bluuebunny/crossref_metadata_2025_split huggingface dataset:
Endogenous S-Nitrosothiol Induced hypertrophic Astrogliosis has potential to maintain CNS homeostasis under pathological Conditions (72.8)
Authors: Paintlia Manjeet , Paintlia Ajaib , Singh Avtar , Singh Inderjit 
Published: January 2023

Abstract
Compelling evidence suggests that reactive astrogliosis exert both detrimental and beneficial effects in response to central nervous system (CNS) inflammatory insult. The exact mechanism behind such diversity in astrocytic functions to maintain CNS homeostasis is not clear. Recent studies explicated the biphasic functions of nitric oxide (NO) in EAE, where reactive astrogliosis is known to play critical role in its pathogenesis. Here, we investigated the effect of endogenous NO donor, S-nitrosoglutathione (GSNO) on astrocytic functions in vitro settings. Interestingly, GSNO induced hypertrophic astrogliosis with increased expression of ciliary neurotrophic factor (CNTF) in treated astrocytes. This effect of GSNO on astrocytes was ascribed to an induction of NO-soluble guanylate cyclase-cGMP activated PKG signaling and activation of p38MAPK and PPAR-g with RhoA inhibition. Also, we found that GSNO-mediated increased GFAP expression in astrocytes was ascribed to CNTF induced JAK2/STAT3 signaling in an autocrine manner. This GSNO-mediated hypertrophic astrogliosis had potential to maintain CNS homeostasis under pathological conditions. Together, these data showed the beneficial functions of GSNO and suggest new therapeutic interventions to protect CNS against Inflammatory insult similar to MS brain.
CD4+CD25+FOXP3+ regulatory T cells and association with survival in epithelial ovarian cancer (74.9)
Authors: Preston Claudia , Maurer Matthew , Oberg Ann , Hartmann Lynn , Kalli Kimberly , Goode Ellen , Knutson Keith 
Published: January 2023

Abstract
Ovarian cancer is an immune reactive tumor with a complex suppressive network that blunts immune eradication. One putative pathway of evasion may be recruitment of CD4 regulatory T cells (Treg) into the tumor microenvironment. This study investigated the association of tumor infiltrating Tregs in ovarian cancer patients with poor survival. Methods: Immunofluorescence of intraepithelial CD4+CD25+FOXP3+ Tregs, CD3+ and CD8+ cells was performed in tumors of 52 ovarian cancer patients. Average and maximum count of cells were associated with survival via Cox regression models and summarized with hazard ratios (HR) and 95% confidence intervals (CIs). Results: The maximum counts of Tregs in these patients suggests an association between higher number of Tregs and poor clinical outcome (HR=1.48, CI=0.97, 2.28), although not statistically significant (p=0.07). Moreover, presence of high CD8/Treg ratio (HR=0.7, CI=0.53, 0.91; p=0.008) as well as high CD8/CD4 ratio (HR=0.76, CI=0.57, 1.0; p=0.05) appear to be associated with good outcome. The CD3 lymphocyte count was not associated with survival (HR=0.9, CI=0.66, 1.24; p=0.52). Conclusion: Although the present pilot study was conducted with a limited number of patients, our data suggests that higher numbers of infiltrating Tregs might be associated with poor survival and low CD8/Treg ratio may be correlated with improved survival in patients with epithelial ovarian carcinoma. We plan to confirm these results in a larger set of patients.
Development and preclinical evaluation of SEL-068, a novel targeted Synthetic Vaccine Particle (tSVP™) for smoking cessation and relapse prevention that generates high titers of antibodies against nicotine (75.11)
Authors: Pittet Lynnelle , Altreuter David , Ilyinskii Petr , Fraser Christopher , Gao Yun , Baldwin Sam , Keegan Mark , Johnston Lloyd , Kishimoto Takashi 
Published: January 2023

Abstract
Novel vaccine-based therapies provide new avenues for the treatment of chronic conditions such as addiction. Here we describe SEL-068, an entirely synthetic, self-assembling nanoparticle vaccine against nicotine for smoking cessation. SEL-068 is a targeted Synthetic Vaccine Particle (tSVP™) comprised of four main components: a biodegradable and biocompatible polymer matrix, a synthetic TLR agonist, a novel universal T cell helper peptide, and nicotine covalently conjugated to the nanoparticle surface (B cell antigen). The nanoparticles are designed to flow freely to the lymph nodes, enabling direct and concomitant delivery of all required components to the responsive cells of the immune system. Encapsulation and controlled release of the TLR agonist in the nanoparticle minimizes the generation of systemic inflammatory cytokines, improving the overall safety profile and enabling the use of novel adjuvants. The novel universal T cell helper peptide used in SEL-068 elicited T cell recall responses in PBMC from a broad panel of human donors. Immunization of mice with SEL-068 led to the generation of high titers of high affinity anti-nicotine antibodies. Rhesus and cynomolgus monkeys immunized with increasing doses of SEL-068 showed a robust and dose dependent induction of high titers of anti-nicotine antibodies with strong affinity to nicotine. No dose-limiting systemic toxicities were detected. SEL-068 is currently being evaluated in humans in Phase 1 clinical trials.
Impact of IgA constant domain on HIV-1 neutralizing function of monoclonal antibody F425-A1g8 (75.12)
Authors: Yu Xiaocong , Duval Mark , Lewis Christopher , Cavacini Lisa 
Published: January 2023

Abstract
With the majority of HIV infections resulting from mucosal transmission of HIV, induction of an effective mucosal immune response would be pivotal in preventing transmission. HIV-specific IgA but not IgG has been detected in genital tract, seminal fluid, urethral swabs, urine and vaginal wash samples of HIV-negative sex workers and HIV-status discordant couples. Purified mucosal and plasma IgA from HEPS individuals neutralize infection and though present at low levels, cross-clade neutralization activity. We generated a CD4i human monoclonal antibody F425-A1g8 and characterized the impact of its isotype variants to HIV neutralizing activity. The result showed that in contrast to little neutralization by F425-A1g8 IgG1 in the absence of sCD4, the IgA1 variant of the antibody displayed significant neutralization activity against a range of HIV clade B isolates in the absence of sCD4. This research clearly suggests that IgA isotype utilizing its unique molecular structure plays an important role in HIV neutralization. The studies of the neutralizing function of IgA isotypes and the functional relationship between epitopes and IgA antibodies may also suggest strategies for intervention in virus transmission or spread within the mucosal of host, as well as serve to inform the design of vaccine strategies that may be more effective at preventing mucosal transmission.
Adjuvanting influenza vaccines for enhanced immunity to seasonal or pandemic influenza virus. (75.9)
Authors: Fairman Jeffery , Carroll Tim , Liu Feng , Hancock Kathy , Katz Jacqueline , Lu Xiuhua , Matzinger Shannon , Miller Chris 
Published: January 2023

Abstract
The efficacy of FDA approved influenza vaccines depends on eliciting neutralizing antibodies that provide protection against antigenically well-matched viruses. To determine if the JVRS-100 adjuvant could enhance vaccine-induced protection against matched and non-matched influenza virus challenge, mice or non-human primates (NHP) were immunized with vaccine +/- the JVRS-100 adjuvant. All animals that received JVRS-100 adjuvanted vaccine had significantly lower levels of virus replication following viral challenge. In matched and cross-clade challenge experiments JVRS-100 adjuvanted H5N1 vaccine elicited enhanced humoral responses, T-cell responses, and protection in mice. In heterosubtypic challenge experiments, mice and nhp immunized with adjuvanted H1N1 vaccine demonstrated enhanced antibody responses, T-cell responses, and protection against H3N2 challenge compared with groups that received nonadjuvanted vaccine. These data demonstrate that JVRS-100 enhances antibody and T-cell responses, resulting in matched, cross-clade, and heterosubtypic immunity for influenza vaccination.
Selective Depletion of CD19+CD24hiCD38hiIL-10high Regulatory B cells (Bregs) in HIV-infected Aviremic Individuals (76.2)
Authors: Siewe Basile , Stapleton Jack , Martinson Jeffrey , Keshavarzian Ali , Kazmi Nazia , Demarais Patricia , French Audrey , Alan Landay 
Published: January 2023

Abstract
HIV-infection associated upregulation of IL-10 contributes to impaired antiviral effector T-cell functions promoting viral persistence. Bregs (CD19+CD24hiCD38hi) IL-10-mediated modulation of T-cell function has been reported in autoimmune disease but yet undefined during HIV infection. In this study, we confirmed high expression of IL-10 by Bregs in healthy controls (HC, n=16) and determined Breg frequency, T-cell immune activation (CD8+HLA-DR+CD38+, CD4+HLA-DR+CD38) and T-cell exhaustion (CD8+PD-1+) in aviremic (HIVavir, n=10) and viremic (HIVvir, n=15) HIV-infected individuals. We found a significant positive correlation between Breg frequency and viral load (r2=0.5166, p=0.0097); HIVavir-individuals exhibited a significant Bregs loss compared to HC (p=0.0004) and HIVvir-individuals (p=0.0122), despite no significant differences in total B-cell frequency. A significant positive correlation was observed between Breg frequency and immune activation (CD8+HLA-DR+CD38+, r2=0.6262, p=0.0041; CD4+HLA-DR+CD38+, r2=0.6298, p=0.0039) as well as T-cell exhaustion (r2=0.8853, p=0.0333). Additionally, we found a significant difference in Breg CD95 (Fas) expression between HIVavir and HIVvir-individuals (p=0.0119) yet no such difference between in HIVavir-individuals and HC. We hypothesize that, Bregs loss during HIV-infection may be due to Fas mediated apoptosis and that activated IL-10-secreting Bregs contribute to T cell impairment hence viral persistence in HIVvir individuals.
A novel human CD1d knock-in mouse model demonstrates potent anti-tumor potential of human CD1d-restricted iNKT cells (P5001)
Authors: Yuan Weiming , Wen Xiangshu , Rao Ping , Cresswell Peter , Porcelli Steven , Kim Seil , Carreño Leandro , Lawrenczyk Agnieszka 
Published: January 2023

Abstract
Despite a high degree of conservation, subtle but important differences exist between the CD1d antigen presentation pathways of humans and mice. These differences may account for the minimal success of Natural Killer T (NKT) cell-based anti-tumor therapies in human clinical trials, which contrast strongly with the powerful anti-tumor effects in conventional mouse models. To develop an accurate model for in vivo human CD1d (hCD1d) antigen presentation, we have generated the first hCD1d knock-in (hCD1d-KI) mouse. In these mice, hCD1d is expressed in a native tissue distribution pattern and supports NKT cell development. Reduced numbers of invariant NKT (iNKT) cells were observed, but at an abundance comparable to that in most normal humans. These iNKT cells predominantly expressed mouse Vβ8, the homolog of human Vβ11, and phenotypically resembled human iNKT cells in their reduced expression of CD4. Importantly, iNKT cells in hCD1d-KI mice exert potent anti-tumor function in a melanoma challenge model. Our results show that replacement of mCD1d by hCD1d can select a population of functional iNKT cells closely resembling human iNKT cells. These hCD1d knock-in mice will allow more accurate in vivo modeling of human iNKT cell responses and will facilitate the preclinical assessment of iNKT cell-targeted anti-tumor therapies.
Therapeutic inhibition of gut antigen trafficking through scavenger receptor A (P5027)
Authors: Raycroft Maury , Mamula Mark 
Published: January 2023

Abstract
We have demonstrated that B cells acquire specific antigen (Ag) through B cell receptor for transfer to both mouse and human macrophages (MOs) and dendritic cells (DCs). Our studies also determined that this process is dependent on the MO/DC surface scavenger receptor A (SR-A). Interestingly, SR-A deficient, lupus-prone mice exhibit reduced autoantibody production and pathology. Thus, this new pathway of Ag trafficking is important in human immunity and/or autoimmunity, leading to current investigations in targeting SR-A for therapeutic intervention. To identify novel modulators of SR-A, we screened bioactive small molecule inhibitor (SMI) libraries and identified candidates that alter Ag transfer from B cells to MO/DCs ex vivo in human cell lines and in vivo in mice. An SR-A-specific and dose-dependent SMI, sennoside B, was effective in reducing autoantibody production and lupus pathology in lupus-prone mice. Interestingly, we have observed drug-mediated reduction in antigen trafficking between APCs in gut Peyer's patches. The data suggest that SRA inhibition alters the gut immune responses, which has recently been linked to autoimmune diseases such as diabetes and arthritis. Finally, several studies indicate that SR-A has regulatory affects on TLR4. We are presently investigating TLR4 and SRA-mediated interactions in mediating antigen trafficking in Peyer's patches and downstream autoimmunity.
GILT expression in medullary thymic epithelial cells is required for presentation of tissue-specific self antigens and deletion of autoreactive T cells (P5037)
Authors: Rausch Matthew , Metzger Todd , Anderson Mark , Hastings Karen 
Published: January 2023

Abstract
Clonal deletion of thymocytes with high avidity for self-peptide:MHC complexes is critical for the maintenance of tolerance and prevention of autoimmunity. We have shown that gamma-interferon-inducible lysosomal thiol reductase (GILT) is required for thymic deletion of T cells specific for a self and melanoma Ag, tyrosinase-related protein 1 (TRP1). To define GILT’s role in central tolerance, we show that GILT expression is enriched in thymic APC capable of mediating deletion, medullary thymic epithelial cells (mTEC) and dendritic cells. TRP1 expression is restricted to mTEC. GILT facilitates the MHC class II-restricted presentation of endogenous TRP1 by pooled thymic APC (TEC and bone marrow (BM)-derived APC). Using BM chimeras, we show that TRP1-specific T cells are deleted in chimeras in which GILT is expressed in both TEC and BM-derived APC or GILT expression is restricted to TEC, indicating that GILT expression in TEC is sufficient for deletion of TRP1-specific thymocytes. In contrast, a large number of TRP1-specific T cells develop in chimeras which lack GILT expression in both TEC and BM-derived APC. Chimeras in which GILT expression is limited to BM-derived APC have intermediate levels of TRP1-specific T cells, demonstrating that GILT expression in TEC is required for efficient deletion of TRP1-specific thymocytes. These findings suggest that GILT operates in mTEC to facilitate the presentation of tissue-specific self Ag and promote deletion of autoreactive T cells.
Inflammation-induced splicing of Deaf1 in the pancreatic lymph nodes during the progression of Type 1 diabetes (P5061)
Authors: Yip Linda , Creusot Remi , Whiting Chan , Taylor Cariel , Akter Rahima , Matsumura Teppei , Fathman C. Garrison 
Published: January 2023

Abstract
Peripheral tolerance is partially controlled by ectopic peripheral tissue antigen (PTA) expression in lymph node stromal cells (LNSCs). We previously showed that the transcriptional regulator deformed epidermal autoregulatory factor 1 (Deaf1) can directly control the transcription and indirectly regulate the processing and/or presentation of PTA genes in LNSCs of the pancreatic lymph nodes (PLN). We showed that Deaf1 is alternatively spliced in the PLN of Non-Obese Diabetic (NOD) mice and Type 1 diabetes (T1D) patients, to form a dominant negative isoform, Deaf1-Var. We previously demonstrated that Deaf1-Var expression was highest in the PLN of NOD mice during the onset of disease, compared to the non-inflamed PLN of healthy age-matched NOD.B10 and NOD.SCID mice. In NOD mice, Deaf1-Var expression in the PLN was found to increase as disease progressed from peri-insulitis to destructive insulitis. In this study, we asked if inflammation induces Deaf1 splicing in the PLN. Deaf1 splicing could be induced in the PLN of NOD.B10 control mice and NOD.SCID mice by i.p. injection of activated splenocytes that home to the PLN. The supernatant of activated splenocytes could also to induce Deaf1 splicing in pancreatic islets, a tissue previously shown to splice &gt;500 genes in response to inflammation. Together, these data suggest that the inflammation that occurs in the PLN during the progression of T1D leads to Deaf1 splicing, loss of Deaf1 function, and reduced PTA expression.
Functionally distinct ERAP1 haplotype combinations define individuals with ankylosing spondylitis (P5047)
Authors: James Edward , Reeves Emma , Colebatch Alexandra , Elliott Tim , Edwards Christopher 
Published: January 2023

Abstract
Ankylosing Spondylitis (AS) is a chronic inflammatory disease which commonly affects the spine occurring in approximately 0.5% of adults of European descent. In addition to HLA-B27, recent genetic linkage studies have identified polymorphisms within the aminopeptidase, ERAP1, associated with the disease. ERAP1 lies on the antigen processing pathway, generating peptide antigens for immune presentation by HLA. Here we show that ERAP1 from individuals are polymorphic and comprise haplotypes that are predominantly made up of multiple SNPs. ERAP1 haplotype combinations found in AS cases are distinct from those in controls. AS case ERAP1 haplotype combinations are unable to trim peptide precursors to generate the final epitope for presentation to T cells and are also unable to reconstitute MHC class I to normal levels as observed with control haplotype combinations. We therefore provide strong evidence that ERAP1 variation predisposes to chronic inflammatory disease via its influence on the antigen processing pathway.
Characterization of human monocyte subsets (P4161)
Authors: Boyette Lisa , Macedo Camila , Elinoff Beth , Metes Diana , Lakkis Fadi 
Published: January 2023

Abstract
Monocytes are an important source of antigen-presenting cells and macrophages in tissues affected by infection or inflammation and in organ transplantation. Monocytes have been grouped into classical (CD14+CD16-), non-classical (CD14dimCD16+), and intermediate (CD14+CD16+) subsets. The biologic significance of these subsets is not completely understood. We phenotyped monocytes in peripheral blood from 25 healthy subjects and performed functional studies on monocyte subsets sorted from leukapheresis products to better characterize these subsets in humans. Monocyte subset frequencies were found to be tightly controlled over time and across healthy individuals: 83.95% classical, 95% CI [81.12, 86.78], 6.65% intermediate [4.32, 8.99], and 9.28% non-classical [7.12, 11.44]. Subsets were distinct in their secretion of TNFα, IL-6, and IL-1β in response to Toll-like receptor agonists, and cytokine secretion by classical and intermediate monocytes was suppressed when all three subsets were recombined in physiologic ratios. After incubation with IL-4 and GM-CSF, classical monocytes acquired dendritic cell markers and morphology; intermediate and non-classical monocytes did not. From these studies we conclude that classical monocytes appear to be the principal source of antigen-presenting dendritic cells, and non-classical monocytes suppress secretion of inflammatory cytokines by the other subsets in mixed cultures, perhaps indicating a regulatory role.
Multiple pathways are employed by TLR and NOD agonists to induce PGE2 production in primary human monocytes and macrophages (P4163)
Authors: Zaitseva Marina , Endo Yukinori , Romantseva Tatiana , Godling Hana 
Published: January 2023

Abstract
Innate immune activators that are potent inducers of fever also up-regulate plasma PGE2. To determine whether TLR and NOD agonists activate production of PGE2 directly or through IL-1β, human monocytes (Mo) and differentiated macrophages (MF) were cultured with TLR agonists with or without Caspase-1 inhibitor ZVAD, which inhibited LPS-induced IL-1β in both cell types. PGE2 production in MF was not affected by ZVAD, while in Mo, ZVAD reduced PGE2 and COX2 expression by 30-60%. Surprisingly, IL-1β alone did not up-regulate PGE2 in Mo. However, in Mo pretreated with low dose of LPS, PGE2 and COX2 mRNA were up-regulated by IL-1β. LPS (but not TLR2 agonist) induced phosphorylation of IRF3 (Ser 396) in Mo. Knock-down of IRF3 or of TRIF by siRNA abrogated IL-1β induced COX2 mRNA expression in LPS-pretreated U937-CD14 cells. MDP did not induce PGE2 in Mo alone. However, COX2 mRNA and PGE2 protein in MDP-activated Mo was significantly augmented by human CD3 bead-purified T cells or T cell supernatant. Treatment of Mo with inhibitors of autophagy including ATG5 or ATG7 siRNA, inhibited MDP-induced PGE2 production. T cells were also required for PGE2 produced in response to flagellin and IE-DAP but not Poly I:C. In summary, Mo employ multiple pathways to produce PGE2, depending on the type of innate receptors involved: IRF3/TRIF pathway is employed in response to TLR4, while autophagy/ROS and T cell-derived signals are involved in PGE2 production after triggering of NOD1/2 and IPAF.
E3 ubiquitin ligase NKLAM positively regulates nitric oxide production and the generation of reactive oxygen species in macrophages (P4167)
Authors: Lawrence Donald , Kornbluth Jacki 
Published: January 2023

Abstract
Stimulated macrophages generate potent anti-microbial reactive oxygen and nitrogen species within their phagosomes. There is mounting evidence that ubiquitination plays a role in modulating macrophage bactericidal functions. Recently, we demonstrated that the ubiquitin ligase NKLAM was a component of the macrophage phagosome. In this present study we investigated the effects of NKLAM on macrophage generation of nitric oxide (NO) and reactive oxygen species. Stimulation with LPS induced wild type (WT) macrophages to produce NO in a time and concentration-dependent manner. NO production was attenuated in NKLAM-knock out (KO) macrophages. We also found that inducible nitric oxide synthase (iNOS) protein expression was lower and that iNOS translocation to the maturing phagosome was defective in KO macrophages. In vivo, intraperitoneal injection of LPS induced a time-dependent increase in serum nitrite (NO metabolite) in both WT and KO mice. However, nitrite levels in KO mice were lower than in WT mice. The generation of reactive oxygen species (ROS) is a potent killing mechanism; however the role of ubiquitination in regulating ROS generation is unclear. To this end, we investigated ROS production in WT and KO macrophages and found that ROS production was defective in KO macrophages stimulated with either PMA or Escherichia coli. Collectively, our data implicate NKLAM as a potent modulator of macrophage oxidative killing functions.