bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel mechanism for IL-23 to up-regulate AICDA and promote pathogenic autoantibodies in lupus
Authors: Hong Huixian , Wu Qi , Yang PingAr , Lou Bao , Li Jun , Li Hao , Cua Daniel J. , Hsu Hui-Chen , Mountz John D. 
Published: January 2023

Abstract
Targeting IL-23 to treat autoimmune disease and chronic inflammation is under development based on its proinflammatory function and pro-Th17 cell effects. IL-23 was previously shown to be important for promoting pathogenic autoantibody production in B6-Faslpr/lpr mice. In contrast, the lack of IL-23 did not influence germinal center (GC) formation and IgG anti-CII autoantibodies in type II collagen immunized DBA/1 mice. We determined if IL-23 regulated development of spontaneous GCs and autoantibody production in lupus-prone BXD2 mice by generating IL-23 deficient BXD2-p19−/− mice. There was significantly decreased total IgG, but increased total IgM and IgM anti-DNA and RF autoantibodies in BXD2-p19−/− mice compared to BXD2 mice. Flow cytometry and confocal imaging analyses showed that both the size and number of GL-7+Fas+ GC B cells were increased, yet there was a decreased development of GC dark zone (DZ) (CD86−CXCR4+) B cells in BXD2-p19−/− mice. Interestingly, the expression of activation-induced cytidine deaminase (AICDA or Aicda) was significantly decreased in GC B cells in BXD2-p19−/− mice. However, the expression of GC program genes (Bach2, Pax5) was increased except Bcl6. In contrast, the expression of key plasma cell program genes Prdm1 in GC B cells was diminished in the absence of IL-23.
Our data suggest a novel pathway for IL-23 to promote IgG autoantibody production by upregulation of AICDA in the GC DZ. Importantly, such pathway is independent of the canonical GC program genes that have been implicated for the induction of AICDA. These results suggest a potential novel action of anti-IL-23 therapies to inhibit adaptive immune responses in addition to its known inhibitory role of innate response in autoimmune diseases.
IL-27 controls allergic airway inflammation via Foxp3+ regulatory T cells
Authors: NGUYEN QUANG TAM , MIN BOOKI 
Published: January 2023

Abstract
Asthma is a chronic inflammatory disease in the lung mediated by allergen reactive Th2 type CD4 T cells producing the Th2 type cytokines. We recently reported that IL-27 plays a key role in Treg functions to control Th17-mediated autoimmune inflammation. However, a role for IL-27 in modulating Treg function during Th2-mediated allergic airway inflammation remains unclear. Here, we report that during cockroach antigen (CA)-induced allergic airway inflammation, IL-27 attenuates inflammatory responses primarily via Foxp3+ Tregs. Sensitizing mice with CA in alum followed by intranasal challenge induces allergic airway inflammation characterized by infiltration of eosinophils and effector CD4 T cells expressing Th2 cytokines in the lung. Recombinant IL-27 administration prior to antigen challenge significantly reduces the inflammation. Accumulation of CD4 T cells expressing inflammatory cytokines is dramatically reduced in the inflamed sites following IL-27 administration. However, IL-27-mediated treatment effects are completely lost when Foxp3+ Tregs are deleted prior to CA challenge. Adoptive transfer of wild type Tregs restores IL-27-mediated inhibition of airway inflammation, suggesting that Tregs may be the direct responders of IL-27. IL-27 stimulation in Tregs increases expression of lymphocyte activation gene 3 (Lag3), a surface molecule implicated in Treg functions. Unlike WT type cells, adoptive transfer of Tregs deficient in Lag3 fails to attenuate airway inflammation even after IL-27 administration. Taken together, these results demonstrate a novel IL-27-dependent Treg function to control allergic airway inflammation
Activation of neonatal mouse lung ILC2s by endogenous IL-33 impacts type 2 responses later in life
Authors: Steer Catherine A. , Shim Brian , Mathae Laura , Takei Fumio 
Published: January 2023

Abstract
Neonatal lung immunity is functionally distinct from adults as responses to environmental antigens are predominantly type 2 biased, and early exposure has been linked to the development of airway allergic diseases. Recently our lab and others have shown that endogenous release of IL-33 due to the stresses of the first few breaths of life leads to expansion and activation of group 2 innate lymphoid cells (ILC2s) from 10 days (D10) to two weeks after birth. However, the amount of IL-33 is lower than in allergen-treated lungs of adult mice, and neonatal lungs do not show signs of type 2 inflammation. Nevertheless, our analyses of IL-33KO and ILC2 deficient pups have suggested that the activation of neonatal lung ILC2s by endogenous IL-33 drives type 2 immunity. As well, neonatal ILC2s labeled with BrdU persist in adulthood. Therefore, we further characterized neonatal lung ILC2s and investigated the effects of neonatal activation on ILC2 functions later in life. Our microarray analysis of neonatal ILC2s showed more similarity to IL-33 activated than naïve adult ILC2s. D10 ILC2s have a high expression of IL-25R, similar to activated adult ILC2s, and can be stimulated by IL-25. We compared IL-33KO and WT ILC2s in adult mice and there was no difference in the phenotype or number of lung ILC2s. However, intranasal (IN) IL-33 injections induced significantly fewer IL-5+IL-13+ ILC2s in IL-33KO mice. We then injected IL-33 into IL-33KO neonates to test the effect on adult ILC2s. Preliminary results show that IN IL-33 into neonatal IL-33KO mice rescue the impaired response of adult IL-33KO ILC2s. These results suggest that the weak activation by the endogenous IL-33 of neonatal lung ILC2s has significant effects on the development of type 2 responses later in life.
Profiling Serum Antibody Specificities in Infants Reveals a Significant Number with Autoreactive Antibodies
Authors: van Oers Nicolai S. , Raj Prithvi , Pichilingue-Reto Patricia , Dozmorov Igor , de la Morena M. Teresa , Wakeland Edward K. 
Published: January 2023

Abstract
The antibody repertoire of an infant develops in response to infections, environmental exposures, and vaccinations. By adulthood, many will produce antibodies that react against self-antigens, often causal to autoimmune diseases such as lupus erythematosus. Limited scientific literature exists as to whether such autoreactive antibodies develop early in infancy. For this reason, the antibody specificities in the serum of healthy infants at 1 and 2 years of age was analyzed. Screening in a cohort of 82 infants revealed that 23 (28%) had moderate to high titered antibodies directed to diverse self-antigens. These numbers are consistent with observed ANA positivity in adults. Ongoing custom targeted DNA sequencing analysis will assess the genetic load of known autoimmune risk alleles in the ANA positive infants. Relationships between the antibody specificities and the genetic risk alleles assembled for each infant will be presented. The comparisons will include clinical information; sex, ethnicity, growth records, vaccination status, infectious history, antibiotic and antiviral treatments, disease status, and family history. Genomic analysis of infants with autoantibodies may facilitate implementation of a new wellness screen to identify antibody positive infants that have significant genetic predisposition and therefore are at risk for developing diverse immune system abnormalities including autoimmune disorders.
Neurokinin 1 receptor-signaling sustains T-cell survival during thymus development and following T-cell activation in secondary lymphoid organs
Authors: Larregina Adriana T. , Sumpter Tina L. , Rojas-Canales Darling M. , Tkacheva Olga A. , Shufesky William J. , Falo Louis D. , Morelli Adrian E. 
Published: January 2023

Abstract
T-cell receptor (TCR)-signaling triggers intracellular Ca2+ increase required for NFAT1/2-mediated IL-2 secretion. This pathway is necessary for thymocyte maturation and survival of activated T cells in secondary lymphoid organs (SLOs). In T cells, cellular Ca2+ levels are regulated by TCR- and G-protein coupled receptor (GPCR)-signaling via the PLCγ and PLCβ subunits, respectively. Nevertheless, the GPCR(s) involved in this phenomenon has not been identified. The neurokinin-1 receptor (NK1R) is a GPCR that induces Ca2+ flux in neurons, and NK1R-signaling by the neuropetides substance P (SP) and hemokinin 1 (HK1) promotes T-cell immunity. We studied the role of NK1R-signaling in T-cell development in the thymus and after T-cell priming in SLOs. By Imagestream, we found that the NK1R and its ligands localize at the site of dendritic cell (DC)-T cell contact. Following CD3-signaling, the NK1R was required for optimal Ca2+flux and NFAT-mediated IL-2 secretion in T cells, effects that were abrogated in NK1RKO or SP/HK1double KO T cells. In the thymus, absence of NK1R resulted in decreased maturation and survival of TCR+ double positive CD4 CD8, single positive CD4, and single positive CD8thymocytes. In SLOs, the NK1R was required for survival of Ag-activated CD4 Th1 and CD8 T cells. In vivo, in a skin model of Th1-DTH induced in NK1RKO T-cell or HK-1/SPdouble KOT-cell bone marrow chimeras, 73±5% of activated CD4 and CD8 T cells died during priming in skin-draining SLOs, and the remaining T cells died in the skin following elicitation. We conclude that the NK1R cooperates with the TCR to increase intracellular Ca2+ necessary for thymocyte maturation and survival of activated T cells in SLOs.
NIH R01 AR068249 and AR071277 to ATL and LDF.
Cell culture oxygen levels affect B cell migration through HIF-1a signaling
Authors: Henn Alicia D. , Garigen Jessica , Cipolla Valentina , Perry Sheldon , Farovitch Lorne , Yerden Randy , Zand Martin Stuart , Hilchey Shannon 
Published: January 2023

AbstractB lymphocytes in vitro encounter oxygen levels far in excess of what they experience in vivo. Room air incubator O2 is not 21%, the accepted level for outdoor O2[1]. High CO2 and humidity drive incubator O2 down (16–19%), with variability from door openings. However, in vivo O2 is far lower even (tissues 0–7%, blood 5–10%). Hypoxia-induced factor 1 alpha (HIF-1a) is a tightly regulated transcription factor responsive to O2 levels [2]. Downstream of HIF-1a are genes tied to the unfolded protein response, metabolism, proliferation, and migration. A therapeutic target in multiple myeloma [3], HIF-1a protein is stabilized at low O2 levels and rapidly degraded at higher O2 levels. We previously showed in primary human B cells and myeloma cell lines, that HIF-1a protein levels stabilize at physioxic 1% and 4% oxygen, but not at 19% oxygen. B cell migration in response to the CXCR4 ligand, CXCL12, was decreased at low O2 levels. Here we seek to add direct evidence to HIF-1a participation in B cell migration changes by using a lentiviral vector to produce stable transfectants expressing a HIF-1a shRNA to genetically ablate HIF-1a. We used C-chamber sub-chambers and Pro-Ox 110 gas controllers to stably control the gas levels inside a standard incubator. At 1% O2, HIF-1a shRNA expression resulted in undetectable HIF-1a protein levels, and a correlating increase in CXCR4-mediated B cell migration as compared to a lentiviral control vector or non-transfected cells. This adds genetic evidence that directly links incubator O2 levels, HIF-1a activity and B cell migration in vitro.
Methyl-CpG binding domain protein-1 polymorphisms impact immune homeostatic levels of IgG
Authors: Hampton Brea K. , Noll Kelsey E. , Whitmore Alan C. , Plante Kenneth S. , Ferris Martin T. , Heise Mark T. 
Published: January 2023

Abstract
Host immunity plays a key role in protection against infection, however, there is significant individual-to-individual variation in the induction and quality of immune response that is driven in part by genetic factors. A growing body of evidence suggests that variation in immune status prior to specific insult may influence subsequent response to vaccination or pathogen challenge. Therefore, understanding the variation in immune status prior to specific insult is necessary for assessing variability in immune response to vaccination or pathogen challenge. The Collaborative Cross (CC) mouse population is a genetically diverse population that was designed to mimic the genetic and phenotypic diversity that is observed in humans and allows us to investigate mechanistically the contribution of genetics to complex phenotypes. Consequently, we asked whether there was variation in antibody levels in 57 CC strains prior to pathogen challenge and if that variation was associated with specific genetic factors. To assess this, we measured antibody levels for IgM, total IgG, IgG1, IgG2a, IgG2b, and IgG2c by ELISA and mapped quantitative trait loci (QTL) associated with variation in antibody concentrations. We observed substantial variation in all antibody isotypes measured. We also identified a suggestive (p=0.1) QTL on chromosome 18. Allele effects show that the C57Bl/6J allele at the locus of interest is linked to higher levels of total IgG. Subsequent analysis of genes under the QTL identified Methyl-CpG binding domain protein-1 (Mbd1) as the most promising candidate gene associated with total IgG variation. Ongoing studies are evaluating the role of Mbd1 in the regulation of total IgG production.
Development and characterization of a bovine monocyte-derived macrophage cell line
Authors: Chitko-McKown Carol G. , Bartens Marie C. , Gibson Amanda J. , Angela Holder , Vats Ashutosh , Brown Emma , Kolakowski Jeannine , Workman Aspen Mikaela , Heaton Michael P. , Kalbfleisch Theodore S. , Werling Dirk 
Published: January 2023

Abstract
Monocytes are components of the innate arm of the immune response and are one of the first lines of defense again invading pathogens. However, they also serve as host cells for intracellular pathogens such as Mycobacteria, Brucella, and Salmonella. Monocytes represent only a small percentage of circulating leukocytes, thus harvesting sufficient quantities of them for in vitro experiments can be costly, time consuming, and can vary among cattle. It was our objective to develop a monocyte-derived macrophage cell line that could be utilized for studies involving macrophages. Monocytes were obtained from the whole blood of a cross-bred steer by density gradient centrifugation, and monocytes purified by adherence. After extended culture, a population emerged spontaneously that proliferated in culture and could be easily detached from the tissue culture vessels using tryspin-EDTA. These proliferating cells were tested for cell surface determinants indicative of monocyte/macrophage lineage, as well as bactericidal and phagocytic activity. Furthermore, whole genome sequencing on both the cell line and whole blood from the donor steer was performed to determine how they align with the bovine whole genome build, and to identify any genetic mutations. This bovine monocyte-derived macrophage cell line has been passaged over 25 times, resembles macrophages in culture, expresses the CD markers CD14/16, C172a, CD11b, is phagocytic and bactericidal, produces IFN in response to TLR/RLR ligands, and appears to fall into the M2 macrophage category. There is a dearth of bovine macrophage cell lines available to veterinary researchers, and this cell line should be a useful tool for the in vitro study of numerous macrophagetrophic pathogens.
Mechanisms Regulating IL-1β Mediated Inflammation during Secondary Bacterial Pneumonia
Authors: Lupfer Christopher R. , Rodriguez Angeline E. , Freeman Thomas , Rippee-Brooks Meagan D. , Abysalamah Hazzar M. , Smith Amber M. , McCullers Jonathan A. , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Viral bacterial coinfections are known to cause severe pneumonia due to enhanced pathogen growth and excessive or altered immune responses. Despite knowing the overall causes for disease severity, current treatments fail to improve disease in many patients. Thus, an improved understanding of the specific immune pathways involved is needed to improve therapeutic treatment design. Interlukine-1β (IL-1β) is an important immune signaling cytokine responsible for inflammation. It exists as an inactive precursor that can be activated by caspase-1 containing inflammasomes. We found that coinfection of Influenza A virus and Streptococcus pneumoniae results in inflammasome activation mainly through the NOD-like receptor protein NLRP3. IL-1β levels are synergistically enhanced during coinfection, but not overall inflammasome activation. Instead, enhanced activation of the transcription factor NF-κB results in increased production of pro-IL-1β, which is subsequently processed into its active form by the inflammasome. In vivo, we discovered that IL-1β regulates lung pathology, and mice treated with a combination of antibiotics and IL-1β neutralizing antibodies have improved weight loss and mortality compared to either treatment individually.
T cell exhaustion during murine cutaneous and visceral leishmaniasis.
Authors: Valadares Diogo Garcia , Davis Richard E. , Kiser Ellen , Chen Yani , Borbon Tiffany , Wilson Mary E. 
Published: January 2023

Abstract
Leishmaniasis is a chronic progressive disease usually controlled by TH1-type IFN-γ producing cells. Human or murine leishmaniasis can progress despite the presence of T cells capable of producing IFN-γ. It is reported that CD8 cells in humans with visceral leishmaniasis (VL) express markers of T cell exhaustion. We hypothesized that myeloid cells play a role in exhausting T cells at the local sites of Leishmania species infection inhibiting proper adaptive responses. We addressed this hypothesis in BALB/c models of cutaneous leishmaniasis (CL) or VL caused by L. major or L. donovani infection, respectively. We examined inhibitory receptors PD1, LAG3, CTLA4 and TIM3 on lymphoid cells, and counter-receptors PDL1, MHCII, and CD80 in myeloid cell by flow cytometry throughout 4 weeks of infection. Both models showed significant increases in PD-1 in circulating dendritic cells after 1 week of infection. However, after 3–4 weeks when infection was chronic, PDL-1 was expressed on neutrophils in the blood and at the local sites of infection (infected ear tissue and draining lymph nodes of L. major infected mice; spleen cells of L. donovani-infected mice). Furthermore, PD1 was expressed on both CD4 and CD8 T cells in blood and lymph nodes of mice infected with L. major at the same time. In contrast, only CD4 T cells, but not CD8 T cells) displayed higher PD-1 in blood and spleens of mice infected with L. donovani. In conclusion, both disease models elicited the exhaustion counter-receptor PDL1 on neutrophils in the blood and the local site of parasite growth. The T cell exhaustion marker PD1 was expressed on T cells at the same sites, although both CD4 and CD8 T cells displayed PD1 in the CL model, whereas PD1 expression was more profound on CD4 T cells in the VL model.
Exogenous induction of type I IFN protects mice from Cryptococcus neoformans and Cryptococcus gattii infections by cellular immunity and iron restriction.
Authors: Davis Michael J. , Moyer Shannon , Sionov Edward , Mayer-Barber Katrin D. , Barber Daniel L. , Cai Hongyi , Walter Peter J. , Jenkins Lisa , Chang Yun C. , Kwon-Chung Kyung J. 
Published: January 2023

Abstract

Rational
Cryptococcus neoformans (Cn) causes deadly mycosis in AIDS patients whereas Cryptococcus gattii (Cg) infects mostly non-HIV patients. As HIV infection induces type I IFN (t1I) we hypothesized that exogenously induced t1I would differentially affect infections by Cn and Cg.

Results
Cn or Cg infected mice were dosed with poly(I:C)LC (PIC) to induce t1I. PIC treated mice were protected from Cn and Cg infection mortality with reduced fungal burdens. In Cn infected mice, PIC protection was mediated by classical Th1 immunity as CD4 T cells and IFNγ were required and Cn mediated Th2 polarization and eosinophilia were reversed by PIC. In contrast, PIC protection from Cg did not require any of the immune factors previously associated with Cn immunity; T cells, B cells, NK cells, IFNγ and monocyte derived myeloid cells were all dispensable for protection. Interestingly, PIC protection depended on β-2-microglobulin (B2m). In addition to its role as a co-factor for several immunological proteins, B2m is involved in iron trafficking suggesting that PIC protection may function by limiting Cg’s access to iron. In support of this, iron supplementation reversed PIC protection and iron chelation mediated similar protection as PIC. PIC also induced expression of several proteins involved in iron trafficking.

Conclusions
These data show that t1I induction protects mice from Cn and Cg but protection was mediated by distinct immune mechanisms. PIC mediated protection from Cn requires CD4 T cells and IFNγ. However, this classical immune pathway to cryptococcosis is not involved in PIC-mediated protection from Cg. Instead, PIC protection from Cg seems to involve iron limitation suggesting a novel role for t1I in regulating host iron in the lungs.
A2B Adenosine Receptor Signaling on Epithelial Cells Promotes Type 2 Immunity Against Helminths
Authors: El-Naccache Darine W. , Chen Fei , Palma Mark , Wu Wenhui , Csoka Balazs , Hasko George , Gause William C. 
Published: January 2023

Abstract
Intestinal helminth infections trigger potent type 2 immune responses, although the mechanisms that initiate these host protective responses against these multicellular parasites remain uncertain. We have previously reported that inoculation with the murine intestinal nematode parasite Heligmosomoides polygyrus (Hp) results in rapid increases in intestinal epithelial surface expression of the ectonucleotidases, CD39 and CD73, which in turn degrade extracellular ATP into adenosine. Adenosine binds to adenosine G-protein-coupled cell surface receptors (A1, A2A, A2B and A3 –AR). Mice deficient in A2BAR have an impaired type 2 immune response and delayed worm expulsion during helminth infection. We now report that mice lacking A2BAR specifically on intestinal epithelial cells (Villin-Cre-A2BARfl/fl), but not on myeloid cells (LysMCre-A2BARfl/fl), have an impaired type 2 memory immune response with delayed worm expulsion and decreased egg production after Hp secondary inoculation. pSTAT6 levels in CD4+ T cells and B cell MHC-II surface expression (an indicator of IL-4 activity) from mesenteric lymph nodes of Villin-Cre-A2BARfl/fl mice were significantly decreased after primary Hp inoculation. Taken together, our data demonstrate a key role for epithelial cell specific A2BAR signaling in the initiation of the protective type 2 immune response to helminth infection. These studies indicate that extracellular adenosine binding to epithelial cell A2BAR acts as an endogenous danger signal that is released following helminth infection, which triggers host protective type 2 immune responses.