bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ccr7 is critical for resistance against experimental Trypanosoma cruzi infection (56.4)
Authors: Roffe Ester , Santiago Helton , Lionakis Michail , Barber Daniel , Murphy Philip 
Published: January 2023

Abstract
The protozoan parasite Trypanosoma cruzi is the etiological agent of Chagas’ Disease, which affects ~12 million people in Latin America despite coordinated international control efforts. Cardiomyopathy due to chronic inflammation and fibrosis is the most common and serious presentation of Chagas’ Disease, but its pathogenesis is poorly understood. Several inflammatory chemokines and chemokine receptors have been implicated in mouse models of T. cruzi infection; however, roles for homeostatic chemokines have not previously been reported. Here, we found that mRNA for the homeostatic chemokine Ccl19 and its receptor Ccr7 were both increased in the heart during the acute phase after infection of WT C57BL/6 mice. Consistent with an important functional role, infection was uniformly fatal during the acute phase in infected Ccr7-/- mice, whereas mortality was only 10% in WT control mice. Death in infected KO mice was associated with uncontrolled parasitemia and increased parasite burden in both heart and liver, whereas infected WT mice controlled parasite burden in all 3 compartments. Leukocyte infiltration was also greater in the hearts of infected KO mice compared to infected WT controls, and was characterized by a lower frequency of CD8+ T cells and a greater frequency of Gr-1hiCD11b+IA- cells. Thus Ccr7 is a critical factor for survival and control of T. cruzi infection in C57BL/6 mice. The mechanisms of resistance conferred by Ccr7 are currently under investigation.
A novel mouse model of Schistosoma haematobium infection to study inflammatory mechanisms of bladder fibrosis (56.5)
Authors: Fu Chi-Ling , Knudsen Giselle , Thathireddy Anuradha , Caffrey Conor Caffrey , McKerrow James , Herbert De’Broski , Thai Kim , Xie Mingqian , Hsieh Michael 
Published: January 2023

Abstract
Chronic inflammatory diseases often result in tissue fibrosis through poorly understand mechanisms. Over 112 million people worldwide are infected with the parasitic worm Schistosoma haematobium. Long-term infection with S. haematobium is associated with parasite egg-induced bladder inflammation and bladder fibrosis. Studying S. haematobium infection may provide clues regarding mechanisms of tissue fibrosis in general. We sought to develop and characterize a more reliable mouse model of S. haematobium infection to study inflammatory mechanisms of tissue fibrosis. BALB/c mice underwent bladder wall injection with S. haematobium eggs and exhibited synchronous bladder granuloma development by 4 days post-injection. Egg-injected bladders began showing fibrosis by 14 days post-injection. Urine was serially sampled to profile the proteome by mass spectrometry and revealed that distinct blood, coagulation, and immune proteins were increased in the urine of egg-injected mice relative to controls. Bladder granulomas grew over time and contained many eosinophils and macrophages. Serum cytokines were analyzed by Luminex analysis and showed early and extended upregulation of IL5, IL1A, and VEGF; and late upregulation of multiple chemokines (KC, MCP1, MCP3, and IP10). We have successfully recapitulated inflammation-associated schistosomal bladder fibrosis in mice. We are now poised to employ genetically modified mice to dissect the inflammatory mechanisms which drive bladder fibrosis.
Characterization of sepsis pathogenesis using molecular biomarkers and drug treatments in the mouse CLP model (56.6)
Authors: Bender Andrew , Wu Yin , Burgess Laurette , Teceno Tyler , Skokanova Eva , Cao Qiongfang , Wilcoxen Keith , Ishizaka Sally , Shirota Hiroshi 
Published: January 2023

Abstract
Sepsis is a severe systemic disease caused by an uncontrolled immune response to an inflammatory stimulus. Sepsis has a high mortality rate and few successful treatments are available. Furthermore, one of the confounding aspects of sepsis treatment is the highly variable and dynamic manifestation of the disease. Thus, there is a need not only for new sepsis treatments but also for biomarkers of sepsis that may guide physicians in patient treatment. The cecal ligation and puncture model (CLP) is a clinically relevant model that replicates the nature of human sepsis after trauma as it has a focus of infection rather than a systemic initiation. Thus, we have chosen to characterize the CLP model and use it to evaluate candidate sepsis biomarkers and therapeutics. We evaluated a variety of potential biomarkers and correlated them with body temperature to determine which ones best reflect the health of the animal. Interestingly, some cytokines and blood chemistry markers highly correlated with health and bacterial load while others did not. To gain further insight into sepsis pathogenesis we tested drugs with differing mechanisms of action. A significant observation was that an antibiotic was highly efficacious while the immunosuppressive dexamethasone was not beneficial; suggesting that bacterial elimination is crucial for survival. In summary, our results suggest potential biomarkers for guiding sepsis treatment and provide insight into sepsis pathogenesis.
Single-cell heterogeneity analysis provides rapid validation of biophysical parameters in living cells and quantifies heterogeneous sensitivity to interleukin-2, -7, and -15. (57.12)
Authors: Coward Jesse , Hathorn Matthew , Karabacak Volkan , Even Dar Orly , Altan-Bonnet Grégoire 
Published: January 2023

Abstract
The single-cell resolution of flow cytometry provides a quantitative measurement of the heterogeneity of protein expression within clonal populations of cells. As the biological significance of this variability is becoming more widely appreciated, there is a need for new tools and methodology to analyze and leverage the effects of endogenous variation. These tools can provide alternatives to genetic methods, such as siRNA knockdown or germline knockout, that are commonly used to manipulate protein expression. We introduce such a new tool, named ScatterSlice, that enables rapid quantification of the correlation between heterogeneity in cellular responses (measured by staining of phospho-proteins) and varied expression of signaling molecules. We demonstrate its application to the validation of biochemical models of cellular responses to IL-2, IL-7, and IL-15 in living, unperturbed cells. In addition, we demonstrate that heterogeneous distribution of receptor subunits within a population of lymphocytes results in gradations of sensitivity to each cytokine. Finally, using experimental measurements of error and uncertainty in combination with biochemical models, we demonstrate the robustness of this analysis to experimental noise, and consider both limits and extensions to its use.
Engineering cell-type selective immune responses using mechanism-based designer IL-4 cytokines. (57.8)
Authors: Creusot Remi , Junttila Ilkka , Bates Darren , Moraga Ignacio , Lupardus Patrick , Fathman C. Garrison , Paul William , Garcia K. Christopher 
Published: January 2023

Abstract
IL-4 supports the differentiation and function of B and T cells, but also induces a response in virtually all cell types. Lymphocytes express mostly the IL-4 type I receptor (IL-4Ra and common gamma (CG) chains), and non-hematopoietic cells express primarily the type II receptor (IL-4Ra and IL-13Ra1 chains; also receptor for IL-13), while other leukocytes express both. We have engineered human IL-4 variants with up to 3-logs higher affinity for CG, or up to 2-logs increased binding to IL-13Ra1. Binding of these variants did not perturb receptor orientation. Selected variants were tested on various human cell lines or peripheral blood cells for their ability to differentially induce phospho-STAT6. The RGA variant, which has the highest affinity for CG and lost 1-log binding affinity to IL-13Ra1, induced significantly more pSTAT6 in the Ramos B cell line and in primary lymphocytes, and less pSTAT6 in epithelial and fibroblastic cells, compared to IL-4. Conversely, the KFR variant, which had the highest affinity to IL-13Ra1, had the opposite effect on these cells. Relative abundance (or scarcity) of CG and IL-13Ra1 on the cell surface dictated the potency of each variant: cells expressing high levels of both receptors were not affected, whereas cells expressing limited CG benefited most from the increased affinity afforded by RGA. These new variants have potential therapeutic application in targeting selective cell types, and thereby reducing toxic effects in vivo.
Lack of direct antigenic processing results in delayed CD8+ T cell response, which fails to control Salmonella typhimurium infection (58.12)
Authors: Tzelepis Fanny , Alcon Valeria , Russmann Holger , Krishnan Lakshmi , Sad Subash 
Published: January 2023

Abstract
CD8+ T cells are activated rapidly to control prolific pathogens, however this is not the case during Salmonella typhimurium (ST) infection. In resistant mice, recombinant ST expressing ovalbumin (ST-OVA) induces a delayed CD8 T cells response and a chronic infection is established. As ST-OVA resides in the phagosomes of infected cells, CD8+ T cells are considered to be activated by the alternative cross-presentation pathway. We wanted to address if the CD8 T cells can be activated through the classical MHC-I antigen presentation in this model, and whether that would improve the control of virulent ST-OVA infection. We modified ST to secrete and translocate the immunodominant antigen OVA directly to the cytosol of infected cells through the bacterial type III secretion system. Our results reveal that this modification induces rapid antigen-presentation, which results in profound CD8+ T cell response and differentiation into memory cells. More importantly, this modification makes the targets susceptible to CD8+ T cell-mediated death, which results in abridgment of ST persistence even in susceptible hosts. Thus, lack of direct antigen-processing in ST infected cells facilitates the virulence of ST and perpetuation of infection.
Innate and adaptive Type 2 cytokines drive mucosal eosinophilia (58.2)
Authors: Nussbaum Jesse , Liang Hong-Erh , Sullivan Brandon , Price April , Locksley Richard 
Published: January 2023

Abstract
Helminth infection, asthma, and allergy share classical hallmarks of Type 2 immunity, in which Type 2 helper T (Th2) cells coordinate antibody production, mucus secretion, and innate effector recruitment via IL-4, IL-5, and IL-13 in affected tissues. Identifying the cells and interactions that drive Type 2 immunity may lead to improved treatments of human diseases. IL-5 is the specific survival and maturation factor for eosinophils and is essential for eosinophilia. Although eosinophils are a key Type 2 effector cell, their precise physiologic role in immunity is controversial. We generated an IL-5 reporter mouse and crossed it to our previously published IL-13 reporter strain to study eosinophil recruitment and tissue maintenance. IL-5-producing cells were found in resting lung, small intestine, and adipose tissue. After helminth infection, lung, liver, and intestinal IL-5-producing cells increased in number and in fluorescence intensity, corresponding with local eosinophilia. Intriguingly, much of the IL-5 signal arose not from Th2 cells, but instead from an “innate lymphocyte” population lacking T cell, B cell, and natural killer markers. Expression of IL-5 and IL-13 was similar but not identical. Innate lymphocytes did not express IL-4, and their distribution in mucosal tissues and their cytokine pattern suggest they may help maintain barrier and/or metabolic homeostasis. Studies are ongoing to define the relationship of these cells to Th2 cells and eosinophils.
Toll-like receptor 2 signaling regulates the tubercle granuloma (58.6)
Authors: McBride Amanda , Salgame Padmini 
Published: January 2023

Abstract
The granuloma developed in the host in response to Mycobacterium tuberculosis (Mtb) is a protective structure that curtails the spread of the pathogen and controls reactivation of latent/persistent Mtb. Despite this knowledge, the molecular pathways that regulate the structural reorganization of immune cells in the lung remains poorly understood. We have found that TLR2KO mice can resist acute infection following low dose aerosol infection with Mtb, but are unable to maintain the granulomatous response and control chronic infection. Consequently, the goal of this study was to identify molecular pathways downstream of TLR2 signaling that may regulate granuloma maintenance in the chronic phase. We therefore examined the transcriptional signature of WT and TLR2KO lungs obtained at several time points following Mtb infection using Affymetrix GeneChip Mouse Genome 430A 2.0 Array. Gene set enrichment analysis (GSEA) indicated that enriched upregulated gene sets in the TLR2KO mice corresponded to Cell cycle, cell proliferation and cell activation related gene sets, while enriched down-regulated gene sets in the TLR2KO mice corresponded to extracellular matrix related gene sets, specifically matrix metalloproteinases (MMPs). Together with earlier reports that MMPs regulate the developing tubercle granuloma, our finding of reduced MMP expression in the lungs of TLR2KO mice indicates a novel role for TLR2 in host defense against Mtb through the control of MMP secretion.
Characterization of B cell responses to Salmonella infection (58.7)
Authors: Nanton Minelva , Lee Seung-Joo , Jackson Amy , Davies D. Huw , McSorley Stephen 
Published: January 2023

Abstract
Infection of susceptible mice with Salmonella typhimurium provides a model for studying immunity to typhoid or disseminated non-typhoidal bacteremia. Although B cells are required for adaptive immunity to Salmonella infection the mechanism of protection is unclear. We immunized IgA-, and polymeric Ig receptor-deficient mice with attenuated Salmonella and examined resistance to secondary infection. Neither of these strains displayed any deficiency in acquired immunity to secondary infection. We also used proteomic microarrays to examine the specificity of B cell responses in mice vaccinated with attenuated Salmonella. These studies have revealed new protein targets that can be used to quantify the antibody response and track Salmonella-specific B cells. Our data suggest that B cells primarily contribute to anti-Salmonella immunity at systemic sites and recognize a diverse array of protein targets in vivo.
Innate stimulation of bacterial-specific Th1 CD4 T cells (58.8)
Authors: O'Donnell Hope , Srinivasan Aparna , McSorley Stephen 
Published: January 2023

Abstract
CD4 T cells are activated in secondary lymphoid organs upon T cell receptor (TCR) ligation with MHC-II bound peptide on antigen presenting cells (APCs). Effector Th1 CD4 T cells traffic to sites of infection, where subsequent interactions with infected APCs can stimulate production of IFNγ. Our laboratory has recently described an alternative mechanism of bacterial-specific Th1 re-activation via innate stimulation in the absence of MHC-II interaction. Using mixed bone marrow chimeric mice we extend these observations and show that CD4 T cell stimulation is dependent upon intrinsic MyD88 and/or TRIF expression. Current work attempts to identify whether innate stimulation occurs directly on CD4 T cells through TLR4 or indirectly by MyD88-dependent cytokine signaling. Understanding innate stimulation of bacterial-specific CD4 T cells may lead to the development of more effective vaccines and therapeutics.
Immunization of heat-killed Klebsiella pneumoniae promotes strong mucosal Th17 responses and protects mice from live infection (59.4)
Authors: Chen Kong , Fisher Candice , Zheng Michael , Weaver Casey , Kolls Jay 
Published: January 2023

Abstract
Bacterial pneumonia is a leading cause of mortality and one major pathogen associated with this disease is Klebsiella pneumoniae (KP). Thus, a vaccine against KP is in great need especially for immunocompromised patients. Here we show that intranasal immunization of mice with heat-killed KP strongly induces IL-17 production in the lung. Using an Il17f reporter mouse, we were able to sort IL-17F positive cells after immunization by FACS. Further analysis reveals that these cells are conventional CD4+TCR-beta+ Th17 cells and they express Il17a, Il17f, Il22, Rora and Rorc. These Th17 cells are antigen specific as they proliferate and produce IL-17 upon re-stimulation with heat-killed KP but not heat-killed Escherichia coli or Staphylococcus aureus. These responses ultimately lead to the protection of immunized mice from live KP infection. This immunization strategy also confers protection in B cell-deficient (mu-MT) mice, demonstrating the vaccination can be antibody independent. We also show that other mucosal routes of vaccination such as intragastric immunization also reduces bacterial burden during pulmonary infection, suggesting that Th17 cells can be primed or recruited from other mucosal sites distal to the lung and oral vaccine can be effective in pulmonary infection. These data suggest that the Th17 pathway can be a target of pulmonary vaccine development through intranasal or intragastric immunization.
The role of Pax5 and E47 on AID expression in B lymphocytes of Aged BALB/c Mice (61.1)
Authors: Landin Ana Marie , Frasca Daniela , Scallan Martina , Harrison Patrick , Hagman James , Blomberg Bonnie 
Published: January 2023

Abstract
Previous results from our laboratory demonstrated that stimulated enriched (95%) B cells from senescent mice (19 to 24 months) have an intrinsic defect in class switch recombination (CSR), and a decrease seen in both Pax5 and E47. The decrease in both Pax5 and E47 is proportional to the decrease in activation induced cytidine deaminase (AID) with aging. Binding sites for both Pax5 and E47 have been identified in the gene for AID, Aicda, and stimulation of B cells results in the recruitment of Pax5 and E47 to the Aicda gene and its expression. Our current results show that overexpression of both E47 and Pax5 rescues levels of AID and CSR in old B lymphocytes to young-like levels. Furthermore, exogenous retroviral E47 mRNA is more stable than the endogenous E47 due to the removal of the 3’UTR. Overexpression of Pax5 leads to up-regulation of E47 in both primary B cells and 293T cells; however, overexpression of E47 up-regulates Pax5 in B cells but not in 293T cells. Nuclear extracts from B cells of senescent mice show lower levels of binding consistent with reduced levels of E47 and Pax5 in aging. Using a DsRed lentiviral vector driven by the Aicda promoter and containing the E47 and Pax5 enhancer region, we show that in 293T cells DsRed is only expressed when the cells are co-transduced with E47 and/or Pax5.
B cell extrinsic MyD88 and FcRγ control contraction of the extrafollicular B cell response (61.12)
Authors: Sweet Rebecca , Cullen Jaime , Shlomchik Mark 
Published: January 2023

Abstract
The extrafollicular (EF) B cell reaction is emerging as a lynchpin of systemic autoimmune responses. External factors that limit this response remain undefined. AM14 transgenic mice have been useful in elucidating how autoreactive Rheumatoid Factor B cells are activated. On a lupus-prone genetic background, AM14 B cells undergo an isotype-switched EF plasmablast response. A similar response ensues when these cells are exposed in vivo to IgG2aa anti-chromatin antibodies, which presumably form immune complexes (ICs) with chromatin shed from dying cells. ICs have Fc that could also stimulate myeloid cells via FcRγ and TLRs, which in turn could influence the EF response. To investigate this, we transferred AM14 B cells into mice lacking FcRγ, MyD88 or both. Initial expansion and differentiation of AM14 B cells was equivalent in all hosts. However, by day 7, whereas the AM14 B cell response contracted in the WT environment, dysregulated expansion continued in the deficient environments. Furthermore, MyD88 and FcRγ are non-redundant, as responses in mice lacking both proteins were larger and more prolonged compared with mice deficient in either factor alone. By d10, the AFC frequency was 40-fold larger in double-deficient compared to WT recipients. These results reveal novel regulatory roles in the EF B cell response for two receptors that are typically considered proinflammatory. Targeting these pathways may provide a way to curtail dysregulated autoreactive B cell activation.
Follicular dendritic cell activation during vaccination correlates with antibody response and germinal center formation (61.14)
Authors: Cantisani Rocco 
Published: January 2023

Abstract
FDCs are stromal cell of the lymphoid organs that play a central role in the induction of humoral immune response. FDCs deliver signal to B cells by trapping intact antigen onto their surface as antigen-antibodies or antigen-complement fragments immune complexes. Thus cognate B cells recognize the antigen presented by FDCs via BCR, while surface proteins or soluble factors expressed by FDCs serve as co-stimulation. FDC activation is pivotal for the origin of germinal center (GC) reaction, and TLR4 engagement appears critical for this purpose. Nevertheless, no studies analyzing the commitment of FDC during vaccination with licensed adjuvants have been performed. Here we show for the first time, that the intramuscular injection of adjuvants approved for human use leads to FDC activation, which occurs late after injection. In addition, we observe that the activation of FDCs upon injection of an adjuvanted antigen takes place only in the follicles where a GC reaction occurs and the number of follicles with activated FDCs strictly correlates with the antigen-specific antibody titers. Moreover, retention of the injected antigen is observed only in the follicles that contain activated FDCs. Thus the extent of the humoral immune response against an antigen correlates with the activation state of FDCs, which can be modulated by the presence of the adjuvants, maybe through an indirect effect. The elucidation of the adjuvant affect on FDCs can help to design more efficacious vaccines.
PARP-14 mediates B cell memory response through the regulation of energy metabolism and gene transcription (61.5)
Authors: Cho Sung Hoon , Trochtenberg Alyssa , McGuinness Owen , Boothby Mark 
Published: January 2023

Abstract
Immunological memory, including both persistent and recall antibody (Ab) responses, enables more efficient and effective protection against recurrent or chronic infection. Although many basic features of memory responses are understood, key molecular mechanisms regulating emergence and persistence of memory B cells and plasma cells remain unknown. Recent evidence suggests that the regulation of energy metabolism might mediate memory T cell differentiation and function. PARP-14, a Stat6-interacting protein is highly expressed in lymphoid organs, influences B cell subset ratios as well as the initial IgA response to antigen, and has intrinsic ADP-ribosyltransferase activity. We now show that PARP-14 is essential for the regulation of energy metabolism in B cells and has been significant role in the increased Ag-specific Ab levels that follow booster immunization. Investigation of the mechanism(s) revealed that PARP-14 mediates at least in part to the formation of long-lived plasma cell formation and regulates IL-4 induction of PRDM1 (Blimp-1), a master regulator of plasma cell differentiation. These results reveal a novel function of a mammalian ADP-ribosyltransferase in memory of B lineage cells and may shed a light on a new target for vaccine development.