bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibody microarray analysis of inflammatory mediator release by human leukemia T cells and human non-small cell lung cancer cells (96.28)
Authors: Hommema Eric , Garcia Bradley , Hargrave Aubrey , Kilmer Greg , Morgan Aric , Narahari Janaki , Webb Brian , Wiese Rick 
Published: January 2023

Abstract
In order to expedite the analysis of samples for multiple cytokines/chemokines, we have developed slide-based ExcelArray™ antibody sandwich microarrays. Each slide consists of 16 subarrays (wells) printed with 12 specific antibodies in triplicate and positive and negative control elements. This 16-well format allows for the analysis of 10 test samples using a six-point standard curve. The array architecture is based on the “sandwich” ELISA, in which an analyte protein is “sandwiched” between an immobilized capture antibody and a biotinylated detection antibody, using streptavidin-linked DyLight™ 649 dye for quantitation. We used the Jurkat cell line as a model for human T cell leukemia and the A549 cell line as a model for human lung cancer. In order to evoke a cytokine/chemokine response, cells were stimulated with Tumor Necrosis Factor alpha (TNFα), Phorbol-12-myristate-13-acetate (PMA, TPA) and Phytohemagglutinin (PHA). Cell supernatants derived from both untreated and stimulated cells were analyzed on 41 unique analytes. Stimulated cells showed an increase in the expression level of many of the test analytes, including IL-8, TNF-α, and MIP-1α, compared to the non-treated controls. Our experiments clearly demonstrate the utility of antibody microarray analysis of cell culture supernatants for the profiling of cellular inflammatory mediator release.
Respiratory Syncytial Virus Vaccine-Enhanced Pulmonary Eosinophilia Requires IL-13, IL-4Rα, and STAT6 (96.6)
Authors: Castilow Elaine , Varga Steven M. 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract disease in children. A formalin-inactivated RSV vaccine administered to children in the 1960′s led to enhanced morbidity and mortality upon natural RSV infection. Histological analysis of the lungs upon autopsy revealed pulmonary eosinophilia. BALB/c mice primed with a recombinant vaccinia virus expressing the attachment (G) protein of RSV (vacG) develop pulmonary eosinophilia and vaccine-enhanced disease mimicking the response of the vaccinated children. Although the G protein elicits a mixed Th1 and Th2 response in these mice, the Th2 cells are known to be necessary for the induction of pulmonary eosinophilia. Here we have investigated role of individual Th2 cytokines in mediating RSV vaccine-enhanced disease. Our work shows that IL-13, the IL-4 receptor α chain (IL-4Rα), and STAT6, are required for the development of pulmonary eosinophilia in vacG-primed mice challenged with RSV whereas IL-4 is not required. We have found that IL-4, IL-13, IL-4Rα, and STAT6 do not contribute to weight loss or clinical illness whereas STAT4 is necessary. Lung histology and viral titers have also been examined in IL-13-, IL-4-, IL-4/13-, and IL-4Rα-deficient mice. Our results demonstrate a critical role for IL-13 in driving RSV vaccine-induced pulmonary eosinophilia.
Critical role of IL-17R signaling in granulocyte recruitment and airway hyperreactivity in an Aspergillus model of asthma (96.7)
Authors: Kapadia Shernaaz B , McKinley Laura , Henry Adam , Ray Anuradha , Kolls Jay K 
Published: January 2023

Abstract
Rationale IL-17 is a proinflammatory cytokine that enhances neutrophil recruitment in response to allergen in murine lung. In an Aspergillus fumigatus (Af)-induced asthma model, wild-type C57BL/6 mice develop bronchial inflammation and hyperreactivity. We hypothesized that, compared to C57BL/6, Af sensitized IL-17R KO mice would have reduced neutrophil recruitment to the lung, airway resistance and hyperreactivity.
Methods C57BL/6 and IL-17R KO male mice, 6–8 wk old, were sensitized intraperitoneally with 40 ug Af extract and 2 mg Al(OH)3 on days 0 and 7, and intratracheally with 25 ug Af on days 21, 22, and 23. Bronchoalveolar lavage (BAL) differential cell counts and cytokine analysis were done at 24 and 72 hr. Flexivent was performed at 24 hr to assess airway physiology.
Results WT mice had significant influx of both neutrophils and eosinophils in response to Af at 24 hr. In contrast, IL-17R KO mice showed reduced neutrophil and eosinophil emigration to the lung which was associated with lower KC and eotaxin levels in BAL fluid. Furthermore, Flexivent data showed lower airway resistance and bronchial hyperreactivity in IL-17R KO mice.
Conclusion Our findings in Af treated IL-17R KO mice of decreased pulmonary neutrophil and eosinophil recruitment at 24 hr suggest that IL-17 promotes granulocyte recruitment as well as airway hyperreactivity.
Funding: NHLBI
A Th2 response to chronic intermittent antigen exposure induces pulmonary vascular muscularization (96.8)
Authors: Grunig Gabriele , Daley Eleen , Emson Claire , Taraseviciene-Stewart Laima , Kurup Viswanath P. , de Waal-Malefyt Rene , Hogaboam Cory , Grunig Ekkehard , Voelkel Norbert F. 
Published: January 2023

Abstract
Pulmonary arterial hypertension is associated with pulmonary vascular remodeling (PVRm) and its incidence is increased in many autoimmune diseases and chronic inflammatory lung diseases. Currently it is not known whether inflammatory and immune responses are cause or consequence of PVRm. In this study immunized mice were intermittently exposed to inhaled soluble antigen over a period of several weeks. The antigen exposure induced dramatic PVRm in a CD4 T cell dependent manner. PVRm was characterized by hyperplasia of smooth muscle cells. To identify pathogenic mediators, IL-4 deficient mice and wild type mice transiently given an IL-13 blocker were examined. In both cases, the mice developed significantly less PVRm relative to immunized, antigen challenged, control wild type mice. Mice given recombinant IL-13 intranasally did not develop severe vascular muscularization indicating that IL-13 and, most likely IL-4 promote PVRm indirectly. In conclusion, our data are the first to provide proof of concept that the adaptive immune response to inhaled antigen can cause PVRm.
Supported by: Flight Attendant Medical Research Institute, Stony Wold Herbert Fund, American Lung Association of the City of New York
Plant polyphenols induce differential regulation of CD11b expression and function on monocytes and γδ T cells (97.11)
Authors: Graff Jill C , Jutila Mark A 
Published: January 2023

Abstract
Leukocyte adhesion and migration is partially mediated by CD11b/CD18 (MAC-1, CR3). Earlier studies have demonstrated a role for green tea polyphenols in down-regulating CD11b on monocytes and CD8+ T cells. We have recently shown a stimulatory effect of unripe apple polyphenols (APP) on γδ T cells. Thus, we compared the effect of APP on γδ T cell and monocyte CD11b expression. Purified bovine monocytes and monocyte-depleted peripheral blood lymphocytes were cultured with APP for 24 hours. CD11b levels decreased on monocytes in response to APP. In contrast, a subset of γδ T cells responded to APP by up-regulating CD11b. Consistant with the changes in surface CD11b expression, APP treated γδ T cells showed increased adherence to plastic, whereas monocyte adhesion was reduced. The affects of APP on monocytes could be inhibited by anti-CD11b mAb, whereas the mAb had no effect on the APP response in γδ T cells, suggesting the two cell types respond to APP via different mechanisms. Polyphenols are typically thought of as anti-inflammatory agents, however, these data, as well as other ongoing studies, indicate they have a pro-inflammatory effect on γδ T cells. In vivo, plant polyphenols may enhance γδ T cell migration to sites of inflammation, where they can enact rapid immunoregulatory and innate-like responses.
Funded by: NIH –NIAID DAIT-BAA-03 -41
ICAM-1 expressed on neutrophils is critically involved in vaccine-induced protection in a murine model of acute pneumonia (97.3)
Authors: Priebe Gregory P , Walsh Rebecca L , Cederroth Terra A , Pier Gerald B 
Published: January 2023

Abstract
Murine neutrophils are not known to express ICAM-1. In studying the mechanism of protection against P. aeruginosa pneumonia following immunization with a live-attenuated vaccine in a model of acute pneumonia, we detected ICAM-1 expression on lung neutrophils after challenge. We hypothesized that neutrophil ICAM-1 elicited homotypic adhesion as well as enhanced bacterial killing. C3H/HeN mice immunized intranasally (IN) with an aroA deletion mutant of P. aeruginosa strain PAO1 (PAO1ΔaroA) or E. coli (as control) at weekly intervals with 108, 5 x 108, and 109 CFU were challenged IN 3 weeks later with a cytotoxic variant of PAO1. Lung leukocytes and BAL cells were analyzed by flow cytometry, and decreased migration due to homotypic adhesion was assessed using Transwells. We also tested the impact of ICAM-1 blockade on bacterial clearance by IN instillation of anti-ICAM-1 antibody prior to challenge of immunized mice. PAO1ΔaroA-immune mice had more ICAM-1+ neutrophils in BAL 18 h after challenge compared with E. coli-immune controls (mean 74% vs. 37%, p &lt; 0.01). Non-immune mice had low levels of ICAM-1+ neutrophils. BAL neutrophils from PAO1ΔaroA-immune mice transmigrated significantly less well, and blockade of ICAM-1 prior to challenge increased bacterial CFU in PAO1ΔaroA-immune mice. These data suggest that vaccine-induced neutrophil ICAM-1 plays an important role in the adaptive immune response in the lung.
Initiation of Adaptive-Innate Immune Interaction Sequences in Contact Sensitivity Responses (98.5)
Authors: Kish Danielle D. , Fairchild Robert L. 
Published: January 2023

Abstract
Contact hypersensitivity (CHS) is a CD8 T cell-mediated response to cutaneous sensitization and subsequent challenge with hapten. Hapten-primed effector CD8 T cell recruitment into the challenged skin to mediate CHS requires prior CXCL1/KC and CXCL2/MIP-2 directed neutrophil infiltration into the challenge site. The mechanisms inducing neutrophil chemoattractant production and the dynamics of neutrophil-CD8 T cell infiltration into the challenge site during the elicitation of CHS to dinitrofluorobenzene (DNFB) were investigated. Following DNFB challenge, CXCL1/KC and CXCL2/MIP-2 were produced within 3–6 h at 10-fold higher levels in sensitized vs. non-sensitized mice. This early chemokine production was dependent on hapten-specific CD8 T cells. At 6 h post-challenge of sensitized but not naïve mice, vascular endothelial cells (EC) in the challenge site stained positively for CXCL1/KC protein as well as for hapten. Although DNFB-primed CD8 T cells induced early CXCL1/KC production, the effector T cells did not infiltrate the parenchymal tissue of the challenge site at 6 h post-challenge but did 12 h later. In contrast, neutrophils infiltrated the challenge site at 6 h post-challenge and this was dependent on hapten-primed CD8 T cell induction of CXCL1/KC production. In vitro, CD8 T cells also stimulated hapten-presenting EC to produce KC. These results indicate hapten challenge to elicit CHS results in EC presentation of hapten to primed CD8 T cells. The T cells stimulate EC production of neutrophil chemoattractants that direct the neutrophil recruitment required for the subsequent infiltration of the effector CD8 T cells to mediate the allergic response.
Migration and expansion of dendritic cells improves cross-presentation of tumor antigens in virotherapy (98.8)
Authors: Edukulla Ramakrishna , Woller Norman , Mundt Bettina , Kuehnel Florian , Kubicka Stefan 
Published: January 2023

Abstract
Triggering of dendritic cell (DC) migration into tumor tissue and subsequent expansion of DCs may be useful to improve cross-presentation of tumor antigens during virotherapy.
Appropriate syngenic mouse models were established for lung carcinoma cells CMT64 and KLN205.Crystal violet staining showed that CMT64 and KLN205 are permissive for replication of the oncolytic telomerase-dependent adenovirus hTert-Ad.
We used adenoviral expression of Macrophage Inflammatory Protein 1α (MIP 1α) to recruit DCs into hTert-Ad infected tumors and Ad-Flt3L to expand DCs within the tumor nodules during virotherapy.
Compared with virotherapy alone or virotherapy with expression of a single CC chemokine, expression of both MIP1α and Flt3L during virotherapy significantly increases infiltration of DCs and T cells, as shown by histochemistry. Interferon-γ, Pentamer staining and CTL assay showed that the combination of cytokines improves cross-presentation of tumor antigens and induces an antitumoral immune response during virotherapy. Combination of DC vaccination and viroimmunotherapy showed nearly complete regression of the subcutaneous tumors
In conclusion mobilizing of DCs to virally infected tumors during virotherapy improves cross-presentation of tumor antigens and induces antitumoral immune response which facilitates an effective viroimmunotherapy of established tumors.
Accumulation of antigen-specific B and T cells in ectopic lymphoid tissue after immunization (99.11)
Authors: Weinstein Jason Scott , Nacionales Dina C , Lee Pui Y , Kelly-Scumpia Kindra M , Sobel Eric , Satoh Minoru , Reeves Westley H 
Published: January 2023

Abstract
Objective: Ectopic lymphoid tissue is associated with autoantibody production. Although reminiscent of authentic secondary lymphoid tissue, it is unclear if it can serve as a site of cognate T-B cell interactions leading to autoimmunity. This question was examined in mice with ectopic lymphoid tissue induced by tetramethylpecadentane (TMPD).
Methods: B6 and BALB/c mice were treated i.p. with TMPD followed 3 mo later by s.c. immunization with NP-KLH or NP-OVA. NP specific B cells were identified by analysis of VH gene usage, flow cytometry, and ELISPOT. OVA transgenic T cells from DO.11 mice were transferred i.v. into BALB/c mice 3 d before immunizing with NP-OVA. OVA-specific T cells were identified by flow cytometry and proliferating B and T cells by BrDU staining.
Results: 12 d after immunizing with NP-KLH, 90% of H chain genes from ectopic lymphoid tissue of B6 mice bore V186.2 or analogous NP-specific VH sequences. B cells stained by NP-PE were identified in ectopic lymphoid tissue by flow cytometry and secretion of anti-NP antibodies shown by ELISPOT. The ectopic lymphoid tissue and draining lymph nodes also had numerous OVA-specific T cells 7 d after immunization with NP-OVA and there was a progressive increase of BrDU+ T and B cells.
Conclusion: The data suggest that following immunization, antigen-specific B and T cells undergo a local germinal center-like reaction within ectopic lymphoid tissue.
Using the chicken feather as a window into cell-mediated tissue responses (99.12)
Authors: Erf Gisela F. , Lockhart Becky , Bowen Olivia T. , Bateman Kristin , Finley ReGina C. 
Published: January 2023

Abstract
Cell-mediated immune (CMI) responses are typically tested by injecting the skin and monitoring the skin reaction. In chickens, various integumental tissues are suitable to measure the CMI reaction, including the wattle, wing web, toe web and foot pad. Using the Smyth line chicken vitiligo model to study the spontaneous autoimmune destruction of pigment cells in feathers, we have found that one growing feather-tip (3 mm diameter by 8 mm length) yields generous amounts of material for down-stream gene-expression-, histological-, and cell population-analyses. Moreover, growing feathers are easily accessible for microinjection, removal of a feather is minimally invasive, and feathers are very suitable for repeated sampling during an immune response. To investigate the potential of the feather as a window into tissue immune responses, time-course studies were conducted comparing leukocyte infiltration in wattles, wing webs and growing feathers in response to injection of vehicle, PHA or LPS, and during the recall response to Mycobacterium butyricum. Based on histology and immunohistochemical staining, leukocyte infiltration profiles over a 72 hour period were similar in all three tissues tested. Hence, the feather provides excellent opportunity to monitor and assess CMI responses and immunomodulation strategies in situ and via down-stream analyses.
Support: Arkansas Experiment Station Animal Health Grant
Abdominal lymphatic pump treatment increases leukocyte count and flux in thoracic duct lymph (99.13)
Authors: Hodge Lisa , King Hollis H , Williams Arthur , Reder Stephanie , Belavadi Tejaswi , Simecka Jerry , Stoll Scott , Downey H. Fred 
Published: January 2023

Abstract
Studies suggest that lymphatic pump techniques (LPT) enhance immunity and resistance to infection, but direct evidence of this has not been documented. In this study, the immediate effects of LPT on thoracic duct lymph flow and leukocyte output was measured. Lymph flow was measured and lymph was collected at baseline and during LPT. The baseline leukocyte count was 4.8 ± 1.7 x 106 cells/ml, and LPT increased leukocytes to 11.8 ± 3.6 x 106cells/ml (P&lt;0.05). While macrophages, neutrophils, total lymphocytes, T cells and B cell numbers were similarly increased during LPT, their relative percentage in lymph was unaltered by LPT; however, IgA antibody forming B cells increased from 5.8% at baseline to 17% during LPT. Furthermore, LPT enhanced lymph flow 4-fold. Leukocyte flux was computed from the product of lymph flow and cell count. LPT enhanced leukocyte flux from 8.2± 4.1 x 106total cells per min to 60 ± 25 x 106cells per min. In summary, LPT significantly increased both thoracic duct lymph flow and leukocyte count, so lymphatic leukocyte flux was markedly enhanced. In addition, our data suggests that LPT can act preferentially on mucosal tissues, revealing a source of these mobilized leukocytes. Enhanced mobilization and lymphatic transport of immune cells during abdominal compression is likely an important mechanism responsible for the increased immune responses of patients treated with LPT.
Developmental and antigen-driven switches in the trafficking receptors of FoxP3+ regulatory T cells (99.2)
Authors: Kim Chang H , Lee Jeeho , Kang Seung G 
Published: January 2023

Abstract
FoxP3+ regulatory T cells play important roles in immune regulation and tolerance. There is an increasing body of evidence that the migration ability of FoxP3+ T cells is important for their regulatory functions at effector tissue sites. We investigated the two different trafficking receptor switches of FoxP3+ T cells occurring in the thymus and secondary lymphoid tissues. The first trafficking receptor switch in the thymus is developmentally programmed: Precursors of FoxP3+ cells undergo the first trafficking receptor switch from CCR8/CCR9 to CXCR4 and then finally to CCR7, generating mostly homogeneous CD62L+CCR7+ FoxP3+ T cells. The recent thymic emigrant CD62L+CCR7+ FoxP3+ T cells are programmed to migrate to secondary lymphoid tissues. The CD62L+CCR7+ FoxP3+ T cells undergo the second switch in trafficking receptors in an antigen-dependent manner. This second switch involves down-regulation of CCR7 and CXCR4 but up-regulation of a number of memory/effector type homing receptors, resulting in generation of heterogeneous FoxP3+ T cell subsets expressing various combinations of trafficking receptors including CCR2, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9 and CXCR5. FoxP3+ cells undergo the second switch to selected non-lymphoid tissue homing receptors at highly accelerated rates. This results in generation of FoxP3+ T cells with unconventionally efficient migratory capacity to major non-lymphoid tissues such as intestinal lamina propria and bone marrow. Importantly, this accelerated switch of FoxP3+ T cells is conserved in both men and mice. The two switches in homing receptors are though to be important for effective distribution and differentiation-dependent effector functions of FoxP3+ regulatory T cells.
Where do blood-borne cells enter the brain? (99.9)
Authors: Lampson Lois A. , Tripp Cara A. 
Published: January 2023

Abstract
The blood-brain barrier stops passive entry of proteins, but not living cells. Both tumor cells and leukocytes can enter the brain, and it is attractive to exploit blood-borne leukocytes to attack blood-borne metastases or pathogens. However, more needs to be known about where different cell types enter, lodge, and grow.
METHODS. In 6u sections of rat brain, inert beads, blood-borne metastases of marked tumor, and T cells were stained histochemically or by common antibodies.
RESULTS.
Tumor, T cells, and inert beads can all enter vessels of the brain proper, choroid plexus (within the ventricles), and meninges. However, the relative importance varies with the cell type.Metastases in the brain proper were common for one tumor line, and rare for the other. Both lines metastasized to the choroid plexus.In contrast, the choroid plexus was not, in general, a favored site of T cell entry. T cells were more likely to enter at cerebral vessels.When T cells did accumulate in the ventricles, they were most concentrated in the narrow foramen of Monro.
DISCUSSION. The traffic patterns of other populations (T cell subsets, B cells, monocytes) must now be compared. In seeking to exploit blood-borne leukocytes against blood-borne tumor or pathogens, the optimal responding cell type may vary with the target site.
Enumeration of double stranded DNA breaks in developing thymocytes in young and aged mice (B1)
Authors: Hesse Jill , Faulkner Matthew F , Durdik Jeannine M 
Published: January 2023

Abstract
Aging at the cellular and molecular level has been shown to involve a decline in the immune system. In spite of this decline, the aged thymus is still functional in both thymocyte development and output of mature naïve T cells producing a small but significant population. During V(D)J recombination double stranded breaks (DSB) are created in the DNA of developing thymocytes. Complete and accurate resolution of the DSB is critical to the proper function and survival of cells. In our studies we utilized gamma phosphorylated-H2A.X(γH2A.X) to estimate the amount of DSB that are created and/or persist in thymocytes. H2A.X phosphorylation is the earliest ATM-dependent response to DSB, and it is speculated that the γH2A.X may help keep broken DNA ends together. For quantitation it is import to note that many H2AX molecules are phosphorylated in response to one DSB. The amplification of the signal allows us to visualize foci. We find cells from aged thymus have more DSB than cells from the young thymus. This increase in breaks may be the result of more breaks being created or breaks persisting due to slow or lack of DSB repair. By utilizing fluorescent staining and confocal microscopy we were able to further investigate the number of cells with DSBs and compare different thymocyte subpopulations. Here we show the enumeration of γH2A.X foci in thymocyte subpopulations in both young and aged mice.