bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting inhibitors of NOD2 signaling to allow productive inflammation in genetic inflammatory disease (157.9)
Authors: Marinis Jill , Homer Craig , McDonald Christine , Abbott Derek 
Published: January 2023

Abstract
The intracellular bacterial sensor, NOD2, coordinates innate immune signaling pathways. Maintaining appropriate activation of NOD2 is paramount in immunologic homeostasis as dysregulation of NOD2 signaling causes a variety of inflammatory diseases including Crohn’s disease (CD). We have identified the atypical TRAF family member, TRAF4, as a key inhibitor of NOD2 signaling. We show that this inhibition occurs through two mechanisms - through IKK phosphorylation and activation of TRAF4 and through direct binding of TRAF4 to NOD2. Further work shows that TRAF4 also binds CD-associated NOD2 variants and we hypothesize that competing TRAF4 from these NOD2 variants could allow a productive inflammatory response. To this end, we have generated cell permeable peptides that mimic the region of NOD2 that binds TRAF4 in hopes of competing TRAF4 from NOD2 in vitro and in vivo to prevent the inhibitory effect of TRAF4. This may partially revert loss-of-function CD-associated NOD2 variants and could serve as a platform for future therapeutics in Crohn’s disease patients harboring these mutations.
Production and characterization of polyclonal antibodies against CD38 (159.3)
Authors: Trujillo Ocampo Abel , Vences Catalan Felipe , De la Cruz Hernandez Hernandez Fidel , Santos Argumedo Leopoldo 
Published: January 2023

Abstract
CD38 is a 42 kDa transmembrane glycoprotein widely expressed in hematopoietic cells. Anti-CD38 agonistic antibodies induce proliferation, migration, and apoptosis of B lymphocytes. Some of the available monoclonal antibodies do not recognize CD38 by Western blot. Then, we decide to prepare antisera that can be equally useful in biochemical and functional assays. To reach this goal, we used the extracellular domain of murine CD38 fused with a V5 epitope and a Histidin tag or fused with GST as antigens. These proteins were characterized by western blot, enzymatic activity and by reactivity with the mAB NIM-R5, which recognizes a conformational epitope. We immunized New Zealand rabbits and a female goat with the CD38-V5-Histidin tag fusion protein. Antibodies from both species stained cells from C57BL/6 but not from CD38 deficient mice. In addition, the polyclonal antibodies recognized in extracts from splenic cells a band of the expected weight. Finally, these antibodies induced B lymphocyte proliferation of mouse splenic cells. These results demonstrated the production of functional reagents that can be used in the analysis of CD38 in B cells.
Multicolor flow cytometric analysis of mouse T cell subsets in aging (159.4)
Authors: Ernst David , Gao Guo-jian , O'Donnell Erika , Elia Jeanne , Wu Xiao-Wei , Chronopoulou Efthalia , Sepulveda Homero , Hingorani Ravi 
Published: January 2023

Abstract
Aging is accompanied by profound changes in the function and subset makeup of peripheral CD4 and CD8 T cells. Using the Violet Proliferation Dye 450, we found significantly fewer splenic CD4 and CD8 T cells from aged mice progressed through cell cycle in response to CD3/CD28 cross linking. However, intracellular cytokine staining showed higher percentages of total T cells in older mice that produced IFN-γ and IL-17 on stimulation with little difference in their signaling pattern. Additionally, the age-associated responses were accompanied by dramatically increased proportions of memory (CD44hiCD62Llo/hi) and decreased fractions of naïve (CD44loCD62Lhi) T cell subsets. To determine if the altered T cell subset proportions could be responsible for functional differences, phospho-specific flow cytometry was used to evaluate alterations in naïve and memory T cells from young vs. old mice. Interestingly, many signaling responses were reduced in memory T cells relative to naïve T cells, including the phosphorylation of STAT1 (pY701) and STAT3 (pY705) in response to stimulation with exogenous IL-6 or IFN-γ and the phosphorylation of SLP-76 (pY128) and AKT1 (pS473) in response to CD3/CD28. However, signaling responses were largely similar within equivalent T cell subsets from old and young mice. Our results suggest that the altered T-cell functions associated with advanced age may primarily result from the dramatic shifts in naïve and memory CD4 and CD8 T cell subset representations.
Nasopharyngeal colonization by Streptococcus pneumoniae elicits long-lasting immune responses that confer protection from invasive lung infections (161.19)
Authors: Maung Nang , Goodwin Aaron , Muñoz-Elías Ernesto , Camilli Andrew , Gerstein Rachel , Leong John 
Published: January 2023

Abstract
Streptococcus pneumoniae can cause life-threatening pulmonary and systemic infection, but resides usually harmlessly in the nasopharynx of many people. Humoral immunity is thought to be the primary defense against serious pneumococcal infection, and we hypothesized that nasopharyngeal colonization of mice may result in the generation of an antibody response that provides long-term protection against lung infection. In fact, we found that intranasal colonization elicited pneumococcal-specific antibody responses and protected from a S. pneumoniae lung challenge that is lethal in naive mice. B cells were required for protection, and transfer of immune sera from colonized mice protected naive mice. CD93-/- mice, which are deficient in long-term antibody secreting cells (ASC), had fewer pneumococcal-specific bone marrow ASC after intranasal colonization and did not survive lung challenge. T cells were required to generate a fully protective response, but elimination of CD4+ T cells just prior to lung challenge did not compromise. These data are consistent with the hypothesis that CD4+ T cells promote the development of post-colonization immunity but are not needed to sustain the response that provides protection from lung challenge. These results stand in contrast to the T-cell independent antibody responses that are sufficient to protect mice from systemic infection, and have implications for development of effective vaccines.
Intestinal epithelial cells modulate CD4 T cell responses via the thymus leukemia antigen (161.8)
Authors: Olivares-Villagómez Danyvid , Scott Algood Holly , Singh Kshipra , Parekh Vrajesh , Ryan Kaitlyn , Piazuelo Maria , Wilson Keith , Van Kaer Luc 
Published: January 2023

Abstract
The thymus leukemia (TL) antigen is a non-classical MHC class I molecule primarily expressed on intestinal epithelial cells (IEC) that interacts with CD8αα, which is primarily expressed by intestinal intraepithelial lymphocytes. The close proximity of cells expressing these two molecules suggests that TL-expressing IEC can regulate CD8αα-expressing lymphocytes. Here, we report that TL influences the cytokine profile and pathogenic potential of CD4+ T cells expressing CD8αα. We show that TL+ IEC regulates IL-17 production by CD4+ T cells. Consequently, TL expression enhances development of colitis in an adoptive transfer model and confers resistance to Citrobacter rodentium infection by allowing the generation of an adequate Th17 response. Thus, TL expressed on IEC modulates CD4+ T cell responses in the intestinal mucosa.
Adenosine synthesized by CD73 and CD39 regulates inflammation during murine Salmonella infection (162.10)
Authors: Alam Mohammad , Gaines Dennis , Kuo Jennifer , Pereira Marion , Bigley Elmer , Ernst Peter , Williams Kristina 
Published: January 2023

Abstract
Salmonella spp. is a leading cause of hospitalization & death in the US associated with foodborne illness. Salmonella causes acute gut inflammation. Adenosine is an antiinflammatory mediator that limits tissue damage during inflammation. Surface enzymes CD39 & CD73 mediate the synthesis of adenosine. We studied the expression of CD39 & CD73 in liver, spleen, & CD4+ Th cells after infection of C57BL/6 mice with Salmonella enterica serotype Typhimurium (ST) & evaluated the role of CD73 in regulating ST-induced inflammation. Proinflammatory cytokine mRNA expression was assayed by RT-PCR arrays & cytokine proteins were tested by FACS & ELISA after ST infection or ST antigen (Ag) treatment. Function of CD73 was tested by examining proinflammatory cytokine production with or without CD73-inhibitor treatment of activated splenocytes exposed to ST Ag. Inflammatory responses to ST infection in CD73-/- mice were evaluated in liver & spleen tissue in H&E sections & in activated CD4+ Th cells stained for intracellular IL17a & IFN-γ. ST Ag-treated & CD73-inhibited splenocytes produced more proinflammatory cytokines (IL17A). Splenocytes from ST-infected CD73-/- mice produced more IL17a cytokine & Th17 cells & infected CD73-/- mice also had greater infiltration of inflammatory cells in the liver. Extracellular adenosine synthesized by CD73 may regulate host inflammation during murine Salmonellosis.
Toll-like Receptor 2 signaling in human endothelial cells (162.5)
Authors: Wilhelmsen Kevin , Mesa Kailin , Hellman Judith 
Published: January 2023

Abstract
Toll-like receptors (TLRs) recognize microbial components and initiate inflammation during infection. TLR2 is the receptor for microbial lipoproteins and other parts of Gram-positive and Gram-negative bacteria, and fungi. The endothelium expresses TLR2 and is important in sepsis-induced organ failure. We found that activation of TLR2 broadly affects endothelial cell (EC) functions, leading to upregulation of cytokine and chemokine production, increased neutrophil trafficking, altered expression of coagulation pathway intermediaries, and increased permeability. We probed the signaling pathways involved in TLR2-dependent activation of the EC by treating human umbilical vein EC (HUVECs) with lipopeptide TLR2 agonists and performed immunoblots, NF-κB EMSA’s, transcription factor arrays, and qRT PCR to assess responses. We found that TLR2 agonists transiently activate p38-MAPK and JNK at early time points and induce a sustained, albeit weak, activation of the transcription factor NF-κB. Through the use of siRNA knockdown and specific inhibitors, we determined that p38-MAPK, JNK, and NF-κB are necessary for TLR2-dependent upregulation of IL-6, IL-8 and TLR2 itself. Additionally, using mRNA human immune arrays, we found that markers of neutrophil trafficking and activation are most robustly upregulated by TLR2-dependent activation of the endothelium. We are currently investigating the mechanisms responsible for neutrophil trafficking and activation responses in more detail.
Ethanol consumption inhibits Th1 but not Th2 skewed contact hypersensitivity responses (163.11)
Authors: Brand Rhonda , Stottlemyer J. Mark , Cline Rachel , Falo, Jr Louis 
Published: January 2023

Abstract
Alcohol abuse is associated with suppression of both of innate and adaptive immune systems. Functionally, several studies suggest that ETOH consumption polarizes immune responses away from Th1 responses toward Th2 skewed immunity. Understanding the mechanisms by which alcohol alters immune function is critical to the development of strategies to reverse these changes. Contact Hypersensitivity reactions are dendritic cell dependent, antigen specific, T cell-mediated immune responses triggered by skin exposure to haptens that become immunogenic after binding to peptides or proteins. We examined the impact of alcohol consumption on the response to Th1 skewing (DNCB) and Th2 skewing (FITC) contact sensitizers. Our studies suggest that ETOH consumption significantly inhibits CHS responses to the Th1 skewing sensitizer DNCB. In contrast, alcohol consumption did not diminish the CHS response triggered by the Th2 skewing sensitizer FITC. Importantly, the differential immune effects of ETOH consumption occurred independently of ETOH associated liver damage, as the results were similar in mice fed alternative alcohol diets that that did or did not induce liver damage. These results demonstrate specificity in ETOH-induced immunomodulation, and suggest strategies to prevent and treat ETOH associated immune dysfunction relevant to vaccination and immune mediated ETOH induced pathologies.
Induction and suppression of allergic diarrhea and systemic anaphylaxis in a mouse model of food allergy. (163.21)
Authors: Kucuk Zeynep , Mahler Ashley , Strait Richard , Hogan Simon , Finkelman Fred 
Published: January 2023

Abstract
We evaluated the requirements for development of allergic diarrhea (AD) and anaphylactic shock (AS) in response to Ag ingestion and whether ingested Ag must be absorbed systemically to trigger these responses. TNP-BSA ingestion induced AS without AD in IgE anti-TNP mAb-primed, immunologically naïve wild-type mice and intestinal IL-9 transgenic mice, even though the latter mice have intestinal mastocytosis, but both AS and AD in IgE anti-TNP mAb-primed mice that had recently been immunized i.p., then p.o. with ovalbumin (Ova). Mice inoculated once i.p. with Ova/alum, then multiple times p.o. with Ova, develop both AD and AS in response to oral Ova challenge; a second i.p. Ova/alum inoculation prevents AS, but not AD. J chain- and pIgR-deficient mice, which have increased serum IgA and decreased gut IgA, are relatively resistant to development of AD. Treatment of IgE anti-TNP mAb-primed, Ova-immunized mice with i.v. IgG anti-TNP mAb inhibits both AD and AS responses to oral TNP-BSA challenge, with AS more sensitive to inhibition. We conclude that: 1) induction of AD, but not AS, by ingested Ag requires local Ag-induced changes to the intestinal immune system beyond mast cell and IgE induction; 2) ingested Ag must be systemically absorbed to induce both AD and AS; and 3) more ingested Ag must be absorbed systemically to induce AS than to induce AD. These observations suggest that induction of a systemic IgG Ab response can suppress established food allergy.
Plasmin plays an essential role in amplification of psoriasiform skin inflammation in mice (164.10)
Authors: Ke Fang , Wang Honglin 
Published: January 2023

Abstract
Background Although increased levels of plasminogen activators have been found in psoriatic lesions, the role of plasmin converted from plasminogen by plasminogen activators in pathogenesis of psoriasis has not been investigated. Methodology/Principal Findings Here we examined the contribution of plasmin to amplification of inflammation in psoriasis. We found that plasminogen was diminished, but that the amount and activity of its converted product plasmin were markedly increased in psoriasis. Moreover, annexin II, a receptor for plasmin was dramatically increased in both dermis and epidermis in psoriasis. Plasmin at sites of inflammation was pro-inflammatory, eliciting production of inflammatory factors, including CC chemokine ligand 20 (CCL20) and interleukin-23 (IL-23), that was mediated by the nuclear factor-κB (NF-κB) signaling pathway and that had an essential role in the recruitment and activation of pathogenic C-C chemokine receptor type 6 (CCR6)+ T cells. Moreover, intradermal injection of plasmin or plasmin together with recombinant MCP-1 resulted in induction of psoriasiform skin inflammation around the injection sites with several aspectsκof human psoriasis in mice. Conclusions/Significance Plasmin converted from plasminogen by plasminogen activators plays an essential role in amplification of psoriasiform skin inflammation in mice, and targeting plasmin receptor - annexin II - may harbor therapeutic potential for the treatment of human psoriasis.
Mammaglobin-A DNA vaccine increases the IFN-γ CD4+ICOS+ T- cells frequency in human breast cancer patients (165.26)
Authors: Tiriveedhi Venkataswarup , Fleming Timothy , Goedegebuure Peter , Naughton Michael , Ma Cynthia , Lockhart Craig , Gao Feng , Gillanders William , Mohanakumar T. 
Published: January 2023

Abstract
Mammaglobin-A (Mam-A), a 10kD secretory protein, is over expressed in 80% of primary and metastatic human breast cancers. Previous studies in our laboratory using a HLA-A2/Human CD8 transgenic murine model have demonstrated that Mam-A DNA Vaccine elicit specific CD8+ cytotoxic T-cell (CTL) responses to Mam-A and induces tumour regression in SCID mice. In our current study we report the activation of T-helper cells with specific expression of ICOS (inducible costimulator), a T-cell specific activation marker, following Mam-A vaccination. Stage IV breast cancer patients were chosen for the study and vaccinated by Mam-A DNA vaccine on 0, 28 and 56 days. Six months following Mam-A vaccination (n=5), flow cytometry analysis demonstrated a increase in CD4+ICOS+ T-cell frequency from 5±2% to 23± 4% with a concomitant drop in the CD4+Foxp3+T-cell (Treg) frequency from 19±6% to 10±5%. However, the frequency of CD4+CD25+ICOS+ did not change following vaccination (23±5%). ELISpot analysis demonstrated that following vaccination, CD4+ICOS+ sorted T-cells demonstrated a cytokine phenotype switch from IL-10 to IFN-γ. We conclude that following Mam-A DNA vaccination, there is a specific activation T-cells with increase in the pro-inflammatory IFN-γ CD4+ICOS+T-cells along with decrease in Treg frequency. We propose that this switch in T-cell frequency following vaccination may be important in tumor regression in breast cancer patients.
A novel NOD.SCID mouse model suggests CD200-mediated regulation may be important for both growth and therapy of CLL (165.36)
Authors: Wong Karrie , Spaner David , Gorczynski Reg 
Published: January 2023

Abstract
Chronic lymphocytic leukemia is characterized by the accumulation of malignant CD5+CD19+ B cells that, like anergic B cells, typically fail to engraft in immunocompromised murine hosts. CD200, a type I transmembrane molecule with potent immunoregulatory functions, is known to be overexpressed on CLL cells. Using a CD200 sandwich-ELISA, we have identified a soluble form of CD200 (sCD200), whose expression is increased by &gt;5-fold in the serum of CLL patients compared to healthy controls. Further analysis of sCD200 levels in CLL patient serum showed a correlation between sCD200 levels and Rai disease stage. Infusion of sCD200hi CLL serum into NOD.SCIDIL-2γ-/- mice receiving 1x108 CLL splenocytes enhanced engraftment of CLL cells, which were detectable in both the spleen and peritoneum of animals, in comparison to mice receiving sCD200lo normal serum. Enhanced engraftment using CLL serum was abrogated by depletion of sCD200 from the serum. When given at 21 days post-CLL splenocyte infusion, anti-CD200mAb was as effective as Rituximab in eliminating engrafted CLL. In vivo T cell depletion with anti-CD3mAb also suppressed CLL engraftment, despite ongoing CLL serum infusion, suggesting that T cells are important to maintain CLL survival and growth. Our studies show that infusion of sCD200hi CLL serum into NOD.SCID mice allows development of a novel xenograft model for CLL, and further suggest that anti-CD200 mAb may have value in treatment of disease.
Low dose γ-radiation can mitigate some deleterious effects of carcinogen injection in A/J mice. (165.42)
Authors: Gonzales Veronica , Gott Katherine , Makvandi Mackey , Scott Bobby , Wilder Julie 
Published: January 2023

Abstract
Low dose radiation (LDR) activates the immune response and may lead to cancer suppression. We investigated the acute effect of LDR and the carcinogen benzo(a)pyrene (BaP) on lung and spleen cell number and phenotype as well as spleen cell cytokine secretion in an A/J mouse model. Mice were given a single gamma radiation dose (10, 100, 1000 mGy) one day prior to intraperitoneal injection with BaP or Tricaprylin vehicle. Lung and spleen tissues were harvested at days (d) 2, 7, and 9 post-irradiation to determine viable cell counts and phenotype. Splenocytes were cultured for 48 hours +/- Concanavalin A (5 μg/ml) and cytokine secretion was measured by Luminex. BaP treatment decreased lung cell (d2) and splenocyte (d7) number while lung neutrophila was increased on d7 and 9. BaP also decreased activated (CD69+) CD4+ and CD8+ cells in both organs by d7. The reduction in activated CD8+ was attenuated in mice treated with 10 mGy radiation. BaP also decreased lung B cell and dendritic cell number. Treatment with 1000 mGy decreased B cells in both the lung and spleen. BaP treatment decreased Th2 cytokine production while increasing IL-1β and IL-17. 10 mGy treatment increased IL-2, IL-4, IL-12p70, VEGF, and IL-17 while IL-6 and IL-10 production was decreased by higher radiation doses on d2 post-radiation. These data suggest that treatment with BaP and LDR significantly affect the immune system with 10 mGy, but not 1000 mGy, treatment often mitigating the deleterious effect of BaP.
Peptide vaccines prevent tumor growth by stimulating a similar T cell repertoire as the native tumor antigen (165.46)
Authors: Slansky Jill , Jordan Kimberly , Buhrman Jonathan , Kappler John , Sprague Jonathan , Moore Brandon , Gao Dexiang 
Published: January 2023

Abstract
A major goal of immunotherapy for cancer is to focus and activate the T cell response to tumor-associated antigens. One strategy to stimulate these T cells is to immunize with specific peptides in combination immunostimulatory modalities. We are using an immunogenic mouse colon cancer model to characterize both the activating peptides and the responding T cells. We have identified peptides that interact with the restricting MHC molecule and a tumor-specific T cell clone that are either non-protective like the natural antigen, or protective against tumor challenge. The T cell repertoires responding to these peptides and the tumor are distinct, although they overlap with the natural antigen. Examination of the Vb regions expressed by the tumor-specific T cells, showed that the responses to the peptide variants were more focused than those to the natural antigen. Further ex vivo analysis showed that the T cells responding to the protective peptides and in the TIL (tumor infiltrating lymphocytes) were selected to express a CDR3b motif, which was also shared by the T cells responding to the natural antigen. We suggest that the increased signal delivered by the peptide variants that target the T cells naturally responding to the tumor such as those found in TIL result in more effective antitumor responses.
The effect of receptor-associated protein level on the immunogenicity of HSP-peptide complexes (165.7)
Authors: Zhou Yu , Binder Robert 
Published: January 2023

Abstract
Heat shock proteins (HSPs) serve as molecular chaperones in every type of cell. As HSPs are released from cells as a result of necrotic death, they are sampled by antigen presenting cells (APCs), which then prime T cell mediated immune responses. CD91, an endocytic receptor on APCs, has been shown to recognize HSPs in the extracellular milieu allowing for cross-presentation of HSP-chaperoned peptides. Various CD91 ligands, such as the ER-resident receptor-associated protein (RAP), compete with HSPs for receptor binding in vitro. Since the HSP-CD91 pathway appears critical for initiation of immune responses, we hypothesize that expression of elevated levels of RAP will negatively affect the immunogenicity of HSP-peptide complex, when RAP is released from necrotic tumor cells. To address this hypothesis, RAP was over-expressed in several tumor cell lines, and those cell lines were then tested for their immunogenicity by tumor challenge experiments. Tumor cells expressing RAP grew with faster kinetics than the non-RAP expressing counterparts in wild type mice despite maintaining identical proliferation rates in vitro. Additionally, the reduced immunogenicity of RAP-expressing cells is tested in prophylaxis of cancer in murine models and receptor competition is examined in vivo. The data suggest a potential novel mechanism of immune evasion for tumors expressing competing ligands for immune receptors.