bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mycoplasma gallisepticum requires ATP to activate the canonical inflammasome in vitro.
Authors: McDaniel Dylan , Karetnyi Anastasia , Caswell Clayton , Allen Irving C 
Published: January 2023

Abstract
Mycoplasma gallisepticum (MG) is a bacteria which lacks a cells wall and colonizes mucosal epithelial cells. This colonization commonly occurs in the respiratory tracts of chickens and turkeys, leading to chronic respiratory disease. MG is an economically significant pathogen in poultry and can cause large economic losses on poultry farms because of decreased egg laying. Although the importance of MG as an economically relevant pathogen is understood, there is a paucity of knowledge in regards to its pathogenesis. One possible mechanism is via the activation of the inflammasome, a multi-protein complex leading to the maturation of IL-1β and IL-18 as well as the inflammatory form of cell death called pyroptosis. Interestingly, Mycoplasma hyorhinis (MH) was shown to activate the NLRP3 inflammasome in THP-1 and mouse bone marrow derived dendritic cells (BMDCs). In addition, HD-11 cells co-cultured with tracheal epithelial cells that were exposed to MG, show increased levels of IL-1β. In this study, we aimed to determine if MG activated the inflammasome in murine and chicken macrophages. We found that MG activates the canonical inflammasome in primary murine macrophages via the induction of IL-1b release. However, we determined that this MG-mediated activation of the canonical inflammasome requires the addition of ATP in vitro. In addition, we also found that MG alone is not sufficient to activate pyroptosis in vitro. Interestingly, MG also initiated the production of IL-6 in these cells, suggesting potential activation of the NF-κB pathway. We are currently working with the HD-11 chicken cell line to determine if MG activates the inflammasome in these cells as well.
Number and location of AID ‘hotspot sequences’ in germline variable genes of diverse species
Authors: Levings Randall 
Published: January 2023

Abstract
Generation of antibody (Ab) diversity is accomplished through five mechanisms: Heavy/Light chain combination, Variable-(Diversity)-Joining gene combination, junctional base addition/deletion, gene conversion, and somatic hypermutation (SHM). Different species have been shown to use the mechanisms to varying degrees. Species with relatively fewer variable (V) genes might be expected to have V genes containing more ‘hotspots’ for activation induced cytidine deaminase (AID), so as to ‘preposition’ the genes for SHM. Germline V sequences identified in the Im Muno Gene Tics (IMGT) database were analyzed for AID target sequences (RGYW, WRCY) relative to framework and complementarity determining regions as designated in the IMGT amino acid numbering system. Examples of functional germline sequences from six classes of vertebrates (only Agnatha was excluded) - including multiple orders of mammals - were analyzed. No clear pattern of AID hotspot number or location was observed to be associated with phylogeny or diversification strategy (when known). However, ‘outlier species/genes’ were noted. The results indicate that caution should be exercised in generalizing or extrapolating between species regarding Ab gene sequences. They also reinforce the importance of conducting gene studies including germline, fetal B lymphocytes, and mature B-cells across many species. Information on the origin and mechanisms of Ab diversity has relevance to the potential breadth of the immune response to infection and vaccination, and possibly to engineering effective Ab for immunotherapy.
Dairy cattle naturally infected with bovine leukemia virus exhibit abnormal adaptive immunity after primary and secondary challenges with keyhole limpet hemocyanin.
Authors: Frie Meredith , Sporer Kelly R. B. , Coussens Paul M. 
Published: January 2023

Abstract
Bovine leukemia virus (BLV) is a retrovirus that is widely prevalent in the US: over 83% of dairy herds and approximately 40% of all dairy cows are infected. While less than 5% of infected cows will develop leukemia or lymphoma as a result of BLV, new research demonstrated that BLV+ cows produce less milk and have shorter life spans than their BLV− herdmates. BLV+ cows likely have impaired immunity, putting them at risk for contracting other infectious diseases that negatively impact production and welfare. To investigate whether BLV+ cows develop abnormal immune responses to a primary infection, BLV+ and BLV− cows were challenged with the protein keyhole limpet hemocyanin (KLH) to elicit a primary immune response and T and B cell responses to KLH were characterized. 56 days after the primary challenge, BLV+ and BLV− cattle were re-inoculated with KLH to elicit a secondary memory immune response and in vivo T and B cell responses to KLH were characterized. BLV+ cows produced less IgM in response to KLH and exhibited abnormal γδ T and B cell populations. T and B cell responses to in vitro stimulation were also characterized. CD8+ and γδ T cells from BLV+ cows produced more IFNγ, and CD8+ T cells from BLV+ cows exhibited atypical IL4 production. In comparison to B cells from BLV− cows, BLV-expressing and BLV-non-expressing B cells from BLV+ cows exhibited altered MHCII expression. Activated B cells also exhibited a much higher proportion of BLV expression in comparison to unactivated B cells. Together, these data show that BLV+ cows exhibit a reduced and abnormal immune response even upon a primary immune challenge.
Epithelial expression of IL-17 mediates a gut-associated inflammatory response in the purple sea urchin larva.
Authors: Buckley Katherine M , Rast Jonathan P 
Published: January 2023

Abstract
Immune response to microbial disturbance in the gut involves coordinated activity of gut epithelial cells, motile immune cells and other cells throughout the organism. Many of these functions are ancient among bilaterians, although it is not yet clear how they relate among animal phyla. As morphologically simple organisms that share an important genetic heritage with vertebrates, sea urchin larvae offer a novel, experimentally tractable system in which to characterize the gut-associated immune response. We have developed a model to characterize gene regulatory control of this response using the marine bacterium Vibrio diazotrophicus. When exposed to Vibrio in the surrounding seawater, bacteria accumulate in the gut and later invade the body cavity. Bacteria that enter the blastocoel are rapidly cleared by a coordinated response of phagocytic and granular immune cells. Peripheral immune cells migrate through the body cavity and accumulate at the gut. RNA-Seq data indicate that the most acutely upregulated gene in the early phase of response are two types of IL17 genes. Whole mount in situ hybridization and BAC-based fluorescent protein reporters indicate that these genes are expressed within gut epithelial cells. Notably, perturbation of IL17 receptor signaling results in reduced levels of tnfaip3 (a negative feedback inhibitor of IL17), nfkbiz (an IL17 target gene in vertebrates), the transcription factors cebpα and cebpγ and soul1 (SOUL domains are evolutionarily widespread and involved in immune responses). These results indicate that the highly regulated IL17 expression in the gut epithelium and signaling through IL17R1 form a central axis of larval gut-associated immunity.
High expression of genes encoding signaling molecules (LAX1, BLK) and transcriptional factors (ICOS, GATA3) characterizes bovine neonatal immunity.
Authors: Kaushik Azad Kumar , Pasman Yfke , Merico Daniele 
Published: January 2023

Abstract
We analyzed 18 RNA-seq libraries from 3 bovine calves at day 0, 7, 14 and 28, together with their dams (day 0 and 7) for cell signaling and transcription factors gene expression. LAX1, lymphocyte transmembrane adaptor 1 is consistently expressed at higher levels post-birth. Similarly, BLK, a cell signaling tyrosine kinase required for from pro- to pre-B cell transition is expressed at high levels 4 weeks post-birth. A trend for increased expression of Igα (CD79A) and Igβ (CD79B) in neonatal B cells is noted day 0 to day 28 (LogFC 0.8 and P-value&lt; 0.05 FDR &lt;0.15). The CD40 expression is also significantly increased day 0 versus day 14 (LogFC 1.05, FDR .00039) to achieve steady levels. A lack of increase in T-cell signaling molecules CD3G and CD3D expression suggests neonatal T cell quiescence, but is higher than the dams (FDR&lt;0.05). However, the ICOS (CD278) expression is increased from day 0 to day 7 post-birth (LogFC 1.11, FDR 0.00027). Similarly, an increased expression of FOXP3, relevant to the development of regulatory T cells, occurs in bovine neonates day 0 vs day 7 (LogFc 1.23, FDR 0.00063). The expression of GATA3 transcription factor, regulator of innate and adaptive immunity associated with TH2 differentiation, is high in neonates as compared to the adults (FDR&lt;0.001). As would be expected, various transcriptional factors and cell signaling molecules contribute towards T and B cell proliferation and expansion, but a lack of increased expression of CD3G and CD3D genes suggests T cell quiescence early during ontogeny. [Supported by NSERC Canada Discovery Grant]
The Actin Cytoskeleton Controls FcgRIIB-Mediated Inhibition of B-Cell Signaling
Authors: Shu Zhou , Lazzaro Michelle , Liu Chaohong , Seeley-Fallen Margaret , Csiszer Ivan , Upadhyaya Arpita , Zhao Xiaodong , Song Wenxia 
Published: January 2023

Abstract
FcgRIIB is a major inhibitory coreceptor of B cell receptor (BCR), and its deficiency leads to B cell autoimmunity. Upon colligated with the BCR by immune complexes (ICs), this coreceptor activates SH2-containing inositol-5 phosphatase (SHIP) and inhibits BCR clustering in lipid rafts. However, the molecular mechanisms underlying FcgRIIB-mediated inhibition of BCR activation remain elusive. This study reveals a critical role of the actin cytoskeleton in FcgRIIB-mediated inhibition. Upon interacting with ICs tethered to lipid bilayers, an actomyosin ring is quickly formed surrounding the B cell membrane region interacting with the lipid bilayer (contact zone). Perturbing the actomyosin ring, by myosin inhibitors or B cell-specific gene knockout of non-muscle myosin IIA (NMIIA) or neuronal Wiskott-Aldrich syndrome protein (N-WASP), releases B cell spreading, BCR clustering and signaling from the inhibition by FcgRIIB. Using conditional gene knockout mice, we further show that NMIIA is recruited to the B cell contact zone in an SHIP-dependent manner, while N-WASP-mediated actin remodeling organizes NMIIA and actin into a ring. Surprisingly, WASP-mediated actin dynamics is required for the FcgRIIB-induced activation of SHIP but not the formation of the actomyosin ring. Our results suggest that FcgRIIB-triggered signaling induces unique actin remodeling involving NMII, WASP, and N-WASP, which limits B cell morphological changes and BCR lateral mobility and enhances the activation of inhibitory signaling. These data potentially provide a mechanistic explanation for the elevated autoantibody levels caused by WASP or N-WASP deficiency.
ITK is a magnesium sensor during T cell activation
Authors: George Alex B , Kanellopoulou Chrysi , Masutani Evan , Chaigne-Delalande Benjamin , Lenardo Michael J 
Published: January 2023

Abstract
Interleukin-2-Inductible T-cell Kinase (ITK), a member of the TEC family of kinases, serves a vital role in the activation and differentiation of T-cells. Following stimulation of the T cell receptor (TCR) and phosphorylation by LCK, ITK phosphorylates PLCy1 and initiates a transduction pathway that results in the release of intracellular Ca2+ stores and the transcription of IL-2. Although ITK’s role in this signal transduction pathway is known, there is still much to be characterized about the regulation of ITK activity. Previous studies have indicated that Mg2+ serves a key role as a second messenger in T lymphocyte activation and that attenuated intracellular Mg2+ levels inhibit the phosphorylation of PLCy1 and impair Ca flux. We propose that ITK serves as a sensor for the Mg2+ flux required for optimal T cell activation. Kinase assays show that ITK activity is enhanced by Mg2+ and suggest a model where two Mg2+ ions occupy the catalytic site. Molecular modeling also suggests that ITK possesses two Mg2+ binding sites in the catalytic domain. We are currently studying the effect of Mg2+binding on ITK kinase activity via site directed mutagenesis and are in the process of generating crystal structures of ITK complexed with Mg2+. Finally, we propose that this ‘second site’ Mg2+ is a general feature of kinases and may highlight the importance of divalent cations in kinase regulation.
ITK signaling via the Ras/IRF4 pathway regulates the development and function of type 1 regulatory T cells
Authors: Huang Weishan , Solouki Sabrina , Koylass Nicholas , Zheng Song-Guo , August Avery 
Published: January 2023

Abstract
Type 1 regulatory T (Tr1) cells differentiate in response to signals engaging T cell receptor (TCR), express high levels of the immunosuppressive cytokine IL-10 but not Foxp3, and can suppress inflammation and promote immune tolerance. Here, using IL-10GFP/Foxp3RFP dual reporter transgenic mouse system, we show that ITK, a key modulator of TCR signaling, is required for the development of Tr1 cells in various organs following TCR activation, and in the mucosal system during parasitic and viral infections. ITK kinase activity is required for mouse and human Tr1 cell fate programming and regulates the balance between IL-10 and IFN-γ production during Tr1 cell differentiation. The requirement for ITK function during Tr1 cell development and suppressive function can be restored by the expression of the transcription factor IRF4. Downstream of ITK, Ras, and MAP kinases ERK1/2, p38 and JNK, are required for Tr1 cell differentiation, but not Th17 cell differentiation. We have previously shown that Th17 differentiation is also dependent on ITK signaling. However, in the absence of ITK, expression of the constitutively active HRasG12V rescued IRF4 expression and Tr1 cell differentiation, but failed to restore Th17 cell differentiation. We conclude that the TCR/ITK signaling through the Ras/MAPK/IRF4 pathway is specifically required for functional development of Tr1 cells, and that targeting these signaling components may be of therapeutic benefit.
Effect of genetics on the development of Hypersensitivity Pneumonitis (HP) using the BXD recombinant inbred mouse reference population
Authors: Fitzpatrick Elizabeth , Yoon Tae-Won , Williams Robert , Yi Ae-Kyung 
Published: January 2023

Abstract
HP is an interstitial lung disease that develops following repeated exposure to a variety of inhaled environmental antigens. The disease is characterized by alveolitis, granulomas and in some patients, develops into a chronic form associated with fibrosis. Not all individuals exposed to HP causative antigens develop disease suggesting that genetic factors play a role in susceptibility; however, the host co-factor(s) that play a role in susceptibility are unknown. We are using the innovative systems genetics tool, the BXD panel of recombinant inbred mice with the Saccharopolyspora rectivirgula (SR) model of HP to identify genetic loci and candidate genes associated with disease development. We exposed the parental strains, C57Bl/6 and DBA/2, and sixteen BXD strains to SR for 3 weeks and analyzed the lungs 18h after the last exposure. Bronchoalveolar lavage (BAL) was performed and cells recovered from the BAL fluid were analyzed by flow cytometry to determine the frequency of infiltrating immune cells. The results reveal variability in the cellular composition of the BAL; some BXD strains exhibited a neutrophilic alveolitis whereas other BXD strains exhibited a more lymphocytic alveolitis. BAL recovered from mice of the BXD45 strain was 68% neutrophils and 7% T lymphocytes, whereas BAL from the BXD73 strain was 92% neutrophils and 2% T lymphocytes. In addition, there were significant differences in the level of activated CD4+ and CD8+ T cells across the strains. H&E staining of lung tissue revealed significant differences in the extent of granuloma formation across strains. These data support the contention that genetic differences critically influence SR responsiveness given that environmental factors were held constant.
Resveratrol improves a murine model of asthma through alterations in the gut microbiome
Authors: Alharris Esraah Sattar , Alghetaa Hasan F , Busbee Philip Brandon , Nagarkatti Mitzi , Nagarkatti Prakash 
Published: January 2023

Abstract
Asthma is a condition characterized by bronchial spasms, inflammation, and mucous hypersecretion which leads to difficulties in respiration. Asthmatic patients are usually presented with recurrent attacks of coughing, wheezing, and shortness of breath which could be life-threatening. More than three million cases of asthma in the United States are diagnosed annually. Resveratrol, a polyphenolic stilbene, is known to be useful in controlling asthmatic attacks via different molecular mechanisms within the lung epithelium and infiltrating immune cells. However, few studies mentioned the effect of resveratrol on the microbiome in ovalbumin-induced asthma mouse model. In this study, we induce asthma in BALB/c mice by injecting ovalbumin (OVA) with aluminum hydroxide intraperitoneally followed by 7 days treatment with resveratrol (100mg/kg) by oral gavage. Intranasal ovalbumin was given on day 8 to induce asthma followed by 7 more days of treatment with resveratrol. We performed 16S rRNA microbial analysis on cecal flushes between disease and treatment groups, noting changes in microbial diversity among disease and treated groups. Specifically, we identified alterations in gut microbial composition, which included significantly higher levels of Prevotella in the disease groups, which was reduced upon treatment with resveratrol. In addition, Bacteroides acidifaciens species were significantly lower in the asthma-induced mice and significantly increased in resveratrol treated.
Supported in part by NIH grants P01AT003961, R01AT006888, R01ES019313, R01MH094755, P20GM103641, MoHESR and R01AI129788
Atypical protein kinase PKCλ/ι on pathogenesis of asthma
Authors: Yang Jun-Qi , Yang Yingying , Dong Panpan , Guo Fukun , Zheng Yi 
Published: January 2023

Abstract
Asthma is a chronic airway inflammation in which Th2 and Th17 cells play critical roles in its pathogenesis. We have reported that atypical protein kinase PKCλ/ι is a new regulator for Th2 differentiation and function. Our recent findings indicate that PKCλ/ι may regulate Th17 cells. The secretion of Th17 effector cytokines, including IL-17, IL-17F, IL21 and IL-22, were inhibited from PKCλ/ι-deficient T cells under non-polarized or Th17-polarized culture conditions. Further studies indicate that impaired Th17 differentiation and function is associated with the down-regulation of Stat3, NFATc1 transcription factors in PKCλ/ι-deficient CD4+ T cells in culture. We developed a model of Th17 and neutrophil-involved allergic airway inflammation by intratracheal inoculation of allergenic extracts of house dust mite (HDM). PKCλ/ι-deficiency significantly inhibited airway inflammations. The infiltrating cells in the lungs and bronchoalveolar lavage (BAL) fluid were significantly reduced in conditional PKCλ/ι-deficient mice. Th17 effector cytokines were reduced in the BAL and lungs at protein and mRNA levels. Our findings suggest that PKCλ/ι might regulate allergic airway inflammation through the control of Th17 cells. The underlying molecular mechanisms are currently under investigation.
An 18-day Residential Lifestyle Program Improves Fitness of Asthma Patients
Authors: Ramirez Francisco E , Nedley Neil , Freed Spencer 
Published: January 2023

Abstract

Rationale
Lifestyle interventions are known to improve many medical conditions. We are documenting the effect that a, 18-day residential medical lifestyle program has on the fitness of asthma patients using metabolic equivalent (MetS), which correlate to exercise capacity.

Methods
The program included 3 daily buffet-like whole food plant-based meals with no snacks, 2 to 3 times a day sessions of exercise (aerobic and anaerobic), massage, hydrotherapy, medical and spiritual interventions. The program is called NEWSTART program and takes place in Weimar, California. NEWSTART means: Nutrition, Exercise, Water, proper Sunlight exposure, fresh Air, Rest and Trust (relational, spiritual, and psychological aspects). Board certified physicians monitored patient progress. Nutritionist, exercise physiologist, psychological, and chaplaincy care is also provided. From 2282 patients that completed the 18-day NEWSTART program in 13 years we found 66 that had the starting diagnosis of asthma. From those 66, 39 had a pre and post stress test to determine MetS. These 39 patients had an average age of 59, SD 14.9.

Results
At baseline, asthmatics had on average a MetS of 8.38, SD 3.3, median 9, mode 10, min 2 and max 16. At the end of the program the stress test reported a MetS of 9.6, SD 3.8, median 10, mode 9, min 2 and max of 18. Asthmatic crises decreased during the program.

Conclusions
The lifestyle Program seems to improve fitness of asthma patients. The end MetS of 9.6 is important since a 10 to 11 MetS is considered good exercise capacity and may decrease mortality.
Experimental infection with human rhinovirus-A16 reveals an amplified Th1 response in allergic asthmatics
Authors: Muehling Lyndsey , Agrawal Rachana , Wright Paul , Kwok William , Heymann Peter , Woodfolk Judith 
Published: January 2023

Abstract
Human rhinovirus (RV) is a major cause of the common cold and is known to induce acute asthma episodes in allergic subjects. Despite established roles for CD4+ T cells in both anti-viral immunity and allergic disease, the T cell mechanisms of RV-induced asthma are poorly understood. Current theory suggests a role for deficient anti-viral responses, as well as pro-allergic Th2 responses. In this study, we leveraged new pMHCII tetramer reagents to specifically probe both RV- and allergen-specific circulating CD4+ T cell responses to experimental RV infection in allergic asthmatic and control subjects. T cells were monitored for 3 weeks to capture acute and convalescent phases. RV-specific cells were readily detected prior to infection in both asthmatics and controls, whereas allergen-specific cells were only detected at appreciable frequencies in asthmatic subjects. While allergen-specific cells displayed predominantly Th2 phenotypes (CCR4+, CRTH2+) in allergic asthmatics, RV-specific cells were Th1-like (CXCR3+, CCR5+). The RV-specific Th1 signature was enhanced during acute infection and persisted into convalescence. A minority of cells of both specificities expressed hybrid Th1/Th2 molecular signatures. As predicted, RV-specific cells expanded and became activated during infection with RV-A16. However, this response was augmented in allergic asthmatics, and was accompanied by increased numbers of allergen-specific cells. Additionally, plasmacytoid dendritic cells (pDCs) increased during acute infection, but only in asthmatics. Taken together, these data suggest that RV-induced asthma is characterized by an augmented anti-viral Th1 response that is mediated by pDCs, and which co-opts allergen-specific Th2 cells.