bluuebunny/crossref_metadata_2025_split huggingface dataset:
Absence of Tec family kinases Itk and Btk significantly impairs FcϵRI dependent mast cell responses (86.26)
Authors: Morales Jose , Iyer Archana , Ojo Folake , Ning Gang , August Avery 
Published: January 2023

Abstract
While the etiology of asthma can be multi factorial, signaling through the IgE receptor (Fcϵ receptor I) on mast cells is critical to its onset. The Tec family tyrosine kinases ITK and BTK serve important roles as signal amplifiers downstream of the FcϵRI, however, their individual contribution to FcϵRI signaling has not been fully established. We have generated Itk and Btk double deficient (DKO) mice in an effort to determine the relative role of Itk and Btk to FcϵRI signaling in mast cells. Analyses of skin mast cells from these mice revealed reduced mast cell granule content and density. However, in vitro cultured bone marrow derived mast cells (BMMCs) had normal gross morphology and granule density. Mast cell histamine release was also compromised in DKO mice, likely associated with elevated endogenous IgE expression in these mice. In vitro growth analysis indicated that Btk negatively regulates BMMC growth as previously published, while the absence of both Itk/Btk did not further contribute to this phenotype. However, there was a significant reduction in FcϵRI mediated calcium responses, accompanied by reduced phosphorylation of PLCγ2 and the mitogen activated protein kinase p38. Lastly, a reduction in the production of several cytokines was also observed in DKO BMMCs. Our results suggest that Itk and Btk have redundant, complementary, and unique functions in mast cell FcϵRI signaling that could be pharmacologically manipulated to treat asthma.
Using mast cells derived from human CD34+ bone marrow stem cells to identify drugs that may produce pseudoallergic reactions (86.3)
Authors: Haskell Keith , Whritenour Jessica , Donahue Christopher , Blake William , Wolfram Thaddeus , He Linyan , Kawabata Thomas , Thiede Mark 
Published: January 2023

Abstract
Drugs such as morphine, vancomycin, radio-contrast dyes and smooth muscle relaxants are known to cause pseudoallergic or anaphylactoid reactions, which are very similar to Type 1 hypersensitivity reactions. These drugs directly stimulate mast cell degranulation by an IgE independent-mechanism and in drug development, there is a critical need to identify compounds that may be pseudoallergens in humans. To this end, an in vitro culture was developed to generate a reproducible supply of human mast cells Immuno-selected CD34+ bone marrow cells (106) were cultured for 63 days with weekly media changes and sequential cytokine additions. Cultures were maintained under hypoxic conditions from days 1-28 and switched to normoxic conditions from days 29-63. On day 63, the cultures were FcER1+, CD45+, CD117+, tryptase+, chymase+ CD34-, and CD123- and released histamine and PGD2 when exposed to anti-IgE, substance P, or compound 48/80. We tested several known pseudoallergens (morphine, tubocurarine, and vancomycin) for their ability to stimulate histamine release by mast cells produced from various donors. All 3 drugs reproducibly stimulated histamine release at 100uM with Morphine = tubocurarine &gt; vancomycin. Based on these studies, it appears that the mast cells derived from CD34+ cells do respond to known pseudoallergens, and may be used to determine if drugs in development have the potential to produce pseudoallergic reactions.
TGFb1 Suppresses the Mast Cell Response In Vitro and In Vivo (86.6)
Authors: Ryan John , Fernando Josephine , Barnstein Brian 
Published: January 2023

Abstract
Mast cell activation is a critical component of allergic disease, and may be regulated by cytokines present in the microenvironment. We recently described the suppressive effects of TGFb1 on IgE signaling in mouse and human mast cells. We have furthered this study, demonstrating that TGF injection or aerosolization reduces mast cell IgE receptor expression in vivo, and suppresses eosinophil recruitment in a mast cell-dependent model of airway hyperresponsiveness. We find that TGFb1 inhibits expression of the key signaling molecules Syk, Fyn, Akt, and Stat5 at the protein level without any change in mRNA expression. In contrast, expression or activation of ERK, JNK, GSK, and p38 was unaffected. Fyn transfection partially reversed TGFb1-mediated suppression of IgE-induced IL-13 secretion. Our data support the theory that TGFb1 possesses potent mast cell suppressive activities both in vitro and in vivo.
Antagonistic Control of IgG-mediated Mast Cell Activation by Lyn and Fyn Kinases (86.8)
Authors: Ryan John , Falanga Yves 
Published: January 2023

Abstract
Mast cells are key players of the innate immune system. In addition to their role in atopic disease, mast cell involvement is strongly suspected in inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, and atherosclerosis. Once activated, they degranulate and produce cytokines, resulting in the recruitment of other immune cells. It is well-appreciated that mast cells are activated via the high-affinity immunoglobulin E (IgE) receptor FcϵRI. In this study we focus on mast cell activation via the immunoglobulin G (IgG) receptor FcγRII/RIII and downstream signaling by Fyn and Lyn kinases. Lyn-deficient mast cells displayed increased FcγR expression, correlating with enhanced IgG-mediated histamine and leukotriene C4 secretion. The mast cells did not show a change in IgG-mediated cytokine production, suggesting a restricted and negative regulatory role for Lyn kinase in mast cell IgG signaling. In contrast, we show that Fyn kinase is rapidly activated by IgG crosslinkage, and is required for both IgG-mediated histamine release and cytokine production in mast cells. We obtained similar results when studying Lyn- or Fyn-deficient basophils and macrophages. Finally, we found that Lyn-deficient mice had exacerbated responses to passive systemic anaphylactic shock elicited by IgG challenge in vivo. These data illustrate the importance of Src family kinases in IgG signaling, a means by which mast cells may be activated in chronic inflammatory diseases.
Potential of cannabinoids to modulate the inflammatory response in colitis (87.1)
Authors: Zaric Violeta , Busbee Brandon , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Recent studies have suggested the role for cannabinoid (CB) type 1 and CB2 receptors in modulating gastrointestinal inflammation in animal models of experimental colitis. However, the immunosuppressive effect of the cannabinoid system has not been fully investigated. Our preliminary studies using delta-9-tetrahydrocannabinol (THC) showed promising results in a 2,4,6-trinitrobenzene sulfonic acid TNBS-induced colitis. 50mg/kg body weight of TNBS/ethanol was administered in 10 female BALB/c mice. Each group received intraperitoneal injections of vehicle or THC (10 mg/kg) once every other day. At day 6, mice were sacrificed and cells from spleen and MLN were analyzed for the expression of surface markers for T-cells, myeloid-derived suppressor cells (MDSC) and regulatory T-cells (Treg) by flow cytometry. The body weight increased in TNBS-treated mice administered THC when compared to vehicle. THC improved the survival up to 60% of mice in TNBS-induced colitis when compared to vehicle treated mice (30%). In the spleen and MLN, the ratio of T-cells is lower in THC-treated mice compared to vehicle-treated group whereas the ratio of Treg is increased in THC-treated mice. THC also reduced the ratio of MDSCs in TNBS-treated mice. THC treatment ameliorated colitis by modulating the immune response towards Treg and altering the T-cells and their subsets. Future experiments will address the potential of other cannabinoids in modulating the inflammatory response in colitis.
H.polygyrus infection induces functionally distinct FOXP3+IL-10+ and FOXP3+IL-10- cell subsets in the Lamina propria of mice (87.15)
Authors: Stoyanoff Korynn , Hang Long , Blum Arthur , Setiawan Tommy , Leung John , Weinstock Joel 
Published: January 2023

Abstract
Background: Infection with the murine intestinal helminth H. polygyrus (Hp) results in the regulation of inflammation in the gut Lamina propria (Lp) of mice. Infection upregulates FOXP3 expression in Lp T cells, but the effect of Hp infection on various FOXP3+ subgroups and their function in unknown. We hypothesized that Hp infection induces one or more FOXP3+ T cell subgroups and results in enhanced regulation. Method: We distinguished several FOXP3+ subgroups using a dual reporter mouse for FOXP3 and IL-10. Spleen (Sp), mesenteric lymph nodes (MLN), ileum Lp, and colon Lp cells from mice were analyzed by FACS for percentages of FOXP3 and/or IL-10 expressing cells. Subsets were also tested for their ability to regulate Sp cell proliferation. Results: The FOXP3+IL-10- subset is dominant in the Sp and MLN, the FOXP3+IL-10+ dominates in the colon Lp, and the subsets are roughly equal in the ileum Lp of infected and uninfected mice. Hp infection increases FOXP3+IL-10+ cells in ileum and colon Lp cells vs. uninfected mice. We tested the ability of FOXP3+IL-10+ cells to regulate the proliferation of Sp T cells in 48hr cultures. FOXP3+IL-10- inhibited proliferation better than FOXP3+IL-10+ cells (67.41% vs. 23.88% inhibition, 1:1 ratio). Discussion: Hp infection increases FOXP3+IL-10+ expression in the Lp. Other regulatory roles for FOXP3+IL-10+ cells are being investigated and may be important in understanding how Hp infection regulates gut inflammation.
Role of Adenosine A2B Receptors in Regulating Inflammatory Responses of TNBS-induced Experimental Colitis (87.2)
Authors: Alam Mohammad , Wilson Jeff , Ernst Peter 
Published: January 2023

Abstract
Background: Adenosine, an endogenous purine nucleoside exerts anti-inflammatory activities on host cells via signaling through the G protein-coupled receptors subtypes referred as A1, A2A, A2B, and A3. Both A2A and A2B adenosine receptors regulate inflammatory responses during experimental colitis. Since intestinal epithelial cells and antigen presenting cells express the A2BAR and contribute an important role in the development of colitis, we studied the effect of A2BAR gene deletion in mice during 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Methods: TNBS pre-sensitized A2BAR-null (A2BAR-/-) & wildtype mice were intrarectally instilled with TNBS and changes in body weight, colonic tissue inflammatory responses and pro-inflammatory cytokine production were compared. Liver/spleen tissues were also cultured for evaluating bacterial translocation. Results: Studies indicated that the severity of colitis was increased in A2BAR-/- mice relative to wildtype controls, as evidenced by more dramatic weight loss and increased colonic tissue damage with histopathology revealing an extensive infiltration of immune/inflammatory cells. Levels of pro-inflammatory cytokines (TNF-a) were elevated in the colons of TNBS-treated A2BAR-/- mice. A2BAR-/- mice also showed increased systemic dissemination of intestinal bacteria in the spleen/liver cultures. Conclusion: Our data suggest A2BAR play a protective, anti-inflammatory role in TNBS-induced colitis.
Growth Detection: A Novel Role for CD4 T-cells (87.20)
Authors: Bewick Sharon 
Published: January 2023

Abstract
One of the central goals of immunology has been to understand the cellular and molecular mechanisms of immune regulation. Progress on the cellular front has developed rapidly with the identification of distinct CD4 T-cell populations with unique functions and patterns of cytokine production. Similarly, the discovery of TLRs in the late 1990s resulted in rapid advances towards an understanding of the molecular mechanisms of immune regulation. Despite the progress being made on both the cellular and molecular fronts, however, many aspects of immune regulation remain unexplained. Two Th-cell populations - induced regulatory T-cells (iTreg) and Th17 cells have recently been characterized. How iTreg and Th17 cells integrate with other Th populations remains an active area of research. Recently, we proposed a novel mechanism of immune regulation based on detection of pathogen growth (Bewick et. al, PLoS ONE, 2009). We then showed how growth detection can arise naturally as a result of signaling interactions and maturation kinetics of iTreg and Th17 cells. Building off of the basic premise of this growth detection model, we now extend the iTreg/Th17 system to include additional immune effector cells. This allows us to show how the immune system is organized in such a way that it can accurately and robustly classify different pathogens based on growth rate.
Immune regulatory roles of microparticles in autoimmune and allergic diseases (87.22)
Authors: Kahraman Tamer , Simsek Ismail , Gursel Mayda , Hakan Erdem , Pay Salih , Dinc Ayhan , Gay Steffen , Kocabas Can , Ozcan Celal , Civelek Ersoy , Gursel Ihsan 
Published: January 2023

Abstract
Microparticles (MP) are small vesicles secreted from immune and non-immune cells and their diameter ranges from 200 to 700nm. Studies demonstrated that MP number and source varies in several disease conditions. Therefore, we plan to characterize the cell source, activation state and levels of MPs in different clinical states of Behcet`s Syndrome (BS) and asthma. Isolated MPs were stained with Annexin V and cell specific CD markers (i.e. CD42a, CD69, CD105, CD31 and CD146). Number of marker specific MP/ml blood was detected by FACS. In the case of BS, blood plasma of 22 healthy, 28 inactive BS patients and 29 active BS patients were examined while blood plasma of 13 healthy, 15 atopic dermatitis (AD) and 15 asthma patients were used for asthma. The number of MPs/ml plasma is increased by &gt;3 fold in active BS (34.1±10.9 x 104) and &gt;2 fold in inactive BS (21.3±8.3 x 104) compared to healthy subjects (9.7±3.8 x 104). We also found that majority of secreted MPs were platelet origin, this was valid for allergic patients too. Surprisingly, MPs (number/ml plasma) were much higher in healthy donors (9.8±1.3 x 105) compared to atopic dermatitis (3.5±2.9 x 105) and asthma (2.4±0.5 x 105) patients. This data suggest that plasma MPs have differential immune regulatory roles in autoimmune and allergic conditions.
A genetically determined, serine-based and phosphorylation-dependent molecular switch regulates the anti-inflammatory potential of human IgA (87.34)
Authors: Ji Chuanyi , Wu Jianming , Gibson Andrew , Edberg Jeffrey , Kimberly Robert 
Published: January 2023

Abstract
Monteiro and colleagues have suggested dual roles for FcαRI in immune regulation through tyrosine-based “inhibitory/activating” mechanisms mediated by the immunoreceptor tyrosine-based activation motif (ITAM) in the Fc receptor γ-chain (FcRγ) [Pasquier et al., Immunity 2005, 22(1)]. We report a second mechanism in which the natural S248G variants of the FcαRI α-chain alter the inflammatory capacity of FcαRI and constitute a serine-based, genetically determined switch for regulation of the anti-inflammatory potential of human IgA. To elucidate the basis for this α-chain mechanism, we sought allele-specific FcαRI-associated molecules. Sab (SH3-domain binding protein that preferentially associates with Btk), a trans-inhibitor of Bruton's tyrosine kinase (Btk), is recruited by the common Ser248 allele in a phosphorylation-dependent fashion, whereas the protein tyrosine kinase Lyn is reciprocally recruited by the Gly248 variant. Compared to FcαRI Gly248, recruitment of Sab by FcαRI Ser248 results in inhibition of Btk activation and suppression of IgA effector functions independent of FcRγ-pairing. Expression of dominant negative Sab releases FcαRI-mediated inhibition in a Ser248-allele specific manner. These findings suggest an immunoreceptor serine-based inhibition motif (ISIM) which functions as a molecular switch for receptor induced activation/inhibition and advances our understanding of the divergent properties of human IgA in inflammation.
Role of the 3’ heavy chain enhancers in germline transcription and switch recombination (88.1)
Authors: Dunnick Wesley , Shi Jian , Fontaine Clint , Zerbato Jennifer , Collins John 
Published: January 2023

Abstract
Class switch recombination (CSR) is preceded by germline transcription and then mediated by a DNA recombination event. To study germline transcription and CSR we utilize transgenic mice with a 230 kb bacterial artificial chromosome that includes a rearranged VDJ gene, the entire heavy chain C region locus, and all known 3’ enhancers. In the intact transgene, germline transcription of and CSR to the γ, ϵ, and α heavy chain genes are regulated in parallel to the endogenous genes. We have reported that deletion of a 28 kb segment, including the 3’ enhancers HS3A, HS1,2, HS3B, and HS4, results in a reduction of CSR to all heavy chain genes to about 1% of the level of the intact transgene. We have derived four new versions of the heavy chain transgene: with 1 kb deletions of (i) HS1,2 and HS3B, (ii) HS1,2, HS3B, and HS4, (iii) HS3A, HS1,2, and HS3B, and (iv) HS3A, HS1,2, HS3B, and HS4. Comparing levels of CSR in these deletion variants to intact transgenes, we find that deletion of more enhancers results in a reduction in CSR for more heavy chain genes, and the reduction is more severe for a given heavy chain gene. Nevertheless, different enhancers have different effects on different heavy chain genes. Finally, there are enhancer deletions for which the reduction in CSR is much greater than the reduction in germline transcription. This suggests that the 3’ enhancers have a role in CSR that is distinct from the enhancement of germline transcription.
Lymphotoxin alpha gene polymorphism influences the cardiomyopathy development in chagas’ disease patients (88.16)
Authors: Rodrigues Virmondes , Pisseti Cristina , Oliveira Rafael , Correia Dalmo 
Published: January 2023

Abstract
Chagas’ disease, caused by Trypanosoma cruzi, is an important parasitic disease in Latin America, affecting more than 8 millions of people. The chronic phase is characterized by commitment of heart or digestive organs and the reason for the development of one clinical form or another is not well known. It is speculated that host and parasite genetic characteristics contribute for it. LT-α is an inflammatory cytokine, important in the organization and maintenance of the inflammatory process by stimulating adhesion molecules, such as VCAM, ICAM and E-selectin. Polymorphism on LT-α gene, position +252, were analyzed on subjects (n= 295) from an endemic area for Chagas’ disease grouped according to serology for T. cruzi and with clinical form (cardiac or indeterminate forms).Expression of LT-α was evaluated in culture supernatants and by flow cytometry. Presence of G allele was significantly more frequent in patients with heart commitment (cardiac form) (p=0.016, OR=2.4). Patients with cardiac form produced significantly more LT-α than individuals with the indeterminate form. Furthermore, infected individuals with G allele produced significantly higher levels of LT-α than those without G allele. CD8+ T lymphocytes were the major source of LT-α in basal condition and after stimulation with specific antigens. This study presents genetic and biological evidence for a role of LT-α in the development of cardiac form of Chagas’ disease.
Identification of a novel deubiquitinating enzyme DUBS6 that regulates Sirt6-mediated suppression of NF-κB in T cells (88.17)
Authors: Fang Deyu , Yang Heeyoung 
Published: January 2023

Abstract
The type III histone deacetylase Sirt6 is known as a regulator of aging in diverse species by regulating NF-kB-dependent gene expression. Here, using an LC-MS/MS approach combined with immunoprecipitation, we have identified a complex of Sirt6-interacting molecules (interactome) that contains about 40 proteins from mouse primary T cells. One of the interactome members is a novel deubiquitinating enzyme (DUB); we thus define this gene as a DUB of Sirt6 (DUBS6). Co-immunoprecipitation and western blotting analysis further validated that DUBS6 interacts with Sirt6 in transiently transfected HEK293 cells as well as in mouse primary T cells. Expression of DUBS6 inhibits Sirt6 ubiquitination and thus stabilizes Sirt6. Unexpectedly, DUBS6 expression, despite increased Sirt6 stability, reversed Sirt6-mediated suppressive function of NF-kB transcription activity. These results suggest that ubiquitination of Sirt6 appears to be required for its functions and that DUBS6 is a novel regulator in Sirt6-mediated NF-kB suppression in T cells.
HuR protein is involved in S49 thymoma cell proliferation and is regulated by cAMP/PKA signaling (88.20)
Authors: LaJevic Melissa , Cohen Rhonna , Chambers Donald 
Published: January 2023

Abstract
The RNA binding protein human antigen R (HuR) participates in the post transcriptional regulation of many mRNAs bearing AU rich and U rich elements by influencing stabilization and translation. Based on the types of mRNAs regulated by HuR, the cellular role of HuR can be identified as pro-survival and pro-inflammatory. Using the S49 thymoma cell line, we recently showed that cAMP treatment, which triggers apoptotic signaling events, leads to a decrease in the effectiveness of HuR function. In this work, we investigate if there is a relationship between cAMP mediated cell death and HuR activity by utilizing small interfering RNA-mediated suppression of HuR in S49 cells. We found that knockdown of HuR protein decreases normal cell growth, and increases cAMP mediated apoptosis. qPCR analysis of mRNAs important in inflammation, cell growth, and apoptosis reveal that knockdown of HuR downregulates TNF-α mRNA and upregulates IL-10 and Bim mRNAs in untreated cells, and upregulates p27, and Bim mRNAs upon treatment with cAMP. Furthermore, cAMP treatment causes a PKA dependent decrease in HuR cytoplasmic levels. These results suggest that a pro-inflammatory and pro-survival role for HuR in S49 thymoma cells exists and may be compromised upon the elevation of cAMP by decreasing cytoplasmic HuR protein and allowing the expression of apoptotic factors.
The V(D)J recombination machinery is associated with the nuclear matrix (88.3)
Authors: Lange Miles , Xie Wanqin , Hong Sang Yong , Yu Zhihong , He Ti , Huang Lin , Yu Yangsheng , Marran-Nichol Kathleen , Swanson Patrick , Lu Runqing , Su Kaihong , Zhang Zhixin 
Published: January 2023

Abstract
Somatic recombination of immunoglobulin or T cell antigen receptor genes occurs through recombination activating gene product (RAG)-mediated cleavage at recombination signal sequences (RSS) and subsequently, the broken DNA ends are repaired by non-homologue end joining (NHEJ) enzymes. Here, we show that RAG1, RAG2, and many NHEJ factors in B lineage cells are associated with the nuclear matrix. The core RAG1 and RAG2 proteins have their own nuclear matrix targeting regions. RAG-mediated double stranded DNA breaks at the Jκ4 RSS can be readily detected in the nuclear matrix fraction in Gleevec treated Abelson transformed murine B cells or mouse bone marrow cells, indicating that the recombination reaction is ongoing on the nuclear matrix. In the HEK 293 cell-based recombination system, artificial recombination substrates are recruited to the nuclear matrix for recombination. Moreover, nuclear matrix proteins purified from 293 cells expressing the core RAG proteins or human B lineage cells expressing endogenous RAG proteins support cleavage of RSS substrates in vitro. Based on these results, we propose that the V(D)J recombination machinery is associated with the nuclear matrix.