receptor Common γ (γc) Chain
Authors: Raskin Natalya , Jakubowski Aniela , Sizing IreneDougas , Olson Dian L. , Kalled Susan L. , Hession Catherine A. , Benjamin Christopher D. , Baker Darren P. , Burkly Linda C. 
Published: December 2022

AbstractThe IL receptor common γ (γc) chain is required for the formation of high affinity cytokine receptor complexes for IL-2, IL-4, IL-7, IL-9, and IL-15, and for signals regulating cell survival, growth, and differentiation. Our current understanding of how γc chain associates with multiple ligands and receptor subunits is drawn largely from its structural homology to the human growth hormone (hGH) receptor and known structure of the hGH/hGH receptor complex. These receptors share distinct features in their extracellular portions and are believed to function by a mechanism of ligand-induced association of receptor subunits. Here, we report the first directed mutational analysis of the human γc chain by alanine scanning conducted across seven regions likely to contain residues required for intermolecular contact. Functionally distinct, neutralizing anti-γc mAbs were employed to define critical residues. One particular mAb, CP.B8, unique in its ability to inhibit IL-2-, IL-4-, IL-7-, and IL-15-induced proliferation and high affinity cytokine binding of normal T cells as an intact mAb and as a Fab fragment, localized critical residues to four noncontinuous stretches, namely residues in loops AB and EF of domain 1, in the interdomain segment, and in loop FG of domain 2. Notably, these residues form a contiguous patch on the γc chain surface in a three-dimensional structural model. These results provide functional evidence for the location of contact points on γc chain required for its association with multiple ligands.
B Cell Responses to a Peptide Epitope. V. Kinetic Regulation of Repertoire Discrimination and Antibody Optimization for Epitope
Authors: Nayak Bishnu P. , Tuteja Renu , Manivel Venkatasamy , Roy Rajendra P. , Vishwakarma Ram A. , Rao Kanury V. S. 
Published: December 2022

AbstractThe influence of imposing various conformational constraints on immune responses to a model epitope within a synthetic peptide immunogen was examined in mice. Although overall immunogenicity was affected, the model epitope (sequence DPAF) remained the predominant recognition site regardless of the conformation in which it was presented. A comparison of anti-DPAF mAbs obtained in response to two analogue peptides, PS1CT3 and CysCT3, in which the DPAF segment was either unconstrained or held within a cyclic loop, respectively, revealed a significant homology in the paratope composition. At one level a subset of anti-PS1CT3 and anti-CysCT3 mAbs was found to share a common heavy chain variable region. In addition, nucleotide sequence homology comparisons of both heavy and light chain variable regions identified the presence of anti-PS1CT3 and anti-CysCT3 mAbs that collectively appeared to derive from a common progenitor, but with nonidentical somatic mutations. Interestingly, however, no bias toward homologous Ag could be discerned on measurement of relative affinities of the mAbs for the two peptides. In contrast, mAb binding on-rates clearly discriminated between peptides representing the homologous vs the heterologous confomer of the DPAF epitope. Thus, it would appear that the kinetics of Ag recognition dominate over equilibrium binding criteria both in epitope-driven repertoire selection and Ab maturation in a humoral response.