bluuebunny/crossref_metadata_2025_split huggingface dataset:
Programming for a CD8 T Cell Memory Response Requires IL-12 and/or Type I IFN (95.23)
Authors: Xiao Zhengguo , Jameson Stephen C , Mescher Matthew F 
Published: January 2023

Abstract
Naïve CD8 T cells proliferate in response to Ag and costimulation, but require signals from IL-12 or IFNα/β to make a productive response. Whether these are the only cytokines with this activity, and their relative contributions, are only beginning to be defined. Response to LCMV infection by CD8 cells lacking the IFNα/β receptor (IFNIR−/−) is reduced &gt; 99%, but response to vaccinia virus (VV) remains largely intact (J.Exp.Med. 202, 637; J. Immunol. 176, 4225). We have studied responses of OT-I T cells to OVA-expressing VV and Listeria monocytogenes (LM), using cells that lack IFNIR, IL-12 receptor (IL-12R−/−), or both (DblKO). Clonal expansion of receptor-deficient OT-I cells to LM-OVA or VV-OVA is comparable to that of wild type (wt) cells, but effector functions are compromised, and their numbers decline more rapidly. IL-12R−/− cells exhibit greater defects than do IFN-IR−/− cells, and DblKO cells are the most compromised. Compared to the wt memory population following VV-OVA infection (&gt; day 30), memory cell numbers are reduced by 27% for IFN-IR−/− cells, 96% for IL-12R−/− cells, and &gt;99% for DblKO cells. For LM-OVA, the reduction in memory is 76% for IFN-IR−/− cells, 97% for IL-12R−/− cells, and &gt;99% for DblKO cells. Thus, both IL-12 and IFNα/β contribute to responses to VV and LM, with IL-12 making the greater contribution, and programming for CD8 T cell memory requires signals from one or the other of these cytokines.
Lymphoid organ specificity in the maintenance of secondary lymphoid tissues by TNF produced by B and T cells (95.26)
Authors: Tumanov Alexei , Grivennikov Sergei I , Kruglov Andrei A , Kuprash Dmitry V , Nedospasov Sergei A 
Published: January 2023

Abstract
Secondary lymphoid organs play an important role in generation of efficient immune responses. Using a genetic approach we revealed distinct contribution of TNF produced by B and T cells in homeostasis of secondary lymphoid organs. In spleen, the development of follicular dendritic cells (FDC) and B cell follicles was controlled primarily by TNF produced by B cells while TNF from T cells provided a distinct complementary signal, similarly to contributions reported earlier for the surface lymphotoxin (LT). Organized marginal zone structure was dependent on TNF from both B-, T- and other cell types. In contrast, in Peyer’s patches, the lymphoid microarchitecture was exclusively dependent on TNF produced by B cells. Furthermore, we show that TNF signals from both B and T cells acted in concert for the expression of TNF dependent genes in lymph nodes, generation of FDC and B cell follicles, and in mounting an efficient humoral immune response to thymus-dependent antigen. Finally, we found that soluble form of TNF provided by B cells was critical for the maintenance of secondary lymphoid organs. Thus, the organization of different secondary lymphoid organs is maintained by distinct contribution of TNF produced by B and T cells, on one hand, and distinct role of soluble versus membrane TNF form, on the other hand.
Immunorestorative effect of intranasal granulocyte/macrophage colony stimulating factor (GM-CSF) on alcoholic lung macrophages is mediated through local release of zinc (96.30)
Authors: Joshi Pratibha C , Guidot David M 
Published: January 2023

Abstract
Our recent studies have shown that chronic alcohol ingestion dampens alveolar macrophage immune function by decreasing GM-CSF receptor expression, and that these chronic defects can be reversed with rGM-CSF treatment in vivo (JI 2005). However, the mechanisms by which alcohol and GM-CSF modulate macrophage GM-CSF receptors are not clear. Zinc is an essential micronutrient that is implicated in many immunostimulatory functions. Since alcoholics are known to be deficient in zinc, we hypothesized that zinc may be responsible for the functional integrity of alveolar macrophages. To explore the impact of zinc deficiency on normal alveolar macrophage immune function, we treated these cells in vitro with or without a zinc chelator and then added zinc acetate before evaluating phagocytosis by flow cytometry. In parallel, we added 0.2% zinc acetate to the liquid diet of alcohol-fed rats. Finally, we measured zinc levels in the bronchoalveolar lavage from these animals using a zinc specific dye FluoZin-3. Zinc chelator treated alveolar macrophages showed a decrease in phagocytosis that was restored by zinc suggesting an impact of zinc deficiency on macrophage function. In in vivo model, zinc upregulated the membrane GM-CSF receptors on alveolar macrophages similar to that reported earlier with the in vivo GM-CSF treatment. The decreased levels of zinc in the lavage of alcohol-fed rats were significantly increased by in vivo zinc and intranasal GM-CSF. These data suggest a novel mechanism of zinc release in the lungs through which GM-CSF restores the immune function of the hyporesponsive alveolar macrophages from alcohol-fed animals.
Supported by P50 AA013757 and 2R01 AA011660-05A1
Gp100 is Chemotactic for Mononuclear Cells and Utilizes Chemokine Receptors (96.4)
Authors: Howard OM Zack , Buzas Krisztina , Dong Huifang , Oppenheim Joost J 
Published: January 2023

Abstract
Based on the observations that some patients vaccinated with melanoma antigens develop autoimmune vitiligo, and other cancer patients develop autoantibodies to tumor antigens, we tested two, melanoma tumor antigens gp100 and, NY-ESO-1 to determine if tumor antigens, like autoimmune antigens, are chemoattractants. We consistently observed a chemotactic response by human iDC, monocytes and T cells to nM concentrations of gp100, but not at all to NY-ESO-1. Using desensitization studies and receptor transfected HEK 293 cells, we showed that human gp100 interacts with human CCR2, the receptor for CCL2 (MCP1), and thereby accounted for its capacity to attract CCR2 expressing monocytes, T cells and iDC, but not mDC. Pathophysiological doses of CCL2 did not activate iDC, however, gp100 at concentrations of 2–10 mg/ml induced low levels of CD40, CD80, MHC Class II and functional CCR7. High levels of IL6, IL10, MDC, TARC and functionally active TGF-b were induced by gp100 stimulation. Based on CCR2 expression, we predict that gp100 can recruit innate, adaptive and regulatory immune cells to sites of tumor induced tissue damage. Consequently, the net effect of the tumor antigen stimulus can potentially result in inflammation, tumor immunity or tolerance depending on how it is presented to these cell types.
*Funded in part by DHHS #NO1-CO-12400
Ligand binding of leukocyte integrin Very Late Antigen-4 induces exposure of sulfhydryl groups and is subject to redox modulation by reactive oxygen species (97.10)
Authors: Liu Si-Yen , Tsai Ming-Yi , Chuang Kuo-Pin , Shieh Chi-Chang 
Published: January 2023

Abstract
Activation of leukocyte integrins is important for selective recruitment of cells into inflammatory tissues. Our previous studies showed that the binding between the integrin Very Late Antigen-4 (VLA-4) and Vascular Cell Adhesion Molecule-1 (VCAM-1) is modulated by reactive oxygen species. In this study we investigated the molecular nature of redox modulation on the activation states of VLA-4. We found that ligand binding of VLA-4 induced exposure of sulfhydryl groups on the α4 peptide. Moreover, low concentration (10μM), but not higher concentrations (≥100μM), of hydrogen peroxide enhanced the ligand binding ability of VLA-4 to VCAM-1. This activating effect may be dependent on S-glutathionylation of α4 subunit. Increased concentrations (up to15μM) glutathione in the medium enhanced the H2O2 induced activation of VLA-4. The redox regulation of the VLA-4 involves outside-in signaling and cytoskeleton rearrangement. In conclusion, our data indicate that ligand binding of VLA-4 involves redox modulation. Redox-sensitive site(s) thus may play a role in regulating the activation of VLA-4 in inflammatory tissues.
The adhesion molecules CD44 and hyaluronan, not physical trapping, are responsible for neutrophil sequestration in inflamed liver sinusoids (97.5)
Authors: McAvoy Erin , McDonald Braedon , Kubes Paul 
Published: January 2023

Abstract
Adhesion molecules important for neutrophil recruitment in many organs during endotoxemia are not involved in recruitment into the sinusoids of the liver. Using a whole body dual radiolabel adhesion molecules screen, we observed that hyaluronan (HA), a CD44 ligand, is disproportionately expressed in the liver (versus other organs) under both basal and endotoxemic conditions. Using intravital microscopy, we observed that blocking CD44-HA interactions reduced neutrophil adhesion in sinusoids but had no effect on rolling or adhesion in post-sinusoidal venules (mediated by α4 and β2 integrins). Inhibition of CD44-HA interactions, but not integrins, reduced the entry of neutrophils into the liver parenchyma and significantly improved LPS-induced hepatic injury. Anti-CD44 antibody rapidly detached adhering neutrophils in sinusoids and improved sinusoidal perfusion in endotoxemic mice, revealing CD44 as a potential therapeutic target in LPS-associated liver disease. In conclusion, the identification of CD44-HA as partners for neutrophil adhesion in sinusoids, disproves the hypothesis that physical trapping is entirely responsible for sequestration of neutrophils in sinusoids.
This work was funded by Alberta Heritage Foundation for Medical Research (AHFMR), and the Canadian Institutes of Health Research (CIHR).
Non-hematopoietic NADPH Oxidase Regulation of Lung Eosinophilia and Airway Hyperresponsiveness in Experimentally-Induced Asthma (98.3)
Authors: Abdala-Valencia Hiam , Earwood Julie , Babcock George , Garvy Beth , Wills-Karp Marsha , Cook-Mills Joan 
Published: January 2023

Abstract
Infiltration of eosinophils into the lung in experimental asthma is dependent on the adhesion molecule vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells. Ligation of VCAM-1 activates endothelial cell NADPH oxidase which is required for VCAM-1-dependent leukocyte migration in vitro. To examine whether endothelial-derived NADPH oxidase modulates eosinophil recruitment in vivo, mice deficient in NADPH oxidase (CYBB mice) were irradiated and received wild type hematopoietic cells to generate chimeric CYBB mice. In response to OVA challenge, the chimeric CYBB mice had increased numbers of eosinophils bound to the endothelium as well as reduced eosinophilia in the lung tissue and bronchoalveolar lavage. This occurred independent of changes in VCAM-1 expression, cytokine/chemokine levels (IL-5, IL-10, IL-13, IFNã, or eotaxin), or numbers of T cells, neutrophils or mononuclear cells in the lavage fluids or lung tissue of OVA-challenged mice. Importantly, the OVA-challenged chimeric CYBB mice had reduced airway hyperresponsiveness (AHR). The AHR in OVA-challenged chimeric CYBB mice was restored by bypassing the endothelium with intratracheal administration of eosinophils. These data suggest that VCAM-1 induction of NADPH oxidase in the endothelium is necessary for the eosinophil recruitment during allergic inflammation.
(supported by NIH HL069428, J.M.C-M.)
T-Cell Induced Colitis Is Associated With Enhanced Myelopoiesis, PMN Infiltration into Secondary Lymphoid Tissue and Modulation of T-Cell Activation (98.6)
Authors: Ostanin Dmitry V. , D’Souza Dwain , Hoffman Jason , Gray Laura , Pruett Stephen , Grisham Matthew B. 
Published: January 2023

Abstract
The objective of this study was to determine how the adoptive transfer of naïve T-cells into lymphopenic recipients affects myelopoiesis during the development of chronic colitis. We found that adoptive transfer of CD4+CD45RBhigh T-cells (5x105 cells) into RAG-1−/− recipients induced chronic colitis that was preceded by a dramatic leukocytosis/myelopoiesis in which total circulating leukocytes increased 5–6 fold compared to unmanipulated RAG-1−/− mice with PMNs representing &gt;85% of the total blood leukocytes. In addition, we found that serum levels of the granulopoietic cytokines IL-17 and G-CSF increased by 4- and 9-fold, respectively compared to the healthy CD45RBlow→RAG-1−/− controls. Corresponding to these dramatic increases in systemic PMN production in the CD45RBhigh→ RAG-1−/− mice, were large and significant increases (30–50-fold) in the presence of granulocytes (Gr-1+/Mac-1+) in the spleen, MLNs and colonic lamina propria. Co-culture of irradiated leukocytes (5 x 103 cells) obtained from the MLNs or spleen (&gt;60% PMNs) of colitic mice with CD4+ T-cells (105 cells) enhanced CD/CD28-induced T-cell proliferation by 2–3 fold. We conclude that the T-cell induced myelopoiesis and infiltration of PMNs into the secondary lymphoid tissue of lymphopenic recipients may enhance T-cell activation thereby modulating the onset and/or severity of colonic inflammation
(Supported DK64023).
In vitro generated antigen specific CD4+CD25+Foxp3+ regulatory T cells control the severity of virus induced immunoinflammatory lesion (99.3)
Authors: sehrawat sharvan , Suvas Susmit , Rouse Barry T 
Published: January 2023

Abstract
A previous model of stromal keratitis (SK) developed in our laboratory was established in which several TCR transgenic (Tg) mice back crossed to SCID or RAG KO were shown to develop typical SK lesions upon ocular infection with HSV. The lesions were judged to be the consequence of a bystander activation of T cells that in turn orchestrated tissue damage. Most studies were done with the CD4 TCR Tg crossed with DO11.10 RAG KO mice which recognize the OVA323-339 peptide. In current studies we evaluated if CD4+CD25+Foxp3+ OVA-specific Tregs were capable of modulating the bystander tissue damage caused by HSV infection. To this end, we generated CD4+CD25+Foxp3+ regulatory T cells with OVA-specificity by in-vitro culturing splenocytes from DO11.10 TCR Tg RAG KO mice for five days in presence of a cocktail TGFβ1(5ng/ml), OVA323-339 peptide (0.62uM) and rIL-2 (25U). A total of 1x106 CD4+CD25+Foxp3+ cells were adoptively transferred i.v. in DO11.10 TCR Tg Rag KO one day prior to ocular infection with 5x105 pfu of HSV I KOS. The SK lesion and angiogenesis were scored every alternate day beginning at day 4. The lesion severity (p=0.05) and the angiogenesis (p=0.005) were significantly reduced in treated mice as compared to untreated mice. The implications of these observations in terms of the necessary specificity characteristics of Treg to control tissue damage will be discussed.
Studying lineage relationships of T cells by cellular barcoding (99.7)
Authors: van Heijst Jeroen W.J. , Schepers Koen , Swart Erwin , Gerlach Carmen , Kerkhoven Ron M. , Arens Ramon , Schumacher Ton N.M. 
Published: January 2023

Abstract
Antigen-specific T cell responses are composed of phenotypically and functionally distinct subsets. However, it is often unclear how these subsets arise from a pool of naive T cells. To address such issues, we have developed a novel technique that allows the study of lineage relationships of hematopoietic cells. In this technique, progenitor cells are labeled with a unique stretch of DNA. Upon cell division, all offspring of a labeled progenitor will inherit this ‘barcode’. By analyzing the barcodes present in separate cell populations, it can be determined whether these populations are derived from a common progenitor.
To demonstrate the value of this technique, we analyzed kinship of CD8+ T cells accumulating at subcutaneous tumor and lung effector sites. Even though distinct T cells are primed in the draining lymph node of each site, the offspring of these cells is found back at both effector sites. This indicates that CD8+ T cells are endowed with the capacity to migrate to distinct effector sites, independent of the site of priming in this model.
In a parallel project, we have used cellular barcoding to determine the number of T cell progenitors recruited into the immune response under conditions of varying antigen dose. We find that at increasing antigen dose both the recruitment of progenitor cells as well as their rate of proliferation are increased, and thus both factors contribute to the size of the CD8+ T cell response.
Reduction of contact sensitivity responses to FITC, but not DNFB in mice lacking the tyrosine kinase, Itk (B103)
Authors: Ralph Kerry L. M. , Raymond Ernie L. , Sellati Rosemarie , Haynes Nancy , Thibodeau Michael S. , Nabozny Gerald H. 
Published: January 2023

Abstract
Itk (inducible T cell kinase) is a member of the Tec kinase family of nonreceptor protein tyrosine kinases that are expressed in T cells, NK cells, and mast cells. Defects in Itk protein lead to impaired T cell activation and effector function. Although mice lacking Itk are impaired in their ability to generate protective Th1 and Th2 responses in vivo, LCMV clearance requiring CD8+ T cells appear to be intact. Contact sensitivity (CS) is a type IV hypersensitivity reaction resulting from secondary skin exposure to a reactive hapten. Murine models of CS using chemical haptens such as DNFB are shown to be predominantly CD8+ T cell mediated with CD4+ T cells acting as modulators. Recently, data from CS models using FITC suggest a dominant role of Th2-type CD4+ T cells. The purpose of our studies was to determine whether Itk plays a role in the development of contact sensitivity responses to FITC or DNFB using Itk knockout mice. Indeed mice lacking Itk have significantly reduced ear swelling and leukocyte infiltration following FITC ear challenge compared to WT mice, which correlates with the lack of FITC-specific cytokine secretion from primed Itk KO LNC in vitro. In contrast, Itk−/− mice were able to generate ear swelling responses comparable to WT mice following DNFB challenge. Together, these data further support the effector role of CD4+ T cells in FITC-CS and the inability of Itk−/− mice to generate effective Th2 responses in vivo.
IL-17 is selectively induced by epicutaneous sensitization and mediates neutrophil recruitment to the lungs following antigen challenge (B104)
Authors: He Rui , Oyoshi Michiko , Geha Raif 
Published: January 2023

Abstract
IL-17 producing effector CD4+ T cells (TH17) have been recently identified to be a new lineage of T helper cells, and thought to be important contributors to tissue inflammation. We here show that IL-17 is selectively induced by epicutaneous (EC) sensitization with ovalbumin (OVA) compared to intraperitoneal (i.p.) immunization. This is evidenced by increased serum IL-17 and increased IL-17 production by OVA stimulated cells from draining lymph nodes and spleen in EC sensitized mice. Significantly higher expression of IL-17 and CXCL-2 and infiltration of neutrophils was found in the BAL fluid and lungs of EC-sensitized mice compared with i.p-immunized mice following inhaled OVA challenge. Blockade of IL-17 prior to inhaled OVA challenge reduced CXCL-2 expression, airway inflammation and responsiveness to methacholine in EC sensitized mice. FITC+ dendritic cells (DCs) collected from draining lymph nodes 24 hrs after painting the skin with FITC, but not DCs from spleen, induced IL-17 secretion by TCR-OVA CD4+ T cells. This was inhibited by intradermal injection of anti- TGFβ neutralizing antibody. These results indicate that IL-17 is selectively induced by EC sensitization, in part due to the presence of TGFβ in the skin, and suggest that IL-17 might play an important role in airway hyper-reactivity and neutrophil infiltration in the lungs when skin is the initial site of antigen sensitization.
Defining the role of BmIL-5 in tropical pulmonary eosinophilia (B110)
Authors: Kwon Sung (Steve) , Gnanasekar Munirathinam , Balakrishnan Anand Setty , Ramaswamy Kalyanasundaram 
Published: January 2023

Abstract
This study describes characterization of a novel pathogen-derived human IL-5 receptor binding molecule designated BmIL-5. This molecule was identified in our laboratory by screening a phage displayed cDNA expression library of the infective larvae of the human lymphatic filarial parasite, Brugia malayi. Soluble human IL-5 receptor (huIL-5R) was used as the bait. This screening procedure yielded a single clone and sequence analyses of the insert from this clone showed a novel protein designated as “Brugia malayi-derived IL-5-like molecule” (BmIL-5). Recombinant BmIL-5 was then expressed and purified. ELISA-based binding assays confirmed binding of rBmIL-5 to huIL-5R. Functional significance of this binding was then evaluated using a human eosinophil cell line, named Hs 454.T. Initial studies showed that BmIL-5 binds to eosinophil surface. To test whether this binding initiates any signaling events, we evaluated STAT1 phosphorylation status in the eosinophil. We also performed a series of experiments to identify the receptor binding region of BmIL-5 by peptide mapping. In these studies, we designed overlapping 20 mer peptides and evaluated the binding pattern of these peptides to huIL-5R. These studies identified peptide 1 and peptide 11 as the domains binding to huIL-5R. An interesting observation was that, both these peptides interfered with the binding of huIL-5 to huIL-5R in vitro. Thus, we believe that BmIL-5 interferes with IL-5 function by preventing IL-5 binding to IL-5R. Our findings demonstrate a novel method of how pathogens potentially manipulate the human immune system.
TSLP-TSLPR interactions regulate CD4+ T cell-dependent immunity and inflammation in the intestine (B112)
Authors: Taylor Betsy C. , Zaph Colby , Troy Amy , Berman-Booty Lisa , Du Yurong , Artis David 
Published: January 2023

Abstract
Thymic stromal lymphopoeitin (TSLP) is an IL-7-like cytokine expressed by epithelial cells that has been implicated in promoting Th2 cytokine-dependent inflammation in the skin and lung. However, the role of IEC-derived TSLP in regulating immunity and/or inflammation in the intestine has not been tested. Following exposure to the intestinal-dwelling nematode pathogen Trichuris, IECs isolated from genetically resistant C57BL/6 mice exhibit an upregulation in expression of TSLP mRNA. To address the functional role of TSLP-TSLPR interactions in the intestine, WT and TSLPR−/− mice were infected with Trichuris. While WT mice exhibited expression of Trichuris-specific Th2 cytokines in the mesenteric LN (mLN) and cleared infection, mLN cells isolated from TSLPR−/− mice secreted increased IFNγ over WT controls and these mice harbored persistent infection. Further, infected TSLPR−/− mice developed infection-induced intestinal inflammation. In vitro studies suggest IEC-derived TSLP can influence DC function. Consistent with this, DC isolated from the gut-associated lymphoid tissue of TSLPR−/− mice exhibited more IL-12/23p40 than WT DC. In addition, conditioning of WT BMDC with rTSLP inhibited LPS induced IL-12/23p40 in vitro. Taken together, these studies implicate a role for TSLP-TSLPR interactions in IEC-DC cross-talk in the intestine that limits the expression of proinflammatory cytokines.