bluuebunny/crossref_metadata_2025_split huggingface dataset:
Depletion of IL-10- and TGF-β-Producing Regulatory γδ T Cells by Administering a Daunomycin-Conjugated Specific Monoclonal Antibody in Early Tumor Lesions Augments the Activity of CTLs and NK Cells
Authors: Seo Naohiro , Tokura Yoshiki , Takigawa Masahiro , Egawa Kohji 
Published: January 2023

AbstractIt has been demonstrated that γδ T cells accumulating in early tumor lesions and those purified from spleen cells of tumor-bearing mice attenuate the activity of CTLs and NK cells. We, therefore, investigated whether depletion of γδ T cells from early lesions of tumors results in restoration of CTL and NK cell activities and subsequent regression of tumors. A daunomycin-conjugated anti-γδTCR mAb UC7-13D5 (Dau-UC7) was prepared to efficiently deplete γδ T cells. An in vitro study revealed that Dau-UC7 specifically lysed γδTCR+ cells and effectively inhibited splenic γδ T cells from tumor-bearing mice to produce cytotoxic cell-suppressive factors. Furthermore, intralesional injections of Dau-UC7 at an early stage of tumor development led to augmentation of tumor-specific CTL as well as NK cell activities and to the resultant regression or growth inhibition of the tumors. On analysis of cytokine profile, γδ T cells transcribed mRNAs for IL-10 and TGF-β, but not IL-4 or IFN-γ, suggesting the T regulatory 1-like phenotype. Finally, a blocking study with mAbs showed that the inhibitory action of γδ T cells on CTLs and NK cells was at least partly mediated by IL-10 and TGF-β. These results clearly demonstrated the novel mechanism by which T regulatory 1-like γδ T cells suppress anti-tumor CTL and NK activities by their regulatory cytokines in early tumor formation.
The CD3ε Subunit of the TCR Contains Endocytosis Signals
Authors: Borroto Aldo , Lama Juan , Niedergang Florence , Dautry-Varsat Alice , Alarcón Balbino , Alcover Andrés 
Published: January 2023

Abstract
Ligand binding to TCR induces its internalization and cell surface down-modulation. These phenomena contribute to the extinction of activation signals. Due to the multicomponent nature of the TCR-CD3 complex, its internalization may be mediated by one or several of its subunits. Although it has been reported that CD3γ and CD3δ contain endocytosis motifs involved in the internalization of the TCR-CD3 complex, other subunits could also be involved in this process. For instance, CD3ε and CDζ display amino acid sequences reminiscent of internalization motifs. To investigate whether CD3ε bears endocytosis signals, we have analyzed the internalization capacity of a panel of deletion and point mutants of CD3ε that were expressed on the cell surface independently of other TCR-CD3 subunits. Here we report that CD3ε displays endocytosis determinants. These data indicate that CD3ε could contribute to the internalization and cell surface down-regulation of TCR-CD3 complexes. Moreover, the existence of endocytosis signals in this polypeptide could serve to retrieve unassembled CD3ε subunits or partial CD3 complexes from the plasma membrane, thus restricting the expression on the cell surface to fully functional TCR-CD3 complexes.
A Single-Chain IL-12 IgG3 Antibody Fusion Protein Retains Antibody Specificity and IL-12 Bioactivity and Demonstrates Antitumor Activity
Authors: Peng Lisan S. , Penichet Manuel L. , Morrison Sherie L. 
Published: January 2023

Abstract
IL-12 is a heterodimeric cytokine with many actions on innate and cellular immunity that may have antitumor and antimetastatic effects. However, systemic administration of IL-12 can be toxic. Tumor-specific Abs provide a means to selectively target a metastatic/residual nodule and deliver therapeutic quantities of an immunostimulatory molecule like IL-12 with lower systemic levels and ideally, toxicity. We report the construction and characterization of an Ab fusion protein in which single-chain murine IL-12 is fused to an anti-Her2/neu Ab at the amino terminus (mscIL-12.her2.IgG3). The use of single-chain IL-12 in the fusion protein simplifies vector construction, ensures equimolar concentrations of the two IL-12 subunits, and may confer greater stability to the fusion protein. SDS-PAGE analysis shows this 320-kDa protein is secreted and correctly assembled. FACS analysis demonstrates that this fusion protein binds to cells transfected with the Her2/neu Ag, thus retaining Ab specificity; this fusion protein also binds to a cell line and to PHA-activated PBMC that express the IL-12R, thus demonstrating cytokine receptor specificity. T cell proliferation assays and NK cytotoxicity assays demonstrate that this fusion protein exhibits IL-12 bioactivity comparable to recombinant murine IL-12. In vivo studies demonstrate that this fusion protein has antitumor activity. These results are significant and suggest that this IL-12 Ab fusion protein can effectively combine the therapeutic potential of IL-12 with the tumor-targeting ability of the Ab and may provide a viable alternative to systemic administration of IL-12.
Constitutive Nuclear Translocation of NF-κB in B Cells in the Absence of IκB Degradation
Authors: Doerre Stefan , Corley Ronald B. 
Published: January 2023

Abstract
Members of the NF-κB/Rel family of transcription factors are involved in many aspects of B lymphocyte development and function. NF-κB is constitutively active in these cells, in contrast with most other cell types. In the inactive form, NF-κB/Rel proteins are sequestered in the cytoplasm by members of the IκB family of NF-κB inhibitors. When activated, NF-κB is translocated to the nucleus, a process that involves the phosphorylation and proteasomal degradation of IκB proteins. Thus, NF-κB activation is accompanied by the rapid turnover of IκB proteins. We show that while this “classical” mode of NF-κB activation is a uniform feature of IgM+ B cell lines, all IgG+ B cells analyzed contain nuclear NF-κB yet have stable IκBα, IκBβ, and IκBε. Furthermore, Iκβε levels are at least 10 times lower in IgG+ B cells than in IgM+ B cells, an additional indication that the regulation of constitutive NF-κB activity in these two types of B cells is fundamentally different. These data imply the existence of a novel mechanism of NF-κB activation in IgG+ B cells that operates independently of IκB degradation. They further suggest that different isoforms of the B cell receptor may have distinct roles in regulating NF-κB activity.
Estrogen Protects Against Cellular Infiltration by Reducing the Expressions of E-Selectin and IL-6 in Endotoxin-Induced Uveitis
Authors: Miyamoto Noriko , Mandai Michiko , Suzuma Izumi , Suzuma Kiyoshi , Kobayashi Kaori , Honda Yoshihito 
Published: January 2023

Abstract
Anterior uveitis associated with Behcet’s disease and ankylosing spondylitis preferentially occurs in adult men, which may suggest the effects of sex hormones on acute anterior uveitis. Recently, estrogen receptors in the vascular endothelium have been reported to be involved in several pathological conditions. In the present study, we examined the gender differences in susceptibility to endotoxin-induced uveitis (EIU) and the effects of estrogen on anterior inflammation. EIU was induced in adult male, female, and ovariectomized female Lewis rats (200 g) by hind footpad injection of 200 μg of LPS. In EIU, cellular infiltration was more marked in male than in female rats, and ovariectomy increased cellular infiltration. Treatment with 10 μg of 17β-estradiol significantly reduced the cell number in male and ovariectomized female rats with EIU. Estrogen receptor immunoreactivity was found in the nucleus of vascular endothelium and in some stromal cells of the iris-ciliary body. Semiquantitative PCR revealed that E-selectin and IL-6 gene expressions were increased in rats following LPS injection, and an overdose of tamoxifen, an estrogen receptor antagonist, reversed the effect of 17β-estradiol on E-selectin, but not its effect on IL-6. These observations suggested that the down-modulation of these inflammatory genes by estrogen may contribute to the reduction in cellular infiltration in acute anterior uveitis.
Mouse Monocyte-Derived Chemokine Is Involved in Airway Hyperreactivity and Lung Inflammation
Authors: Gonzalo Jose-Angel , Pan Yang , Lloyd Clare M. , Jia Gui-Quan , Yu Gary , Dussault Barry , Powers Christine A. , Proudfoot Amanda E. I. , Coyle Anthony J. , Gearing David , Gutierrez-Ramos Jose-Carlos 
Published: January 2023

Abstract
The cloning, expression, and function of the murine (m) homologue of human (h) monocyte-derived chemokine (MDC) is reported here. Like hMDC, mMDC is able to elicit the chemotactic migration in vitro of activated lymphocytes and monocytes. Among activated lymphocytes, Th2 cells were induced to migrate most efficiently. mMDC mRNA and protein expression is modulated during the course of an allergic reaction in the lung. Neutralization of mMDC with specific Abs in a model of lung inflammation resulted in prevention of airway hyperreactivity and significant reduction of eosinophils in the lung interstitium but not in the airway lumen. These data suggest that mMDC is essential in the transit/retention of leukocytes in the lung tissue rather than in their extravasation from the blood vessel or during their transepithelial migration into the airways. These results also highlight the relevance of factors, such as mMDC, that regulate the migration and accumulation of leukocytes within the tissue during the development of the key physiological endpoint of asthma, airway hyperreactivity.
Altered Memory T Cell Differentiation in Patients with Early Rheumatoid Arthritis
Authors: Skapenko Alla , Wendler Jörg , Lipsky Peter E. , Kalden Joachim R. , Schulze-Koops Hendrik 
Published: January 2023

Abstract
The chronic immune response in rheumatoid arthritis (RA) might be driven by activated Th1 cells without sufficient Th2 cell differentiation to down-modulate inflammation. To test whether disordered memory T cell differentiation contributes to the typical Th1-dominated chronic inflammation in RA we investigated differentiation of resting CD4+ memory T cells in patients with early (6 wk to 12 mo) untreated RA and in age- and sex-matched healthy controls in vitro. No difference in cytokine secretion profiles of freshly isolated memory T cells was detected between patients and controls. A cell culture system was then employed that permitted the differentiation of Th effectors from resting memory T cells by short term priming. Marked differences were found in response to priming. Th2 cells could be induced in all healthy controls by priming with anti-CD28 in the absence of TCR ligation. By contrast, priming under those conditions resulted in Th2 differentiation in only 9 of 24 RA patients. Exogenous IL-4 could overcome the apparent Th2 differentiation defect in seven patients but was without effect in the remaining eight patients. In all patients a marked decrease in IL-2-producing cells and a significant increase in well-differentiated Th1 cells that produced IFN-γ but not IL-2 were evident after priming with anti-CD3 and anti-CD28. The data suggest that CD4+ memory T cells from patients with early untreated RA manifest an intrinsic abnormality in their ability to differentiate into specific cytokine-producing effector cells that might contribute to the characteristic Th1-dominated chronic (auto)immune inflammation in RA.
Recognition of a Shared Human Prostate Cancer-Associated Antigen by Nonclassical MHC-Restricted CD8+ T Cells
Authors: Housseau Franck , Bright Robert K. , Simonis Toni , Nishimura Michael I. , Topalian Suzanne L. 
Published: January 2023

AbstractTo identify prostate cancer-associated Ags, tumor-reactive T lymphocytes were generated using iterative stimulations of PBMC from a prostate cancer patient with an autologous IFN-γ-treated carcinoma cell line in the presence of IL-2. A CD8+ T cell line and TCR αβ+ T cell clone were isolated that secreted IFN-γ and TNF-α in response to autologous prostate cancer cells but not to autologous fibroblasts or lymphoblastoid cells. However, these T cells recognized several normal and malignant prostate epithelial cell lines without evidence of shared classical HLA molecules. The T cell line and clone also recognized colon cancers, but not melanomas, sarcomas, or lymphomas, suggesting recognition of a shared epithelium-associated Ag presented by nonclassical MHC or MHC-like molecules. Although Ag recognition by T cells was inhibited by mAb against CD8 and the TCR complex (anti-TCR αβ, CD3, Vβ12), it was not inhibited by mAb directed against MHC class Ia or MHC class II molecules. Neither target expression of CD1 molecules nor HLA-G correlated with T cell recognition, but β2-microglobulin expression was essential. Ag expression was diminished by brefeldin A, lactacystin, and cycloheximide, but not by chloroquine, consistent with an endogenous/cytosolic Ag processed through the classical class I pathway. These results suggest that prostate cancer and colon cancer cells can process and present a shared peptidic Ag to TCR αβ+ T cells via a nonclassical MHC I-like molecule yet to be defined.
Effects of recombinant vectors expressing Chlamydia pneumoniae’s major outer membrane protein (MOMP) on humoral and T cell-mediated immunity. (129.1)
Authors: Elkadri Rehab Ahmed shafik , Grobler Anne-Katrien , Bermudez-Fajardo Alexandra , Puolakkainen Mirja , Stewart Graham , Oviedo-Orta Ernesto 
Published: January 2023

Abstract
Chlamydia pneumoniae is an obligatory intracellular pathogen. C. pneumoniae's major outer membrane protein (MOMP) is a 40 kDa porin and represents. The antigenic and immunomodulatory properties of this protein remain controversial.our aim is to assess the effect of recombinant vectors expressing MOMP on the induction of humoral and T cell-mediated immunity. : We used Mycobacterium vaccae and pcDNA3.1 expressing MOMP. Plasmids containing untruncated MOMP and truncated MOMP were created and used to express MOMP in M. vaccae.. In this study, we found that the signal sequence of MOMP effect on the viablity of M. vaccae. rMOMP expressed either by M. vaccae or by DNA plasmid is capable of significantly induce regulatory Treg of both CD4+CD25+FoxP3+ and CD8+CD25+FoxP3+phenotype. DNA immunisation induced a significant up-regulation of IL-4 and IL-10 in mice indicating a shift towards a predominant Th2 immune response. Additionally, DNA immunisation was also capable of significantly suppres the secretion of IFN-gamma. We also found that M. vaccae expressing MOMP significantly increases IL-10 secretion but without having a significant effect on IFN-γ and IL-4 secretion. We conclude that recombinant vectors expressing MOMP have the ability to induce predominantly T cell-mediated immunosuppressive responses which shed light on the effect of this protein on host immune responses that may explain Chlamydia's long term survival and also be responsible for the pathogen's immune evasion mechanisms.
Development of a non-living vaccine against Burkholderia mallei (129.2)
Authors: Deeraksa Arpaporn , Qazi Omar , Whitlock Gregory C , Judy Barbara M , Torres Alfredo G 
Published: January 2023

Abstract
Burkholderia mallei is a gram-negative highly pathogenic bacterium, responsible for glanders. B. mallei is considered potential bioterrorism agents, classified as such in list B by the Centers for Disease Control and Prevention. Currently there is no vaccine available against B. mallei. In this study, we aimed to identify the protective proteins of B. mallei. The candidate proteins were selected based on the previous data of proteomics and linear expression library (LEE) whole-genome screens coupled with known structures or functions of the proteins in other bacteria. The selected proteins were BMA-4, BMA-8, BMA-9, BMA-10, BMA-11 and BMA-12. The genes encoding each protein were cloned into pCDNA3.1 and pET28a. The His-tag proteins were then purified. BALB/c mice were vaccinated with DNA encoding each protein and then boosted 3 weeks later with the recombinant proteins. Two weeks after boosting, mice were infected with B. mallei ATCC23344. The result showed that, 19 days after infection, there was 100% survival in the BMA2821 and BMAA0768 groups, however only 78% survival in the control group. The study needs to be repeated and the result will be discussed.
Anti-viral CD8+ T cell Effector Activity Is Regulated by Hematopoietic Cells in the Influenza-Infected Lung Parenchyma (130.16)
Authors: Hufford Matthew Thomas , Sun Jie , Kim Taeg , Braciale Thomas J 
Published: January 2023

Abstract
CD8+ T cell effector activity is instrumental in the resolution of influenza infection, as well as additional respiratory viral infections, by their ability to eliminate infected cells and to release potent cytokines. CD8+ T cells localized to the lung airways are believed to be responsible for this anti-viral activity because CD8+ T cells located in the lung parenchyma have limited access to the respiratory epithelium, the principal site of influenza replication. To test this hypothesis, we utilized the detection of CD8+ T cell IFN? production ex vivo and in vivo as a convenient means to identify influenza-specific CD8+ T cells and to define the location of antigen presentation within the influenza-infected lungs. Surprisingly, influenza-specific CD8+ T cells collected from the airways produced limited amounts of IFN? in vivo. In contrast, significant numbers of influenza-specific CD8+ T cells in the lung parenchyma were actively producing IFN?. Because IFN? production by CD8+ T cells is principally TCR dependent, data indicated antigen engagement occurs in the lung parenchyma, not the airways. Because the lung parenchyma is enriched in hematopoietic (CD45+) cells compared to the airways, it was hypothesized that these cells were the primary antigen presenting cells in the lungs. To test this hypothesis, H-2Kb-/-Db-/- mice were irradiated and reconstituted with H-2Kb+/+Db+/+ bone marrow. Despite the absence of MHC class I on respiratory epithelial cells, CD8+ T cells in the lung parenchyma produced significant amounts of IFN? in vivo. These observations challenge conventional knowledge on the mechanism by which CD8+ T cells clear virally-infected cells in the respiratory epithelium.
CD4 memory T cells regulate mediators of lung damage during respiratory infections (130.8)
Authors: Jelley-Gibbs Dawn M , McKinstry K. Kai , Dibble John P , Roberts Alan D , Woodland David L , Swain Susan L 
Published: January 2023

Abstract
Activated T cells play a central role in controlling respiratory virus infections. However, these T cells also mediate considerable pulmonary immunopathology and lung damage. The level of immunopathology tends to be much less in secondary compared to primary infections, but the underlying reasons for this difference are unclear. To determine if CD4 memory T cells regulate pulmonary immunopathology, we transferred influenza-specific CD4 memory T cells into naïve mice challenged with influenza virus and assessed (i) inflammation, (ii) antigen presentation, and (iii) de novo T cell stimulation in the lung environment. The presence of CD4 memory T cells during influenza infection greatly reduced the expression of pro-inflammatory cytokines and chemokines in the lungs, as well as the ability of alveolar macrophages and pulmonary dendritic cells to stimulate naïve CD4 and CD8 T cell responses directly ex vivo. CD4 memory T cells also inhibited the accumulation of newly primed CD4 and CD8 T cells in the lungs, without reducing virus titers or de novo T cell responses in the periphery. We propose that CD4 memory T cells are capable of limiting pulmonary immunopathology by preventing the influx and retention of newly primed T cells in the lungs during respiratory infections. We are currently assessing whether the effective induction of CD4 memory populations through vaccination protects against excessive pulmonary immunopathology and secondary bacterial pneumonia. Funded by NIH AI057158 NBC-Lipkin and Trudeau Institute.
Cationic Liposome-DNA Complex Adjuvant (JVRS-100) Combined with Heat-Inactivated Whole Virion Influenza Vaccine Confers Protection against Homotypic and Heterosubtypic Lethal Challenge (132.1)
Authors: Chang Stella , Lay Marla , Callejo Bernadette , Liang Lily , Fairman Jeff 
Published: January 2023

Abstract
Background: Current vaccine strategies for influenza are highly dependent on matching of hemagglutinin, which is susceptible to periodic antigenic drift. A vaccine that elicits a robust antibody and T-cell response would be less susceptible to primary vaccine failure. Methods: Balb/c mice were vaccinated with heat-inactivated PR/8/34 (H1N1) or HKx31 (H3N2) with or without the JVRS-100 adjuvant. Mice were assessed for antibody and T-cell responses as well as challenged with a lethal dose of PR/8/34. Mice were monitored for clinical signs of infection, changes in body weight, and mortality. Results: Mice vaccinated with heat-inactivated virus combined with JVRS-100 adjuvant, showed increased HAI (450 vs. 30, P=0.007) and T-cell responses based on splenocyte restimulation (7700 vs. 30 pg/ml, P&lt;0.001) when compared with unadjuvanted virus. Mice vaccinated with homologous virus with adjuvant exhibited no weight loss at six days post infection (+5.4% vs. -15.3%) compared with unadjuvanted virus following 6xLD50 challenge with PR/8/34. Mice vaccinated with JVRS-100-adjuvanted heterologous virus also showed less weight loss (-7% vs. -26.7%, P=0.004) compared with unadjuvanted virus following 6xLD50 challenge with PR/8/34. Conclusion: Heat-inactivated whole influenza virus adjuvanted with JVRS-100 showed increased immunogenicity and greater protection from homotypic and heterosubtypic lethal challenge. Adoptive transfer studies suggest that the cross-protection is due at least in part to a cross-reactive antibody response.
Topical patch vaccines target antigen to cutaneous dendritic cells efficiently inducing potent cell mediated immune responses (132.19)
Authors: Erdos Geza , Toussaint Helene , Williams Melissa , Ozdoganlar Burak O. , Falo, Jr. Louis D. 
Published: January 2023

Abstract
The skin contains a high density of accessible dendritic cells making it an attractive target for vaccine delivery. However, the remarkable barrier function of the skin continues to present a formidable physical obstacle to cutaneous vaccine design. To selectively overcome this obstacle, we evaluated the capacity of dissolving microneedle arrays (MNAs) to deliver protein vaccines to the skin and skin DCs. In initial experiments, biocompatible solid MNAs were fabricated with integrated particulate tracers. Image analysis demonstrated that these MNAs effectively penetrated both mouse and human skin. Further, following patch immunization labeled particulates were identified in dendritic cells in human and murine skin, and in the draining lymph nodes of immunized animals. To evaluate immunogenicity, we incorporated OVA as a soluble (sOVA) or particulate (pOVA) model protein antigen into fabricated patches, and evaluated CTL responses in immunized animals. Patch delivery of low doses of either sOVA or pOVA efficiently induced potent CTL responses comparable to those generated by optimized gene gun based immunization. Taken together, these results demonstrate the feasibility of antigen delivery for cutaneous immunization, and support the development of topical patch vaccines as an efficient and potent strategy for protein antigen based immunization. Funded by NIH.