bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of human B cells bearing highly charged and highly hydrophobic amino acids in CDR-H3 at the pre B cell receptor stage (HEM2P.267)
Authors: Khass Mohamed , Blackburn Tessa , Saad Ayman , Lamb Lawrence , Burrows Peter , Schroeder Harry 
Published: January 2023

Abstract
Successive checkpoints in B cell development including differentiation in the bone marrow and the peripheral lymphoid organs control the composition and reactivity of the antibody repertoire. One of the first key checkpoints is testing of the efficiency of the heavy chain (HC) by forming a pre B cell receptor (pre BCR) composed of the newly generated μHCs and invariant surrogate light chains. Similar to mouse, human B cells with highly charged or highly hydrophobic antigen binding sites are rare in the mature B cell repertoire. Suppression of highly hydrophobic antigen binding sites in mouse reflects, in part, reduced use of RF2 due to Dμ protein containing preBCRs, but the mechanism(s) by which these clones are suppressed in human is unknown. We cloned and sequenced HC transcripts from early to late pre B cells in bone marrow samples obtained from healthy human donors and then used our bioinformatics approach to dissect and analyze the diversity of the third complementary determining region of the HC (CDR-H3). The prevalence of both highly hydrophobic and highly charged CDR-H3s dropped in the transition from early to late pre B cells. This suggests a novel mechanism of selection in human, where the pre BCR controls the passage of B cells based on their CDR-H3 amino acid content. These findings can help explain the diminished production of HIV neutralizing antibodies with abnormal highly charged or highly hydrophobic CDR-H3s due to their loss at the pre BCR stage.
Age-associated B cells (ABC) cause reduced surrogate light chain expression and loss of B cell precursors in old age (HEM2P.269)
Authors: Riley Richard , Ratliff Michelle , Alter Sarah , Frasca Daniela , Wright Jacqueline , Khan Wasif , Blomberg Bonnie 
Published: January 2023

Abstract
Age-associated B cells (ABC, CD21/35- CD23-) accumulate in spleen and bone marrow of ~2 yrs. old mice and induce TNF-α-dependent apoptosis of pro-B cells. The remaining B cell precursors in aged mice exhibit reductions in surrogate light chain (SLC) and compromise of the preBCR checkpoint. Notably, the residual pro-B cells in aged bone marrow are resistant to ABC/TNF-α-mediated apoptosis. SLC is associated with cadherin 17 on the surface of pro-B cells. Like SLC-low aged pro-B cells, pro-B cells from λ5 SLC-deficient young adult mice were also resistant to TNF-α-induced apoptosis. This implies that expression of the SLC/cadherin 17 complex is required for normal susceptibility to apoptosis in pro-B cells. Resistance to apoptosis in aged pro-B cells and λ5-deficient young pro-B cells coincided with increased phosphorylation of the pro-apoptotic protein Bim, a modification that tags Bim for degradation. We hypothesize that, in old mice, expanded pro-inflammatory ABC within the bone marrow cause apoptosis of pro-B cells. However, this cell death occurs preferentially in pro-B cells which express relatively high levels of SLC. The remaining pro-B cells in aged mice are consequently skewed to lower SLC expression. We suggest that ABC, by inducing the loss of “high SLC” pro-B cells, deviate B lymphopoiesis into an ”SLC low pathway” with inefficient B cell production and altered B cell antibody repertoire.
Chronic TLR4 stimulation selectively cripples human lymphopoiesis (HEM3P.286)
Authors: Wang Yujuan , Liu Ailing , Ding Ying , Payne Kimberly , Borghesi Lisa 
Published: January 2023

Abstract
Long-term hematopoietic stem cells (LT-HSCs) must replenish all immune cells lost due to natural turnover, inflammation or injury throughout lifetime. Our recently published data demonstrate that following chronic, low-dose TLR4 agonist stimulation, murine HSCs hyperproliferate, preferentially undergo myeloid-specific differentiation and exhibit compromised self-renewal. However, the impact to human LT-HSC function in vivo has not been established. Here we reconstituted NSG mice with human CD34+ HSCs derived from 3 different donors, and tracked hematopoiesis. Following chronic, low-dose exposure to the TLR4 agonist lipopolysaccharide (LPS), human bone marrow CD34+ HSCs were decreased ~2-fold while downstream CLPs and pro-B cells were dramatically reduced about 90%. In spleen and peripheral blood the percentage of CD19+ B cells was decreased 2-4-fold while CD56+ NK and CD3+ T were proportionately increased. Myeloid cells were largely unaffected. These findings are potentially important because TLR4 on HSCs is activated by two ligands, LPS and dietary fatty acids that are commonly elevated in diseases of chronic inflammation as well as in diet-induced obesity. Our studies may provide a previously unrecognized mechanism of human HSC exhaustion and/or compromised hematopoiesis in patients suffering from these chronic conditions.
Plasma gelsolin as a novel biomarker for cardiac surgery associated acute kidney injury in infants and young children (HUM1P.311)
Authors: Shu Qiang , Shi Shanshan , Fan Jiajie , Chen Chi , Zhao Dongyan , Liu Xiwang , Cheng Baoli , Wu Shuijing , Lin Ru , Tan Linhua , Fang XiangMing 
Published: January 2023

Abstract
Objective Cardiac surgery associated acute kidney injury (CS-AKI) strongly correlated with postoperative morbidity and mortality, especially in pediatric population. The purpose of this study was to investigate the role of plasma gelsolin (pGSN) in patients ≤3 years old, and determine whether pGSN was associated with the occurrence and severity of CS-AKI. Methods 77 patients ≤3 years of age performed on cardiac surgery were enrolled. Plasma gelsolin, creatinine, cystatin C and total protein were measured at 7 perioperative time points. Results Normalized pGSN (pGSNN) of AKI group dropped immediately after CPB (p=0.029), and still remained at lower level at 48 hours after CPB (p=0.036); PGSNN of AKI group at 2 hours after CPB was significantly lower than that of non-AKI group (p=0.002). The earliest increase of normalized pCys C (pCys CN) in AKI patients was 24 hours after CPB (p=0.008); PCys CN of AKI group at 24 hours after CPB was much higher than that of non-AKI group (p=0.002). PGSNN at 2 hours CPB was the only independent predictor of CS-AKI (p=0.032), and negatively correlated with the severity of CS-AKI (r=-0.488, p=0.002). Conclusions This study suggested that CS-AKI patients underwent a rapid and persistent drop of pGSNN after surgery; Compared with pCys CN, pGSNN appeared to be a more sensitive predictor of CS-AKI in infant and young children; PGSNN at 2 hours CPB was the only independent predictor of CS-AKI, and negatively correlated with the severity of CS-AKI.
Defining pathogenic T cells in experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS) (HUM1P.318)
Authors: Wagner David , Waid Dan , Vaitaitis Gisela , Carter Jessica , Corboy John 
Published: January 2023

Abstract
The etiology of MS is unclear, but autoimmunity plays a significant role. The autoimmune components are defined in the EAE model yet defining a biomarker remains problematic. We described a cell subset, Th40 cells, which proved highly auto-aggressive in diabetes. We compared Th40 levels in 90 MS subjects including clinically isolated syndrome (CIS), relapsing/remitting (RRMS) and secondary progressive (SPMS) patients to non-autoimmune controls. Diagnosed MS subjects have significantly increased percentages of Th40 cells in PBL compared to controls. Th40 cells produce pro-inflammatory cytokines. HLA haplotypes of MS patients were diverse although many carry DR2 (DR15). Th40 cells were elevated in MS patients regardless of HLA haplotype. Th40 cells responded to CNS antigens in an HLA dependent fashion. TRAV 8.1.3+ cells were expanded within the Th40 population in MS subjects. We used the EAE model to better define Th40 cells. EAE induction drastically expanded Th40 cell levels prior to symptoms. Th40 cells produce IFNg and IL-17 and infiltrate both brain and spinal cord. Th40 cells transfer EAE but surprisingly CD4+CD40- T cells also did but significantly (p &lt; 0.001) more slowly and with less severity. CD4+CD40- cells became CD40+; suggesting the disease process induces CD40 on T cells. CD40flox.CD4.Cre mice did not develop EAE even after multiple disease induction attempts. These data suggest an important role for CD40 in T cell pathogenesis of MS and EAE.
RGC-32 regulates TGF- β extracellular matrix expression in reactive astrocytes (HUM1P.320)
Authors: Tegla Cosmin , Cudrici Cornelia , Boodhoo Dallas , Martin Alvaro , Danoff Jacob , Sugarman Adam , Rus Violeta , Rus Horea 
Published: January 2023

Abstract
Excessive extracellular matrix (ECM) deposition in active demyelinating multiple sclerosis (MS) lesions may impede axonal regeneration and modify immune reactions. In this study we investigated first the expression of collagen types I-V, decorin and RGC-32 in MS lesions. Collagens I, III, IV and V and decorin were expressed in the perivascular space and parenchyma, and collagens I, IV and V were expressed by astrocytes that also expressed RGC-32. Since RGC-32 is involved in mediating some of TGF-β effects we investigated its role in TGF-β-induced ECM expression. In cultured astrocytes, both fibronectin and collagens I, IV and V were significantly induced after 18 h of stimulation with TGF-β. Silencing RGC-32 expression by transfecting astrocytes with siRGC-32 caused a significant reduction in the TGF-β-induced expression of collagen I (p&lt;0.01), collagen IV (p&lt;0.02), collagen V (p&lt;0.05) and fibronectin (p&lt;0.05) when compared to treatment with control siRNA. We also found that α-smooth muscle actin (α-SMA) was significantly reduced after RGC-32 silencing (p&lt;0.05). The effect on α-SMA suggests that RGC-32 is also required for the transition of astrocytes to a reactive state. These data indicate that RGC-32 plays an important role in TGF-β-mediated induction of ECM expression in astrocytes. RGC-32 may therefore represent a useful new target for therapeutic intervention in MS.
B cells from relapsing remitting multiple sclerosis patients modulate T cell behavior in a neuroantigen-specific manner (HUM1P.322)
Authors: Ireland Sara , Guzman Alyssa , Davis Laurie , Cowell Lindsay , Remington Gina , Greenberg Benjamin , Frohman Elliot , Monson Nancy 
Published: January 2023

Abstract
The role of B cells in the pathoetiology of relapsing remitting multiple sclerosis is not well understood. B cells may contribute to MS by antibody secretion, cytokine production and antigen presentation. Antigen-specific B cells are highly efficient antigen presenting cells in the context of their cognate antigens. Since antibodies from multiple sclerosis patients bind to neuroantigens we hypothesized that neuroantigen specific memory B cells promote disease in multiple sclerosis patients by serving as antigen presenting cells for auto-reactive T cells. We observed that B cells from a subset of multiple sclerosis patients induce naïve and memory T cell proliferation in the presence of recombinant neuroantigens, myelin oligodendrocyte glycoprotein and myelin basic protein, but not MOG 35-55 peptide, in B-T cell co-cultures. Similarly, in the presence of neuroantigen proteins cells from multiple sclerosis patients secreted greater levels of IFN-γ, IL-6, -17A, -17F, -21, -22, -31, -33, TNF-α and soluble CD40L than healthy donors, while levels of IL-4 and IL-25 were similar. These data support a role for human B cells as antigen presenting cells in multiple sclerosis and highlight the role of B cells in the context of autoimmune disease beyond antibody production.
Matrine suppresses glutamate excitotoxicity in experimental autoimmune encephalomyelitis (HUM1P.323)
Authors: Xiao-Jian Zhang , Ju-Kui Song 
Published: January 2023

Abstract
Multiple sclerosis (MS) is T cell-mediated inflammatory diseases characterized by lymphocyte infiltration and demyelination. Matrine (MAT) is a quinolizidine alkaloid derived from the herb Radix Sophorae Flave and has long been used in the treatment of hepatitis B in clinical without obvious side effects. Previous reports have revealed that MAT restrained the central inflammation after experimental autoimmune encephalomyelitis (EAE), the most common animal model of MS due to similar symptoms and mechanisms; however whether MAT is effective to inhibit glutamate excitotoxicity is still unclear. In this study, we provide evidence that MAT attenuated the EAE disease severity, accompanied by down-regulated glutamate level, increased GABA level, as well as promoted the expressions of two dependent glutamate transporters (GLT-1 and GLAST). In addition, MAT treatment significantly decreased the content of the NMDA- and AMPA- glutamate receptor in EAE rats. Together, these data indicate that MAT treatment inhibited glutamate excitotoxicity after EAE, and suggest that MAT may be a novel choice in MS therapy.
Amelioration in generalized anxiety disorder is associated with decreased Treg number and function (HUM1P.327)
Authors: Akimova Tatiana , Negorev Dmitri , Luthra Lalita , Berton Olivier , Hancock Wayne , Lohoff Falk 
Published: January 2023

Abstract
In preclinical models, chronically stressed and highly anxious animals displayed impaired protective immunity and associated increased risks of cancer progression. These observations may reflect stress-linked increases in Treg numbers or function. We therefore assessed the number and function of FOXP3+ T-regulatory (Treg) cells in 9 patients with generalized anxiety disorder (GAD). These patients had Hamilton Anxiety Scale (HAM-A) scores of &gt;20, and provided blood samples for Treg evaluation at baseline and after 8 weeks of treatment with the antidepressant, Desvenlafaxine, or placebo. After treatment, patients with significant improvement of HAM-A scores (i.e. responders) showed decreased Treg suppressive function in comparison with non-responders (23.8±9.8 vs. 83.4±50 units for CD4+ responder cells, p=0.035). Decreased HAM-A scores correlated with decreased suppression of CD4 cell proliferation (Spearman r=0.745, p=0.021) and with decrease in Treg numbers (Spearman r=0.775, p=0.041). Treg ability to suppress CD8 cell proliferation after treatment was also inversely correlated with decreases in HAM-A scores (Spearman r=-0.75, p=0.02). Conclusion: clinical improvement in GAD is accompanied by normalization of Treg numbers and function, which may have long-term impact on patient health and, potentially, risk of cancer progression.
Monocyte aggregates and intermediate (CD14++CD16+) cells that are associated with cardiac events are increased in human psoriatic blood and CD11b +Ly6C high cells and aggregates are increased in the KC-Tie2 psoriasiform mouse model (HUM1P.307)
Authors: Golden Jackelyn , Groft Sarah , Wang Yunmei , Diaconu Doina , Simon Daniel , McCormick Thomas , Ward Nicole , Cooper Kevin 
Published: January 2023

Abstract
Psoriasis confers an increased risk for developing and dying of cardiovascular disease (CVD). We found increased circulating CD14++CD16+ intermediate monocytes (IntMo), a cell type associated with increased cardiovascular risk, in psoriasis patients compared to healthy controls (N=42, 16.5%±2.7 vs. 11.9%±1.4, P=.056). Amnis imaging flow cytometry of the psoriatic IntMo cells revealed increased doublets and aggregates. Overall, CD14++ doublets are increased in psoriasis patients compared to controls (2.72%±0.58 vs. 1.09%±0.20; P=0.006). Murine psoriasiform skin models (KC-Tie2) exhibit increased vascular thrombosis and monocyte doublets, indicating a skin inflammation-driven process. KC-Tie2 mice are pro-thrombotic, develop aortic root vascular lesions, and have increased splenic CD11b +Ly6C high (Ly6Chigh) monocytes (33.8%±3.8, n=7) compared to controls (8.05%±0.55, n=9; P=0.0002), suggesting that chronic skin inflammation generates Ly6Chighcells which mediate vascular inflammation. KC-Tie2 mice also show an increase in Ly6Chighcells in skin-draining lymph nodes compared to controls (8.83%±0.77, n=18, vs. 1.94%±0.8, n=3, P=.002). Crossing IL-6 KO mice with KC-Tie2 decreases these splenic Ly6Chighcells, reverses thrombosis and aortic root vascular lesions, and eliminates skin inflammation. Psoriasis-driven accumulation, increase, and aggregation of monocytes in circulation suggests a novel mechanistic linkage between psoriatic skin disease and its CVD comorbidity.
Do particular antibody genes recognize gray matter cellular targets? (HUM2P.338)
Authors: Monson Nancy , Ligocki Ann , Rounds William , Li Min , Henson Paul , Rivas Jacqueline , Guzman Alyssa , Graves Donna , Greenberg Benjamin , Frohman Elliot , Ward Sally , Stowe Ann 
Published: January 2023

Abstract
My lab’s recent work on the antibody genetics of multiple sclerosis (MS) patients has led to the astonishing discovery that patterns of mutation accumulation into the antibody genes of B cells in the central nervous system (CNS) of MS patients are unique and can be used to predict clinically isolated syndrome (CIS) conversion to MS with 94% accuracy. This Antibody Gene Signature (AGS) is a novel, potentially diagnostic biomarker that is currently in clinical trials and heralds the advent of antibody genetics as a means to categorize patients early in their disease course in order to provide prompt treatment and prevent extensive CNS damage. Our goal for this project was to determine what antibodies expressed by these B cells from MS patients reacted against. We cloned and expressed more than 30 antibodies that contain the AGS-MS from cerebrospinal fluid derived single B cells of MS and CIS patients, and tested their capacity to bind to brain tissue. We found that these AGS-MS enriched antibodies bind to neurons and astrocytes that reside in the brain gray matter tissue with high specificity. Some of these antibodies recognize both neurons and astrocytes in the gray matter, whereas others are specific for a single cell type. We hypothesize that AGS-enriched rhAbs and B cells expressing these antibodies could be participating in gray matter disease pathology in both early CIS and later MS stages.
Loss of AMPKα1 in neuroinflammatory phenotype of X-linked adrenoleukodystrophy (HUM2P.335)
Authors: Singh Jaspreet , Giri Shailendra , Suhail Hamid , Deshpande Mandar 
Published: January 2023

Abstract
X-linked adrenoleukodystrophy (X-ALD) is the most common inherited neuroinflammatory demyelinating leukodystrophy. All X-ALD patients present with mutations in the ABCD1 gene, which encodes a peroxisomal adenosine triphosphate (ATP)-binding cassette transporter protein (ABCD1). X-ALD is a complex disease where the same mutation in the ALD gene (ABCD1) can lead to clinically very distinct phenotypes; a fatal neuroinflammatory childhood cerebral ALD (ALD) or the relatively benign disease of adrenomyeloneuropathy (AMN). Further, no relationship could be established between genotype and severity of the disease as same mutation is known to give different phenotypes, even within a family. We recently documented a role for decreased AMP kinaseα1 (AMPKα1) levels in initiating severe inflammatory and demyelinating phenotype in the CNS of AMPKα1-knockout mice. Abundant data is also available that implicates loss of AMPK in generating spontaneous inflammatory response in several cell types including immune cells. Our data using healthy control, AMN and ALD patient-derived skin fibroblasts and lymphocytes documents, for the first time, loss of a metabolic gene- AMPKα1 in ALD patient-derived fibroblasts and lymphocytes. Consistent with this, the basal expression of inflammatory cytokines (iNOS and IL-6) were increased in ALD patient-derived lymphocytes. Together these observations suggest that AMPKα1 may be involved in the progression of benign AMN to neuroinflammatory ALD phenotype.
Fluvastatin enhances IL-33-mediated cytokine production in mouse mast cells (HYP3P.410)
Authors: Taruselli Marcela , Kolawole Elizabeth , Ryan John 
Published: January 2023

Abstract
The HMG-CoA reductase inhibitor Fluvastatin is a commonly used treatment for hypercholesterolemia. This drug recently has been shown to have immunological suppressive effects. Interleukin 33 (IL-33), an alarmin, has been suggested to contribute to allergies and anaphylaxis by activating mast cells. Due to the suppressive functions of Fluvastatin, we investigated the role of Fluvastatin on interleukin IL-33-stimulated mast cell function. Unlike previous studies, we have found that Fluvastatin increased production of the pro-inflammatory cytokines IL-6 and TNF-α in IL-33 stimulated mast cells. Furthermore the increase in cytokine production appeared to be stem cell factor (SCF)-dependent, since in the absence of SCF, Fluvastatin decreased cytokine production. Investigating the role of Fluvastatin in IL-33-activated mast cells will reveal a better understanding of how innate immunity may be altered by a widely-prescribed class of medications.
Preliminary investigation on the role of micro-RNAs in IL-10 resistance in mast cells (HYP3P.412)
Authors: Abdul Qayum Amina , McLeod Jamie , Ryan John 
Published: January 2023

Abstract
Mast cells are immune cells that play a critical role in allergic diseases both as effector and immunoregulatory cells. A well understood mast cell activation pathway is the binding of IgE to its high affinity IgE receptor (FcεRI). Possible regulators of mast cell homeostasis are regulatory cytokines produced by the immune response. We have shown that IL-10 hinders mast cell cytokine production by selectively targeting the Fyn-Stat5/Akt cascade proteins activated downstream of FcεRI. Interestingly, these effects are altered by genetic background, as cytokine suppression is observed in mast cells from Th1-prone C57BL/6 but not Th2-prone 129/Sv mice. We hypothesize that IL-10 responsiveness is mediated by induction of MicroRNAs (miRNAs), and that a failure to induce these miRNAs results in IL-10 non-responsiveness. miRNAs could potentially be targeting Akt, Stat5B, and Fyn to antagonize cytokine production. We demonstrate that miR-135a-5p expression increases in IL-10-treated BMMC derived from Th1-prone C57BL/6 mice, but not BMMC from Th2-prone 129/Sv mice. Unraveling how mast cell homeostasis is maintained will shed light on the predisposition to numerous inflammatory diseases.
IgE receptor induces Stat5b serine phosphorylation in mast cells (HYP3P.346)
Authors: Kiwanuka Kasalina , Ryan John 
Published: January 2023

Abstract
Signal Transducers and Activators of Transcription (STATs) are latent transcription factors that mediate a number of cellular responses including but not limited to cell survival, proliferation and differentiation. This protein family consists of seven members, STAT1-6 including two closely related molecules, STAT5a and STAT5b, that show 96% amino acid sequence homology and are critical for lymphoid, myeloid, and erythroid cell development and function. We recently identified Stat5b as the critical regulator of IgE-mediated cytokine production in mast cells. We found that Fyn-mediated Stat5 tyrosine phosphorylation is critical for Stat5-dependent effects. Our recent work demonstrates that Stat5b is also phosphorylated on serine residues during IgE signaling in mast cells. We find that STAT5b serine phosphorylation occurs through a Src-independent pathway requiring PI3-kinase function. Understanding the pathways leading to Stat5b serine phosphorylation and its role in the inflammatory response will yield new targets for mast cell-associated diseases.