bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of intermolecular disulfide bonds on assembly of mouse MHC class I molecules (130.25)
Authors: Simone Laura , Wang Xiaojian , Tuli Amit , Solheim Joyce 
Published: January 2023

Abstract
Within the major histocompatibility (MHC) class I peptide-loading complex, tapasin and ERp57 are joined by a stable disulfide bond which is necessary for the efficient assembly of MHC class I molecules. In our investigation of murine tapasin, we identified a large covalently bound complex involving wild type mouse tapasin and ERp57. This complex, which migrates at ~150 kDa, is noticeably larger than the 110 kDa tapasin-ERp57 complex previously identified in human cells. Additional data suggest that the mouse MHC class I molecule Kd may be a part of this ~150 kDa complex. In order to investigate the importance of tapasin-ERp57 disulfide bonding for the assembly of murine MHC class I molecules, we introduced a tapasin C95S mutant into tapasin knockout mouse fibroblasts. Although this tapasin mutant is unable to disulfide bond with ERp57, mouse tapasin C95S was found to maintain a non-covalent association with ERp57. Mutation of tapasin C95 impaired the maturation and stability of MHC class I Kd molecules to a greater extent than was previously observed for the human MHC class I molecule, B*4402. Furthermore, the maturation and stability of mouse MHC class I Kb molecules was also impaired in cells expressing mouse tapasin C95S. In total, these results highlight inter-species differences in the MHC class I assembly pathway and emphasize the importance of studying diverse MHC class I allotypes.
Adjuvanted Flu Vaccination Using a Novel High-Efficiency Hollow Microprotrusion Skin Patch (130.37)
Authors: Chang Stella , Kaspar Roger , Fairman Jeff , Speaker Tycho 
Published: January 2023

Abstract
Influenza spreads around the world in seasonal epidemics, resulting in the deaths of hundreds of thousands worldwide annually, and millions in pandemic years. Hypodermic needle-based administration of current vaccines to large underserved populations is hampered by refrigeration requirements, painful injections, and infectious sharps waste disposal issues. We have developed a prototype micro-protrusion array device (PAD), comprising water-soluble polymer microneedles loaded with a dry-stable adjuvanted flu antigen vaccine for topical application as a patch. Three groups of mice were tested to evaluate the efficacy of PAD vaccination: Non-treatment, Fluzone (Flz) and Flz adjuvanted with JVRS-100. PADs were applied to anesthetized mice with a primary dose, followed by a second boost dose applied two weeks later. Four weeks after the primary dose, the animals were challenged with 1x LD50 of HKx31 (H3N2) Flu virus. Serum antibody levels were assessed before the challenge. Total IgG of Anti-Flz was 6 fold higher in the Flz/JVRS-100 than Flz. Control and Flz mice had 60% mortality while the Flz/JVRS100 group had a 100% survival rate. We conclude that the dry PAD adjuvanted vaccine format may provide a powerful new approach to transdermal vaccine delivery. Immunization potency is significantly increased through adjuvanted antigen which generating a robust immune response that was protective against lethal influenza virus challenge.
Enhanced IL-2 signaling renders IL-12 primed CD8+ T-cells superior in adoptive immunotherapy. (131.20)
Authors: Lisiero Dominique , Soto Horacio , Liau Linda , Prins Robert 
Published: January 2023

Abstract
Sustained persistence and function of adoptively transferred T-cells are critical parameters for effective anti-tumor activity. To address this, our group has explored the mechanisms by which TLR agonist-induced cytokines, such as IL-12, are capable of efficiently priming CD8+ T-cells. IL-12 primed CD8+ T-cells upregulate the expression of the high affinity IL-2 receptor (CD25) and STAT5. Enhancement of these signaling components increases IL-2 mediated signaling events at low concentrations of IL-2. Blockade with anti-CD25 antibody effectively terminates this enhanced IL-2 signaling in IL-12 primed CD8+ T-cells. More importantly, IL-12 priming of CD8+ T-cells significantly increases in vivo proliferation as measured by CFSE dilution, and ex vivo recovery from lymphoid organs. Utilization of in vivo bioluminescent imaging of firefly luciferase transduced IL-12 primed T-cells demonstrated their ability to traffic into the tumor area, through lymph nodes and bone marrow. Finally, adoptive transfer of IL-12 primed CD8+ T-cells decreases tumor burden in a subcutaneous melanoma model and increases survival in an intracranial tumor model. This anti-tumor activity can be abrogated by addition of an anti-CD25 blocking antibody. These results demonstrate the importance of pro-inflammatory signals in the tumor setting, and their ability to render CD8+ T-cells functional and responsive to proliferative signals.
Vaccination with tumor cell lysate plus CpG oligodeoxynucleotides elicits tumor-specific T cells with potent antitumor efficacy in adoptive immunotherapy (131.31)
Authors: Shindo Maki , Chen Wei 
Published: January 2023

Abstract
This study investigated whether tumor-specific T cells can be elicited by vaccination with tumor cell lysate plus CpG oligodeoxynucleotides (CpG/lysate) for adoptive immunotherapy. Vaccination of C57BL/6 mice with syngeneic EL-4 tumor cell lysate mixed with a CpG-B ODN (CpG 685) significantly increased the number of T cells and activated DCs in the draining lymph nodes (DLN). Mice vaccinated with CpG/lysate developed protective immunity against EL-4 tumor challenge with a significant longer median survival time (P&lt;0.01). The CpG/lysate-primed DLN T cells were expanded in a 5-day culture with anti-CD3 mAb and IL-2, and adoptively transferred into EL-4 tumor-bearing mice following cyclophosphamide (CY) treatment. Therapy with CY plus CpG/lysate-primed T cells induced an 80% tumor-free survival rate whereas EL-4 mice in the control groups all died within 30 days. When the mice cured by CpG/lysate-primed T cells were re-challenged with EL-4 tumor 2 months later, all the mice (100%) survived without tumor growth, suggesting the persistence of T cell-mediated antitumor immunity. The efficacy of CpG/lysate-primed T cells was comparable to DC/lysate vaccine-primed T cells in adoptive immunotherapy. Cell-depletion studies showed that CD4 T cells play a pivotal role in CpG/lysate vaccine-induced antitumor effects. The results suggest that CpG/lysate vaccine provides an effective approach to induce host antitumor immunity and tumor-reactive T cells for adaptive immunotherapy.
Investigating the involvement of Foxo1 in T cell ageing (132.1)
Authors: Gubbels Bupp Melanie 
Published: January 2023

Abstract
Advanced age is associated with increased susceptibility to and mortality from infectious diseases, especially influenza and pneumonia; and vaccination responses in the elderly are diminished. Declining resistance to infection in aging individuals is correlated with a progressive decline in the size and functionality of the naïve T cell pool. A better understanding of the mechanisms underlying age-related deterioration of T cell immunity could inform therapies that enhance general immune function and vaccination effectiveness in this vulnerable population. The forkhead transcription factor, Foxo1, has recently been identified as a major regulator of naïve T cell homeostasis. Moreover, a Foxo1 homolog, DAF-16, promotes longevity in C. elegans. Therefore, these studies documented the decline of the naïve T cell pool in mice selectively deficient for Foxo1 in the T cell compartment (Foxo1 cKO mice) to determine the involvement of Foxo1 in ageing T cells. Our studies indicate that naïve CD8+ CD122- T cells decline at a more rapid rate in the blood of Foxo1 cKO mice as compared to WT controls. Together, these observations suggest the intriguing possibility that in mice, Foxo1 may be involved in retarding age-related deterioration of T cell immunity.
Upregulation of B220 expression correlates with sensitivity to arsenic trioxide (As2O3)-induced cell death in different leukemic T-cell lines (132.11)
Authors: Bobé Pierre , Benbijja Mohcine 
Published: January 2023

Abstract
As2O3 kills a variety of leukemia cells, but with different efficacy. Although cellular redox status is considered essential for As2O3 sensitivity, we show here that murine and human leukemic T-cell lines display dramatic differences in As2O3 sensitivity independently of their GSH content and O2− production. Unexpectedly, As2O3 differently upregulated expression of phosphatase B220 on these T cells in dose-dependent manner. Since we previously reported that As2O3 selectively eliminates pathogenic B220+ T cells in autoimmune MRL/lpr mice (Bobé et al, BLOOD, 2006, 108:30967), we have studied the possible correlation between B220 expression and As2O3-induced T-cell death in leukemic T-cell lines. In the presence of As2O3, we found low (HPB-ALL), intermediate (Jurkat) and high (EL-4, BW5147) levels of both B220 expression and cell death. Ca2+ ionophore A23187 also upregulated B220 expression and cell death, but with opposite efficiencies in the T-cell lines compared to As2O3. A23187 only upregulated the T-cell activation marker CD69 before B220 expression and cell death, suggesting that it kills by an activation-induced cell death mechanism. In L1210 cells, we found constitutive B220 expression. L1210 cells were the most resistant to cell death among As2O3-treated T-cell lines whereas they were the most sensitive to A23187. As2O3 and A23187 probably trigger different signaling pathways through B220 leading to T-cell death, suggesting a checkpoint role for B220 in death pathways.
An evaluation of techniques to efficiently induce in vitro T-cell expansion. (132.29)
Authors: Langsdorf Chris , Bantly Andrew , Bradford Jolene , Buller Gayle , Liu Jixiang 
Published: January 2023

Abstract
The current study evaluated three in vitro methods of inducing proliferation in human T lymphocytes. The first method involved treatment with mouse anti-human CD3 antibody and interleukin-2. The second approach used the lectin Concanavalin A, a mitogen which preferentially activates T cells. The third technique made use of mouse anti-human CD3 and CD28 antibodies immobilized on the surface of superparamagnetic polystyrene microspheres. Human lymphocytes were isolated from fresh whole blood and stained with CellTrace™ Violet, a novel cell proliferation tracing dye with a mode of action similar to CFSE. Samples of stained cells were stimulated and cultured for 7 days. T cell subsets induced to proliferate with each treatment were characterized in multicolor flow cytometry. Cells treated with anti-CD3/IL-2 or Concanavalin A showed a moderate, heterogeneous level of stimulation. Microsphere-bound antibodies provided a much stronger, more uniform stimulus. This strong, homogeneous stimulus is useful when massive numerical expansion of T cells is desired for subsequent research or adoptive transfer in vivo, but does not appear to be as useful when a population containing several discrete generations of cells is desired. The more moderate stimuli resulting from treatment with anti-CD3/IL-2 or Concanavalin A result in a less efficient numerical expansion, but provide an excellent tool to evaluate the phenotype and pedigree of cells as they proliferate through several generations.
Influence of partial agonist expression on CD8 T cell responses for non-self antigens (132.35)
Authors: Ryu Su Jeong , Sproule Tom , Christianson Gregory , Roopenian Derry , Choi Eun Young 
Published: January 2023

Abstract
Differences in affinity of TCR for peptide/MHC complex induced by single amino acid substitutions in the peptide dramatically alter recognition by T cell in the periphery. With H60 as an antigenic model, we investigated the influence of transgenic expression of natural H60 antigen (H60N Tg) or of H60 antigen with single amino acid substitution at position 4 to histidine (H60H Tg) on H60H- or H60N-specific CD8 T cell responses, respectively. Immunization of B6 female mice with splenocytes from male H60NTg or H60H Tg mice induced strong CD8 T cell responses against each epitope, H60N or H60H, in CD4 T cell help dependent ways. While the immunization with splenocytes from male H60H Tg induced proliferation of H60-specific CD8 T cell responses in the female H60N Tg mice, CD8 T cell response for H60H induced in female H60N Tg showed significant reduction in the magnitude of H60H-tetramer-binding CD8 T cells at the peak time. And differences in TCR V beta usages of H60H-reactive CD8 T cells were also found between in the female B6 female and female H60N Tg hosts. Our data suggest that expression of proteins with amino acid changes influences the antigen-specific CD8 T cell responses and repertoire of TCRS involved in the responses.
Regulation of Surface Free Thiol Levels During CD8+ T Cell Activation, Proliferation and Differentiation (132.7)
Authors: Pellom Samuel , Michalek Ryan , Grayson Jason 
Published: January 2023

Abstract
Upon both B and T cell activation the level of reactive oxygen intermediates (ROI) increases. Prior studies in the literature demonstrated that activated lymphocytes have increased total levels of surface free thiols, potentially as a compensatory mechanism to prevent surface oxidation. In this study, we determined how surface free thiol levels were regulated during the activation, proliferation, and differentiation of naïve CD8+ T cells both in vitro and in vivo. In vitro, we found that the levels of surface free thiols increased within 24 hours regardless of the activation stimulus and were maintained for at least three divisions. To understand how levels were modulated in vivo, we examined antigen-specific CD8+ T cells using lymphocytic choriomeningitis virus (LCMV) infection of mice and MHC Class I tetramers. On day 5 postinfection, the levels of surface free thiols were 8-fold higher on effector cells compared to those found on naïve CD8+ T cells. By day 8 postinfection this had decreased to 4-fold. After the infection was cleared, memory CD8+ T cells had 1.5-fold higher levels of surface free thiols than naïve cells. This contrasted with the results during chronic LCMV-Clone 13 infection. On day 8 postinfection surface free thiol levels were 11-fold higher and remained elevated until day 35 postinfection. These results suggest that surface free thiol levels are elevated most highly in rapidly proliferating cells and do not correlate with lymphocyte function.
STAT3 controls the neutrophil migratory response to CXCR2 ligands by direct activation of G-CSF-induced CXCR2 expression and via modulation of CXCR2 signal transduction (133.2)
Authors: Nguyen-Jackson Hoainam , Panopoulos Athanasia , Zhang Huiyuan , Li Haiyan , Watowich Stephanie 
Published: January 2023

Abstract
Neutrophil mobilization, the release of neutrophils from the bone marrow reserve into circulating blood, is important to increase peripheral neutrophil amounts during bacterial infections. Granulocyte colony-stimulating factor (G-CSF) and chemokines, such as MIP-2 (CXCL2), can induce neutrophil mobilization, but the mechanism(s) they employ remain unclear. The transcription factor STAT3 is the principal intracellular signaling molecule activated upon G-CSF-ligation of its receptor. Using a murine model with conditional STAT3 deletion in bone marrow, we demonstrated previously that acute G-CSF-responsive neutrophil mobilization and MIP-2-dependent neutrophil chemotaxis is mediated by STAT3. In this study, we show that STAT3 is also necessary for MIP-2-elicited neutrophil mobilization. STAT3 appears to function by regulating the amplitude of Raf/MEK/Erk activation, a signaling pathway that is important for MIP-2-mediated chemotaxis. In addition, we demonstrate that G-CSF stimulates the expression of the MIP-2 receptor via STAT3-dependent transcriptional activation of Il8rb. G-CSF treatment also induces changes in bone marrow chemokine expression levels in a STAT3-dependent manner, which may further affect neutrophil retention and release. Taken together, our study demonstrates that STAT3 regulates multiple aspects of chemokine and chemokine receptor expression and function within the bone marrow, indicating a central role in the neutrophil mobilization response.
Atypical protein kinase C (aPKC) isoforms ζ and λ/ι synergically control Th2 development and function (134.13)
Authors: Yang Jun-Qi , Diaz-Meco Maria , Moscat Jorge 
Published: January 2023

Abstract
The protein kinase C (PKC) family consists of three subfamilies: classical, novel, and atypical PKCs. The atypical PKC (aPKC) consists of two isoforms, PKCζ and PKCλ/ι. We have recently presented in vitro, ex vivo, and in vivo genetic evidence that PKCζ is necessary for optimal activation of the IL-4 signaling cascade upstream of Jak1. Since whole-body genetic deletion of PKCλ/ι is lethal, we have used a conditional gene knockout system by loxP-Cre recombination to generated conditional PKCλ/ι KO mice to selectively delete PKCλ/ι in activated T cells. With this system, we found that PKCλ/ι was necessary for Th2 cytokine production. Upon anti-CD3/CD28 stimulation, IL-4 and IL-13 production from CD4+ T cells was significantly reduced in conditional KO mice as compared to their WT littermates. The activation of the transcription factors NFAT and NF-κB was impaired in PKCλ/ι-deficient activated T cells. Furthermore, deficiency of PKCλ/ι inhibited OVA-induced allergic airway inflammation in vivo. Recently, we have introduced the activated T cell-specific conditional PKCλ/ι deficiency into a PKCζ-/- background to generate mice with PKCζ and PKCλ/ι double knockout (DKO) in activated T cells. From these T cells, the secretion of Th2 cytokines, IL-4, IL-5 and IL-13, was more dramatically reduced and the proliferation was more significantly impaired compared to T cells with single PKCζ or PKCλ/ι deficiency. PKCλ/ι, like PKCζ, emerges as a new critical regulator of Th2 cell activation.
Dendritic cells in the skin of autoimmune mice display an inability to migrate: a novel mechanism and role in skin inflammation (135.12)
Authors: Eriksson Anna , Kim Peter , King Jennifer , Okereke Chetachi , Singh Ram 
Published: January 2023

Abstract
Tissue-resident DC carry antigens to tissue-draining lymph nodes (LN) to induce immunity. To investigate the role of this DC function in the pathogenesis of autoimmune disease, we analyzed the migration of skin-resident DC, namely Langerhans cells (LC) and dermal DC, in lupus dermatitis. LC and dermal DC, but not blood-derived DC, from lupus-prone MRL-lpr and MRL+/+ mice exhibited defects in their ability to emigrate from the epidermis, to migrate through dermal lymphatics, and to immigrate into LN (JI 2008). Since conventional DC migration assays require DC activation, we used Lang-EGFP knock-in (KI) mice to visualize LC at steady-state. EGFP+ cells (LC) were lower in LN of KI MRL mice than of KI B6 mice, thus confirming what we found using inbred mice. Importantly, skin DC migration defect precedes disease and correlates with the severity of dermatitis. Conversely, treatment with αGalCer restores skin DC migration and ameliorates dermatitis. Surprisingly, αGalCer-mediated increase in skin DC migration does not require NKT cells, but is associated with epidermal γδ T cell (eγδT) expansion in a CD1d-dependent manner. Finally, eγδT are reduced in MRL mice, and deficiency of CD1d or eγδT, but not of NKT, reduces skin DC migration and worsens dermatitis. Thus, we elucidate a novel mechanism, whereby CD1d-dependent eγδT normally facilitate skin DC migration. This regulatory mechanism is impaired in lupus dermatitis, providing evidence for a novel mechanism of autoimmune disease.
Dominant epitope p21-40 of snRNP D protein recognized by both autoreactive T and B cells in lupus-prone MRL lpr/lpr mice (135.22)
Authors: Ding Chuanlin , Cai Yihua , Yan Jun 
Published: January 2023

Abstract
Aberrant T and B cell responses to multiple nuclear and cytoplasmic Ags such as small nuclear ribonucleoparticles (snRNPs) are a hallmark of systemic lupus erythematosus. Although Th1 response is considered to be predominant in lupus pathogenesis, the identification of Th17 cells may challenge Th1 paradigm. To investigate the respective roles of Th1, Th17, and Treg cells in lupus development, we analyzed different T cell subsets in spleen and kidney of lupus-prone MRL lpr/lpr mice. With increasing age and disease severity, the IFN-gamma-producing T cells were significantly increased. No significant change was observed in the IL-17-secreting CD4 T cells. Furthermore, the ration of Th1/Treg was significantly increased when mice aged while Th17/Treg ratio remained unchanged. To determine the antigenic epitopes recognized by autoreactive T and B cells, synthesized peptides of autoAg snRNP D protein were used. Two epitopes were recognized by autoreactive T cells while three epitopes were recognized by autoantibodies in MRL lpr/lpr mice. Interestingly, peptide 21-40 of snRNP D protein was recognized by both autoantibodies and T cells. Furthermore, one of T cell hybridomas generated from snRNP D-immunized B10.A Tg mice was also responsive to peptide 21-40. Taken together, our studies demonstrate that Th1 response remains dominant in lupus-prone MRL lpr/lpr mice and p21-40 of snRNP D protein appears to be a pathogenic epitope that can be recognized by both autoreactive B and T cells.
Chronic Stimulation of the DNA Repair Enzyme DNA-PKcs Impairs T-Cell Survival in Rheumatoid Arthritis (RA) (135.7)
Authors: Shao Lan , Goronzy Jörg , Weyand Cornelia 
Published: January 2023

Abstract
Patients in RA have premature aging of the immune system, with accumulation of pro-inflammatory and tissue-destructive T cells. Accelerated immune senescence has been attributed to chronic T cell loss. Naïve T cells in RA have a defect in repairing damaged DNA. We have examined whether chronic stimulation of DNA repair mechanisms affects T cell survival in RA. Naïve CD4+CD45RA+ T cells from RA patients had a higher load of double-stranded DNA breaks (p=0.0005) and spontaneous apoptosis rates (p=0.03) than in healthy controls. High DNA damage failed to upregulate the ATM-p53 axis. Instead, the transcript and protein levels of the DNA repair enzyme DNA-PKcs were elevated. Induction of DNA-PKcs was disease-specific and not found in patients with SLE. Knocking down DNA-PKcs transcription or inhibiting enzyme activity with a kinase inhibitor normalized T cell apoptosis rates in patients. Activation of DNA-PKcs was correlated with stimulation of SAPK/JNK pathway. Inhibition of either DNA-PKcs or JNK suppressed the pro-apoptotic protein Bim and protected T cells from apoptotic death. T cells in RA patients accumulate damaged DNA, causing chronic induction of repair activity. DNA double-strand breaks are recognized by a ATM-independent pathway involving DNA-PKcs. Due to the activation of the JNK-Bim death signaling cascade, RA T cells are apoptosis sensitive. Therapeutic blockade of DNA-PK-dependent cell death machinery could help rejuvenate the immune system of RA patients.