bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of Jak/STAT signaling synergizes with CTLA4-Ig to promote transplant acceptance (IRC2P.454)
Authors: Raimondi Giorgio , Khalifian Saami , Oh Byoung , Lee W.P. Andrew , Brandacher Gerald 
Published: January 2023

Abstract
Transplant tolerance induction through conventional costimulation blockade (CoB) remains an elusive goal. Recent evidence suggests that inflammatory cytokines (IC) contribute to the activation of alloreactive T cells in a CD28- and CD40-independent manner. We aimed to understand the mechanisms underlying this process and delineating the possible synergism between CD28 blockade and inhibition of IC signaling via Jak/STAT inhibition. Murine T cell activation was assessed by quantification of IL-2 secretion (at 24h post TCR/CD28 engagement). CTLA4-Ig diminished the proportion of IL-2+ cells. However, the concomitant addition of the supernatant (MATSup) from maturing dendritic cells completely abolished this effect. This suggested an early impact of IC on T cell activation. In a CFSE-based proliferation assay, MATSup counteracted the anti-proliferative effect of CTLA4-Ig on both CD4 and CD8 T cells. Inhibition of Jak/STAT signaling through addition of Tofacitinib (Tofa) restored CTLA4-Ig regulation in presence of MATSup. Finally, in a BALB/c to B6 skin transplant model a short course of Tofa (d0-12) synergized with CTLA4-Ig doubling skin allograft survival (MST untreated: 9.5d, CTLA4-Ig only: 13d, Tofa+CTLA4: 18d). Overall, our results indicate that the stimulatory effect of IC counteracts the efficacy of CoB. However, their redundant activity can be neutralized through concomitant inhibition of Jak signaling - a promising strategy for promotion of transplant acceptance.
Anti-TIM-4 mediates long-term engraftment of islet allografts by promoting IL-10 expression by TIM-1+ Bregs and inhibiting IFNγ expression by proinflammatory “Be1” B cells (IRC3P.464)
Authors: Ding Qing , Yeung Melissa , Xiao Sheng , Najafian Nader , Kuchroo Vijay , Rothstein David 
Published: January 2023

Abstract
We previously defined TIM-1 as an inclusive marker for IL-10+ regulatory B cells (Bregs). Moreover, α-TIM-1 mAb prolongs islet allograft survival (GS) through induction of TIM-1+ Bregs. We recently noted that TIM-4, a TIM-1 ligand, identifies B cells enriched for IFNγ (Be1). We therefore hypothesized that TIM-4 blockade should also prolong GS and that this may involve B cells. In fact, α-TIM-4 (RMT4-53) treatment resulted in long-term GS in BALB/c recipients of B6 islets (MST &gt;100d). Moreover, α-TIM-4 increases Th2 cytokines (IL-4, IL-10) and Foxp3+Tregs, while reducing both Th1 cytokines (IFNγ) and CD4 proliferation. However, these α-TIM-4-mediated changes and long-term GS were B cell dependent, and did not occur in either B cell deficient (JHD) or depleted (α-CD20) recipients. To address whether α-TIM-4 directly targets (TIM-4+) Be1 cells, we showed that adoptive transfer of WT, but not TIM-4-/- B cells, into μMT recipients, reconstitutes long-term GS mediated by α-TIM-4. We also found that α-TIM-4 inhibits IFNγ expression by TIM-4+ B cells by 40%. Bregs were also required for α-TIM-4 induced transplant tolerance. Transfer of B cells from TIM-1Δmucin (mucin domain deletion) mice, which exhibit a defect in Bregs, is unable to prolong GS in B deficient mice. In wt mice, α-TIM-4 increases TIM-1+ IL-10+ Bregs by ~100%. Taken together, our data reveal that targeting TIM-4 enhances IL-10 expressing Bregs and also reduces inflammatory Be1 B cells to promote allograft tolerance.
CD86-dependent FoxP3+ regulatory T cells promote resolution of disease after influenza virus infection (IRC4P.492)
Authors: Moser Emily , Hufford Matthew , Braciale Thomas 
Published: January 2023

Abstract
Influenza A virus (IAV) infection in the respiratory tract triggers a robust immune response, resulting in both virus clearance and lung inflammation and injury. After virus clearance, resolution of lung disease occurs during a distinct recovery period. The costimulatory molecule CD86 can modulate T cell activity during the initiation and effector stages of the host response to IAV infection, but the role of CD86 during recovery is unknown. We found that antibody-mediated CD86 blockade after virus clearance led to a delay in recovery, characterized by increased numbers of lung neutrophils and innate cytokines in the bronchial alveolar lavage fluid (BAL), but no change in conventional IAV-specific T cell responses. However, CD86 blockade led to decreased numbers of FoxP3+ regulatory T cells (Tregs), and adoptive transfer of Tregs into α-CD86 treated mice rescued the effect of the blockade, supporting a role for Tregs in promoting recovery after virus clearance. Direct depletion of Tregs late after infection using DEREG mice mimicked the CD86 blockade phenotype, confirming a role for Tregs during recovery after virus clearance. Finally, we identified neutrophils as a target of Treg suppression, since neutrophil depletion in Treg-depleted DEREG mice reduced excess BAL cytokines. These results show that CD86-dependent Tregs contribute to the resolution of disease after IAV infection, in part by suppressing neutrophil-driven cytokine release into the airways.
Development of a new mouse/human FoxP3 antibody using rabbit recombinant antibody technology (IRC4P.477)
Authors: Goetz Christine , Bonnevier Jody , Bjordahl Ryan , Houchins J. , Munshi Cyrus , Aggeler Birte 
Published: January 2023

Abstract
Tregs play a key role in immune system suppression during autoimmunity and tumor development. Tregs are classified as CD4+CD25+FoxP3+, and exist as either nTregs originating from the thymus, or iTregs derived from CD4+ effector T cells in the periphery. Reduction in Treg numbers leads to autoimmunity; this is clearly shown in scurfy mice where FoxP3 has been deleted, Treg numbers are severely reduced, and mice succumb to autoimmunity early in life. In contrast, Tregs play a debilitating role in cancer biology when the Treg numbers are elevated, and the immune response is dampened allowing cancer cells to evade the immune system. Thus, the correct balance of Tregs is essential for maintaining immune homeostasis. FoxP3 has been widely used as a marker of Tregs, and many antibodies are available on the market that distinguish the Treg population in human or mouse cells. We developed a novel FoxP3 antibody and Fix and Perm buffer system that is comparable to current clones, but recognizes both mouse and human FoxP3 equally well, eliminating the need for separate antibodies. The ms/hu FoxP3 antibody was generated using our Rabbit Recombinant antibody technology, which allows for selecting high specificity, high affinity antibodies, lot to lot consistency and detection of both mouse and human nTregs and iTregs. This allows researchers to extend their mouse models into clinical studies without a change in the antibody clone eliminating a variable and streamlining the process.
DNA methylation profiles at the human Th2 locus in BAC transgenic mice point to novel putative regulatory elements (IRM6P.720)
Authors: Pivniouk Vadim , Rosenbaum David , Pivniouk Oksana , Herrell Amanda , Miller Susan , Sprissler Ryan , Lyons Eric , Vercelli Donata 
Published: January 2023

Abstract
Most cis-regulatory elements in the mouse Th2 cytokine locus were initially identified based on their evolutionary conservation. In contrast, the regulatory landscape of the human Th2 locus, which is ~40 kB larger than the mouse syntenic region, remains poorly defined because recently arisen putative regulatory elements are less readily identifiable by bioinformatics and cannot be functionally interrogated in humans. To address these limitations we created BAC transgenic (TG) mice carrying 160 kB of the human Th2 locus including RAD50, IL13, and IL4. These mice appeared normal and demonstrated faithful transcriptional regulation of human Th2 cytokine genes. Because dynamic changes in chromatin architecture typically mark functional non-coding sequences, we used next generation bisulfite sequencing to compare CpG methylation of DNA isolated from TG naïve CD4+ T cells and in vitro polarized Th2 or Th1 cells. All major regulatory elements previously characterized in mice were clearly identifiable by their CpG methylation profile in the human TG Th2 locus. Moreover, we identified at least three other non-coding regions (conserved highly in primates and moderately in species of the clade Laurasiatheria) that were differentially methylated in Th2, Th1 and naïve T cells, and are located in the RAD50/IL13 and IL13/IL4 intergenic areas. Our data suggest these regions contain novel cis-regulatory elements involved in the transcriptional regulation of human Th2 cytokines expression.
Type I interferon negatively regulates acute IL-5 and IL-13 expression in human memory Th2 cells (IRM6P.717)
Authors: Gonzales-van Horn Sarah , Huber Jonathan , Farrar David 
Published: January 2023

Abstract
IFN-α/β is a potent antiviral cytokine that induces the expression of hundreds of interferon-sensitive genes (ISGs). ISGs are positively regulated by IFN-α/β signaling and collectively interfere with various aspects of viral replication. Although the positive regulatory aspects of IFN-α/β signaling through STAT activation have been studied at length, less is known about its role as a negative regulator. Previously, we demonstrated that IFN-α/β suppresses Th2 development in human naïve CD4+ T cells by negatively regulating GATA3 expression, leading to the suppression of Th2 cytokines. In the current study, we found that IFN-α/β also suppressed Th2 cytokine expression from fully committed Th2 cells responding to recall antigen stimulation. Human memory CD4+ T cells acutely stimulated in the presence of IFN-α showed suppressed IL-5 and IL-13 gene expression, however, IL-4 expression was unaffected. Nuclear run-on studies demonstrated that IFN-α signaling suppressed de novo transcription of IL-5 and IL-13, which did not require new protein synthesis. Our data suggest that IFN-α/β was able to acutely repress specific genes by directly inhibiting their transcription. We are assessing direct STAT2 binding by ChIP-Seq, which will identify cis-regulatory regions necessary for gene repression at these cytokine promoters. Taken together, these results demonstrate that IFN-α/β is a key negative regulator of the Th2 response in humans.
The generation of peptide: MHCII-specific TH17 cells in response to intranasal bacterial infection requires IL-6 and TGF-β1, produced from a distinct mucosal dendritic cell subset. (LYM3P.740)
Authors: Linehan Jonathan , Jenkins Marc , Dileepan Thamotharampillai , Cleary Paul 
Published: January 2023

Abstract
Intranasal infections preferentially generate TH17 cells while intravenous infections induce TH1 cells. We explored the basis for these anatomic preferences by tracking endogenous polyclonal CD4+ T cells specific for a p:MHCII ligand derived from the mucosal pathogen Streptococcus pyogenes (Sp). Sp p:MHCII-specific CD4+ T cells were first activated in the cervical lymph nodes (CLN) following intranasal inoculation and then migrated to the spleen and nasal-associated lymphoid organs. The effector cell population generated in the CLN consisted of TH17 cells and TFH cells. Sp-induced TH17 formation was associated with IL-6 and TGF-β1 production by Batf3- CD103- CD301b+ dendritic cells that were located in the CLN but not the spleen. Thus, the tendency of intranasal infection to induce TH17 cells is related to cytokine production by specialized dendritic cells that drain this site.
The coexpression of Tim-3 and PD-1 leads to CD8 T cell exhaustion in aged female mice (LYM4P.760)
Authors: Shin Kwnag-Soo , Lee Kyoo-A , Kim Ga-Young , Kang Chang-Yuil 
Published: January 2023

Abstract
T cells, which are major components of adaptive immune system, are dysfunctional in aged individuals due to numerous intrinsic and extrinsic changes of aged T cell. Recently, several studies have shown that the expression of inhibitory receptors is increased on CD8 T cells from aged mice, which is also observed on exhausted CD8 T cells in chronic infection model, by which effector T cells are negatively regulated. In this study, we described that Tim-3+ PD-1+ CD8 T cells increased in aged mice compared to those of young mice. The more extensive investigations on the functional characteristics of aged Tim-3+ PD-1+ CD8 T cell revealed that these cells had defects in cytokine (IFN-γ, TNF-α) production and proliferation, consistent with features of exhausted CD8 T cell in a chronic infection model. Aged Tim-3+ PD-1+ CD8 T cell produced more immune regulatory cytokine, IL-10, than PD-1 single positive or Tim-3 and PD-1 double negative CD8 T cells. Furthermore, there was more diminished homeostatic proliferation of aged Tim-3+ PD-1+ CD8 T cells than other subsets in lymphopenic conditions, indicating that aged Tim-3+ PD-1+ CD8 T cell had cell-intrinsic defect in proliferation capacity. Interestingly, adoptively transferred aged Tim-3+ PD-1+ CD8 T cells in naïve B6 hosts survived longer than other subsets. Collectively, our findings on aged Tim-3+ PD-1+ CD8 T cell would provide a important insight for understanding the defect in the immune senescence.
Spatial and temporal T cell compartmentalization over the human lifetime: a quantitative, system wide analysis (LYM4P.745)
Authors: Yudanin Naomi , Thome Joseph , Ohmura Yoshiaki , Farber Donna 
Published: January 2023

Abstract
The efficacy of memory T cells in protective immunity has been shown in mouse models, but their lifelong compartmentalization and maintenance in humans remains unclear. To systematically probe lifelong memory T cell generation and maintenance, we have analyzed their distribution and function among 9 tissues acquired through from over 40 healthy donors aged 3 to 73 years old. We statistically analyzed data derived from FACS analysis of these donors, and generated a quantitative map of T cell compartmentalization and maintenance over the human lifespan. Our analyses revealed that differentiated effector memory subsets primarily resided in peripheral tissues while central and naïve subsets remained in circulatory sites. Moreover, expression of the resident memory marker CD69 effectively predicts compartmentalization in peripheral tissues, indicating the need for increased activation to enter the periphery. CD127 and CD28 expression revealed differential maintenance of the subsets in distinct sites, with more cytokine and antigen-driven maintenance prevalent in TEM and TEMRA subsets localized to the periphery. We found that the distribution and maintenance patterns of these subsets are established early in life, and vary little with increasing age. These findings suggest that tissue localization is the primary driver of human T cell maintenance and compartmentalization, and remains stable over the lifetime.
Immunomodulatory effects of IL-10 in tuberculosis granulomas of non-human primates (MPF1P.772)
Authors: Wong Eileen , Gideon Hannah , Flynn JoAnne 
Published: January 2023

Abstract
Tuberculosis (TB), caused by M. tuberculosis (Mtb), continues to be a leading cause of deaths from infectious diseases worldwide. Active TB disease has been associated with suppressed T cell responses and increased production of immunosuppressive cytokines, particularly IL-10. However, published research has focused on the systemic immune response to Mtb as measured in blood. Little is known about the local immune response within individual granulomas. Using a cynomolgus macaque model of Mtb infection, we quantified IL-10 production in granulomas and determined immunomodulatory effects of IL-10 on granuloma immune cells by flow cytometry. Although all granulomas from Mtb infected macaques were found to produce IL-10, the percent of CD3+ T cells that produced this cytokine varied between granulomas, even within an individual host. Neutralization of IL-10 in granulomas ex vivo increased IL-2 and IL-17 production, suggesting IL-10 may normally suppress T cell production of IL-2 and IL-17 in the granuloma. Other cytokines critical for control of TB, such as IFN-γ and TNF, were not affected by IL-10 neutralization. Proliferation was also increased by neutralization of IL-10. Thus, IL-10 may be locally suppressing some, but not all, T cell functions in the granulomas of TB infection. This may be important in either controlling the pathology during infection, or in preventing elimination of the bacilli in granulomas.
Pretreatment of epithelial cells with live streptococcus pneumoniae had no detectable effect on influenza A virus replication in vitro (MPF1P.774)
Authors: Gourapura Renukaradhya , Ouyang Kang , Woodiga Shireen , Dwivedi Varun , Marion Carolyn , Singh Anirudh , Binjawadagi Basavaraj , Hiremath Jagadish , Manickam Cordelia , Schleappi Rose , Khatri Mahesh , King Samantha 
Published: January 2023

Abstract
Influenza A virus (IAV) and Streptococcus pneumoniae (pneumococcus) are two major upper respiratory tract pathogens responsible for exacerbated disease in coinfected individuals. Information on the direct effect of S. pneumoniae on IAV replication in humans is unknown. Such an investigation is important because S. pneumoniae is a commensal of the human upper respiratory tract. The aim of this study was to determine whether treatment of epithelial cells with S. pneumoniae affects IAV replication, determined with the help of a standard immunofluorescence assay (IFA). For this study, four IAV permissive epithelial cell lines including two human-derived cell lines, 12 pneumococcal strains including recent human clinical isolates which represent different genetic backgrounds and serotypes, and six IAV strains of varying genetic nature and pathogenic potential including the pandemic 2009 H1N1 virus were used. Our results suggested that pretreatment of MDCK cells with 7.5×10^6 colony-forming unit (CFU) of live S. pneumoniae resulted in gradual cell-death in a time-dependent manner (1/2 hr to 4 hr). However, pretreatment of cells with 7.5×10^5 and lower CFU of S. pneumoniae had no detectable effect on either the morphology of epithelial cells or on the IAV replication. However, the effect of pneumococci on IAV replication may be different during a coinfection in vivo in the human upper respiratory tract.
Role of IL-17 in chronic pulmonary mycobacterial infection. (MPF1P.775)
Authors: Umemura Masayuki , Fukui Masayuki , Fukui Chiho , Nakae Susumu , Iwakura Yoichiro , Matsuzaki Goro 
Published: January 2023

Abstract
IL-17 family cytokine is comprised of six members, IL-17A to IL-17F. Recent studies using cytokine- and receptor-deficient mice showed that IL-17A and IL-17F were required for responses to extracellular bacterium K. pneumonia in the lungs and C. rodentium in the colon, respectively. However, the involvement of IL-17A and IL-17F in protective immunity was well not clearly demonstrated in mycobacterial infected lung. In this study, we analyzed role of IL-17A and IL-17F in host defense against chronically infected M. tuberculosis. using IL-17A- and IL-17F-KO mice. The IL-17A-KO mice showed significantly decreased survival rates compared with the wild-type (WT) mice during 250-day observation period. In contrast, survival rate of the IL-17F-KO mice were similar to that of the WT mice. Bacterial burdens of various organs of the IL-17F-KO mice on the day 250 were nearly the same as that in the WT mice. In the infected lungs, the IL-17A-KO mice produced less IFN-γ, TNF and IL-6 in comparison to those from the WT mice, while cytokine production of the IL-17F-KO mice were similar to that of the WT mice. This result indicated that the generation of Th1 cells was impaired in the IL-17A-KO mice but not in the IL-17F-KO mice infected with M. tuberculosis. These data strongly support the notion that the lack of IL-17F neither suppress nor enhances protective immunity in the lung after mycobacterial infection.
Role of STAT1 and STAT6 molecules during toxocara canis infection (MPF1P.801)
Authors: Faz Berenice , Ledesma-Soto Yadira , Martínez-Saucedo Diana , Callejas Blanca , Terrazas Luis I. 
Published: January 2023

Abstract
Toxocariasis is a worldwide zoonotic parasitic disease caused by Toxocara canis. Infection is caused by accidental ingestion of embryonated eggs, which hatch and the liberated larvae migrate to different organs. In murine models causes transitory hemorrhagic pulmonary lesions associated with strong TH2 responses and heavy parasite burdens. Alternatively activated macrophages (aaM), through STAT6 signaling, are involved in tissue repair, but its role in T. canis infection has not been determined yet. On the other hand, STAT1 has been clearly related to protection in protozoan infections, but its role on helminths is poorly known. Here we tried to recognize the role of STAT1 and 6 molecules in this infection by using STAT6-/- and STAT1-/- BALB/c mice in determining the outcome of experimental toxocariasis. Mice were infected, and specific antibodies, serum cytokines, parasite burden and lung pathology were analyzed at different times. STAT6+/+ and STAT1-/- mice displayed a Th2 immune response and their lung pathology were less evident compared to STAT6-/- mice, which displayed enhanced Th1 response but more damage in the lung. These differences in lung pathology were associated with the presence of aaM. In the other hand, parasite burden in STAT1-/- and STAT6-/- mice decreased as compared with wild type mice. Our data suggest that the absence of one or another response would contribute to eliminate this parasite, but the presence of aaM is necessary to control tissue damage.
Regulatory B cell induction by Toxoplasma gondii infection implicate in host immune escape and suppression of allergic disease (MPF1P.770)
Authors: Jeong Young-Il , Hong Sung-Hee , Lee Won-Ja , Cho Shin-Hyeong , Lee Sang-Eun 
Published: January 2023

Abstract
Toxoplasma gondii is a worldwide obligate intracellular protozoan parasite that infects humans and animals by either congenital or postnatal routes. Modulation of the immune system by infection with parasites is proposed to suppress the allergic inflammation. In this study we investigated whether experimental infection with T. gondii could induce the immune regulatory cells, including IL-10 producing CD1dhighCD5+ regulatory B cells (Bregs) and CD4+CD25+Foxp3+ regulatory T cells, and whether Breg induction was relevant to parasite immune escape during T. gondii infection. In vivo depletion studies demonstrated an important role for Bregs in T. gondii survival. Furthermore, adoptive transfer of Bregs isolated from T. gondii-infected mice enhanced cyst formation in brain of T. gondii-infected recipient mice. The present study was also investigated whether the development of allergic atopic dermatitis (AD) could be suppressed by T. gondii infection in murine. T. gondii infection substantially suppresses the development AD-like lesions induced by allergen Dermatophagoides farinae in NC/Nga mice, a model of human AD. Collectively, these results indicate that induction of Breg and Treg during T. gondii infection are necessary for evasion of host immunity, and, as the bystander suppressive effect, these cells could suppress allergic inflammation.