bluuebunny/crossref_metadata_2025_split huggingface dataset:
Real-time imaging of TCR dynamics in vivo reveals rapid TCR clustering and internalization in response to antigen (33.13)
Authors: Friedman Rachel S. , Beemiller Peter J. , Sorensen Caitlin M. , Krummel Matthew F. 
Published: January 2023

Abstract
Real-time in vivo imaging has given insight into the cellular dynamics of T cell activation, but less is known about receptor and signaling dynamics in vivo. Live imaging of subcellular signaling complexes has been challenging in living tissues due to low physiological molecular densities, inherent limitations of fluorophores, and strong autofluorescence of tissues. Our goal was to characterize T cell receptor (TCR) dynamics in naïve T cells in the lymph node during their first encounter with dendritic cells (DCs) bearing cognate antigen. To do so we developed a mouse expressing a GFP tagged OT-I TCR to track surface and intracellular TCR, and used image processing facilitated by co-labeling of the T cells to isolate specific fluorescence. In the absence of antigen, the TCR was mainly confined to the cell surface without consistent polarity; however, a small percentage of T cells transiently clustered their TCR. Within 30 min of antigen recognition, many T cells stopped crawling and clustered their TCR. Surprisingly, detectable antigen-dependent TCR clustering duration was short (7.1±5.1 min) and as TCR clustering declined, TCR internalization increased. Thus, naïve T cells in the lymph node can signal through the TCR in response to their first interaction with DCs bearing antigen and these contacts lead to rapid TCR clustering and internalization.
This work was funded by the Larry L. Hillblom Foundation and NIH R21.
Identification through ENU genome-wide mutagenesis of a novel mutant mouse that expresses a non-functional induced T cell kinase (Itk) with 8-amino acid insertion (33.30)
Authors: Kung John T 
Published: January 2023

Abstract
With the human genome sequenced, the race is on to understand the functions of the 30,000 or so genes. Ethyl-nitrosourea (ENU) was used to treat C57BL/6 (B6) male mice, followed by a 3-generation breeding scheme to generate G3 mice that were screened for immunological phenotypes under recessive genetic control. The primary phenotypes of one mutant, P-135, were increased CD44hi representation among both CD4 and CD8 T cells and highly elevated serum IgE. Recessive mode of inheritance was confirmed by mating an affected G3 male to wildtype B6 females to generate F2 offspring. Affected G3 mice were outcrossed to generated F2 offspring and the affected F2 mice were subjected to linkage analysis by haplotype interval mapping (Neuhaus and Beier, 1998. Mammalian Genome 9, 150-154). The mutant gene was found to be linked to chromosome 11. Through DNA sequencing of genes located within the linked region, a point mutation (T to C) in the 12th intron was identified. This point mutation resulted in the creation of a new splice donor site which in turn resulted in a 24-bp (8 amino acids, RVRAGMCR) in-frame insertion. P-135 mutant mice thus carries a non-functional Itk protein with 8 amino acid insertion and may be useful in defining structural-functional relationships of Itk.
Pregnancy primes an antigen specific maternal regulatory T cell response to fetal male antigen (33.6)
Authors: Kahn Daniel A , Baltimore David 
Published: January 2023

Abstract
Maternal CD4+ T cells awareness of fetal antigens is not well studied. T regulatory (Treg) cells, a powerful subset of CD4+ T cells, are capable of limiting robust immune responses. Tregs are implicated in maternal tolerance of the fetus.
Methods
Virgin female 6 week old C57BL/6 mice bred with male littermates or immunized with KLH/CFA. Splenocytes from virgin, parous, and pregnant mice (day post coitus 6.5, 12.5, and 18.5) and from KLH/CFA immunized mice on day 7. Proliferative response to CD4+ T cell peptide epitope of the H-Y antigen (DBY), control peptide (LSO), or KLH determined in co-cultures. Treg isolation by magnetic bead separation of CD25+ cells.
Results
Pregnant and parous splenocytes proliferate in response to DBY. Virgin splenocytes do not respond to DBY. Depletion of Tregs augments response to DBY in the context of pregnancy; not in virgin mice. Earlier in pregnancy, more male fetuses are required for a detectable response than later in pregnancy. Tregs isolated from pregnant mouse splenocytes co-cultured with Treg depleted splenocytes from KLH/CFA immunized mice are able to suppress proliferative response to KLH only in the presence of DBY.
ConclusionsPregnancy leads to maternal CD4+ T cell awareness of the fetus in proportion to the antigenic dose and is associated with a regulatory T cell response. Tregs are functional (suppressive) only in the presence of the antigen to which they were developed.
Critical role of CIN85 in Cbl-mediated regulation of B cell function after antigen receptor stimulation (34.11)
Authors: Niiro Hiroaki , Jabbarzadeh Tabrizi Siamak , Aizawa Kumiko , Inoue Yasushi , Arinobu Yojiro , Akashi Koichi 
Published: January 2023

Abstract
Upon B-cell receptor (BCR) ligation by autoantigens, B cells normally undergo deletion (apoptosis), inactivation (anergy), or receptor editing. These processes are finely tuned by positive and negative signals downstream of B cell receptor (BCR). Aberrant regulation of BCR signals may thus lead to activation and expansion of autoreactive B cells.
B cell adaptors function as molecular conduits to effectively link upstream signals to specific downstream branches. Cbl is an E3 ubiquitin ligase that is highly expressed in self-tolerant lymphocytes. Notably, B cell-specific ablation of Cbl in mice manifests SLE-like autoimmune disease, suggesting a pivotal role of Cbl in BCR-mediated tolerance induction. A novel Cbl-interacting adaptor termed CIN85 (c-Cbl interacting protein of 85 kDa) was recently identified, however little is known about a role of CIN85 in B cell function.
In this study we demonstrate that upon BCR stimulation, CIN85 strongly associated with Cbl as well as BLNK. This association required the SH3 domain of CIN85. The experiments using CIN85-overexpressing and CIN85 knockdown B cells showed that CIN85 is critical for Cbl phosphorylation, which in turn regulates BCR-induced calcium flux and activation of Syk and PLCγ2. CIN85 knockdown in human primary B cells resulted in their enhanced survival and proliferation by upregulating expression of bcl-2 family and cyclin genes following BCR stimulation. Together, these findings suggest a critical role of CIN85 in Cbl-mediated regulation of B cell function after BCR stimulation.
Tissue-Specific Cues Direct Fate Decisions of Mucosal Memory T Cells (39.21)
Authors: HUANG Yujun , Wang-Zhu Yiran , Schoenberger Stephen , Kronenberg Mitchell , Cheroutre Hilde 
Published: January 2023

Abstract
Mucosal memory CD8 T cells are characterized by their location at peripheral interfaces and the ability to immediately exhibit effector functions. By contrast, lymphoid memory T cells reside within lymphoid tissues and require further differentiation to become effectors. Both memory subtypes contribute to protective immunity, but the accumulation of mucosal effector memory T cells at pathogen portals forms a central component of protective immunity. The process that drives the compartmentalization and functional differentiation of this memory subset is poorly understood. Here we show that the thymus leukemia (TL) antigen, a ligand for the TCR co-repressor, CD8αα, serves a key role in the molecular mechanism that determines the fate of mucosal CD8αβ+ effector T cells. TL specifically controls the survival of CD8αα+CD8αβ+ effectors, but eliminates CD8αα negative CD8αβ effectors. This TL-CD8αα-mediated process allows the intestinal environment to selectively preserve the "fittest" effector cells to become mucosal memory T cells that form the first line of immune protection.
Alcohol consumption induces regional immune dysfunction (39.22)
Authors: Brand Rhonda M , Donahue Cara , Miller Megan , Plowey Jeff , Falo Louis D 
Published: January 2023

Abstract
Substantial clinical evidence suggests that alcohol abuse suppresses both innate and adaptive immune responses leading to more infections, greater susceptibility to cancer, and delayed recovery from trauma. The mechanisms underlying alcohol induced immune dysfunction are largely unknown and potentially include effects on dendritic cell function. Though published studies have focused on alcohol effects on systemic immune responses, it is well established that alcohol ingestion results in regional differences in ethanol concentration. This suggests the hypothesis that regional differences in ethanol induced DC dysfunction may lead to differences in regional immunity. Using the well defined Meadows-Cook model of alcohol ingestion, we immunized ethanol and control fed mice either cutaneously or hepatically with plasmid DNA encoding the model antigen ovalbumin. CTL and DTH responses were significantly reduced in ethanol fed mice compared to controls after liver targeted immunization. In contrast, minimal differences were seen between ethanol fed mice and controls after skin targeted immunization. These data suggest that the immunosuppressive effects of alcohol ingestion may be limited to regions of high ethanol exposure, and that intact immune systems in ethanol spared regions such as the skin could be used to induce effective immunity in individuals with significant alcohol ingestion. Funded by NIH.
IL-17 is required for protective immunity to nasal Brucella infections in an IFN-γ-dependent fashion (39.23)
Authors: Skyberg Jerod A. , Thornburg Theresa , Pascual David W. 
Published: January 2023

Abstract
Th1 and Th17 cells are the principal mediators of the inflammatory pathways. While the role of Th1 cells have been extensively studied for Brucella infections, there is little information on the function of IL-17 in response to Brucella infections. To evaluate such role, mice functionally deficient in IL-17 or IL-17 receptor (IL-17R-/-) mice were more susceptible to nasal Brucella infection than immunocompetent mice as evidenced by increased splenic weights and splenic CFUs. Interestingly, inhibition of IL-17 failed to enhance the IFN-γ response, but instead RT-PCR analysis revealed greater lung IL-13 mRNA transcripts than lungs from IL-17-sufficient mice. Splenic mononuclear cells from Brucella-infected mice depleted of IL-17 produced more IL-10, but less IFN-γ and IL-17 upon subsequent restimulation with heat-killed brucellae than the same cells from IL-17-sufficient mice. Interestingly, splenic mononuclear cells from infected IFN-γ-/- mice produced less IL-17, indicating that mice blocked in either IL-17 or IFN-γ pathways are impaired in their ability to produce either cytokine when infected with Brucella. These results implicate IL-17 is important for protection against nasal Brucella infections, and that IL-17 in combination with IFN-γ modulates the host immune responses to Brucella. This work is supported by USDA 2007-0612 and 2008-03776.
Expansion of CD4loCD14+CD40+ LKLF Hi CD4+ T cells correlates with acute pulmonary inflammation and provides a mechanism for impaired pathogen clearance in Cystic Fibrosis (39.24)
Authors: Pennock Nathan D , Vaitaitis Gisela M , Waid Dan M , Saavedra Milene T , Wagner, Jr David H 
Published: January 2023

Abstract
Cystic Fibrosis (CF) is characterized by chronic infection of the lung by bacterial pathogens with a subsequent robust but ineffectual neutrophilic immune response. As a result, patients' airways sustain irreversible injury, with respiratory failure accounting for &gt;80% of deaths from the disease. Examination of lymphocyte/monocyte subsets in the peripheral blood of CF patients revealed a greatly expanded CD4+ T cell subset characterized by a very high expression level of the cell survival/quiescence transcription factor Lung Kruppel Like Factor, LKLF. Furthermore, cells exhibited co-expression of CD40 as well as the pattern recognition coreceptor CD14. Importantly, this observation was mirrored in airway submucosa and pulmonary lymph nodes isolated from CF lung transplant recipients. In a majority of patients, this expanded T cell subset from the peripheral blood was found to produce high levels of IL-4, which coincides with previous observations of Th-2 bias in CF patients infected with Pseudomonas aeruginosa. In vitro assays illustrated that simultaneous cross-linking of CD40 and CD3 on this T cell subset results in the elevation of LKLF. Paired with previous observations of elevated circulating soluble CD40 ligand in CF patients that correlate with worse pulmonary function, this data suggests a mechanism for the persistence of the deleterious Th-2 cytokine producing T cells that hamper the effectual clearance of lung pathogen. Importantly alterations in cell number within this T cell subtype from the peripheral blood correlate with observed severity of pulmonary inflammation and provide a means of evaluating efficacy of treatment. Research support;NIH:DDK (Wagner),Cystic Fibrosis Foundation(Saavedra)
A novel nanosize protein-carrier for the development of adjuvant- free nasal vaccine (39.33)
Authors: Yuki Yoshikazu , Nochi Tomonori , Mejima Mio , Kataoka Nobuhiro , Takahashi Haruko , Sawada Shinichi , Kohda Tomoko , Kazaki Shunji , Akiyoshi Kazunari , Kiyono Hiroshi 
Published: January 2023

Abstract
Most of inactivated or subunit type vaccines are poor immunogens to both systemic and mucosal immune compartments when given mucosally without mucosal adjuvant. We developed an effective intracellular vaccine delivery system with self-assembled nanosize cationic hyrogels (nanogel), which composed of a polysaccharide with ethylendiamine group of cholesteryl group-bearing pullulan. When hold protein antigen in a segregated nanomatrix, the nanogel acts as artificial chaperones, which protect against the aggregation of denature proteins and assist in the refolding of protein after release. In this study, we demonstrated that nasally administered nanogel holding a subunit type of C. botulinum antigen (BoHc-nanogel) was effectively taken up by nasophalynx-associated lymphoid tissue and induced brisk levels of antigen-specific systemic and mucosal immune responses even in the absence of mucosal adjuvant. In contrast, nasal administration with commercialized cationic liposome (Pro-ject) holding the same amount of BoHc without adjuvant induces antigen-specific low systemic and no mucosal antibody immune responses. In addition, mice nasally immunized with BoHc-nanogel protected not only systemically challenged but also nasally challenged botulinum toxin. These data suggest that nanogel-based mucosal vaccination is a novel strategy for the induction of protective immunity without the use of mucosal adjuvant.
Alveolar macrophages control the quality of CD4+ T lymphocytes in the respiratory tract upon recombinant adenoviral mucosal vaccination (39.34)
Authors: Acosta-Ramirez Elizabeth , Xing Zhou , Schwendener Reto , Wang Jun 
Published: January 2023

Abstract
Infectious diseases are acquired through mucosa surfaces making mucosal vaccination important for their control. Previously, using a recombinant adenoviral-based (Ad) vaccine we have demonstrated that a single mucosal but not parenteral immunization confers protection against intracellular bacterial challenge in the lung. Here, we examined the mechanisms regulating induction of OVA-specific CD4 T-cell responses at mucosal and systemic compartments in Balb/C mice after intranasal (i.n.) or intramuscular (i.m.) vaccination with an Ad expressing OVA. We found that i.n. vaccination induced local OVA-specific T cell activation whereas i.m. vaccination induced both local and systemic T-cell activation despite of the dosage used for vaccination. Remarkably, i.n. immunization induced a characteristic proliferative kinetic different from the one induced by i.m. vaccination. Furthermore, i.n. vaccination induced a highly polarized Th1 response while i.m. immunization induced a mixed Th1/Th2 response. Of importance, in vivo depletion of alveolar macrophages led to similar T-cell proliferation kinetics and comparable cytokine production in mice receiving i.n. and i.m. vaccination. Our data suggests that alveolar macrophages provide a special microenvironment at the mucosal site, which control the quantity and quality of CD4 T memory cells and protective immunity against mucosal infectious challenges.
Effects of growth phase and substrate on immunomodulatory activity of lactobacilli (39.37)
Authors: Cooper William , Masotti Adriana , Wagar Lisa , Champagne Claude , Buckley Nicole , Forrester Sean , Green-Johnson Julia 
Published: January 2023

Abstract
Immunomodulatory effects of lactic acid bacteria vary with strain and may vary with growth phase and medium. The ability of different lactobacilli strains (Lactobacillus helveticus R0052, L. rhamnosus R0011, L. rhamnosus GG) at different growth phases to modulate macrophage and intestinal epithelial cell cytokine production following a proinflammatory challenge was examined. Soy and milk ferments were also prepared with strains shown to grow well in soy milk (Bifidobacteria longum R0175 and Lactobacillus helveticus R0052). Modulation of cytokine production by human macrophage cell lines (U937 and THP-1) and intestinal epithelial cells (HT-29) induced by Tumor Necrosis Factor αwas assayed by ELISA for interleukin-8 (IL-8) production. Strain-dependent differences were observed in the ability of spent MRS broths from log versus stationary growth phase, and of soy and milk ferments to down-regulate IL-8 production by challenged U937 cells. Overall, variation in immunomodulatory activity of these lactic acid bacteria reflects not only strain variation but potentially also differences in growth phase and substrate.
Funded by NSERC, Lallemand Inc. and the Canadian Space Agency.
Effects of dietary algal supplementation on bovine immunity (39.50)
Authors: Green-Johnson Julia M , Johnson Kathy , Masotti Adriana , Wagar Lisa , Fillmore Sherry , Glover Kathleen , MacLaren Leslie , Enright Cathy , Fredeen Alan 
Published: January 2023

Abstract
Marine algae are primary producers of long chain omega-3 fatty acids and can serve as novel dairy feed supplements with the added potential to influence bovine immunity. Our objective was to assess the immunomodulatory impact of dietary supplementation with different algal types (macroalgae and microalgae). Effects of dietary algal supplementation were examined using a crossover design and a 28 day feeding period with each algal type. Primary immunization with Keyhole Limpet Hemocyanin (KLH) was carried out 8 days into the trial, with secondary immunization 14 days later. KLH-specific serum IgG responses were significantly higher in cows receiving macroalgae-supplemented feed than in cows receiving microalgae-supplemented feed. In contrast, cows receiving microalgae-supplemented feed showed significantly higher levels of macrophage oxidative burst activity in response to PMA challenge in vitro. These results suggest that dietary supplementation with algal products has immunomodulatory potential and also indicates that the effects on cellular and humoral immunity are dependent on the type of algae used.
Funded by the Atlantic Innovation Fund.
Stratification of antigen-presenting cells within the normal cornea (39.57)
Authors: Knickelbein Jared E. , Watkins Simon C. , Hendricks Robert L. 
Published: January 2023

Abstract
The composition and location of professional antigen presenting cells (APC) varies in different mucosal surfaces. The cornea, long considered an immune privileged tissue devoid of APCs is now known to host a network of bone marrow-derived cells. Here, we utilized transgenic mice that express EGFP from the CD11c promoter in conjunction with immunohistochemical staining to demonstrate an interesting stratification of APCs within non-inflamed murine corneas. CD11c+ DCs reside in the basal epithelium, many seemingly embedded in the basement membrane. Most DCs express MHC class II on at least some dendrites, which extended up to 50 µm in length and traversed up 20 µm tangentially nearly to the apical surface of the epithelium. The DC density diminished from peripheral to central cornea. Beneath the DCs and adjacent to the stromal side of the basement membrane reside CD11c-CD11b+ presumed macrophages that expressed low levels of MHC class II. Finally, MHC class II- CD11b+ cells filled the remainder of the stroma. This highly reproducible stratification of bone marrow-derived cells is suggestive of progression from APC to barrier function.
Expression of indoleamine 2,3-dioxygenase (IDO) in multiple myeloma promotes the generation of regulatory T cells: Investigations into the role of hepatocyte growth factor (40.1)
Authors: Rutella Sergio , Bonanno Giuseppina , Corallo Maria , Mariotti Andrea , Procoli Annabella , Majolino Ignazio , De Cristofaro Raimondo 
Published: January 2023

Abstract
Multiple myeloma (MM) is a plasma cell (PC) malignancy with a unique ability to evade immunosurveillance through the induction of APC dysfunction and the expansion of regulatory T cells (Treg). Hepatocyte growth factor (HGF) is elevated in MM and confers an unfavorable prognosis. We have previously shown that HGF induces the tryptophan (Trp)-degrading enzyme indoleamine 2,3-dioxygenase (IDO) in human monocytes.
In this study, IDO-1 was detected in OPM-2, MOLP-8 and LP-1 MM cell lines but not in HUNS-1 cells. Kynurenine (KYN) production and Trp depletion were evidenced in supernatants of IDO-1+ MM cells, suggesting that IDO-1 may be functional. When challenged with exogenous HGF, IDO- HUNS-1 cells up-regulated IDO-1 mRNA 6-fold on average. IDO-1+ MM cells promoted the conversion of allogeneic CD4+CD25- T cells into bona fide CD4+FoxP3+ Tregs.
We also detected IDO-1 expression in 6 out of 10 PC samples from patients with MM. Higher percentages of CD4+FoxP3+ Tregs were measured in IDO-1+ compared with IDO-1- MM. Intriguingly, serum KYN levels were higher in IDO-1+ compared with IDO-1- MM, suggesting that IDO-1 may mediate the in vivo expansion of Tregs through immunosuppressive KYN. Finally, serum HGF was over-represented in IDO-1+ compared with IDO-1- MM, providing indirect clues in favor of in vivo HGF effects on IDO-1 expression.
Collectively, these data point to HGF as a potential inducer of IDO-1 in MM. Whether this will impact on the ability of the immune system to control disease progression remains to be prospectively investigated in larger cohorts of patients.
Activation of anti-tumor NK cells by gamma delta T lymphocytes through 4-1BB engagement (40.26)
Authors: Chapoval Andrei , Maniar Amudhan , Zhang Xiaoyu , Gastman Brian R , Pauza C. David , Strome Scott E 
Published: January 2023

Abstract
Emerging evidence suggests that gamma delta (γd) T cells play a critical role in bridging innate and adaptive immune responses, yet the mechanisms responsible for these effects are uncertain. In this study we show that isopentenyl pyrophosphate (IPP) activated γd T lymphocytes induce cytolytic function of Ig stimulated NK cells. These "costimulated" NK cells up regulate the CD54 and CD69 markers of cellular activation and also display enhanced cytotoxicity against squamous cell carcinoma of the head and neck (SCCHN). Based on our findings that "co stimulation" of Ig primed NK cells by γδ T lymphocytes requires cell-to-cell contact, we performed a comprehensive expression analysis of TNFSF members expressed on the surface of IPP activated γδ T lymphocytes, and found that these cells express high levels of 4-1BB ligand. Stimulation of Ig primed NK cells with 4-1BBL but not mock transfected P815 cells, induces up regulation of CD54 and CD69 and blockade of 4-1BB/4-1BBL interaction by soluble 4-1BB fusion protein decreased activation of NK co-cultured with γδ T lymphocytes. In addition, we observed that NK cells cultured with Ig/γδ T lymphocytes up regulate NKG2D and anti-NKG2D mAb blockade partially abrogates the cytolytic effects of these "costimulated" NK cells against SCCHN. Taken in concert, our data indicate that γδ T lymphocytes induce activation of Ig primed NK cells through coordinated 4-1BB -4-1BBL interactions and up regulation of targeted cell death pathways.