bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of TNFR1 and TNFR2 in TNF-Induced Platelet Consumption in Mice
Authors: Tacchini-Cottier Fabienne , Vesin Christian , Redard Mireille , Buurman Wim , Piguet Pierre F. 
Published: January 2023

AbstractAn injection of TNF in mice induced profound thrombocytopenia, due to an increase of platelet consumption, that was evident after 1 h and lasted for 3 days. This process was evident in mice that were genetically deficient in TNFR2 (p75) but not in mice lacking TNFR1 (p55), indicating that the process is mediated by TNFR1-bearing cells. To explore the site of action of TNF, labeled platelets from TNFR1 −/− or +/+ donors were transferred to TNFR1 −/− or +/+ recipients. TNF induced the consumption of platelets from TNFR1 −/− donors when injected into +/+ recipients, while platelets from +/+ donors were not consumed when present in TNFR1 −/− recipients; this finding indicates that TNF acts on the TNFR1 of host cells but does not act on platelets. The expression of TNFRs is consistent with this interpretation, since TNFRs were not detected on platelets by flow cytometry. In megakaryocytes, the expression of TNFR1 was detected by immunohistochemistry. These results indicate that TNF induces platelet consumption by acting not on platelets directly but on the TNFR1 of other cells, presumably increasing the release of factors with agonist activity for platelets.
Neutrophils Are Major Contributors to Intraparenchymal Lung IL-1β Expression After Hemorrhage and Endotoxemia
Authors: Parsey Merdad V. , Tuder Rubin M. , Abraham Edward 
Published: January 2023

Abstract
Acute lung injury and the acute respiratory distress syndrome (ARDS) are significant causes of morbidity and mortality following sepsis and hemorrhage. Increased IL-1β production in the lung is important in the development of acute inflammatory lung injury. Although neutrophils are an important component of the inflammatory response that characterizes acute lung injury, there is little information to suggest that they are capable of initiating cytokine-mediated immune responses in the lung. To explore the role of neutrophils in the early stages of acute lung injury, we examined IL-1β production by mouse lung neutrophils after hemorrhage and endotoxemia. There was a significant increase in IL-1β expression among intraparenchymal pulmonary neutrophil/mononuclear cells (IPNMC) 1 h after hemorrhage or endotoxemia. IL-1β was detected only in a neutrophil-rich fraction of the IPNMC, but not in T and B lymphocytes positively selected from the IPNMC. Cyclophosphamide (CTX)-treated neutropenic mice expressed significantly less IL-1β in IPNMC after hemorrhage or endotoxemia compared with CTX-untreated controls. Immunohistochemical analysis of lung sections from mice after hemorrhage or endotoxemia revealed IL-1β expression in infiltrating neutrophils. These data indicate that IL-1β-producing neutrophils traffic to the lungs rapidly in response to hemorrhage or endotoxemia and support the concept that proinflammatory cytokine production by lung neutrophils may contribute to the development of lung injury after blood loss and sepsis.
Requirements for αd in IgG Immune Complex-Induced Rat Lung Injury
Authors: Shanley Thomas P. , Warner Roscoe L. , Crouch Larry D. , Dietsch Gregory N. , Clark Darcey L. , O’Brien Maggie M. , Gallatin W. Michael , Ward Peter A. 
Published: January 2023

Abstract
αd is a newly cloned adhesion molecule that forms a heterodimer with CD18. The requirement for αd in IgG immune complex-induced lung injury in rats has been evaluated by the use of blocking polyclonal and monoclonal antibodies to rat αd. Using whole lung extracts, Northern and Western blot analyses have revealed up-regulation of mRNA and αd protein in inflamed lungs. Immunostaining has revealed the presence of αd in lung tissue and in alveolar macrophages as early as 1 h after initiation of the inflammatory reaction. When polyclonal rabbit Ab to rat αd was coinstilled into lung together with Ab to BSA, lung injury (as determined by leakage of [125I]albumin into lung parenchyma) was significantly diminished. In parallel, there was reduced accumulation of neutrophils recoverable in bronchoalveolar lavage (BAL) fluids. These findings were associated with reduced levels of TNF-α as well as NO2−/NO3− in BAL fluids. A hamster mAb to rat αd was also protective in this lung injury model. Anti-αd inhibited in vitro production of NO2−/NO3− by rat alveolar macrophages (stimulated with LPS and IFN-γ) by approximately 60%. These data suggest that, in the lung inflammatory model employed, αd up-regulation occurs in lung macrophages and is necessary for expression of TNF-α, recruitment of neutrophils, and full development of lung injury.
Development of Murine Allergic Asthma Is Dependent Upon B7-2 Costimulation
Authors: Keane-Myers Andrea M. , Gause William C. , Finkelman Fred D. , Xhou X.-d. , Wills-Karp Marsha 
Published: January 2023

AbstractAllergic asthma is thought to be mediated by CD4+ T lymphocytes producing the Th2-associated cytokines, IL-4, and IL-5. Recently, the costimulatory molecules B7-1 and B7-2, which are expressed on the surface of APC, have been suggested to influence the development of Th1 vs Th2 immune responses. We examined the in vivo role of these costimulatory molecules in the pathogenesis of Th2-mediated allergen-induced airway hyperresponsiveness in a murine model of asthma. In this model, OVA-sensitized A/J mice develop significant increases in airway responsiveness, pulmonary eosinophilia, and pulmonary Th2 cytokine expression following aspiration challenge with OVA as compared with PBS-control animals. Strikingly, administration of anti-B7-2 mAb to OVA-treated mice abolished allergen-induced airway hyperresponsiveness, pulmonary eosinophilia, and elevations in serum IgG1 and IgE levels. Anti-B7-2 treatment of OVA-treated mice reduced both total lung IL-4 and IL-5 mRNA and bronchoalveolar lavage fluid IL-4 and IL-5 protein levels, with no significant changes in IFN-γ message or protein levels. In contrast, treatment with anti-B7-1 mAbs had no effect on allergen-induced airway hyperresponsiveness, IgE production, or cytokine production, however, it significantly suppressed pulmonary eosinophilia. We conclude that B7-2 provides the necessary costimulatory signal required for the development of in vivo allergic responses to inhaled allergen exposure.
Cutting Edge: Cytokine-Dependent Abortion in CBA × DBA/2 Mice Is Mediated by the Procoagulant fgl2 Prothombinase
Authors: Clark David A. , Chaouat Gerard , Arck Petra C. , Mittruecker Hans Willi , Levy Gary A. 
Published: January 2023

AbstractSpontaneous resorption in the CBA × DBA/2 model is attributed to NK cells, macrophages, and Th1-type cytokines. In vivo depletion of NK cells by anti-asialoGM1 Ab or macrophage depletion by silicon dioxide treatment reduced abortion rates, which could no longer be boosted by injecting TNF-α (which activates NK cells) or IFN-γ (which activates macrophages). TNF-α + γ-IFN coadministration aborted &gt;80% of the embryos whether or not NK cells or macrophages had been depleted or estradiol + progesterone was injected to correct potential reduction in ovarian function by cytokines. The cytokines also aborted IRF1+/+ C57BL/6 but not IRF1−/− females pregnant by IRF1+/+ DBA/2. Both spontaneous and cytokine-boosted abortions in CBA × DBA/2 were blocked by Ab to fgl2 prothombinase expressed by cytokine-stimulated vascular endothelial cells and monocytes; in vivo Ab depletion of granulocytes also prevented TNF-α + IFN-γ-induced abortions. Cytokine-triggered thrombotic/inflammatory processes in maternal uteroplacental blood vessels causes abortion.
CD40 Ligand Exerts Differential Effects on the Expression of Iγ Transcripts in Subclones of an IgM+ Human B Cell Lymphoma Line
Authors: Ford Gregory S. , Yin Chun Hui , Barnhart Bryan , Sztam Kevin , Covey Lori R. 
Published: January 2023

AbstractThe CD40:CD40 ligand (CD40L) interaction plays a critical role in T cell-dependent isotype switching. To elucidate the role of CD40 signaling in the activation of γ germline transcription and as an extension, in targeting Cγ regions for isotype switching, an IgM+ Burkitt lymphoma cell line (Ramos 2G6) was assayed for the up-regulation of germline γ transcripts after CD40L stimulation. Independent Ramos 2G6 subclones that either expressed (Iγ+) or did not express (Iγ−) basal levels of Iγ transcripts were assessed for their transcriptional response to CD40L signaling by contact with either a Jurkat T cell line (D1.1) or a transfected CD40L-expressing epithelial cell line (293/CD40L) in the presence or absence of IL-4. Both Iγ− and Iγ+ Ramos 2G6 subclones cultured with IL-4 and CD40L markedly up-regulated germline transcription predominantly from the γ1, γ2, and γ3 subclasses over levels obtained with IL-4 alone. In addition, these two signals were required to obtain de novo switch recombination. However, incubation with CD40L alone resulted in a substantial increase in germline transcription only in the Iγ+ and not the Iγ− subclones. Observed basal transcription at the γ1 locus also correlated with the ability of not only the γ1 locus, but also the γ2 and γ3 loci, to up-regulate germline transcripts in response to CD40 signaling. These data are consistent with CD40:CD40L contact up-regulating germline transcription only after the B cell has received a signal that alters the transcriptional state of the heavy chain locus.
Natural Killer Cell Development and Function Precede αβ T Cell Differentiation in Mouse Fetal Thymic Ontogeny
Authors: Carlyle James R. , Michie Alison M. , Cho Sarah K. , Zúñiga-Pflücker Juan Carlos 
Published: January 2023

AbstractNatural killer (NK) cells mediate MHC-unrestricted cytolysis of virus-infected cells and tumor cells. In the adult mouse, NK cells are bone marrow-derived lymphocytes that mature predominantly in extrathymic locations but have also been suggested to share a common intrathymic progenitor with T lymphocytes. However, mature NK cells are thought to be absent in mouse fetal ontogeny. We report the existence of thymocytes with a mature NK cell phenotype (NK1.1+/CD117−) as early as day 13 of gestation, approximately 3 days before the appearance of CD4+/CD8+ cells in T lymphocyte development. These mature fetal thymic NK cells express genes associated with NK cell effector function and, when freshly isolated, display MHC-unrestricted cytolytic activity in vitro. Moreover, the capacity of fetal thymic NK cells for sustained growth both in vitro and in vivo, in addition to their close phenotypic resemblance to early precursor thymocytes, confounds previous assessments of NK lineage precursor function. Thus, mature NK cells may have been inadvertently included in previous attempts to identify multipotent and bipotent precursor thymocytes. These results provide the first evidence of functional NK lymphocytes in mouse fetal ontogeny and demonstrate that NK cell maturation precedes αβ T cell development in the fetal thymus.
Presence of Activated Antigen-Binding B Cells During Immunization Enhances Relative Levels of IFN-γ in T Cell Responses
Authors: Pasare Chandrashekhar , Morafo Vivian , Entringer Maureen , Bansal Pratima , George Anna , Bal Vineeta , Rath Satyajit , Durdik Jeannine M. 
Published: January 2023

AbstractTo examine the influence of Ag presentation by B cells on immune responses, we have used mice transgenic for an Ig heavy chain from a monoclonal anti-azobenzenearsonate (Ars) Ab to deliver Ag to B cells during immunization. A large proportion of transgene-expressing B cells in these mice binds Ars, while transgenic serum Ig shows poor Ars binding. Transgenic B cells present Ars proteins better than their nonhaptenated counterparts. This is associated with an increase in the proliferative responses of transgenic T cells to Ars protein immunization. Although B cell numbers in the transgenic mice are lower, many B cells in them show an activated phenotype, as identified by altered surface levels of peanut agglutinin reactivity, CD23, CD24, CD44, CD62L, and CD86. Even against nonhaptenated immunogens, transgenic responses show significant enhancement in the relative proportions of the Th1 cytokine IFN-γ over the Th2 cytokines IL-4 and IL-10. Haptenated immunogens further enhance the predilection of transgenic mice to produce relatively more IFN-γ. Consistent with this, there is an increase in IgG2a/IgG1 ratios in serum Abs in response to haptenated immunogens in transgenic mice. Adoptive transfer of primed hapten-specific secondary B cells into nontransgenic mice also induces an increase in relative levels of IFN-γ in response to haptenated immunogens. Thus, presentation of immunogen in vivo by activated Ag-binding B cells contributes to enhanced immunogenicity and a Th1 cytokine bias.
Role of Proteasomes in T Cell Activation and Proliferation
Authors: Wang Xin , Luo Hongyu , Chen Huifang , Duguid William , Wu Jiangping 
Published: January 2023

Abstract
The role of proteasomes in T cell activation, proliferation, and apoptosis was investigated using a proteasome-specific inhibitor lactacystin (LAC). Inhibition of the proteasome activity by LAC repressed the mitogen-induced T cell proliferation. The proteasome activity was definitively required for the T cells to progress from the G0 to S phase. It was necessary to optimize the progress from the G1/S boundary to the G2/M phase, but not for the progress from the G2/M phase to the next G1 phase. Probably as a result of a blockage of cell cycle progress, the cycling, but not the resting, T cells underwent apoptosis when treated with LAC. Mechanistically, we have found that cyclin-dependent kinase-2 (CDK2) and the cyclin E-associated kinase (largely CDK2), but not CDK4, in the G1 phase were strongly inhibited by LAC. This could be an important mechanism for the proteasome to regulate the cell cycle. The degradation of cyclin E in the late G1 and early S phases was dependent on the proteasome, although it was unlikely that this accounted for the observed inhibition of T cell proliferation. There was a reduced decay of p27Kip1 in the late G1 phase when the proteasome activity was suppressed, and this might be a contributing mechanism for the observed inhibition of CDK2 activity. Interestingly, p21Cip1 was up-regulated during the G1 phase, and the up-regulation was inhibited by LAC. Our study shows that the proteasome plays pivotal roles in regulating T cell activation and proliferation, and its effect is probably exerted through multiple mechanisms.
Separation of Function Between the Domains of Toxic Shock Syndrome Toxin-1
Authors: Wahlsten Jennifer L. , Ramakrishnan S. 
Published: January 2023

AbstractToxic shock syndrome toxin-1 (TSST1) is a superantigenic exotoxin produced by certain strains of Staphylococcus aureus. Structurally, TSST1 is composed of two domains: residues determined by crystallography to directly interact with MHC II molecules reside within the N-terminal domain, while TSST1 residues critical for superantigenicity are within the C-terminal domain. In this study, we expressed the individual N- and C-terminal domains of TSST1 in Escherichia coli and studied their biologic activities. The TSST1 N-terminal domain (TSST(1–87)) did not induce proliferation of human PBLs or release of TNF-β, but did induce TNF-α release. However, TSST1-elicited proliferation and release of both TNF isoforms were inhibited by a molar excess of TSST(1–87). The TSST1 C-terminal domain (TSST(88–194)) did not bind MHC II molecules, yet it elicited production of TNF-α and TNF-β, and induced TCR Vβ-specific proliferation similarly to intact TSST1. When covalently cross-linked to tumor cells, TSST(88–194) elicited a local in vivo antitumor response indistinguishable from TSST1. Although intact TSST1 causes lethal shock in vivo, the individual domains of this molecule may have therapeutic potential: the N-terminal domain to antagonize lymphocyte activation and TNF release during acute TSST1-precipitated toxic shock syndrome, and the C-terminal domain to stimulate antitumor responses without MHC II binding.
Distinct Phospholipases A2 Regulate the Release of Arachidonic Acid for Eicosanoid Production and Superoxide Anion Generation in Neutrophils
Authors: Tithof Patricia K. , Peters-Golden Marc , Ganey Patricia E. 
Published: January 2023

Abstract
Arachidonic acid (AA) released from membrane phospholipids by phospholipase A2 (PLA2) is important as a substrate for eicosanoid formation and as a second messenger for superoxide anion (O2−) generation in neutrophils. Different isoforms of PLA2 in neutrophils might mobilize AA for different functions. To test this possibility, we sought to characterize the PLA2s that are activated by the neutrophil stimuli, Aroclor 1242, a mixture of polychlorinated biphenyls, and A23187, a calcium ionophore. Both Aroclor 1242 and A23187 caused release of [3H]AA; however, O2− production was seen only in response to Aroclor 1242. Eicosanoids accounted for &gt;85% of the radioactivity recovered in the supernatant of A23187-stimulated cells but &lt;20% of the radioactivity recovered from cells exposed to Aroclor 1242. Omission or chelation of calcium abolished A23187-induced AA release, but did not alter AA release in Aroclor 1242-stimulated neutrophils. AA release and O2− production in response to Aroclor 1242 were inhibited by bromoenol lactone (BEL), an inhibitor of calcium-independent PLA2. BEL, however, did not alter A23187-induced release of AA. Cell-free assays demonstrated both calcium-dependent and calcium-independent PLA2 activity. Calcium-independent activity was inhibited &gt;80% by BEL, whereas calcium-dependent activity was inhibited &lt;5%. Furthermore, calcium-independent, but not calcium-dependent, PLA2 activity was significantly enhanced by Aroclor 1242. These data suggest that Aroclor 1242 and A23187 activate distinct isoforms of PLA2 that are linked to different functions: Aroclor 1242 activates a calcium-independent PLA2 that releases AA for the generation of O2−, and A23187 activates a calcium-dependent PLA2 that mobilizes AA for eicosanoid production.
Fever-Range Hyperthermia Enhances L-Selectin-Dependent Adhesion of Lymphocytes to Vascular Endothelium
Authors: Wang Wan-Chao , Goldman Lorin M. , Schleider David M. , Appenheimer Michelle M. , Subjeck John R. , Repasky Elizabeth A. , Evans Sharon S. 
Published: January 2023

Abstract
The L-selectin leukocyte adhesion molecule plays an important role in controlling leukocyte extravasation in peripheral lymph nodes and at sites of tissue injury or infection. Although febrile responses during infection and inflammation are associated with enhanced immune activity, the contribution of fever-range temperatures to controlling lymphocyte recruitment to tissues has not been previously examined. In this report we provide evidence that direct exposure of lymphocytes to fever-range temperatures (38–41°C) in vitro for 9 to 24 h resulted in a &gt;100% increase in L-selectin-dependent adhesion of these cells to lymph node high endothelial venules (HEV). Moreover, culture of lymphocytes under hyperthermia conditions markedly enhanced the ability of these cells to traffic in an L-selectin-dependent manner to peripheral lymph nodes, mesenteric lymph nodes, and Peyer’s patches. In contrast, febrile temperatures did not increase LFA-1 function as assessed by measuring lymphocyte adhesion to ICAM-1–3T3 transfectants. Fever-range hyperthermia further did not increase L-selectin surface density on lymphocytes or L-selectin-dependent recognition of soluble carbohydrate substrates; however, a marked increase in ultrastructural immunogold-labeling of L-selectin was observed in response to thermal stimuli. These results suggest that elevated temperatures enhance L-selectin adhesion and/or avidity through the regulation of L-selectin conformation or organization in the plasma membrane. Finally, the observed thermal effects on L-selectin adhesion were attributed to soluble factors in the conditioned medium of heat-treated cells. Taken together, these data provide new insight into the potential physiologic role of the febrile response in enhancing lymphocyte recruitment to tissues through the regulation of L-selectin adhesion.
Cutting Edge: Constitutive Activation of TCR Signaling Molecules in IL-2-IndependentHerpesvirus saimiri-Transformed T Cells
Authors: Noraz Nelly , Saha Kunal , Ottones Florence , Smith Susan , Taylor Naomi 
Published: January 2023

AbstractBoth human T cell leukemia virus type I and simian Herpesvirus saimiri (HVS) transform human T cells in vitro. Although IL-2-independent growth in human T cell leukemia virus type I-transformed T cells is associated with constitutive phosphorylation of JAK/STAT kinases, we now demonstrate that different mechanisms may be responsible for the ability of HVS-transformed T cells to proliferate in the absence of exogenous cytokines. The IL-2 independence of an HVS-transformed cell line correlated with constitutive activation of protein tyrosine kinases known to be induced following TCR engagement. Thus, in these cells we observed increased phosphotransferase activity of Lck as well as constitutive tyrosine phosphorylation of the TCR-associated ZAP-70 kinase and expression of the related Syk protein tyrosine kinase. While Syk is generally not expressed in activated T cells, its introduction has been shown to enhance TCR responsiveness. These results suggest that distinct signal transduction cascades can participate in the transition of T cells to IL-2 independence.
Cutting Edge: Apoptosis Induced by a Chimeric Fas/FLICE Receptor: Lack of Requirement for Fas- or FADD-Binding Proteins
Authors: Memon Sarfraz A. , Hou Jing-zhou , Moreno M. Belen , Zacharchuk Charles M. 
Published: January 2023

Abstract
Current models for Fas (CD95)-mediated apoptosis suggest that FLICE/caspase-8 is recruited and activated, which results in cell death. However, the role of additional molecules in Fas signaling and FLICE activation is not clear. A chimeric Fas/FLICE (F/F) receptor, containing the extracellular/transmembrane portion of Fas and the caspase region of FLICE, mediated anti-Fas apoptosis. FLICE protease subunits were generated from the F/F precursor. Killing induced by Fas, but not F/F, was blocked by a dominant negative FADD. Apoptosis triggered through Fas and F/F was inhibited by coexpression of CrmA and p35, but not Bcl-xL. F/F bypassed Fas resistance in COS-7 cells and blocking by the death effector domain (DED)-containing viral protein MC159. These results show that: 1) F/F induces cell death, indicating that FLICE activation is sufficient for apoptosis and does not require additional Fas- or FADD-binding proteins; and 2) F/F bypasses proximal defects in Fas signaling that prevent FLICE recruitment or activation.
Antigen-Specific IgG Responses from Naive Human Splenocytes: In Vitro Priming Followed by Antigen Boost in the SCID Mouse
Authors: Brams Peter , Nguyen Mai-Lan , Chamat Soulaima , Royston Ivor , Morrow Phillip R. 
Published: January 2023

AbstractHigh titers of Ag-specific human IgG were consistently achieved in SCID mice reconstituted with human splenocytes that had been primed with Ag in vitro and then boosted with Ag after engraftment into SCID mice. Specific human IgG titers in the hu-SPL-SCID mice reached approximately 1:4 × 105 when the mice were immunized with a neo-antigen, whereas titers reached 1:2 × 106 when recall responses were induced. Booster immunizations with Ag 21 days after the initial in vivo boost further enhanced this response, and specific human IgG titers of 1:17 × 106 were achieved. This represented an essentially monospecific IgG population. These responses were CD4+ T cell dependent. In addition, affinity maturation of the human Ab responses was observed. Spleens of hu-SPL-SCID mice with Ag-specific titers ≤1:1 × 106 were often significantly enlarged and often displayed visible tumors. Fourteen of sixteen B cell tumors removed from spleens of five such hu-SPL-SCID mice, produced Abs that were specific for the immunizing Ags. From such tumor, cloned cell lines were established. One such mAb, MLN-7 (γ1,κ), was raised to tetanus toxoid and had no identified cross-reactivity.