bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mice Lacking NK Cells Develop an Efficient Th1 Response and Control CutaneousLeishmania majorInfection
Authors: Satoskar Abhay R. , Stamm Luisa M. , Zhang Xingmin , Satoskar Anjali A. , Okano Mitsuhiro , Terhorst Cox , David John R. , Wang Baoping 
Published: January 2023

AbstractNK cells are believed to play a critical role in the development of immunity against Leishmania major. We recently found that transplantation of wild-type bone marrow cells into neonatal tgε 26 mice, which are deficient in T and NK cells, resulted in normal T cell development, but no or poor NK cell development. Using this novel model we analyzed the role of NK cells in the development of Th1 response and control of cutaneous L. major infection. Mice selectively lacking NK cells (NK−T+) developed an efficient Th1-like response, produced significant amounts of IL-12 and IFN-γ, and controlled cutaneous L. major infection. Administration of neutralizing IL-12 Abs to NK−T+ mice during L. major infection resulted in exacerbation of the disease. These results demonstrate that NK cells are not critical for development of protective immunity against L. major. Furthermore, they indicate that IL-12 can induce development of Th1 response independent of NK cells in NK−T+ mice following L.major infection.
C5a Suppresses the Production of IL-12 by IFN-γ-Primed and Lipopolysaccharide-Challenged Human Monocytes
Authors: Wittmann Miriam , Zwirner Jörg , Larsson Vivi-Ann , Kirchhoff Konstanze , Begemann Gabriele , Kapp Alexander , Götze Otto , Werfel Thomas 
Published: January 2023

Abstract
IL-12 is a key mediator of the immune response, skewing T lymphocytes toward a type 1 cytokine pattern. Priming with IFN-γ or GM-CSF is required for expression of IL-12p70 by cells in which IL-12 is inducible by bacterial products such as LPS. We here show for the first time that the production of bioactive IL-12 by human monocytes can be significantly suppressed by C5a if applied to IFN-γ-primed monocytes before LPS stimulation. There was a dose-dependent suppression by IL-12 (p70) on the levels of intracellular cytokine production and cytokine secretion. mRNA studies consistently showed a reduction of IL-12p40 and IL-12p35 expression by stimulation in the presence of C5a. The results of several different experimental approaches suggest that IL-12 down-regulation was not due to endogenous IL-10, IL-4, or PGE2 production induced by C5a. Moreover, stimulation of IFN-γ-primed monocytes with C5a did not lead to a down-regulation of the CD14 Ag, which is an LPS receptor. These findings show that the anaphylatoxin C5a has the capacity to directly interact with the complex regulation of IL-12.
Mechanisms of IL-10 Production in Human Microglia-T Cell Interaction
Authors: Chabot Sophie , Williams Gary , Hamilton Mark , Sutherland Garnette , Yong V. Wee 
Published: January 2023

Abstract
IL-10, a cytokine with important anti-inflammatory properties, is generated within the CNS during neuroinflammation. The mechanism for its production is poorly understood. Since infiltrating lymphocytes come into close proximity with the macrophage-like cells of the CNS, the microglia, we have used an in vitro human microglia-T cell coculture system to address the mechanisms of IL-10 production. We demonstrate that microglia or activated T cells alone secrete negligible amounts of IL-10, but that their coculture results in significant IL-10 production, which was effected by both cell types. IL-10 generation was cell contact dependent, and treatment with anti-CD40, CTLA-4-Fc, or anti-CD23 decreased the IL-10 content in microglia-T cell cocultures. The combination of anti-CD40 and CTLA-4-Fc reduced IL-10 levels to the negligible amounts seen with T cells or microglia in isolation. By also measuring TNF-α levels, specificity of cytokine regulation was observed; while anti-CD40 and CTLA-4-Fc reduced IL-10 and TNF-α levels, anti-CD23 did not affect TNF-α while attenuating IL-10 generation. Anti-very late Ag-4, which decreased TNF-α levels, did not affect IL-10. These results implicate the CD40, B7, and CD23 pathways in IL-10 production following microglia-T cell encounter and have relevance to the regulation of an anti-inflammatory response within the CNS.
Cyclosporin A-Induced Autologous Graft-Versus-Host Disease: A Prototypical Model of Autoimmunity and Active (Dominant) Tolerance Coordinately Induced by Recent Thymic Emigrants
Authors: Wu Dianna Y. , Goldschneider Irving 
Published: January 2023

AbstractCyclosporin A (CSA)-induced autologous graft-vs-host disease (autoGVHD) is an autoimmune syndrome initiated by autoeffector T cells presumed to be exported from the thymus during CSA treatment. The appearance of noncytotoxic immunoregulatory T cell activity after cessation of CSA treatment is also thymus dependent. In the present study, we have tested the hypothesis that both autoeffector and immunoregulatory T cells in CSA-treated rats are recent thymic emigrants (RTEs). Local syngeneic graft-vs-host reaction (synGVHR) and timed thymectomy (Tx) assays revealed that autoeffector T cells appear initially in the thymus and are promptly exported to lymph nodes (LN) during the first week of CSA treatment. In contrast, immunoregulatory thymocytes are first detectable by local synGVHR inhibition assays during the second week of CSA treatment but are not exported to LN until ∼4 days post-CSA. Both the autoeffector and immunoregulatory T cells in LN express Thy-1, a selective marker for RTEs in the rat. However, the autoeffector RTEs have a CD4+8+ phenotype, whereas the immunoregulatory RTEs have a CD4+8− phenotype. Thus, the coordinate formation in and release from the thymus cortex and medulla of autoeffector and immunoregulatory T cells in CSA-treated rats directly demonstrates that centrally induced, nondeletional tolerance can serve as a fail-safe mechanism by which clones of autoeffector T cells that have escaped intrathymic negative selection for self-MHC class II Ag can be suppressed postthymically.
Treatment of Breast Cancer with Fibroblasts Transfected with DNA from Breast Cancer Cells
Authors: de Zoeten Edwin , Carr-Brendel Victoria , Markovic Dubravka , Taylor-Papadimitriou Joyce , Cohen Edward P. 
Published: January 2023

Abstract
This investigation was based on the hypothesis that weakly immunogenic, breast cancer-associated Ags, the products of mutant or dysregulated genes in the malignant cells, will be expressed in a highly immunogenic form by semiallogeneic IL-2-secreting fibroblasts transfected with DNA from breast cancer cells. (Classic studies indicate that transfection of genomic DNA can stably alter both the genotype and the phenotype of the cells that take up the exogenous DNA.) To investigate this question, we transfected LM mouse fibroblasts (H-2k) modified to secrete IL-2 with genomic DNA from a breast adenocarcinoma that arose spontaneously in a C3H/He mouse (H-2k). To increase their nonspecific immunogenic properties, the fibroblasts were also modified before transfection to express allogeneic MHC determinants (H-2Kb). Afterward, the IL-2-secreting semiallogeneic cells were cotransfected with DNA from the spontaneous breast neoplasm, along with a plasmid (pHyg) conferring resistance to hygromycin. Pooled colonies of hygromycin-resistant cells were then tested in C3H/He mice for their immunotherapeutic properties against the growth of the breast neoplasm. The results indicated that tumor-bearing mice immunized with the transfected cells survived significantly longer than mice in various control groups. Similar beneficial effects were seen in C57BL/6 mice injected with a syngeneic breast carcinoma cell line (EO771) and semiallogeneic, IL-2-secreting fibroblasts transfected with DNA from EO771 cells. The immunity was mediated by CD8+ T cells since immunized mice depleted of CD8+ cells failed to resist tumor growth.
The p65 Subunit of NF-κB Is Redundant with p50 During B Cell Proliferative Responses, and Is Required for Germline CH Transcription and Class Switching to IgG3
Authors: Horwitz Bruce H. , Zelazowski Piotr , Shen Yi , Wolcott Karen M. , Scott Martin L. , Baltimore David , Snapper Clifford M. 
Published: January 2023

Abstract
B cells lacking individual NF-κB/Rel family members exhibit defects in activation programs. We generated small resting B cells lacking p65 or p50 alone, or lacking both p50 and p65, then evaluated the ability of these cells to proliferate, secrete Ig, and undergo Ig class switching. B cells lacking p65 proliferated well in response to all stimuli tested. However, these cells demonstrated an isolated defect in switching to IgG3, which was associated with a decrease in γ3 germline CH gene expression. Whereas, previously reported, B cells lacking p50 alone had a severe proliferative defect in response to LPS, a moderate defect in response to CD40 ligand (CD40L), and normal proliferation to Ag receptor cross-linking using dextran-conjugated anti-IgD Abs (αδ-dex), B cells lacking both p50 and p65 exhibited severely impaired proliferation in response to LPS, αδ-dex, and CD40L. This defect could be overcome by simultaneous administration of αδ-dex and CD40L. In response to this latter combination of stimuli, B cells lacking both p50 and p65 secreted Ig and underwent isotype switching to IgG1 as efficiently as B cells lacking p50 alone. These data demonstrate a role for the p65 subunit of NF-κB in germline CH gene expression as well as functional redundancy between p50 and p65 during proliferative responses.
Fas Ligand Induction in Human NK Cells Is Regulated by Redox Through a Calcineurin-Nuclear Factors of Activated T Cell-Dependent Pathway
Authors: Furuke Keizo , Shiraishi Mitsuhiro , Mostowski Howard S. , Bloom Eda T. 
Published: January 2023

AbstractFas ligand (FasL) on cytotoxic lymphocytes is important for mediating apoptosis of activated lymphocytes and other target cells. We have reported that NK cell functions, such as proliferation, cell death, and killing activity, are subject to regulation by cellular redox status. Here, we report that expression of FasL protein and mRNA in activated NK cells is also regulated by redox. Ligation of CD16 on IL-2-preactivated NK cells resulted in reduction of intracellular peroxide level as well as induction of FasL expression. This CD16-induced FasL expression was suppressed by oxidative stress, including thiol deprivation or treatment with hydrogen peroxide (H2O2). Addition of thiol-reducing compounds, such as l-cystine, 2-ME, or N-acetyl cysteine, restored FasL expression. These data suggest that CD16 stimulation requires cellular reducing status for FasL induction in NK cells. Because FasL gene activation following CD16 cross-linking is regulated by the NF of activated T cells (NFAT), we examined the effect of oxidative stresses on NFAT activation. Electrophoretic mobility shift assays revealed that both thiol insufficiency and H2O2 treatment suppressed DNA-binding activity of NFAT and that addition of thiol-reducing compounds reversed or even enhanced it. Furthermore, these oxidative stresses inhibited activity of calcineurin, a serine/threonine phosphatase that regulates NFAT activation. These results suggest that suppression of calcineurin and NFAT activation is a mechanism by which oxidative stress inhibits FasL induction in activated NK cells and further support the hypothesis that thiol-reducing compounds might be required for maintenance of optimal NK functions under physiologic oxidative conditions.
Human Effector Memory T Cells Express CD86: A Functional Role in Naive T Cell Priming
Authors: Jeannin Pascale , Herbault Nathalie , Delneste Yves , Magistrelli Giovanni , Lecoanet-Henchoz Sybille , Caron Gersende , Aubry Jean-Pierre , Bonnefoy Jean-Yves 
Published: January 2023

AbstractThe glycoprotein CD86 expressed on APCs provides a costimulatory signal necessary for an efficient activation of naive T cells. In contrast, there is controversy about the condition of expression and the function of CD86 on T cells. In this study, we have analyzed the phenotype and the biological activity of CD86+ T cells generated from human PBMC. Results show that CD86 expression on T cells is induced by long term stimulation via CD3 and IL-2R and is down-regulated as the cells become quiescent. The CD86-expressing cells are memory effector T cells: 1) they express CD45RO and high levels of the activation markers CD25, CD54, and HLA-Dr; 2) they selectively express CD30, CD40-ligand, and CD70; and 3) in response to stimulation, most of them produce IFN-γ before dying by apoptosis. We then analyzed whether CD86 expressed on T cells is functional. Results show that paraformaldehyde-fixed CD86+ T cells enhance the proliferation and production of IFN-γ by anti-CD3 mAb-stimulated naive T cells and induce proliferation of resting allogenic T cells. All these effects are prevented by neutralizing anti-CD86 mAbs. In contrast, we report no autocrine effect of CD86 in CD86+ T cell activation. In conclusion, these data show that human memory effector T cells express a functional form of CD86 that can costimulate naive T cell responses.
β1 Integrin Cross-Linking Inhibits CD16-Induced Phospholipase D and Secretory Phospholipase A2 Activity and Granule Exocytosis in Human NK cells: Role of Phospholipase D in CD16-Triggered Degranulation
Authors: Milella Michele , Gismondi Angela , Roncaioli Paola , Palmieri Gabriella , Morrone Stefania , Piccoli Mario , Frati Luigi , Cifone Maria Grazia , Santoni Angela 
Published: January 2023

AbstractRecent data indicate that integrin-generated signals can modulate different receptor-stimulated cell functions in both a positive (costimulation) and a negative (inhibition) fashion. Here we investigated the ability of β1 integrins, namely α4β1 and α5β1 fibronectin receptors, to modulate CD16-triggered phospholipase activation in human NK cells. β1 integrin simultaneous cross-linking selectively inhibited CD16-induced phospholipase D (PLD) activation, without affecting either phosphatidylinositol-phospholipase C or cytosolic phospholipase A2 (PLA2) enzymatic activity. CD16-induced secretory PLA2 (sPLA2) protein release as well as its enzymatic activity in both cell-associated and soluble forms were also found to be inhibited upon β1 integrin coengagement. The similar effects exerted by specific PLD pharmacological inhibitors (2,3-diphosphoglycerate, ethanol) suggest that in our experimental system, sPLA2 secretion and activation are under the control of a PLD-dependent pathway. By using pharmacological inhibitors (2,3-diphosphoglycerate, wortmannin, ethanol) we also demonstrated that PLD activation is an important step in the CD16-triggered signaling cascade that leads to NK cytotoxic granule exocytosis. Consistent with these findings, fibronectin receptor engagement, by either mAbs or natural ligands, resulted in a selective inhibition of CD16-triggered, but not of PMA/ionomycin-induced, degranulation that was reversed by the exogenous addition of purified PLD from Streptomyces chromofuscus.
Serial TCR Engagement and Down-Modulation by Peptide:MHC Molecule Ligands: Relationship to the Quality of Individual TCR Signaling Events
Authors: Itoh Yasushi , Hemmer Bernhard , Martin Roland , Germain Ronald N. 
Published: January 2023

AbstractIn the present study, we examined the relationships among quantitative aspects of TCR engagement as measured by receptor down-modulation, functional responses, and biochemical signaling events using both mouse and human T cell clones. For T cells from both species, ligands that are more potent in inducing functional responses promote TCR down-modulation more efficiently than weaker ligands. At low ligand density, the number of down-modulated TCR exceeds the number of available ligands by as much as 80–100:1 in the optimal human case, confirming the previous description of serial ligand engagement of TCR (Valitutti, et al. 1995. Nature 375:148–151). A previously unappreciated relationship involving TCR down-modulation, the pattern of proximal TCR signaling, and the extent of serial engagement was revealed by analyzing different ligands for the same TCR. Functionally, more potent ligands induce a higher proportion of fully tyrosine phosphorylated ζ-chains and a greater amount of phosphorylated ZAP-70 than less potent ligands, and the number of TCR down-modulated per available ligand is higher with ligands showing this full agonist-like pattern. The large number of receptors showing partial ζ phosphorylation following exposure to weak ligands indicates that the true extent of TCR engagement and signaling, and thus the amount of sequential engagement, is underestimated by measurement of TCR down-modulation alone, which depends on full receptor activation. These data provide new insight into T cell activation by revealing a clear relationship among intrinsic ligand quality, signal amplification by serial engagement, functional T cell responses, and observable TCR clearance from the cell surface.
Identification of Residues in the First Domain of Human Fcα Receptor Essential for Interaction with IgA
Authors: Wines Bruce D. , Hulett Mark D. , Jamieson Gary P. , Trist Halina M. , Spratt Joanne M. , Hogarth P. Mark 
Published: January 2023

AbstractThe FcR family contains multiple receptors for Igs, of which the most distantly related (∼20%) is the IgA receptor (human FcαR), being more homologous (∼35%) to another family of killer-inhibitory receptor-related immunoreceptors with a 19q13.4 chromosomal location in humans. This study of the FcαR demonstrated that, like several IgG receptors, FcαR is a low affinity receptor for Ab (Ka ∼ 106 M−1). Rapid dissociation of the rsFcαR:IgA complex (t1/2 ∼ 25 s) suggests that monomer IgA would bind transiently to cellular FcαRs, while IgA immune complexes could bind avidly. Mutagenesis of histidyl 85 and arginyl 82, in the FG loop of domain 1, demonstrated that these residues were essential for the IgA-binding activity of FcαR, while arginyl 87 makes a minor contribution to the binding activity of the receptor. This site is unusual among the Fc receptors (FcγRII, FcγRIII, and FcεRI), in which the ligand binding site is in domain 2 rather than domain 1, but like FcαR, the FG loop comprises part of the ligand binding site. The putative F and G strands flanking the FcαR ligand binding site are highly homologous in the other killer-inhibitory receptor-related immunoreceptors, suggesting they comprise a conserved structural element on which divergent FG loops are presented and participate in the specific ligand interactions of each of these receptors.
Position Effects of Variable Region Carbohydrate on the Affinity and In Vivo Behavior of an Anti-(1→6) Dextran Antibody
Authors: Coloma M. Josefina , Trinh Ryan K. , Martinez Alexander R. , Morrison Sherie L. 
Published: January 2023

Abstract
IgG is a glycoprotein with an N-linked carbohydrate structure attached to the CH2 domain of each of its heavy chains. In addition, the variable regions of IgG often contain potential N-linked carbohydrate addition sequences that frequently result in the attachment of V region carbohydrate. Nonetheless, the precise role of this V region glycan remains unclear. Studies from our laboratory have shown that a naturally occurring somatic mutant of an anti-dextran Ab that results in a carbohydrate addition site at Asn58 of the VH has carbohydrate in the complementarity-determining region 2 (CDR2) of the VH, and the presence of carbohydrate leads to an increase in affinity. However, carbohydrate attached to nearby positions within CDR2 had variable affects on affinity. In the present work we have extended these studies by adding carbohydrate addition sites close to or within all the CDRs of the same anti-dextran Ab. We find that carbohydrate is attached to all the novel addition sites, but the extent of glycosylation varies with the position of the site. In addition, we find that the position of the variable region carbohydrate influences some functional properties of the Ab, including those usually associated with the V region such as affinity for Ag as well as other characteristics typically attributed to the Fc such as half-life and organ targeting. These studies suggest that modification of variable region glycosylation provides an alternate strategy for manipulating the functional attributes of the Ab molecule and may shed light on how changes in carbohydrate structure affect protein conformation.
Mycobacterium bovisBacille Calmette-Guérin Enhances Pathogenicity of Simian Immunodeficiency Virus Infection and Accelerates Progression to AIDS in Macaques: A Role of Persistent T cell Activation in AIDS Pathogenesis
Authors: Zhou Dejiang , Shen Yun , Chalifoux Laura , Lee-Parritz David , Simon Meredith , Sehgal Prabhat K. , Zheng Leiya , Halloran Matilda , Chen Zheng W. 
Published: January 2023

AbstractIt has recently been proposed that Mycobacterium tuberculosis may enhance the pathogenicity of HIV infections and accelerate the course of HIV disease. This hypothesis has been tested in the present study using a simian immunodeficiency virus of macaques (SIVmac)/Mycobacterium bovis bacille Calmette-Guérin (BCG)-coinfected macaque model. Naive and chronically SIVmac-infected monkeys were evaluated. Following BCG inoculation, the SIVmac-infected monkeys exhibited the dominant responses of TCR-β complementarity-determining region 3-restricted T cell subpopulations. This BCG-driven T cell activation correlated with a marked increase in viral loads in SIVmac-infected monkeys. Moreover, the prolonged T cell activation coincided with the enhanced decline of CD4+ PBL counts and the accelerated progression to clinical AIDS in the coinfected monkeys, suggesting that Mycobacterium-driven T cell activation may be the mechanism underlying the enhanced pathogenicity of AIDS virus infection in the coinfected individuals. Within 2 to 7 mo after BCG coinfection, all chronically SIVmac-infected monkeys died from SIV-induced AIDS including tuberculosis-like disease. Surprisingly, the naive monkeys manifested a T cell activation-related toxic shock syndrome and a profound depletion of CD4+ lymphocytes 2 wk after simultaneous SIVmac/BCG inoculation. These naive animals died 2 mo after SIVmac/BCG inoculation, with the evidence of the persistent SIV p27 antigenemia and SIVmac-induced disease. In contrast, the normal monkeys not infected with SIVmac survived BCG infection; the control SIVmac-infected animals showed a natural course of chronic SIV infection. Thus, results from this SIV/BCG coinfection model strongly support the hypothesis that active coinfection with HIV and Mycobacterium can impact remarkably on the AIDS virus-induced disease.