bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lentivector immunization stimulates potent CD8 T cell responses against melanoma self antigen tyrosinase related protein 1 and generates antitumor immunity in mice (41.10)
Authors: He Yukai , Liu Yanjun , Peng Yibing , Munn David 
Published: January 2023

Abstract
Recombinant lentivector immunization has been demonstrated to induce potent CD8 T cell responses in vivo. In this study, we investigated whether lentivector delivering a self/tumor Ag, tyrosinase related protein 1 (TRP1), could stimulate effective antitumor T cell responses. We found that immunization with lentivector expressing mutated TRP1 (muTRP1) Ag elicited potent CD8 T cell responses against multiple TRP1 epitopes. Importantly, the activated CD8 T cells effectively recognize wild type (wt) TRP1 epitopes. At peak times, as many as 10% of CD8 T cells were effector cells against TRP1 Ag. These cells killed wtTRP1 peptides-pulsed target cells in vivo and produced IFN-gamma after ex vivo stimulation. The CD8 T cell responses were long-lasting (3-4 weeks). Immunized mice were protected from B16 tumor cell challenge. In a therapeutic setting, lentivector immunization induced potent CD8 T cell responses in tumor bearing mice. The number of infiltrating T cells and the ratio of CD8:CD4 were dramatically increased in the tumors of immunized mice. The tumor infiltrating CD8 T cells (TIL) were functional and produced IFN-gamma. The potent CD8 T cell responses stimulated by lentivector immunization eliminated small 3 day subcutaneous B16 tumors and strongly inhibited the growth of more established 5 day tumors. These studies demonstrate that genetic immunization with lentivector expressing mutated self/tumor Ag can generate potent CD8 T cell immune responses and antitumor immunity that prevent and inhibit B16 tumor growth, suggesting that lentivector immunization has the potential for tumor immunotherapy and immune prevention.
Enhancing HER2/neu Specific T-cell Therapy by Targeting the Tumor Endothelium (41.11)
Authors: Lai Vy Phan , Buckanovich Ronald J , Gad Ekram , Bigdeli Laleh , Disis Mary L 
Published: January 2023

Abstract
Success of adoptive T cell therapy for cancer can be hindered by the tumor stroma, which consists of factors that not only promote tumor growth but also prevent egress of tumor-specific T cells into the tumor parenchyma. We recently demonstrated that T cell transit through tumor vasculature is regulated by the endothelin B receptor (ETBR). In addition, the endothelin axis plays a major role in tumor cell growth, apoptosis, angiogenesis and metastasis. The purpose of this study was to determine if, in breast cancer, targeting endothelin-1 via ETBR blockade might abrogate the endothelial barrier and increase T cell homing to tumors, thereby augmenting the efficacy of HER2/neu antigen specific Type 1 helper (Th1) cell therapy of spontaneous HER2/neu+ mammary tumors. By reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC), we found overexpression of ETBR in peritumoral stroma of spontaneous mammary tumors derived from HER2/neu transgenic mice. We treated tumor-bearing mice with HER2/neu specific T cells alone or HER2/neu specific T cells + BQ-788, a specific ETBR inhibitor peptide. The addition decreased tumor growth by 84%. Thus, in this model of HER2/neu mediated breast cancer, targeting the tumor endothelium can significantly enhance the clinical efficacy of HER2/neu specific Th1 cells. Studies are underway to evaluate the biological mechanisms induced by endothelin targeting that cooperate with T cell therapy in the resulting significant success of the combined treatment. The data provide a rationale for combining tumor antigen specific T cell therapy with endothelin blockade to induce an effective anti tumor response in breast cancer.
Generation of long-term protection from acute lymphoblastic leukemia (ALL) by an immune stimulant in a syngeneic murine model (41.21)
Authors: Seif Alix Eden , Barrett David M. , Brown Valerie I. , Grupp Stephan A. , Reid Gregor S.D. 
Published: January 2023

Abstract
ALL is the most common childhood cancer, and relapsed ALL remains a major cause of mortality. The success of stem cell transplant for relapsed ALL suggests immune therapy may be effective. We previously showed CpG ODN, which stimulate immune activity through TLR9, enhance allogeneic Th1 responses and reduce in vivo leukemic burden of primary human ALL xenografts. We are now evaluating anti-leukemia activity induced by CpG ODN in a pre-B ALL minimal residual disease (MRD) model. In mice given 106 syngeneic lymphoblasts, CpG ODN treatment universally induced remissions for &gt;6 months; control mice died of leukemia at a median of 35 days (p=0.0001). CpG ODN treatment of Rag-1-deficient mice significantly decreased peripheral disease at day 28 (p&lt;0.05) but did not induce long-term remissions (p&lt;0.01). T cell-depletion after CpG ODN similarly abrogated protection from ALL, resulting in a median survival of 39 days (p&lt;0.0001). Mice in remission after CpG ODN were protected from rechallenge with ALL blasts, while age-matched leukemia-naïve controls rapidly died of disease (p=0.0005). A protective effect is also noted in mice treated with CpG ODN versus PBS following standard ALL chemotherapy (p&lt;0.03). To our knowledge, this is the first demonstration that CpG ODN induce a durable remission in an ALL MRD model by triggering an adaptive immune response, suggesting this treatment may have clinical utility in children with MRD.
Adoptive therapy of a fast growing tumor with reactivated memory T cells combined with anti-4-1BB eliminates tumors through effects of anti-4-1BB on transferred but not host cells (41.37)
Authors: Lin Gloria Hoi Ying , Liu Yuanqing , Ambagala Thanuja , Kwon Byoung S , Watts Tania H 
Published: January 2023

Abstract
The ability to expand memory T cells with cytokines ex vivo has greatly increased the practicality of adoptive immunotherapy for cancer. Central memory T cells generated in IL-15 have the advantage of longevity after subsequent in vivo transfer, whereas effector cells have the advantage of more immediate tumor cell killing. Here we report that in adoptive immunotherapy against a pre-established EG.7 tumor, a greater proportion of tumor bearing animals were cured with reactivated memory T cells as compared to animals that received central memory cells. Although central memory cells showed an initial survival advantage in the host, reactivated memory cells expanded more rapidly in the tumor, draining lymph node and spleen, resulting in increased accumulation over time. Co-administration of reactivated memory T cells with anti-4-1BB agonist antibody further potentiated the therapeutic effect. Anti-4-1BB therapy resulted in expansion of host NK, NKT, CD11c+ cells, CD4 and CD8 T cells as well as the adoptively transferred T cells. However, use of 4-1BB-deficient hosts showed that the expression of 4-1BB on adoptively transferred T cells was sufficient for the therapeutic effect. Thus, the combination of reactivated memory T cells and stimulatory anti-41BB antibody represents a superior immunotherapy for a rapidly growing cancer, largely through effects of anti-4-1BB on transferred effector T cells.
A novel form of 4-1BBL demonstrates better immunomodulatory activity than an agonistic anti-4-1BB Ab without Ab associated severe toxicity (41.4)
Authors: Schabowsky Rich-Henry , Elpek Kutlu G , Madireddi Shravan , Sharma Rajesh K , Yolcu Esma S , Bondura-Morgan Laura , Mittler Robert S , Shirwan Haval 
Published: January 2023

Abstract
Costimulatory molecules of the CD28 and TNFR superfamilies modulate innate, adaptive, and regulatory immune responses, and therefore agonistic ligands to these receptors have potential to serve as effective immunomodulatory components of therapeutic vaccines. However, the use of costimulatory agonists for vaccines in humans requires the generation of agonists that transduce appropriate stimulatory signals without toxic side effects. We hypothesized that signaling by natural ligands may have better efficacy and safety compared to agonistic Abs. Our laboratory recently developed a novel form of soluble 4-1BB ligand (SA-4-1BBL) which forms functional tetramers/oligomers. SA-4-1BBL demonstrated superior immunostimulatory activity than an agonistic 4-1BB Ab (3H3), and importantly lacked Ab associated toxicity as manifested by greatly enlarged spleen and LNs, non-specific T cell proliferation, hepatitis, and increased T cell and systemic inflammatory cytokine production. Ab associated toxicity did not involve Fc receptor or complement activation, and adoptive transfer studies demonstrate that CD8+ and CD4+ T cells were the direct targets of Ab mediated toxicity. Our findings support further development of this novel form of soluble 4-1BBL as an immunomodulatory component of therapeutic vaccines.NIH (R43 AI071618, R41 CA121665, R44 AI071618, and R43AI074176), KLCRP, and W.M. Keck Foundation.
Autophagosome vaccine cross-protects and breaks tumor immunology paradigm: a p62-dependent mechanism? (41.41)
Authors: Twitty Chris G , Jensen Shawn , Hu Hong-Ming , Fox Bernard A. 
Published: January 2023

Abstract
Vaccination with MCA sarcoma cells protects from a subsequent challenge with the same, but not other MCA sarcomas. This observation of unique tumor-specific protection is a well-established paradigm (Prehn and Main 1957). We postulated that all MCA sarcomas overexpress common mutated gene products with a short half-life (SLiPs), but only the unique tumor-rejection antigens are stable enough to be cross-presented and induce anti-tumor immunity. We recently reported that with proteosomal blockade, SLiPs could be isolated in autophagic-containing (LC-3) vesicles (termed DRibble) that were efficient for cross-presentation of antigens (Li, Y. et al). Here we examined if vaccination with DRibbles from one MCA sarcoma would cross-protect from a challenge with an antigenically distinct MCA sarcoma. While whole tumor cell vaccines provide protection to unrelated tumor in 0 of 9 comparisons, DRibble vaccines provided significant (p&lt;0.05) protection for 8 of 9 tumors (n=10-25 mice/group). Using a model system we showed that SLiPs play a critical role in providing antigen to DRibbles and that siRNA knock down of p62 reduced SLiPs and the capacity of DRibble to stimulate T cells. These results have shaped a model where polyubiquitinated SLiPs spared degradation by the proteasome, are picked up by p62 and delivered to autophagic vesicles. Used as a vaccine, these vesicles can cross-present neo-antigens capable of inducing cross-protection of chemically induced sarcomas. Supported by CA80964 (BAF), CA107243 (HH) the Chiles Foundation and the Murdoch Trust.
Design of artifical tumor cells to induce Natural Killer cell proliferation (42.13)
Authors: Huppert Volker , Müller Sabine , Turkiewicz Anna , Fahrendorf Melanie , Schmitz Jürgen 
Published: January 2023

Abstract
Natural Killer (NK) cells respond to tumor cell contact by cellular cytotoxicity, cytokine release and proliferation upon recognition of the tumor cells by an array of co-activating receptors such as Natural Cytotoxicity Receptors (NKp30, NKp44, NKp46), NKG2D, CD2, NKp80 and CD16 (binding to tumor cell reactive antibodies). Responsiveness of NK cells to tumor cells depends on complicated parameters of the individual's immune system and, if evaluating NK cell function in coculture of NK cells with tumor cell lines, requires maintenance culture of those cell lines and will cause further variation of experimental conditions.
We designed artifical tumor cells to tightly control human NK cell culture conditions. Tumor cells are mimicked by large magnetic spheres ("MACSiBead Particles") coated with ligands to activating NK cell receptors. Combinations of antibodies to NKp46 (CD335), NKG2D (CD314), Nkp80, CD2 have been evaluated as ligands and were feasible to induce a median 30 to 100 fold expansion of MACS isolated human NK cells within 2 to 3 weeks.
We demonstrated that cellular cell culture components such as tumur cell lines or allogeneic peripheral blood mononuclear cells (PBMC) are not required to induce significant NK cell proliferation and that artifical tumor cells (cell sized antibody coated magnetic particles) can be used instead and may provide a tool for NK cell proliferation for either diagnostic / analytical or NK cell therapy production purposes.
Qdot® nanocrystal conjugates in multispectral flow cytometry (42.14)
Authors: Godfrey William L. , Zhang Yu-Zhong , Jaron Shulamit , Buller Gayle M. 
Published: January 2023

Abstract
Qdot® nanocrystal-antibody conjugates allow researchers to detect more parameters in a sample with manageable spectral overlap compensation between fluorophores. These semiconductor nanocrystals optimally excite with ultraviolet or violet light, and provide symmetrical emission peaks with up to 400 nm Stokes shifts. In this study, we review the use of Qdot® nanocrystal and conventional fluorophore conjugates with particular attention to filter selection to minimize the within-laser and cross-laser compensations. All experiments were performed using standard staining protocols with human peripheral blood leucocytes and analyzed using a BD LSR II flow cytometer. Example results: Qdot® 655 and Qdot® 605 nanocrystal conjugates require less than 5% compensation, and the Qdot® 655 nanocrystal can require &lt;20% compensation versus APC. Qdot 705 nanocrystal requires &gt;100% compensation versus RPE-Cy5.5, but filter selection can reduce compensation versus RPE-Alexa Fluor® 700 conjugates to &lt;30%. By optimizing individual channels, 8 to 12 color experiments have been run with good resolution of cell populations using 2 to 4 Qdot® nanocrystal conjugates. The unique spectral characteristics of Qdot® nanocrystals can dramatically facilitate multicolor experimental design when performing multicolor flow cytometry experiments.
Supported by Life Technologies Corporation.
Validation of Bio-Plex Pro™ Human Cytokine Assays on magnetic microspheres (42.26)
Authors: Zhou Hui , Ma Li , Fedynyshyn Joe , Tan Woei , Wang Qian-Shu 
Published: January 2023

Abstract
Bio-Plex Pro(tm) cytokine, chemokine, and growth factors assays are magnetic bead-based multiplex assays designed to measure these cell signaling proteins in diverse matrices like serum, plasma and tissue culture supernatants. The abstract describes the development and validation of a Bio-Plex Pro(tm) Human Cytokine Assay Group II Panel. The assays are based on the Luminex® xMAP® technology capable of simultaneously quantifying 23 targets. Performance characteristics, including assay sensitivity, working ranges, precision, accuracy, linearity of dilution and parallelism are reported. Three assays were compared with commercially available ELISA kits using human serum samples (N=25). A good correlation was observed between the Bio-Plex Pro(tm) assays and the commercially available ELISA kits. Cross-panel-plexing between Bio-Plex Pro(tm) Human Cytokine Assay Group II and Group I panel was also investigated. The study showed that 16 of the 23 targets in Group II panel could be multiplexed with the 27-plex Group I panel. Analysis of normal and disease human serum samples using the Bio-Plex Pro(tm) human cytokine assays reported data consistent with published literatures.
Contribution of CD137 signaling to Hepatic Fibrosis via T cell and Macrophage Mediated Chronic Liver Inflammation (43.5)
Authors: Wang Shengdian , Wang Shengdian 
Published: January 2023

Abstract
It is accepted that viral clearance and disease pathogenesis during HBV infection are mediated by the immune responses. Molecular mechanisms of the emergence or regulation of the immune response in this process remain largely unknown. Here we characterized that CD137 signaling acted as a potential pathogenic factor in progression of chronic hepatitis B. CD137 signaling successively stimulated by agonist anti-CD137 mAb could induce persistent hepatic infiltration and IFN-¦Ã production of CD8 T cells in HBV tansgenic mice. Through secreted IFN-¦Ã, the activated CD8 T cells subsequently recruited macrophage into liver to produce pro-inflammatory cytokines, which caused chronic liver inflammation and hepatic fibrogenesis, and promoted hepatocarcinoma development. Moreover, expression of CD137L was indeed up-regulated in peripheral monocytes of CHB patients and closely correlated with liver cirrhosis and the plasma level of IL-8, but not with ALT. Stimulation of monocytes from cirrhotic patients with recombinant human CD137 ligand induced IL-8 secretion in vitro, a chemokine closely associated with enhanced liver fibrosis in chronic HBV patients.
These resutls support an important role of CD137 signaling in liver inflammation and fibrosis, indicating that CD137 pathway may be a target for immunotherapy of chronic hepatitis B. Our findings also suggest that IFN-¦Ã,secreted by CD8 T cells, plays pivotal roles in mediating chronic liver inflammation and fibrosis. Further elucidation on this question will be of great significance for the immunotherapy of chronic hepatitis B.
Effector T-cells control lung inflammation during acute respiratory virus infection by producing interleukin-10 (43.7)
Authors: Sun Jie , Madan Rajat , Karp Christopher L , Braciale Thomas J. 
Published: January 2023

Abstract
Activated antigen-specific T cells produce a variety of effector molecules for clearing infection, but also contribute significantly to inflammation and tissue injury. Here we report an anti-inflammatory property of anti-viral CD8+ and CD4+ effector T cells (Te) in the infected periphery during acute pulmonary virus infection. We find that, during acute influenza and other respiratory virus infection, IL-10 is produced in the infected lungs at high levels -- exclusively by infiltrating virus-specific Te, with CD8+ Te contributing a larger fraction of the IL-10 produced. These Te in the periphery simultaneously produce IL-10 and proinflammatory cytokines, and express lineage markers characteristic of conventional Th/c1 cells. Importantly, blocking the action of the Te-derived IL-10 results in enhanced pulmonary inflammation and injury associated with the viral infection. Our results demonstrate that anti-viral Te exert regulatory functions --
that is, fine-tune the extent of lung inflammation/injury associated with respiratory virus infection by the production of an anti-inflammatory cytokine.
Plasmacytoid dendritic cells control influenza-specific CD8 T cells numbers through regulated FasL expression during lethal influenza virus infections (44.3)
Authors: Langlois Ryan A , Legge Kevin L 
Published: January 2023

Abstract
Previously we have demonstrated that the difference in survival of high versus low dose conventional influenza infections is linked to a severe T cell lymphopenia in high dose infections similar to what has been described in lethal H5N1 infections. This difference in T cell numbers is mediated by a novel regulatory mechanism where lymph node dendritic cells (LNDC) eliminate influenza-specific CD8 T cells in a FasL/Fas dependent manner within the LN in high but not low dose infections. Currently, it is unknown which LNDC subsets and what additional factors are required for driving this CD8 T cell apoptosis. Our results demonstrate that high dose pDC directly induce apoptosis of influenza-specific CD8 T cells ex vivo. This elimination occurs in the absence of influenza antigen presentation as peptide pulsing of pDC abrogates the induction of apoptosis. Further pDC downmodulate FasL expression, a process that appears to be dependent upon rDC migration and reduced rDC production of IL-12 p40 homodimer during low dose infections, therein allowing influenza-specific CD8 T cells to escape pDC induced apoptosis. Our results detail a heretofore undescribed role for pDC during influenza virus infections whereby pDC control the overall magnitude of the influenza-specific CD8 T cell response within the LN and ultimately, therein contribute to the outcome of influenza virus infections.
Novel IL-17–producing memory cells are key to vaccine-based protective immunity against M.tuberculosis challenge (45.12)
Authors: Lin Yinyao , Guglani Lokesh , Strawbridge Heather , Ritchea Shane , Logar Alison , Khader Shabaana 
Published: January 2023

Abstract
Interferon-gamma (IFNγ) is essential for limiting Mycobacterium tuberculosis infection. Using the mouse model, we have recently shown that vaccination triggered accelerated Interleukin-17 (IL-17) and IFNγ responses by CD4+ T cells in the lung during M. tuberculosis aerosol challenge. We propose that vaccination induces IL-17- producing CD4+ T cells that populate the lung and, after challenge, trigger the production of chemokines. The induction of chemokines results in recruitment of IFNγ-producing CD4+ T cells from the lymphoid compartment and ultimately restricts bacterial growth. The differential ability of IL-17-producing memory cells to populate the lung compared to the IFN-γ-producing cells suggests that these two cell types differ in their ability to migrate in response to chemokines. In support of this, our data using adoptive transfer models and chemotaxis assays suggests that differential responsiveness to chemokines and retention in different organs may provide the basis for the differential tissue distribution of these two cell types. During progression from effector to memory T cells, we also show differential expression of activation markers on IL17-producing and IFNγ-producing T cells. This information will lead us to determine whether altering the nature of the lung-resident IL-17 producing population impacts protective efficacy of the recall response to M.tuberculosis challenge.
This work was supported by AI075106-01 and Children's Hospital of Pittsburgh.
The Lifespan of Naïve CD4 T cells is a Critical Determinant for the Cellular Aging in Periphery (46.2)
Authors: Tsukamoto Hirotake , Clise-Dwyer Karen , Huston Gail E. , Buck Amanda L. , Duso Debra K. , Haynes Laura , Swain Susan L. 
Published: January 2023

Abstract
Cell intrinsic functional defect in aged naive CD4 T cells is a key cause of an age-associated increase in susceptibility to infectious diseases. However the causes of the CD4 T cell defects and how they develop are largely unknown. We have suggested that many defects develop during the sojourn of the naïve CD4 T cells in the periphery. We report here that aged naïve CD4 T cells have a longer lifespan than young cells even when transferred to young intact hosts. In vitro studies suggest that their increased longevity is regulated by an intrinsic property for survival. Consistent with this, aged naïve CD4 T cells express reduced level of a pro-apoptotic molecule, Bim, and the reduction of Bim in naïve CD4 T cells is sufficient for longer lifespan in vivo. This longer lifespan of aged naïve T cells could contribute to the maintenance of T cell homeostasis in aged mice. On the other hand, shutdown of new T cell emigration from the thymus by thymectomy leads to long-term persistence of remaining naïve T cells in the periphery and accelerates development both of the functional defects and of increased lifespan in naive CD4 T cells. Neither of these is observed when new naïve T cells are derived from aged hematopoietic stem cells. Thus, these results suggest that development of functional defects in aged naive CD4 T cells is associated with long-term persistence of aged naïve T cells due to their increased lifespan in periphery. The authors have no financial conflict of interest. This work was supported by National Institutes of Health Grants AG025805, AG021600 and the Trudeau Institute.