bluuebunny/crossref_metadata_2025_split huggingface dataset:
A developmental maturation of a dendritic cell subset controls the Th2 bias of neonatal immunity through IL-12 production and downregulation of IL-13Rα1 (95.2)
Authors: Lee Hyun-Hee , Hoeman Christine , Hardaway J.C. , Jain Renu , Divekar Rohit , Ellis Jason E. , Nast Danielle , Haymaker Cara L. , Guloglu F.Betul , Zaghouani Habib 
Published: January 2023

Abstract
Neonates are susceptible to microbial infections and allergic reactions due to lack of Th1 immunity and skewing of Th2 response. Analysis of primary response in neonates showed that Th1 and Th2 cells develop upon priming with antigen on the day of birth. However, Th1 cells display up-regulated expression of IL-13Rα1 that was found to associate with IL-4Rα and provide a receptor that can be used by both IL-4 and IL-13. Upon rechallenge with antigen, IL-4 produced by Th2 cells utilizes IL-13Rα1/ IL-4Rα receptor to drive apoptosis of Th1 cells hence bias of neonatal immunity towards Th2. Here, we demonstrate that up-regulation of IL-13Rα1 is due to inability of neonatal antigen presenting cells to produce IL-12. In fact, supply of exogenous IL-12 during priming with antigen minimizes IL-13Rα1 expression, leading to inhibition of apoptosis and restoration of Th1 secondary immunity. Interestingly, lack of IL-12 is due to delayed maturation of CD8α+CD4− subset of dendritic cells. However, by day 6 after birth CD8α+CD4− DCs accumulate and produce significant amounts of IL-12 that diminishes IL-13Rα1 expression, leading to inhibition of Th1 apoptosis and restoration of an adult-like Th1 immunity. Thus, developmental maturation of DC subsets is responsible for bias of neonatal immunity towards Th2 providing susceptibility to microbial infections and allergic reactions.
Research supported by grant R01 AI 48541 from NIH.
Cloning and functional analysis of an IL-13 splice variant lacking exon 2 which has inflammatory activity (95.22)
Authors: Yu Jiayi , Tse Harley , Lu Hong , Li Jinzhu , Shaw Michael 
Published: January 2023

Abstract
Interleukin 13 is a Th2 type (anti-inflammatory) cytokine produced by activated T cells. We have cloned a naturally occurring mRNA splice variant of IL-13 in which the second exon is omitted (termed δE2). We have developed RT-PCR assays for quantitation and discrimination of δE2/IL-13 gene expression, and cloned each of the molecules as HIS-tagged proteins for use in in vivo and in vitro studies.
We find that unlike IL-13, δE2 is expressed predominantly under Th1 conditions: δE2 is expressed by spleen cells stimulated with polyclonal activators and in the lymph nodes of mice immunized with CFA as adjuvant. Addition of exogenous δE2 to cultured antigen presenting cells induces phenotypic changes characteristic of an inflammatory (Th1) response, such as increased inflammatory cytokine secretion (TNF-α and IL-12). Addition of exogenous δE2 to antigen specific T cell cultures results in an increase in TNF-α producing cells as determined by ELIspot. Treatment of mice with exogenous δE2 at the time of immunization with CFA and antigen results in significant increases in the expression of inflammatory genes, including IL-12, CD28, and CD5. Lastly, we find that this splice variant is produced primarily by activated (CD19+, CD86+) B cells
Given the expression pattern of δE2 and its ability to modulate the expression of inflammatory cytokines, we hypothesis that δE2 is a unique cytokine involved in inflammatory immune responses
A critical role for IL-6 secretion by dendritic cells promoting Th2 and limiting Th1 response (95.24)
Authors: Krishnamoorthy Nandini , oriss Timothy , Paglia Melissa , Ray Anuradha , Ray Prabir 
Published: January 2023

Abstract
The mechanisms by which dendritic cells (DCs) induce differentiation of naïve CD4+ T cells along the Th2 lineage is not well understood given that DCs themselves do not produce IL-4. In the present study we undertook a microarray approach to identify genes involved in the induction of Th2 differentiation by DCs treated with a Th2-skewing adjuvant cholera toxin (CT). In the microarray analysis, murine bone marrow derived immature DCs were treated with CT. The analysis showed that CT upregulated IL-6 but not IL-12p70 both at the RNA and protein levels. DCs stimulated with CT and antigen preferentially upregulated GATA-3 expression in CD4+ T cells. This was associated with production of high levels of the Th2 cytokines IL-4 and IL-13. This suggested that low IL-12 and high IL-6 cytokine profile in the CT-treated DC cultures had a Th2-stimulatory capacity. To further investigate the role of IL-6 in Th2 response in vitro, IL-6 was neutralized with anti-IL-6 in DC –T cell co-cultures. This resulted in a decrease in Th2 and an increase in Th1 response as evident from the cytokine profile of IL-13 and IFN-&#61543;. We also tested the ability of house dust mite (HDM) extract, an allergen known to induce asthma in humans, to induce IL-6 secretion from DCs. BMDC-stimulated with HDM also secreted high levels of IL-6, but low levels of IL-12p70. IL-6-deficient mice when administered HDM intranasally showed diminished Th2 response as evidenced by decreased IL-13 production. Our ongoing studies are focused on elucidating the role of specific differentially regulated genes in CT-induced IL-6 gene expression. Taken together, these studies will help identify pathways in DCs that participate in Th2 induction via IL-6 upregulation.
(Supported by NIH grants RO1 HL60207 to P.R. and RO1 HL77430 to A.R.)
Constitutive expression of IL-27 in neuro-retina negatively regulates THIL-17 cell development (95.25)
Authors: Yu Cheng-Rong , Amadi-Obi Ahjoko , Mahdi Rashid M. , Liu Xuebin , Lee Yunsang , Gery Igal , Egwuagu Charles 
Published: January 2023

Abstract
Ocular immune privilege is maintained by constitutive expression of inhibitory molecules such as TGF-b, FAS and FAS-ligand. Here, we describe a model of immune privilege for the eye based on IL-27-induced suppression of Th17 responses. We show that Th17 cells are elevated in retina of mice during experimental autoimmune uveoretinitis (EAU) and treatment with anti-IL17 Abs reduces EAU severity. It is of note that the increase and subsequent trafficking of Th17 cells into the retina during EAU is accompanied by corresponding increases in IL-27 and IFN? in the retina. We further show that IL-27 is constitutively expressed in human or mouse retina and that IFN? upregulates its expression in retinal cells. Of particular interest is inhibition of the proliferation of uveitogenic Th17 cells by IL-27, suggesting that IFN? may mitigate uveitis by antagonizing Th17 expansion through induction of IL-27 in the target tissue. In a co-culture system the expansion of Th17 cells (propagated under Th17 polarizing conditions) is inhibited by primary retina cells. However, addition of anti-IL-27 antibody into the culture overrides the inhibitory effects of the retinal cells on the expansion of Th17 cells. The data presented suggests that inhibitory effects on inflammatory cells by the retina are mediated in part by endogenously produced IL-27 and establishes mechanistic links between IL-27 expression and anti- Th17 effects in the eye.
Therapeutic efficacy of immunomodulatory polysaccharides may be explained by synergism with interferon-γ (95.29)
Authors: Sun-A Im , Young-Ran Lee , Young-Hee Lee , Kyungjae Kim , Chong-Kil Lee 
Published: January 2023

Abstract
Immunomodulatory polysaccharides have been implicated for the therapeutic efficacy of many plants or mushrooms. The absorption efficacy of immunomodulatory polysaccharides has been a matter of concern due to the high molecular weight nature. The present study shows that low in vivo concentration of polysaccharides may be enough to exert strong immunomodulatory activity in a microenvironment where a small amount of IFN-γ is present. Immunomodulatory polysaccharides, termed SHP, were isolated from Salicornia herbacea and examined their macrophage-activating activity alone and in combination with interferon (IFN)-γ. The combination of SHP and IFN-γ synergistically inhibited the growth of the mouse monocytic cell line, RAW 264.7, inducing further differentiation to strongly adherent macrophages. The differentiation-inducing activity of SHP alone and in combination with IFN-γ was confirmed by changes in the expression of differentiation antigens such as CD11b, CD18 and CD24. In addition, the combination of SHP and IFN-γ synergistically activated RAW cells to produce cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-1β, and nitric oxide (NO). The synergistic activity of SHP was more prominent when SHP concentration was low. Increased production of TNF-α, IL-1β and NO was correlated with an increased level of their respective transcripts. These results confirm that immunomodulatory polysaccharides exert synergistic monocyte-activating activity with small doses of IFN-γ.
IL-1β, not IL-18, is the primary IFNγ inducing factor released by activated monocytes (95.30)
Authors: Raices Raquel Marie , Sarkar Anasuya , Wewers Mark 
Published: January 2023

Abstract
IFNγ and caspase-1 have a reciprocal relationship. INFγ induces caspase-1 expression but caspase-1 is critical to the activation of the INFγ inducing factor IL-18. To investigate the INFγ inducing function of caspase-1 we characterized the INFγ inducing activity produced in response to caspase-1 activation (LPS + ATP) of human monocytes. Preliminary experiments demonstrate that IL-18 is not alone as an INFγ inducing product of caspase-1. Purified monocytes were stimulated with LPS/ATP and supernatants analyzed for IL-1β and IL-18 release, as well as for their ability to induce IFN-γ release by KG-1 cells. Monocytes released IL-1β and IL-18, as well as an IFN-γ inducing factor. The IFN-γ inducing activity was enhanced with ATP added to LPS and suppressed by a caspase-1 inhibitor, consistent with a caspase-1 regulated factor. However, the IFN-γ inducing activity was neutralized by IL-1β, not IL-18, blocking agents. In contrast, KG-1 cells released IFN-γ in response to rIL-18, not rIL-1β. When supernatants were fractionated by gel filtration, IL-18 eluted at a high molecular weight peak, indicating that it may be complexed to an inhibitory protein. IL-1β eluted at a single molecular weight peak corresponding to its mature form. However, the IL-1β and IL-18 peaks had minimal IFN-γ inducing activity. Taken together, these findings suggest that IL-1β is required, but not sufficient for the induction of IFNγ production.
Support: NIH R01 HL 746767 and HL 40871, F31 AI 061828-02
Epstein-Barr virus-induced gene 3 is a negative regulator of IL-17 and protective immunity against Listeria monocytogenes (95.4)
Authors: Yang Jianfei , Yang Min , Htut Tin Min , Hanidu Adedayo , Li Xiang , Sellati Sosemarie , Jiang Huiping , Zhang Shu , Li Hongxing , Ting Adrian T , Mayer Lloyd , Unkeless Jay C , Labadia Mark E , Hodge Martin , Li Jun , Xiong Huabao 
Published: January 2023

Abstract
The product of the Epstein-Barr virus-induced gene 3 (EBI3) associates with p28, an IL-12p35-related protein, to form IL-27. EBI3 also associates with IL-12p35 to form a distinct heterodimer. The function of EBI3 in innate as well as adaptive immunity to intracellular bacteria has not been established. Here we demonstrate that EBI3 deficient mice exhibit a reduced bacterial load following an acute challenge with Listeria monocytogenes or a re-challenge of animals previously immunized with L. monocytogenes, suggesting EBI3 plays a negative role in both innate and adaptive immunity. The enhanced protective immunity in EBI3-deficient mice is not due to modulation of the Th1 immune response since the production of IFNγ and TNFα was not significantly altered. Importantly, spleen cells from EBI3-deficient mice immunized with L. monocytogenes produced significantly higher levels of IL-17 than wild type cells. Furthermore splenocytes as well as Th17 cells from EBI3-deficient mice secreted increased levels of IL-17. Elevated numbers of IL-17-producing cells were observed when EBI3 deficient CD4+ T cells were cultured under conditions to induce Th17 cell differentiation. Taken together, these data provide direct evidence that EBI3 is an important negative regulator of protective immunity against L. monocytogenes as well as IL-17 expression.
Exposure to LPS induces the generation of immunosuppressive myeloid cells (95.7)
Authors: Arora Meenakshi , Ray Anuradha , Ray Prabir 
Published: January 2023

Abstract
The biologic effects of lipopolysaccharide (LPS) may have different consequences on the expression of asthma dependent upon whether the exposure to LPS occurs during the establishment of the atopic status, or whether the exposure to LPS occurs after the asthma phenotype is established. It has been shown that exposure to LPS during early childhood is associated with a lower incidence of asthma later in the life. Dendritic cells (DCs) are the primary antigen presenting cells of the immune system and play critical role in the induction of peripheral tolerance and regulation of T-cell responses. The diverse functions of DCs in immunity and tolerance depend on their lineage and maturation stage. We have observed that exposure of LPS can generate a population of immature myeloid cells that have potent immunosuppressive activity on T cells. These CD11c− cells are largely MHC IIlow/MHC II- and have poor antigen presenting capacity. These cells poorly stimulate proliferation of naïve allogenic T cells and are unable to stimulate T cell differentiation along the Th1 or Th2 lineage. However, these cells are capable of suppressing the proliferation and differentiation of activated CD4+ T cells both in vitro and in vivo. Our data shows that LPS reduces DC driven T cell development through the induction of cells with suppressive activity, which could be a potential strategy to suppress harmful T-cell responses in patients with allergic diseases.
(RO1 HL60207 to P.R. and RO1 HL77430 to A.R.)
Preferential Chemotaxis and Fugetaxis of CD4−CD8− Thymocytes in Response to CXCL16 (95.8)
Authors: Manilay Jennifer O. , McLelland Bryce 
Published: January 2023

Abstract
Directed migration of developing thymocytes to specific anatomic regions of the thymus can influence normal T cell development. However, the mechanisms which influence migration of early CD4−CD8− (double negative: DN) thymocytes are incompletely understood. We investigated the role of chemokine CXCL16 and its receptor, CXCR6, in thymocyte migration. Gene expression analysis showed that CXCR6 is expressed only in DN thymocytes, but not at later stages of development, whereas CXCL16 is expressed on stromal cells throughout the thymus. The ability of DN thymocytes to migrate in response to CXCL16 was tested with in vitro transwell assays. At concentrations of 100 and 10ng/ml, DN thymocytes demonstrated specific migration toward CXCL16, whereas migration of CD4+CD8+ thymocytes was not significantly different from controls. We also tested the fugetactic ability of DN thymocytes from CXCL16, and observed that DN thymocytes selectively migrated away at concentrations of 10 and 1ng/ml. These data demonstrate that DN thymocytes migrate differentially to CXCL16 in a dose-dependent fashion. Thus, CXCR6 and CXCL16 may be involved in localized cell migration in the thymus. ADAM10 and Notch influence cell migration in other systems; thus experiments to test their roles in the control of CXCR6-mediated migration of DN subsets in the thymus are ongoing.
Supported by UCM Faculty Startup Funds
Characterization of chicken TNFSF15 and it’s role in coccidiosis (96.10)
Authors: Lillehoj Hyun Soon , Park Soon Soon 
Published: January 2023

Abstract
A full-length cDNA encoding chicken tumor necrosis factor superfamily 15 (TNFSF15) was isolated and its functional role was investigated. TNFSF15 transcripts were primarily expressed in spleen, liver, intestinal intraepithelial lymphocytes (IEL), peripheral blood lymphocytes and bursa. In vitro infection of HTC macrophages with three species of Eimeria sporozoites induced TNFSF15 gene expression. In vivo experiments revealed that TNFSF15 gene was highly increased following primary infections with E. acervulina or E. maxima. In contrast, no consistent changes in transcript levels were seen following primary infection with E. tenella, or following secondary infection with any of the three Eimeria species. Following infection with E. acervulina and E. maxima, TNFSF15 transcripts were primarily expressed in intestinal CD4+ and TCR2+ IEL, respectively. A dose-dependent cytotoxic effect of recombinant TNFSF15 protein was observed on HTC and LSCC-RP9 tumor cells. These results indicate that TNFSF15 plays an important role in the local inflammatory response to Eimeria.
Massive colonic cytokine dysregulation in IL-10−/− mice revealed by multiplex analysis (96.17)
Authors: Klein John R. , Hicks John , Montufar-Solis Dina 
Published: January 2023

Abstract
IL-10−/− mice develop enterocolitis resulting from a failure to control the expression of proinflammatory and other immune response analytes. To better understand changes that occur in regulatory and effector cytokines in IL-10−/− mice during the progression of intestinal inflammation, multiplex analysis of 23 analytes was evaluated in colonic intraepithelial cell culture supernatants from 25 IL-10−/− and 6 control mice ranging from 7–52 weeks or age. In IL-10−/− mice, widespread cytokine dysregulation was evident across the period of the study for 18 analytes (IL-1α, IL-1β, IL-6, IL-9, IL-12p40, IL-12p70, IL-13, eotaxin, G-CSF, GM-CSF, IFN-γ, KC/CCXCL1, MCP-1, MIP-1α, MIP-1β, CCL5/RANTES, TNFα, and IL-17), whereas no activity was detected for IL-2, IL-3, IL-4, IL-5, or IL-10. Of analytes with elevated activities, 15 had a low-to-medium correlation with the degree of colonic pathology (r = 0.2649 to r = 0.6494); IL-9 and IL-13 had very high correlations with pathology (r = 0.8026 and r = 0.8137, respectively); and IL-17 had an extremely strong correlation with pathology (r = 0.9885). These findings demonstrate that the lack of IL-10 results in a complex, massive, and early process of intestinal cytokine dysregulation with IL-17 strongly linked to disease overall.
TNF-α promotes CD4 T cell infiltration of the central nervous system (CNS) by altering the polarity of CXCL12 expression at the blood-brain barrier (BBB) (96.18)
Authors: McCandless Erin Elizabeth , Piccio Laura , Cross Anne , Russell John , Klein Robyn 
Published: January 2023

Abstract
CXCL12 is a secondary lymphoid chemokine that localizes lymphocytes within lymph node zones and promotes CD4 T cell help. We have previously shown that expression of CXCL12 at the BBB displays basolateral polarity and localizes leukocytes to the perivascular space, similar to its function within lymphoid tissue. Induction of experimental autoimmune encephalomyelitis (EAE) leads to redistribution of CXCL12 across the BBB and exit of myelin-specific T cells from the perivascular space. In the current study we have extended our findings to human CNS specimens and examined the molecular triggers for CXCL12 redistribution at the BBB. We observed that a similar loss in CXCL12 polarity occurs within active Multiple Sclerosis (MS) lesions, suggesting that altered BBB expression of CXCL12 plays a role in the pathogenesis of MS. Using TNFR1−/− mice, we demonstrate that while loss of TNFR1 signaling does not alter peripheral immune responses, CXCL12 expression in TNFR1−/− mice remains polarized at the BBB and is associated with the accumulation of leukocytes within the perivascular space. Comparisons of myelin-specific CD4 T cells recovered from vessel-associated versus parenchymal sites revealed that only T cells that have successfully infiltrated the CNS parenchyma express both CXCR4 and TNF-α. These results suggest that TNFR1 signaling promotes leukocyte entry via redistribution of CXCL12 at the BBB.
Potential role of CCR9 in gastrointestinal involvement of ATL cells (96.21)
Authors: Nagakubo Daisuke , Jin Zhe , Hieshima Kunio , Nakayama Takashi , Shirakawa Aiko-Konno , Yoshie Osamu 
Published: January 2023

Abstract
Adult T-cell leukemia (ATL) is a mature CD4+ T-cell malignancy etiologically associated with human T-lymphotropic virus type 1 (HTLV-1). Previously, we have shown that ATL cells from the majority of cases are strongly positive for CCR4, which may explain the frequent skin invasion of ATL. Although ATL is also known to frequently invade the gastrointestinal tract, the chemokine receptors possibly involved in the process have not been determined yet. In this study, we examined whether ATL cells express CCR9, which is known to be involved in T-cell homing to the gastrointestinal tract. While ATL cells freshly isolated from peripheral blood hardly expressed CCR9, ATL cells cultured for 1 day consistently upregulated CCR9 in parallel with the upregulation of Tax, the HTLV-1-encoded transcriptional activator. A luciferase reporter gene under the control of the CCR9 promoter was expressed by co-transfection of an expression vector for Tax. Immunohistochemical staining demonstrated that ATL cells infiltrating gastrointestinal tract were frequently positive for CCR9. Thus, CCR9 is inducible by Tax and may play a role in the gastrointestinal involvement of ATL.
This work was supported by grants from Ministry of Education, Culture, Sports, Science and Technology, Japan and Japan Science and Technology Corporation.
Response to SDF-1 defines a subset of CD38+/ZAP-70+ CLL patients with a common genetic signature and aggressive behavior (96.23)
Authors: Deaglio Silvia , Aydin S , Vaisitti T , Bergui L , Omede’ P , Bonello L , D’Arena G , Chiorino G , Efremov D , Malavasi F 
Published: January 2023

Abstract
Chronic lymphocytic leukemia (CLL), traditionally considered a disease of failed apoptosis, is now seen as characterized by a different propensity to grow according to microenvironmental signals. CD38 (surface receptor) and ZAP-70 (cytoplasmic molecule) independently predict unfavorable outcome in CLL patients. We reported that CD38 is a receptor mediating interactions with nurse-like cells (NLC) and transducing proliferation signals. The production by NLC of the attractant SDF-1α and the presence of ZAP-70 in CXCR4 pathway suggested that CD38 and ZAP-70 control CLL cell migration. We show that CD38+/ZAP-70+ patients feature enhanced ability to respond to SDF-1α. The functional link between the two molecules was confirmed by showing that i) CD38 engagement leads to ZAP-70 phosphorylation in CLL cells and in ad hoc modified B line expressing a ZAP-70-myc construct. Further, ii) CD38 signals are effective only in presence of ZAP-70, as seen in 22 molecularly characterized patients.
Lastly, a separation of the patient cohort based on migration, brought to the identification of 23 differentially expressed genes: 12 are involved in cell motility, migration and neoplastic transformation.
This work provides biological evidence explaining why the combination of CD38 and ZAP-70 in routine clinics results in a more efficient identification of CLL patients with aggressive disease.
Sponsors: CLL-GRF, AIRC, FIRMS.