bluuebunny/crossref_metadata_2025_split huggingface dataset:
The elucidation of the oral/respiratory axis in inducing protective immunity against Francisella tularensis (161.22)
Authors: Signarovitz Aimee , Yu Jieh-Juen , Ray Heather , Guentzel M. Neal , Williams Ifor , Yagita Hideo , Berton Michael , Klose Karl , Arulanandam Bernard 
Published: January 2023

Abstract
Using a Francisella tularensis subspecies novicida live attenuated vaccine strain (U112ΔiglB), we have shown that oral vaccination of C57BL/6 mice induces significant protective immunity against pulmonary challenge with the highly human virulent F. tularensis strain SCHU S4. This protection is primarily mediated by antigen-specific CD4+ T cells and IFN-γ production and is dependent on mucosal IgA expression. We are examining the mechanism(s) by which the oral/respiratory axis induces optimal protective immunity against F. tularensis. Intestinal M-cells play an important role in oral vaccination by trafficking antigens from the lumen to the lamina propria and in presentation of antigens to underlying APCs for the subsequent induction of mucosal immunity. We have shown that mCherry-labeled U112ΔiglB co-localizes to intestinal mouse M-cells by 90 minutes following vaccination. Localization to M-cells was further verified with mouse ileal loops. Translocation of U112ΔiglB has also been demonstrated through M-cell-like-cells in an in vitro transwell co-culture system similar to M-cell trophic enteric bacteria. Current studies are examining the functional importance of intestinal M-cells in induction of mucosal immunity for the rational development of an effective oral vaccine against pulmonary tularemia. These studies will provide information for the general design of oral vaccines using live attenuated bacterial strains.
Immediate induction of enhanced CD8 suppressive function following glatiramer acetate therapy in multiple sclerosis (164.20)
Authors: Ayers Chris , Mendoza Jason , Greenberg Benjamin , Frohman Elliot , Karandikar Nitin 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an immune-mediated, demyelinating disorder of the CNS. Glatiramer acetate (GA/Copaxone) is an FDA-approved treatment for relapsing-remitting MS that induces functional changes in antigen-presenting cells (APC) and CD4 and CD8 T-cells. We have previously shown that treatment-naïve MS patients have deficient CD8 T-cell responses to GA, which are restored after GA therapy. This is associated with enhanced cytotoxic/suppressive function in these cells. In this study, we assessed the immediate immunologic changes induced by GA therapy in treatment-naïve MS patients by multiparametric monitoring of T-cell and APC subsets prior to and at 4, 12, 24, 72 hrs of treatment. In untreated healthy control subjects quantitative and functional readouts showed stable dynamics over time. MS patients showed greater proportions of memory/effector CD4 and CD8 populations at baseline, which did not vary. Similarly, cytokine production by APC subsets was also stable. Interestingly, as early as 4 to 12 hrs after the initial GA injection, we observed significantly enhanced suppressive ability in CD8 T-cells. This rapid functional change consistently occurred in GA, as well as an IFN-b-treated control cohort. These studies show that the induction of T-cell suppressive function is an important early event during therapy. Future studies will delineate whether these changes are predictive of sustained longterm immunologic and clinical responsiveness.
Macromolecular prodrug conjugate P-Dex attenuates nephritis in lupus-prone (NZBxNZW)F1 mice (164.5)
Authors: Nelson Richard , Yuan Fang , Wang Dong , Gould Karen 
Published: January 2023

Abstract
Corticosteroids are used in the treatment of nephritis, a severe manifestation of systemic lupus erythematosus. Untreated, nephritis results in chronic glomerular inflammation, loss of renal function, albuminuria, and ultimately leads to renal failure and death. Unfortunately, current therapeutic strategies are problematic, as the prolonged systemic exposure of corticosteroids yields severe side effects, including hypertension, osteoporosis, and immunosuppression. To remedy this, we developed a polymeric macromolecular prodrug (P-Dex) that targets inflamed glomeruli and other sites of inflammation by sequestering the corticosteroid dexamethasone via an acid-labile bond. In this study, we evaluated the efficacy of P-Dex in treating nephritis in lupus-prone (NZBxNZW)F1 mice. After cessation of treatment, the P-Dex treatment group displayed a lower incidence of albuminuria and greater histological preservation of glomerular structure than control groups. Further, P-Dex mitigated the side effects associated with the use of corticosteroids. Compared to control groups, mean arterial pressure was significantly lower in the P-Dex group, approaching normal levels. Finally, in contrast to mice treated with free Dex, mice treated with P-Dex displayed no significant loss of bone mineral density. Together, these data suggest that P-Dex may be a more effective therapy in treating nephritis, while simultaneously minimizing side effects associated with prolonged exposure to corticosteroids.
Inhibition of differentiation, amplification and function of human Th17 cells by intravenous immunoglobulin (164.7)
Authors: Maddur Mohan , Janakiraman Vani , Hegde Pushpa , Lacroix-Desmazes Sebastien , Kaveri Srini , Bayry Jagadeesh 
Published: January 2023

Abstract
Th17 cells play a critical role in the pathogenesis of several autoimmune and inflammatory diseases. Intravenous immunoglobulin (IVIg), a therapeutic preparation of polyclonal IgG, increasingly used in the treatment of diverse autoimmune and inflammatory diseases, may target Th17 cells to exert therapeutic effects. Therefore, we examined whether IVIg interferes with the development and function of human Th17 cells. To address this, Th17 cells were differentiated from naïve human CD4+ T cells in the presence of TGFβ and IL-21. Th17 cells were amplified by stimulating memory CD4+ T cells in the presence of IL-1β and IL-6. The effect of IVIg was examined on differentiation and amplification of Th17 cells, expression of Th17 lineage-specific transcription factor RORC, secretion of Th17 effector cytokines and phosphorylation of STAT3, a transcription factor that plays an important role in Th17 development and function. IVIg inhibits the differentiation and amplification of human Th17 cells, as well as the production of their effector cytokines IL-17A, IL-17F, IL-21 and CCL20. The inhibitory effects of IVIg on Th17 cells are F(ab')2-dependent and involve interference with the expression of RORC and activation of STAT3. Also, IVIg significantly enhanced Foxp3+ regulatory T cells (Treg) among the memory CD4+ T cells. These results reveal a novel mechanism of action of IVIg in achieving therapeutic effect in autoimmune and inflammatory diseases.
Targeting of pre-existing and induced breast cancer stem cells with HER2-specific immunotherapeutics and NK cells (165.18)
Authors: Wischhusen Jörg , Diessner Joachim , Becker Kathrin , Hönig Arnd 
Published: January 2023

Abstract
Trastuzumab (Herceptin®) has substantially improved the overall survival of patients with mammary carcinoma. While recruitment of NK cells and antibody-dependent cell-mediated cytotoxicity (ADCC) contribute to this therapeutic effect, immunoselection of cultured breast cancer cells with trastuzumab and NK cells resulted in selective survival of individual sphere-forming cells. Moreover, epithelial to mesenchymal transition was induced which transformed previously differentiated cancer cells into cells with a CD44highCD24low “cancer stem cell-like” phenotype. Immunoselected breast cancer cells thus showed increased clonogenicity and differentiation potential in vitro and enhanced tumorigenicity in vivo. Their insufficient immunotherapeutic targeting might be due to low HER2 expression and the virtual absence of ligands for the activating NK cell receptors NKG2D and DNAM 1. Susceptibility towards NK cells and trastuzumab, however, was restored upon differentiation. Since relapse after antibody therapy could be due to such trastuzumab- and NK cell-refractory tumor initiating cells, we explored if treatment of these cells with the novel HER2-targeting drugs T-DM1 or the HER2 dimerisation inhibitor pertuzumab could improve NK cell mediated killing or counteract induction of the stem cell-like phenotype. This translational approach might help to identify the most promising drug candidate for the targeted second and third line treatment of HER2-positive metastasized breast cancer.
IFNγ mediates the antitumor effects of radiation therapy (165.34)
Authors: Gerber Scott , Cron Kyle , Lord Edith 
Published: January 2023

Abstract
Radiation (IR) therapy has long been an effective cancer treatment acting primarily through the induction of tumor cell damage at a molecular level. However, a new concept is emerging suggesting that the immune system plays a pivotal role in determining the effectiveness of IR therapy. We have previously demonstrated that IR therapy promotes antitumor immunity through the release of tumor antigens and induction of a pro-inflammatory tumor microenvironment. Our work here has identified IFNγ as an essential cytokine which mediates the efficacy of IR therapy. Local IR (15Gy) to mice bearing colon38, a colon adenocarcinoma, resulted in decreased tumor burden in wild type animals. Interestingly, IR therapy had no effect on tumor burden in IFNγKO mice. We further determined that intratumoral (i.t.) levels of IFNγ spiked two days following IR which directly correlated with a decrease in tumor burden that was not a result of direct cytotoxic effects of IFNγ on tumor cells. CD8+ T cells and NK cells were the predominant producers of IFNγ, however both radiosensitive populations were depleted by IR; a paradox not yet completely understood. IR treated tumors could be further divided into responders and non-responders. Responders were shown to possess higher IFNγ levels, and i.t. treatment with IFNγ further enhanced the effectiveness of IR therapy. These data suggest that IFNγ may play a pivotal role in mediating the antitumor effects of IR therapy.
Early Treg suppression by a Listeriolysin-O-expressing E. coli vaccine in heterologous prime-boost vaccination against cancer (165.49)
Authors: Dai Ming-Shen , You Ren-In , Hsieh Yu-Feng , Lo Kai-Yu , Sytwu Huey-Kang , Vassaux Georges , Chao Tsu-Yi 
Published: January 2023

Abstract
[Background] Earlier studies have shown that higher CD8+ T-cell response and better tumor protection can be achieved by heterologous prime-boost vaccination in mice. We previously demonstrated that a Listeriolysin-O (LLO)-expressing E. coli vaccine can enhanced CD8-cytotoxic T cell (CTL) responses by mitigating regulatory T cells (Treg)-mediated suppression. We herein employed this Treg-inhibiting vaccine into the prime/boost immunization strategy. [Methods] Bacteria E. coli-LLO expressing Ovalbumin (OVA) and plasmid pcDNA—encoding OVA were used as specific antigen immunization. C57B6 mice and syngenic melanoma cell line B16-OVA were used in tumor challenge model. CD25 monoclonal antibody (PC-61) was used for Treg depletion. [Results] Higher OVA-specific CD8+ T-cell responses and superior tumor inhibition were seen in the bacterial-prime/plasmid-boost setting than homologous or reversed sequence. This tumor protection effect from heterologous prime/boost remained significant in the therapeutic model. When examining the Treg effect during the prime/boost immunization, we found that only early Treg-suppression/depletion could lead to better antigen-specific CTL and tumor response. [Conclusion] Our studies offer the first evidence that a Listeriolysin-O E. coli vaccine can induce superior anti-tumor effect in conjunction with DNA in a heterologous prime-boost protocol and proposed that early Treg inhibition is crucial to a successful immunization against cancer.
Severe herpesvirus-induced pulmonary pathology post-syngeneic bone marrow transplant is related to effects of transforming growth factor-beta (169.39)
Authors: Coomes Stephanie , Farmen Sara , Wilke Carol , Laouar Yasmina , Moore Bethany 
Published: January 2023

Abstract
Pulmonary infections and pneumonitis are frequent complications of hematopoietic stem cell transplant. We intranasally infected syngeneic bone marrow transplant (BMT) mice at 5 weeks post-BMT and non-transplanted control mice with the murine gammaherpesvirus, γHV-68. At d7 post-infection, BMT mice have increased levels of lytic virus compared to control. By d21 post-infection, the virus is latent and there is no difference in viral load between BMT and control mice. However, BMT mice develop virus-induced pneumonitis characterized by fibrosis, peripheral inflammation, and hyaline membranes. Alveolar macrophages from BMT mice are foamy and express increased levels of both alternative and classical activation markers. BMT mice have reduced oxygen saturation and an infiltration of neutrophils and T cells in the alveolar space, as well as increased levels of hydrogen peroxide, nitrite, and TGFβ. Using mice expressing the dominant negative TGFβRII transgene (DN-TGFβRII) as BMT donors allowed us to determine the effects of TGFβ in the development of pneumonitis on multiple cell types. T cell DN-TGFβRII BMT mice had a drastic reduction in pulmonary pathology, while CD11c DN-TGFβRII BMT mice had only a modest improvement. These data suggest that development of herpesvirus-induced pneumonitis post-BMT can be significantly reduced by blocking TGFβ signaling, especially in T cells.
Induction of PP2A Bβ, a novel regulator of IL-2 deprivation-induced T cell apoptosis, is deficient in patients with systemic lupus erythematosus. (44.17)
Authors: Crispin Jose , Apostolidis Sokratis , Finnell Melissa , Tsokos George 
Published: January 2023

Abstract
The activity and specificity of the ubiquitously expressed phosphatase PP2A is determined by the type of regulatory (B) subunit that couples to the catalytic/scaffold core of the enzyme. We determined that the Bβ subunit (PPP2R2B) -considered brain specific- is expressed in T cells during late phases of activation (48 to 72 h). To determine its function, we cloned and expressed it in non-stimulated human T cells where it triggered apoptosis. To identify which conditions promote the expression of PP2A Bβ, we induced apoptosis in T cells through Fas ligation, restimulation, or IL-2 deprivation. Transcription of PP2A Bβ was induced exclusively by IL-2 deprivation. Since T cell apoptosis is known to be altered in T cells from patients with systemic lupus erythematosus (SLE), we analyzed the regulation of PP2A Bβ in this autoimmune disease. We found that levels of PP2A Bβ did not increase upon IL-2 deprivation in 2/3 of the patients. Remarkably, this defect was accompanied by resistance to apoptosis. Moreover, kinetics of cell death were normal in cells that upregulated PP2A Bβ. We have identified a novel role for the phosphatase PP2A, particularly the holoenzyme formed by PP2A Bβ. Bβ appears to trigger apoptosis of T cells in the absence of IL-2 and probably contributes to the termination of a no-more-needed immune response. We propose that in SLE patients, defective apoptosis contributed by faulty upregulation of PP2A Bβ contributes to longer survival of autoreactive T cells.
Innate memory phenotype CD8+ T cells exhibit prompt and potent antigen specific response (46.2)
Authors: Huang Weishan , August Avery 
Published: January 2023

Abstract
Innate memory phenotype CD8 T cells (IMP CD8+CD44hiCD122int) develop in thymus and carry abundant preformed mRNA for cytokines such as interferon-γ (IFN-γ). These cells rapidly produce IFN-γ when stimulated with PMA/Ionomycin or IL-12/IL-18. Whether they can also contribute to antigen specific responses is unclear, as it is difficult to distinguish them from “true” memory cells. We developed a novel method to derive antigen specific IMP CD8, using murine bone marrow chimeras to separate naïve and IMP antigen specific CD8 T cells recognizing the model antigen ovalbumin (OVA). WT or OTI bone marrow transplanted into irradiated MHCI-/- mice gave rise to predominantly IMP CD8 T cells. By contrast, MHCI-/- or OTIMHCI-/- bone marrow transplanted into irradiated WT mice gave rise to predominantly naïve CD8 T cells. IMP CD8 T cells thus derived significantly up-regulated IFN-γ 6 days post OVA restimulation in vitro, following primary exposure to transgenic Listeria-OVA, compared to analogously derived naïve CD8 T cells alone or WT controls. More importantly, OTI IMP CD8 T cells exhibited significantly enhanced early response to OVA or OVA257-264 peptide without primary stimulation. Our findings indicate that the dynamics of the antigen specific response of IMP CD8 T cells is enhanced compared to antigen specific naïve CD8 T cells. Furthermore, this work provides a promising murine model for future investigation of IMP CD8 T cell memory development and antigen response.
Verification of immunophenotyping markers for naïve & memory mouse T cells (46.22)
Authors: Hingorani Ravi , Ernst David 
Published: January 2023

Abstract
The immune system responds with greater intensity upon antigen re-exposure. This forms the basis of immunological memory. Mouse memory T cells form a heterogeneous group of long-lived cells that express high CD44 levels. Naïve and central memory T cells express CD62L and recirculate through secondary lymphoid tissues while effector memory cells do not express CD62L and do not recirculate through the lymphoid tissues. Effector memory T cells express homing receptors enabling their entry into nonlymphoid tissues and are prevalent during certain infections. Clear understanding of mechanisms that mediate immunologic memory is essential for effective vaccine development. Tremendous effort has been devoted to characterizing markers on the surface of T cells to help identify cells responding to vaccines. To date no single marker that segregates T cell subsets involved in immunological memory has been described. Based on the literature we developed a 4-color fluorescent antibody panel that identifies naïve, memory and effector mouse T-cells. The panel consists of antibodies towards CD4, CD3, CD44 and CD62L. This panel enables identification of core subsets and provides flexibility to incorporate additional probes/antibodies labeled for the FL1 channel. Optimal set up procedures and gating strategies for different flow cytometers will be presented. The panel was used to differentiate naïve and memory T cells from old and young mice, in different tissues and different mouse strains.
Intrinsic interferon signature in myeloid dendritic cells from lupus-prone mice precedes disease onset (47.1)
Authors: Gallucci Stefania , Varghese Linda , Bennett Heather , Jog Neelakshi , Shivers Debra , Ning Yue , Behrens Edward , Caricchio Roberto , Sriram Uma 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) patients over-express Type I Interferon (IFN) responsive genes but the cellular source of this IFN Signature is unclear. We found that ex vivo dendritic cells (DCs) from Sle1,2,3 lupus-prone mice express the IFN Signature. To test if the IFN Signature was intrinsic to DCs, we grew myeloid DCs in vitro from bone marrow in GM-CSF medium (BMDCs), and found that IFN responsive genes, analyzed by real-time RT-PCR, are constitutively over-expressed in Sle1,2,3 BMDCs as compared to non-autoimmune C57BL/6 BMDCs: expression of IFN-b, signaling molecules IRF-7, STAT1 and STAT2, and IFN responsive genes ISG-15, IL-15, Oas-3 and Mx1, was significantly higher in lupus BMDCs than in C57BL/6 BMDCs in absence of any stimulation. The IFN Signature was similar in BMDCs from mice negative for autoAbs and from mice with high autoAb titers, suggesting that the IFN Signature precedes disease onset and is not affected by the autoimmune process. Sle1,2,3 BMDCs were hyper-sensitive to nucleic acids CpGs and R848, confirming a role for TLR7 and TLR9 in lupus pathogenesis. However, treatment with oligonucleotide sequences inhibitory for TLR7/TLR9 did not suppress the constitutive over-expression of IFN responsive genes, indicating that DC IFN Signature is independent from chronic exposure to TLR7/9 ligands. We suggest that myeloid DCs are a source of IFN Signature in Sle1,2,3 mice and have an intrinsic dysregulation in the IFN signaling/response.
CDR3 analysis of the TCRVβ repertoire in individual pancreatic lymph nodes of nonobese diabetic mice: evidence for public clonotypes throughout diabetes development. (47.12)
Authors: Marrero Idania , Vong Allen , Dai Yang , Davies Joanna 
Published: January 2023

Abstract
The aim of this study was to test the hypothesis that the natural T cell response during chronic spontaneous Type 1 Diabetes (T1D) is part of an ordered sequence of events that results in the expansion of a predetermined TCR repertoire. We analyzed the TCRVβ repertoire ex-vivo in the pancreatic lymph nodes (PLN) of 4 and 10-week-old pre-diabetic female NOD mice, as well as in NOD mice at the onset of the disease, using CDR3 length spectratyping. The TCRVβ repertoire was compared with the repertoire found in diabetes-resistant (NOR) mice and in NOD mice after cyclophosphamide (CYP) treatment. We show that public clonotypes are common in NOD mice at all stages tested and that the responding public T cell repertoire is dynamic and changes during disease development giving a different signature for prediabetic and diabetic mice. Also, we show that public clonotypes are not restricted to disease susceptible mice. In addition, acute diabetes induced in pre-diabetic mice with CYP is associated with a shift in the responding public T cell repertoire to resemble that of mice with spontaneous T1D. Expansions in T cells expressing TCRVβ5.2, and Vβ12 families are strongly associated with diabetes onset, whether disease is acute or chronic. In conclusion, we show that the progression of T1D is associated with the generation of public T cell clonotype signatures that are specific to early prediabetic, late prediabetic and diabetes onset.
Expansion of peritoneal B1a cells in NZM2410 lupus-prone mice is regulated by the cell cycle regulator gene Cdkn2c (47.3)
Authors: Potula Hari Hara , Xu Zhiwei , Morel Laurence 
Published: January 2023

Abstract
Here, we identified the cyclin-dependent kinase inhibitor Cdkn2c (p18) gene in the Sle2c1 NZM2410-derived lupus susceptibility locus by the combination of genetic mapping and candidate gene analysis, that is responsible for an expansion of the B1a cell compartment. A novel -74 T/C SNP in the p18 promoter is associated with a significantly reduced p18 expression in the splenic B cells and peritoneal cavity (Pc) B1a cells from Sle2c1-carrying mice, as evidenced by micro array, RT-PCR and immunoblot analysis. Compared to B6 controls, this reduced p18 expression leads to a defective G1 cell cycle arrest in splenic B cells that impairs plasma cell function, and to an increased proliferation of Pc B1a cells which results to an age-dependent expansion of the B1a cell compartment. To determine the function of the -74 T/C SNP in p18 expression, we have cloned and characterized the proximal promoter region of the mouse p18 gene in reporter vector. Functional analysis of the 5' flanking region by sequential deletions revealed crucial elements between -300 and +1, confirming the in silico prediction, that two conserved YY-1 sites located in the Sle2c1 allele negatively regulates the p18 expression in SLE prone mouse. EMSAs and ChIP analysis demonstrated that YY-1 interacts in vitro and in vivo with the p18 promoter. These results provide evidence that p18 plays a critical role in B1a cell self-renewal, and its impaired expression leads to high autoreactive accumulation of B1a cells.