bluuebunny/crossref_metadata_2025_split huggingface dataset:
UltraSensitive Human Cytokine 10-Plex for the Luminex xMAP™ Platform (B200)
Authors: Bhakta Sheetal , Liu Nina , Fu Yubin , Brodey Mary Michael , Wang Jimin 
Published: January 2023

Abstract
Profiling low concentrations of Th1/Th2 and proinflammatory cytokines can yield valuable information about immune system status in clinical and subclinical disease states. To facilitate cytokine profile analysis, Invitrogen has recently developed an UltraSensitive Cytokine 10-Plex Assay for the Luminex xMAP™ Platform. Analytes include human GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNF-α. This assay system is validated for high content, high sensitivity serum, plasma, and tissue culture medium applications. Through careful choice of antibody pairs and use of proprietary buffer formulations, we are able to achieve an improvement in sensitivity of approximately 10-fold compared to our conventional cytokine assays for the Luminex xMAP™ platform. Sensitivities for all 10 analytes are 1 picogram per milliliter or less. Other performance characteristics, including assay precision, recovery, linearity of dilution, and calibration to NIBSC reference preparations, are similar to our conventional assays.
Co-expression of apoptosis-related molecules on activated CD8+CD38+ T-cells is associated with HIV-1 disease progression (B213)
Authors: Ríos-Olivares Eddy , Rodríguez José W. , Rodríguez Luis A. , Nair Madhavan , Hunter Robert 
Published: January 2023

Abstract
HIV-1 disease progression has been associated with a remarkable increase of CD8+CD38+ T-cells. It has been documented that a significant distribution of HIV-specific CD8+ T-cells resides in the CD8+CD38+ T-cell sub-population. The failure of HIV-specific CD8+CD38+ T-cells to control HIV-1 infection has been attributed to several mechanisms including apoptosis. However, the relationship between the CD38 expression and molecular events involved in CD8+ T-cell apoptosis is not well understood. We evaluated the expression of two membrane-associated apoptosis-related molecules: CXCR4 and CD95, and two cytoplasm-associated apoptosis-related molecules: Bcl-2 and active caspase-3 in 41 HIV-1 positive patients and 15 HIV-1 negative individuals. HIV-1 infection alters the level of expression of CD38, CD95, CXCR4, Bcl-2, and active caspase-3. A positive correlation was established between CD95, CXCR4, and active caspase-3 expression with low CD4 count and high plasma viremia and CD38 expression. The majority of activated CD8+CD38+ T-cells were apoptotic since they expressed active caspase-3 and the rest of these cells were susceptible to become apoptotic since they co-expressed CD95 and CXCR4. We suggest that one of the most likely HIV-mediated apoptosis mechanisms is via CD95 and CXCR4 induction through the caspase cascade despite Bcl-2 expression. This provides another explanation of why HIV-1 infection is not fully contained by HIV-specific CD8+CD38+ T-cells.
Supported by NIH/RCMI Grant # G12-RR03035.
The role of MHC-II in the maintenance of memory B cell function (B22)
Authors: Shimoda Michiko , Koni Pandelakis Andreas 
Published: January 2023

Abstract
By using conditional, B cell-restricted MHC-II-deficient mice (IA-B mice), we previously showed that antigen-specific memory B cells could be established but they then lost MHC-II expression due to ongoing cd19Cre-mediated deletion of the conditional iab allele, without significant loss in memory cell number. In this study, we examined whether or not MHC-II deficient memory B cells are functionally competent to differentiate into plasma cells. IA-B mice carrying a 4-hydroxy-3-nitro-phenyl-acetyl (NP) hapten-specific VH transgene (QM) were immunized with NP-coupled chicken gamma globulin plus alum. QM/IA-B mice generated NP+IgG+ memory B cells (~0.1% of total spleen cells), which subsequently lost MHC-II during maintenance. When cultured in the presence of 3T3 feeder cells with cytokines and various stimuli, both MHC-II(+) and MHC-II(−) NP+ memory B cells purified from QM/IA-B mice produced anti-NP IgG antibody upon LPS stimulation. However, anti-NP IgG production of MHC-II(−) NP+ memory B cells upon polyvalent B cell receptor stimulation with NP−Ficoll was significantly reduced compared to that of MHC-II(+) NP+ memory B cells. These results suggest that expression of MHC-II on memory B cells is required for maintenance of their capacity to differentiate into plasma cells in response to antigen stimulation.
This work was supported by NIAMS AR052470 (M.S.) and NIAID AI47379 (P.A.K.).
Immunoglobulin mutation traits in humans with AID deficiency (B220)
Authors: Longo Nancy S. , Satorius Colleen , Durandy Anne , Lipsky Peter E. 
Published: January 2023

Abstract
Germinal center (GC) B cells express high levels of activation induced cytidine deaminase (AID) which targets WRC (W=A/T, R=A/G) motifs generating a uracil:guanidine mismatch. During somatic hypermutation (SHM) 76% of the mutations are attributed to AID and error-prone repair utilizing polymerase eta (POL η). To characterize the remainder background mutations, 754 nonproductive V(D)J rearrangements from three AID−/− patients were analyzed. The mutation frequency was 20-fold less than normals (0.09% vs 2%) but 5x10e4-fold more than non-B cells. Reduced RGY (6% vs 27%) and WRC (6% vs 15%) motif mutations and decreased replacements in complementarity determining regions were attributed to the lack of AID. Reduced WA motif mutations (12% vs 23%) suggested POL η activity was decreased. An increased percentage of C&gt;T and G&gt;A transitions suggested that uracil DNA-glycosylase (UNG) activity was reduced even though UNG is upregulated in normal GC B cells and large GCs occur in AID−/− patients. The mutation pattern suggests that AID-independent C substitution contributes a small proportion of SHM but lacks the targeting provided by AID. Finally, low level of SHM clearly can develop in the absence of AID, however this mechanism is inefficient at altering the binding capacity of Ig heavy and light chain genes.
IRP, NIAMS, NIH
Apoptosis of Gr-1+ granulocytes in the lung promotes hypersensitivity pneumonitis by inducing dendritic cell maturation (B24)
Authors: hwang sujin , Chung Doohyun 
Published: January 2023

Abstract
Hypersensitivity pneumonitis (HP) is an interstitial lung disease that develops following repeated exposure to inhaled particulate antigens. HP is characterized with lymphocytic alveolitis, granuloma formation and fibrosis, and categorized as a Th1 disease. However, it remains unclear whether apoptosis in the lung contributes to the development of HP.
To investigate the functional role of the Fas-mediated apoptosis in HP, Fas(lpr) or FasL(gld)-deficient and B6 mice were challenged with Saccharopolyspora rectivirgula (SR) intranasally. lpr and gld mice attenuated HP in terms of histological alteration, CD4/CD8 ratio in bronchoalveolar lavage fluids (BALF), and SR-specific immune responses compared with B6 mice. The blockade of apoptosis in B6 mice using caspase inhibitor attenuated HP. In tunnel assay, the apoptosis was not found in respiratory epithelial cells whereas the apoptosis of immune cells, especially Gr-1+ granulocytes in BALF were prominent. Moreover, the depletion of Gr-1+ granulocytes attenuated SR-induced HP in B6 mice. B6 mice produced high levels of IFN-g, TNF-a and TGF-b whereas lpr or gld mice showed high levels of IL-4 production in the lung compared with B6 mice. The apoptotsis of Gr-1+granulocytes promoted pulmonary dendritic cell (DC) maturation and the production of MIP-1a and MCP-1 by DC, which contributed to recruiting inflammatory cells in the lung during SR-induced HP.
Taken together, these results suggest that the apoptosis of Gr-1+ granulocytes promotes SR-induced HP by inducing DC maturation and chemokine production. The blockade of apoptosis in the lung may be a useful therapeutic modality for HP.
Adjuvant Regulates Antigen-Specific Helper T Cell Evolution (B27)
Authors: Malherbe Laurent , McHeyzer-Williams Louise J , Panus Joanne F , McHeyzer-Williams Michael G 
Published: January 2023

Abstract
Antigen-specific helper T cells are the cognate regulators of multiple developmental checkpoints in adaptive immunity. Importantly, TCR-pMHCII interactions also control the evolution of antigen-specific Th cells in ways that remain poorly understood. Here, we demonstrate that typical depot producing adjuvants drive clonal dominance through the action of their adjuvant vehicle. In contrast, non-depot adjuvants re-set the selection threshold to propagate clones expressing high affinity TCR. Furthermore, oil in water formulations that readily disperse antigen-adjuvant aggregates in vivo also impact the magnitude of clonal expansion and substantially exaggerate the penetrance of high affinity clones. Finally, reducing antigen dose does not impact the affinity threshold for clonal selection arguing against a mechanism driven by inter-clonal competition in vivo. These findings reveal vaccine adjuvants as critical regulators of a TCR affinity-based clonal selection mechanism that underpins the evolution of adaptive immunity.
Signaling from the B cell antigen receptor (BCR) induces VH replacement in human immature B cells (B33)
Authors: Liu Jing , Fan Run , Yu Zhihong , Raikwar Himanshu P , Su Kaihong , Zhang Zhixin 
Published: January 2023

Abstract
VH replacement occurs through RAG-mediated secondary IgH recombination between a cryptic RSS (cRSS) from a previously rearranged VH gene and a 23-bp RSS from an upstream VH gene. Our previous studies have shown that VH replacement occurs in human bone marrow immature B cells and contributes to about 5% of the primary B cell repertoire. However, how VH replacement is regulated is not clear. The human EU12 cells, which undergo spontaneous VH replacement, are sensitive to BCR stimulation. BCR crosslinking strongly induces double stranded DNA breaks at the VH3 cRSS border in the μHC+ EU12 cells as measured by semi quantitative ligation-mediated PCR (LM-PCR) and real time LM-PCR, suggesting the induction of VH replacement. The VH3 to VH1 VH replacement excision circles are also induced upon BCR crosslinking. In addition, ongoing VH replacement was found in the newly emigrated immature B cells from peripheral blood of three healthy donors, which can be further induced by BCR crosslinking. These results provide the first evidence that VH replacement in human immature B cells can be regulated by BCR mediated signaling.
Transcriptional repression of the MHC-class I-related Fc receptor for IgG by interferon-gamma through activation of signal transducer and activator of transcription (Stat)-1 (B34)
Authors: Liu Xindong , Ye Lilin , Zhu Xiaoping 
Published: January 2023

Abstract
Expression of the MHC genes has been previously shown to be enhanced at the transcriptional or posttranscriptional level following exposure to cytokine IFN-gamma. However, we here reported that IFN-gamma down regulated the constitutive expression of the FcRn that functions to transport IgG across polarized epithelial cells and protect IgG from degradation. Intestinal epithelial cell, endothelial cell, and macrophage-like THP-1 or fresh isolated human monocytes exposure to IFN-gamma resulted in significantly-decreased human FcRn expression, as assessed by real-time PCR and western blot. Chromatin immunoprecipitation and gel mobility shift assays identified that Stat-1 bound to an IFN-g activation site (GAS) in FcRn promoter region. Functional analysis of luciferase reporter gene under the control of FcRn promoter showed that IFN-gamma had no effect on luciferase expression in JAK- and Stat-deficient cell lines. The unresponsive to IFN-gamma repression was rescued by over-expression of JAK or Stat1, revealing that Stat1-Jak signaling pathway was necessary and sufficient to mediate the repression of IFN-gamma. Furthermore, the inhibitory effects of IFN-gamma responsiveness were selectively reversed or blocked by site-directed mutagenesis of a cis-acting GAS element, and by over-expression of the Stat1 inhibitor, PIAS1, or of the dominant –negative Stat-1 Y701F containing a mutation at Tyr-701. IFN-gamma stimulation dampened down bidirectional transport of IgG across polarized Calu-3 lung epithelial monolayer. Taken together, our results indicate that down-regulation of FcRn gene expression by IFN-gamma may involve direct interaction of activated Stat-1 with the general transcriptional machinery that drives constitutive FcRn expression.
Defective development of follicular, marginal zone and B1 B cells in interferon regulatory factor 4 deficient mice (B4)
Authors: Pathak Simanta , Ma Shibin , Trinh Long , lu runqing 
Published: January 2023

Abstract
Interferon regulatory factor 4 (IRF4) is an immune system specific transcription factor. Previous studies have demonstrated that IRF4 and its closest relative IRF8 control light chain locus activation and are critical in the pre-B cell development. In this report, the role of IRF4 in the development of follicular, marginal and B1 B cells was further investigated. Our results show that the number of follicular B cells is significantly reduced while the marginal zone and B1 B cell population are dramatically increased in the IRF4 deficient mice. Transplantation study supports a B cell intrinsic role of IRF4 in regulating FO and MZ B cell development. Kinetic study reveals that the distorted FO and MZ B cell development in the absence of IRF4 is a result of accelerated MZ B cell production and reduced FO B cell generation. Molecular analysis further reveals that transitional B cells deficient in IRF4 express lower level of Bcl2. Ectopic expression of a Bcl2 transgene partially restore the normal development of follicular and marginal zone B cells in the IRF4 deficient mice. Thus, IRF4 maybe a novel transcriptional regulator of follicular, marginal zone and B1 B cell development and homeostasis.
Auranofin, immunosuppressive drug, inhibits the MHC class I and MHC class II pathway of antigen presentation in dendritic cells (B41)
Authors: Kim Kyungjae , Han Shinha , Kim Hyunyul , Kim Kwanghee , Kwon Jeunghak , Lee Chong-Kil 
Published: January 2023

Abstract
Auranofin (AF) is an orally administered, gold-based, anti-arthritic agent that has been used as a drug for the treatment of rheumatoid arthritis since the late 1970s. The precise mechanism of its anti-arthritic action is unclear. Therefore, this study examined the ability of AF to immunomodulate the antigen presenting cells (APCs), dendritic cells (DCs). The DCs were cultured in the presence of AF and examined for their ability to present exogenous antigens in association with the major histocompatibility complexes (MHC). This study examined whether or not AF (5 ~ 40 μM/ml) can inhibit the cross-presentation of DCs. DC2.4 cells (H-2Kb) or bone marrow-derived DCs (BM-DCs) generated from the BM cells of C57BL/6 mouse (H-2Kb) were cultured in the presence of AF with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by T cell hybridoma, B3Z, that recognizes the OVA (257–264 : SIINFEKL)-(H-2Kb) complex and expresses β-galactosidase. The inhibitory activity of AF appeared to be due not only to its ability to inhibit the phagocytic activity of DCs but also to suppress the expression of MHC molecules on DCs. AF decreased the level of IL-2 production when the effects of AF on CD4+ T cells were examined. The secretion of TNF-α, IL-1β and IL-6 was measured in cell supernatants using ELISA when RAW264.7 was exposed to AF for 24 hrs. AF decreased production and pro-inflammatory cytokine, TNF-α, IL-1β, IL-6 and nitric oxide (NO) in macrophages. These results provide an understanding of the mechanism of immunosuppressive activity and anti-inflammatory effects of AF in relation to their actions on APCs.
Expression and regulation of IL-22 in the IL-17-producing CD4+ T lymphocytes (B46)
Authors: Chung Yeonseok , Yang Xuexian , Chang Seon Hee , Dong Chen 
Published: January 2023

Abstract
IL-22 is a novel cytokine in the IL-10 family that functions to promote innate immunity of tissues against infection. Although CD4+ helper T lymphocytes (TH) were found as a source of IL-22, the regulation of this cytokine has been poorly understood. Here, we show that IL-22 is expressed at both mRNA and protein levels by a novel subset of TH cells that also makes IL-17. IL-22 and IL-17 were found to be coordinately regulated by TGFbeta and IL-6 during TH differentiation by real-time PCR as well as ELISA analysis. However, IL-22 does not regulate TH differentiation; exogenous IL-22 or an IL-22 antagonist had no effect on TH differentiation. These data demonstrate a novel cytokine expressed by IL-17-producing T cells, and suggest interaction and synergy of IL-22 and IL-17 signaling pathways in tissue inflammation and autoimmune diseases.
This work was in part supported by National Institutes of Health (to C.D.). S.H.C receives a post-doctoral fellowship from the Arthritis foundation and C. D. is a Cancer Research Institute Investigator and a MD Anderson Cancer Center Trust Fellow.
IL-10 and natural regulatory T cells: two independent anti inflammatory mechanisms in HSV induced ocular immonopathology (B63)
Authors: Sarangi Pranita Pragnyadipta , Suvas Susmit , Sehrawat Sharvan , Kim Bumseok , Rouse Barry T 
Published: January 2023

Abstract
Our past studies have shown a protective role for both CD4+CD25+Foxp3+ natural regulatory T cells (nTregs) as well as IL-10 in Herpes Simplex Virus (HSV) induced Stromal Keratitis (SK). Among several other mechanisms, IL-10 has been shown to be involved in the nTreg mediated suppression. We have shown in vitro that blocking of IL-10 could partially block the Treg mediated suppression of activated effectors isolated from SK lesions. But it is not known whether in the in vivo conditions, IL-10 and nTreg influence the SK pathology independently or in concert. Interestingly, IL-10−/− animals depleted of nTregs prior to ocular HSV infection had more severe SK lesion as compared to undepleted IL-10−/− animals. These results show the presence of two independent anti inflammatory mechanisms in SK. Along this line, by using mice expressing GFP-Foxp3 we could not show CD4+GFP-Foxp3+IL-10+ cells in the DLNs and spleens of infected wild-type (WT) mice although animals depleted of nTregs had higher levels of IL-10 in the DLNs post infection. Furthermore, by using WT, IL-10−/− and nTreg depleted WT and IL-10−/− animals we have compared the relative importance of IL-10 and nTregs in reducing SK pathology. Although disease severity was significantly higher in IL-10−/− and both the nTreg depleted groups as compared to WT, highest severity was found in the IL-10−/− mice depleted of nTregs. Intracellular cytokine staining with UV-HSV and polyclonal stimulation of the cells isolated from infected corneas, DLN and spleens revealed that IL-10 has more profound anti inflammatory effects in SK as compared to nTregs and that the later could act in an IL-10 independent manner.
TGF-β Induces Microvascular Endothelial Cell Migration and PP-2A Inactivation (B65)
Authors: Walsh Jarrett Elbert , Young M. Rita I. 
Published: January 2023

Abstract
An increasing number of anti-tumor therapies have focused on angiogenesis pathways. Targeting various aspects of angiogenesis has been shown to disrupt tumor growth and reduce tumor progression and recurrence. Our studies show that treatment of microvascular endothelial cells with TGF-β leads to the reduction in activity of the protein phosphatase PP-2A, which results in increased motility. The mechanism by which TGF-β signaling results in PP-2A inactivation remains unknown. Our studies will show that neither change in mRNA nor protein levels of PP-2A subunits is associated with this decreased activity. However, TGF-β induced changes in ceramide levels may play a role in reduced activity. Furthermore, the role of TGF-β signaling on the protein inhibitors of PP-2A, I1PP2A and I2PP2A, will be demonstrated. The goal of these studies is to identify mechanisms by which TGF-β signaling induces PP-2A inactivation and, in turn, increased cellular motility in the tumor microenvironment. Elucidation of these pathways will provide novel cellular targets to interfere with tumor angiogenesis and metastasis.
TGF-β-induced TCR-Tg/NOD regulatory T cells suppress both autoimmune diabetes transfer and islet graft destruction in an antigen dependent manner (B75)
Authors: Tonkin Daniel , Bradley Brenda , Barbour Gene , Haskins Kathryn 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are understood to fall into two categories: natural Tregs and induced Tregs. Induced Tregs can be produced by activating CD4 T cells in the presence of TGF-β. Such induced Tregs are of great promise for immunotherapy, but it has been unclear if they require stimulation by antigen in order to suppress in vivo. We have investigated the limits of induced Tregs in the NOD mouse model of type I diabetes. We produced TGF-β-induced Tregs from the 6.9 TCR-Tg/NOD mouse that are specific for islet autoantigen. In some experiments we used a congenic mouse, the NOD.C6, that specifically lacks the antigen for the 6.9 TCR-Tg T cells but not for other islet-specific T cells. These Tregs express Foxp3, and produce much less IFN-γ than non-Tregs. We have shown that induced Tregs can prevent transfer of diabetes by effector T cells in NOD mice, but fail to protect in NOD.C6 mice that lack the Treg antigen. These Tregs decrease inflammatory cytokine production and macrophage infiltration in the pancreas. In human patients with diabetes, induced Tregs might be used to protect islet grafts from immune destruction, and we have demonstrated that our induced Tregs can prevent islet graft destruction in NOD mice. In addition, we have grafted islets from the NOD.C6 mouse. By using these antigen-free islets, we have conclusively demonstrated that induced Tregs require activation by their antigen in the islet graft in order to protect.
TCRbeta formation: Vbeta accessibility is not the limiting or regulated step (B8)
Authors: Carpenter Andrea C , Bassing Craig H. 
Published: January 2023

Abstract
The T cell receptor β (TCRβ) locus contains three gene segment families, variable (V), diversity (D) and joining (J), each of which contributes one segment to form a TCRβ exon. In DN2 stage thymocytes, a Dβ segment joins a Jβ segment on both alleles, while in the DN3 stage Vβ segments join with previously formed DJβ complexes. Formation of a productive (protein encoding) VDJβ complex on one allele is theorized to prevent Vβ segments from joining with DJβ complexes on the other allele, leading to TCRβ allelic exclusion. To elucidate mechanisms that regulate Vβ joining and TCRβ allelic exclusion, we generated and analyzed mice with a recombination reporter cassette consisting of the Dβ1 and Jβ1.1 gene segments inserted into the endogenous Vβ14 segment. We found Vβ14DJrecombination initiated concurrent with Vβ14 rearrangement, suggesting local chromatin environment determines the developmental timing of Vβ14 rearrangement. Our analysis of Vβ14DJ/WT and Vβ14DJ/DJ αβ T cell hybridomas revealed that Vβ14DJrecombination occurs at a substantially higher frequency than Vβ14 joining. Moreover, our initial findings indicate that Vβ14DJrecombination is not inhibited by expression of a productive VDJβ complex. Collectively, these data suggest that modulation of Vβ14 recombinational accessibility is neither the rate-limiting or regulated step in joining of Vβ14 segments with DJβ complexes.
Training Grant: 5-T32-Al055428