bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Cytomegalovirus Binding to Human Monocytes Induces Immunoregulatory Gene Expression
Authors: Yurochko Andrew D. , Huang Eng-Shang 
Published: January 2023

Abstract
To continue our investigation of the cellular events that occur following human CMV (HCMV) infection, we focused on the regulation of cellular activation following viral binding to human monocytes. First, we showed that viral binding induced a number of immunoregulatory genes (IL-1β, A20, NF-κB-p105/p50, and IκBα) in unactivated monocytes and that neutralizing Abs to the major HCMV glycoproteins, gB (UL55) and gH (UL75), inhibited the induction of these genes. Next, we demonstrated that these viral ligands directly up-regulated monocyte gene expression upon their binding to their appropriate cellular receptors. We then investigated if HCMV binding also resulted in the translation and secretion of cytokines. Our results showed that HCMV binding to monocytes resulted in the production and release of IL-1β protein. Because these induced gene products have NF-κB sites in their promoter regions, we next examined whether there was an up-regulation of nuclear NF-κB levels. These experiments showed that, in fact, NF-κB was translocated to the nucleus following viral binding or purified viral ligand binding. Changes in IκBα levels correlated with the changes in NF-κB translocation. Lastly, we demonstrated that p38 kinase activity played a central role in IL-1β production and that it was rapidly up-regulated following infection. These results support our hypothesis that HCMV initiates a signal transduction pathway that leads to monocyte activation and pinpoints a potential mechanism whereby HCMV infection of monocytes can result in profound pathogenesis, especially in chronic inflammatory-type conditions.
Role of the C-C Chemokine, TCA3, in the Protective Anticryptococcal Cell-Mediated Immune Response
Authors: Doyle Hester A. , Murphy Juneann W. 
Published: January 2023

AbstractActivated T lymphocytes play a crucial role in orchestrating cellular infiltration during a cell-mediated immune (CMI) reaction. TCA3, a C-C chemokine, is produced by Ag-activated T cells and is chemotactic for neutrophils and macrophages, two cell types in a murine CMI reaction. Using a gelatin sponge model for delayed-type hypersensitivity (DTH), we show that TCA3 is a component of the expression phase of an anticryptococcal CMI response in mice. TCA3 mRNA levels are augmented in anticryptococcal DTH reactions at the same time peak influxes of neutrophils and lymphocytes are observed. Neutralization of TCA3 in immunized mice results in reduced numbers of neutrophils and lymphocytes at DTH reaction sites. However, when rTCA3 is injected into sponges in naive mice, only neutrophils are attracted into the sponges, indicating TCA3 is chemotactic for neutrophils, but not lymphocytes. We show that TCA3 is indirectly attracting lymphocytes into DTH-reactive sponges by affecting at least one other chemokine that is chemotactic for lymphocytes. Of the two lymphocyte-attracting chemokines assessed, monocyte-chemotactic protein-1 and macrophage-inflammatory protein-1α (MIP-1α), only MIP-1α was reduced when TCA3 was neutralized, indicating that TCA3 affects the levels of MIP-1α, which attracts lymphocytes into the sponges. TCA3 also plays a role in protection against Cryptococcus neoformans in the lungs and brains of infected mice, as evidenced by the fact that neutralization of TCA3 results in increased C. neoformans CFU in those two organs.
Gene Expression and Production of the Monokine Induced by IFN-γ (MIG), IFN-Inducible T Cell α Chemoattractant (I-TAC), and IFN-γ-Inducible Protein-10 (IP-10) Chemokines by Human Neutrophils
Authors: Gasperini Sara , Marchi Martina , Calzetti Federica , Laudanna Carlo , Vicentini Lucia , Olsen Henrik , Murphy Marianne , Liao Fang , Farber Joshua , Cassatella Marco A. 
Published: January 2023

AbstractMonokine induced by IFN-γ (MIG), IFN-inducible T cell α chemoattractant (I-TAC), and IFN-γ-inducible protein of 10 kDa (IP-10) are related members of the CXC chemokine subfamily that bind to a common receptor, CXCR3, and that are produced by different cell types in response to IFN-γ. We have recently reported that human polymorphonuclear neutrophils (PMN) have the capacity to release IP-10. Herein, we show that PMN also have the ability to produce MIG and to express I-TAC mRNA in response to IFN-γ in combination with either TNF-α or LPS. While IFN-γ, alone or in association with agonists such as fMLP, IL-8, granulocyte (G)-CSF and granulocyte-macrophage (GM)-CSF, failed to influence MIG, IP-10, and I-TAC gene expression, IFN-α, in combination with TNF-α, LPS, or IL-1β, resulted in a considerable induction of IP-10 release by neutrophils. Furthermore, IL-10 and IL-4 significantly suppressed the expression of MIG, IP-10, and I-TAC mRNA and the extracellular production of MIG and IP-10 in neutrophils stimulated with IFN-γ plus either LPS or TNF-α. Finally, supernatants harvested from stimulated PMN induced migration and rapid integrin-dependent adhesion of CXCR3-expressing lymphocytes; these activities were significantly reduced by neutralizing anti-MIG and anti-IP-10 Abs, suggesting that they were mediated by MIG and IP-10 present in the supernatants. Since MIG, IP-10, and I-TAC are potent chemoattractants for NK cells and Th1 lymphocytes, the ability of neutrophils to produce these chemokines might contribute not only to the progression and evolution of the inflammatory response, but also to the regulation of the immune response.
Endothelial Targeting and Enhanced Antiinflammatory Effects of Complement Inhibitors Possessing Sialyl Lewisx Moieties
Authors: Mulligan Michael S. , Warner Roscoe L. , Rittershaus Charles W. , Thomas Lawrence J. , Ryan Una S. , Foreman Kimberly E. , Crouch Larry D. , Till Gerd O. , Ward Peter A. 
Published: January 2023

Abstract
The complement inhibitor soluble complement receptor type 1 (sCR1) and a truncated form of sCR1, sCR1[desLHR-A], have been generated with expression of the selectin-reactive oligosaccharide moiety, sialyl Lewisx (sLex), as N-linked oligosaccharide adducts. These modified proteins, sCR1sLex and sCR1[desLHR-A]sLex, were assessed in the L-selectin- and P-selectin-dependent rat model of lung injury following systemic activation of complement by cobra venom factor and in the L-selectin-, P-selectin-, and E-selectin-dependent model of lung injury following intrapulmonary deposition of IgG immune complexes. In the cobra venom factor model, sCR1sLex and sCR1[desLHR-A]sLex caused substantially greater reductions in neutrophil accumulation and in albumin extravasation in lung when compared with the non-sLex-decorated forms. In this model, increased lung vascular binding of sCR1sLex and sCR1[desLHR-A]sLex occurred in a P-selectin-dependent manner, in contrast to the absence of any increased binding of sCR1 or sCR1[desLHR-A]. In the IgG immune complex model, sCR1[desLHR-A]sLex possessed greater protective effects relative to sCR1[desLHR-A], based on albumin extravasation and neutrophil accumulation. Enhanced protective effects correlated with greater lung vascular binding of sCR1[desLHR-A]sLex as compared with the non-sLex-decorated form. In TNF-α-activated HUVEC, substantial in vitro binding occurred with sCR1[desLHR-A]sLex (but not with sCR1[desLHR-A]). This endothelial cell binding was blocked by anti-E-selectin but not by anti-P-selectin. These data suggest that sLex-decorated complement inhibitors have enhanced antiinflammatory effects and appear to have enhanced ability to localize to the activated vascular endothelium.
NF-κB Regulation by IκB Kinase in Primary Fibroblast-Like Synoviocytes
Authors: Aupperle Karlfried R. , Bennett Brydon L. , Boyle David L. , Tak Paul-Peter , Manning Anthony M. , Firestein Gary S. 
Published: January 2023

Abstract
NF-κB is a key regulator of inflammatory gene transcription and is activated in the rheumatoid arthritis (RA) synovium. In resting cells, NF-κB is retained as an inactive cytoplasmic complex by its inhibitor, IκB. Phosphorylation of IκB targets it for proteolytic degradation, thereby releasing NF-κB for nuclear translocation. Recently, two related IκB kinases (IKK-1 and IKK-2) were identified in immortalized cell lines that regulate NF-κB activation by initiating IκB degradation. To determine whether IKK regulates NF-κB in primary cells isolated from a site of human disease, we characterized IKK in cultured fibroblast-like synoviocytes (FLS) isolated from synovium of patients with RA or osteoarthritis. Immunoreactive IKK protein was found to be abundant in both RA and osteoarthritis FLS by Western blot analysis. Northern blot analysis showed that IKK-1 and IKK-2 genes were constitutively expressed in all FLS lines. IKK function in FLS extracts was determined by measuring phosphorylation of recombinant IκB in vitro. IKK activity in both RA and osteoarthritis FLS was strongly induced by TNF-α and IL-1 in a concentration-dependent manner. Activity was significantly increased within 10 min of stimulation and declined to near basal levels within 80 min. Activation of IKK in FLS was accompanied by phosphorylation and degradation of endogenous IκBα as determined by Western blot analysis. Concomitant activation and nuclear translocation of NF-κB was documented by EMSA and immunohistochemistry. Transfection with a dominant negative IKK-2 mutant prevented TNF-α-mediated NF-κB nuclear translocation, whereas a dominant negative IKK-1 mutant had no effect. This is the first demonstration that IKK-2 is a pivotal regulator of NF-κB in primary human cells.
Inhibition of CCR5 Expression by IL-12 Through Induction of β-Chemokines in Human T Lymphocytes
Authors: Wang Jinhai , Guan Ennan , Roderiquez Gregory , Norcross Michael A. 
Published: January 2023

AbstractIL-12 induces initiation of the differentiation of naive CD4+ T lymphocytes into Th1 cells and is important for the control of cell-mediated immunity. β-Chemokines serve to attract various types of blood leukocytes to sites of infection and inflammation. The specific receptor for the β-chemokines (macrophage-inflammatory protein (MIP)-1α, MIP-1β, and RANTES), CCR5, also functions as the primary coreceptor for macrophage-tropic isolates of HIV-1. IL-12, but not IL-4, IL-10, or IL-13, now has been shown to down-modulate the surface expression of CCR5 induced by IL-2 on both CD4+ and CD8+ T lymphocytes. Decreased CCR5 surface expression was not secondary to transcriptional inhibition, given that CCR5 mRNA was enhanced in cells cultured in IL-12/IL-2 compared with those cultured in IL-2 only. The effect of IL-12 in down-modulation of CCR5 surface expression was shown to be mediated by soluble factors secreted from the T cells. Rapid and transient intracellular Ca2+ mobilization was induced in monocytes by IL-12-induced supernatants, which desensitized the response of monocytes to MIP-1α, but not their response to stromal cell-derived factor-1α. Neutralization with specific Abs identified these factors as MIP-1α and MIP-1β from most donors. IL-4, IL-10, IFN-γ, and IL-18 primarily inhibited MIP-1β secretion and also weakly suppressed MIP-1α secretion. HIV-1 replication was inhibited in IL-2/IL-12-containing cultures that correlated with chemokine and chemokine-receptor levels. These data suggest that the effects of IL-12 on β-chemokine production and chemokine-receptor expression may contribute to the immunomodulatory activities of IL-12 and may have potential therapeutic relevance in controlling HIV-1 replication.
Emt/Itk Associates with Activated TCR Complexes: Role of the Pleckstrin Homology Domain
Authors: Ching Keith A. , Kawakami Yuko , Kawakami Toshiaki , Tsoukas Constantine D. 
Published: January 2023

AbstractExpressed in mast and T-cells/inducible T cell tyrosine kinase (Emt/Itk) is a protein tyrosine kinase required for T cell Ag receptor (TCR)-induced activation and development. A physical interaction between Emt/Itk and TCR has not been described previously. Here, we have utilized laser scanning confocal microscopy to demonstrate that Ab-mediated engagement of the CD3ε chain induces the membrane colocalization of Emt/Itk with TCR/CD3. Removal of the Emt/Itk pleckstrin homology domain (ΔPH-Emt/Itk) abrogates the association of the kinase with the cell membrane, as well as its activation-induced colocalization with the TCR complex and subsequent tyrosine phosphorylation. The addition of a membrane localization sequence to ΔPH-Emt/Itk from Lck restores all of these deficiencies except the activation-induced tyrosine phosphorylation. Our data suggest that the PH domain of Emt/Itk can be replaced with another membrane localization signal without affecting the membrane targeting and activation-induced colocalization of the kinase with the TCR. However, the PH domain is indispensable for the activation-induced tyrosine phosphorylation of the kinase.
Selection of CTL Escape Mutants in Mice Infected with a Neurotropic Coronavirus: Quantitative Estimate of TCR Diversity in the Infected Central Nervous System
Authors: Pewe Lecia , Heard Stephen B. , Bergmann Conni , Dailey Morris O. , Perlman Stanley 
Published: January 2023

AbstractVariant viruses mutated in the immunodominant cytotoxic T cell epitope surface (S) glycoprotein S-510-518 are selected in mice chronically infected with mouse hepatitis virus, strain JHM. We determined whether this selection occurred in the presence of an oligoclonal or polyclonal T cell response using soluble MHC/peptide tetramers in direct ex vivo analyses of CNS-derived lymphocytes. A total of 42% (range, 29–60%) of CD8 T cells in the CNS of mice with acute encephalitis recognized epitope S-510-518. A total of 34% (range, 18–62%) of cells from mice with hind limb paralysis (and chronic demyelination) were also epitope specific, even though only virus expressing mutated epitope is detected in these animals. Sequence analysis of the β-chain CDR3 of 487 tetramer S-510-518-positive cDNA clones from nine mice showed that a majority of clonotypes were identified in more than one mouse. From these analyses, we estimated that 300–500 different CD8 T cell clonotypes responsive to epitope S-510-518 were present in each acutely infected brain, while 100–900 were present in the CNS of each mouse with chronic disease. In conclusion, a polyclonal CD8 T cell response to an epitope does not preclude the selection of T cell escape mutants, and epitope-specific T cells are still present at high levels even after RNA-encoding wild-type sequence is no longer detectable.
Compromised Virus Control and Augmented Perforin-Mediated Immunopathology in IFN-γ-Deficient Mice Infected with Lymphocytic Choriomeningitis Virus
Authors: Nansen Anneline , Jensen Teis , Christensen Jan Pravsgaard , Andreasen Susanne Ørding , Röpke Carsten , Marker Ole , Thomsen Allan Randrup 
Published: January 2023

AbstractTo define the role of IFN-γ in the control of acute infection with a noncytopathogenic virus, mice with targeted defects of the genes encoding IFN-γ, perforin, or both were infected i.v. with two strains of lymphocytic choriomeningitis virus differing markedly in their capacity to spread in wild-type mice. Our results reveal that IFN-γ is pivotal to T cell-mediated control of a rapidly invasive stain, whereas it is less important in the acute elimination of a slowly invasive strain. Moreover, the majority of mice infected with the rapidly invasive strain succumb to a wasting syndrome mediated by CD8+ effector cells. The primary effector mechanism underlying this disease is perforin-dependent lysis, but other mechanisms are also involved. Wasting disease can be prevented if naive CD8+ cells from mice transgenic for an MHC class I-restricted lymphocytic choriomeningitis virus-specific TCR are adoptively transferred before virus challenge, indicating that the disease is the result of an unfortunate balance between virus replication in internal organs, e.g., liver and spleen, and the host response; resetting this balance by increasing host responsiveness will again lead to a rapidly controlled infection and limited tissue damage. Thus, the presence or absence of IFN-γ determines whether CTLs will clear infection with this noncytopathogenic virus or induce severe immunopathology.
Human RNA Helicase A Is a Lupus Autoantigen That Is Cleaved During Apoptosis
Authors: Takeda Yoshihiko , Caudell Patricia , Grady Giuia , Wang Grace , Suwa Akira , Sharp Gordon C. , Dynan William S. , Hardin John A. 
Published: January 2023

Abstract
Proteolytic cleavage by caspases is the central event in cells undergoing apoptosis. Cleaved proteins are often targeted by autoantibodies, suggesting that the cleavage of self Ags enhances immunogenicity and is prone to induce an autoimmune response. We found autoantibodies that immunoprecipitated a 140-kDa RNA-associated protein, provisionally designated Pa, in 11 of 350 patient sera that were positive for antinuclear Abs in an immunofluorescence test. The Pa protein gave rise to three fragments with m.w. ranging from 120–130 kDa during anti-Fas-activated apoptosis. Pure caspase-3 cleaved the Pa protein into a 130-kDa fragment corresponding to the largest of these three products. Peptide sequence analysis of a tryptic digest from immunoaffinity-purified Pa showed 100% identity to human RNA helicase A (RHA). The identity of Pa with RHA was further confirmed by immunoblotting with rabbit anti-RHA Ab using anti-Pa immunoprecipitates as substrates. All 10 anti-RHA-positive patients who were clinically analyzed were diagnosed as having systemic lupus erythematosus, and 7 of them had lupus nephritis. RHA is a multifunctional protein with roles in cellular RNA synthesis and processing. Inactivation of RHA by cleavage may be an important part of the process leading to programmed cell death. The cleaved RHA fragments that are produced during apoptosis may trigger an autoimmune response in systemic lupus erythematosus.
Determination of Glutamic Acid Decarboxylase 65 Peptides Presented by the Type I Diabetes-Associated HLA-DQ8 Class II Molecule Identifies an Immunogenic Peptide Motif
Authors: Herman Ann E. , Tisch Roland M. , Patel Salil D. , Parry Sarah L. , Olson Jennifer , Noble Janelle A. , Cope Andrew P. , Cox Brett , Congia Mauro , McDevitt2 Hugh O. 
Published: January 2023

AbstractParticular HLA class II allelic sequences are associated with susceptibility to type I diabetes. To understand the mechanism, knowledge of the molecular nature of the specific TCR/peptide/class II interactions involved in the disease process is required. To this end, we have introduced the diabetes-associated human class II HLA-DQ8 allele (DQA1*0301/DQB1*0302) as a transgene into mice and analyzed T cell responses restricted by this molecule to an important Ag in human diabetes, human glutamic acid decarboxylase 65. Hybridomas were used to determine the particular peptides from this Ag presented by HLA-DQ8 to T cells and to map the core minimal epitopes required for T cell stimulation. Analysis of these core epitopes reveals a motif and relevant features for peptides that are immunogenic to T cells when presented by HLA-DQ8. The major immunogenic epitopes of glutamic acid decarboxylase 65 do not contain a negatively charged residue that binds in the P9 pocket of the HLA-DQ8 molecule. PBMC from HLA-DQ8+ diabetic and nondiabetic individuals respond to these peptides, confirming that the mouse model is a useful tool to define epitopes of autoantigens that are processed by human APC and recognized by human T cells.
Rapid formation of T cell extended processes (TCEPs) in CNS infiltrating virus specific CD8 T cells (128.21)
Authors: McDole Jeremiah R. , Danzer Steve C. , Pun Raymond Y.K. , Pirko Istvan , Johnson Aaron J. 
Published: January 2023

Abstract
Expansion of an effective antiviral CD8 T cell response is critical for protection from Theiler's murine encephalomyelitis virus (TMEV) induced demyelinating syndrome. However, the specific CNS cell types engaged by these protective CD8 T cells in TMEV resistant strains remain unknown. We employed confocal microscopy to visualize the morphology, migration and specific cellular interactions of GFP+ CD8 T cells specific for the Db:VP2 121-130 epitope. These epitope specific, adoptively transferred CD8 T cells polarized CD8 and granzyme B towards TMEV infected neurons. Furthermore, epitope specific CD8 T cells presented with atypical cellular morphologies, including long cellular processes with varying thickness. Using live tissue imaging we determined these T cell extended processes (TCEPs) formed rapidly and could be partly attributed to T cell migration through tissue. This study is also the first to document and measure the rapid formation of TCEPs on antiviral CD8 T cells using real time imaging in the CNS.
Reconsidering the evolutionary significance of viral immune evasion (128.33)
Authors: Hill Ann B. 
Published: January 2023

Abstract
As do other herpesviruses, cytomegaloviruses (CMVs) all encode genes that interfere with the MHC class I pathway of antigen presentation to CD8 T cells. An extraordinary variety of molecular mechanisms have evolved to interfere with this pathway, and abundant experimental evidence indicates that these genes effectively impair the ability of CD8 T cells to recognize and kill virus-infected cells. The ubiquity of these genes argues that the virus is under strong selective pressure to keep them, and hence that they must provide a real evolutionary advantage. Paradoxically, however, viruses lacking MHC I immune evasion genes appear almost completely unimpaired in their ability to establish acute infection, and to persist for the life of the host. I propose that the apparent paradox results from considering immune evasion in terms of an evolutionary struggle between viruses and their hosts. Most individuals are infected with multiple strains of CMV. I suggest that immune evasion has evolved because it confers selective advantage to CMV strains in their intense evolutionary competition with other strains for access to an ultimately limiting resource. CD8 T cells provide some impediment to accessing this resource, but in most circumstances are not limiting. Hence, avoiding CD8 T cells is primarily of selective advantage when there is competition for access to resources. This concept leads us to focus on identifying limiting resources, and quantifying the degree of hindrance imposed by immune elements, when modeling viral pathogenesis.
The Role of Complement Receptor CD21/35 in a Murine Model of Chronic Wasting Disease (129.21)
Authors: Michel Brady Allen , Zabel Mark 
Published: January 2023

Abstract
Prion accumulation and replication occur in lymphoid follicles or inflammatory foci containing follicular dendritic cells (FDCs). FDCs accumulate immune complexes through their complement receptors CD21/35 and Fc receptors. These receptors, specifically CD21/35, have also been shown to play a critical role in peripheral prion pathogenesis in mouse scrapie models. In this study we examined feces from mice at 0, 1, 3, 6, and 8 days after oral inoculation for PrPCWD to determine whether prion retention and shedding depend on complement receptors CD21/35. We also plan to evaluate if CD21/35 can co-localize with CWD prions in the spleen at time points 0, 1, 2, 4, and 16 hours post i.p. inoculation using conventional Western Blotting and confocal microscopy.