bluuebunny/crossref_metadata_2025_split huggingface dataset:
The thymus leukemia antigen controls intraepithelial lymphocyte function and inflammatory bowel disease (39.53)
Authors: Olivares-Villagomez Danyvid , Mendez-Fernandez Yanice V , Parekh Vrajesh V , Lalani Saif , Vincent Tiffaney L , Cheroutre Hilde , Van Kaer Luc 
Published: January 2023

Abstract
Intestinal intraepithelial lymphocytes (IEL) bear a partially activated phenotype that permits them to rapidly respond to antigenic insults. However, this phenotype also implies that IEL must be highly controlled to prevent misdirected immune reactions. It has been suggested that IEL are regulated through the interaction of the CD8alpha-alpha homodimer with the thymus leukemia (TL) antigen expressed by intestinal epithelial cells. We have generated and characterized mice genetically deficient in TL expression. Our findings show that TL expression plays a critical role in maintaining IEL effector functions. Moreover, TL deficiency accelerated colitis in a genetic model of inflammatory bowel disease. These findings reveal an important regulatory role of TL in controlling IEL function and intestinal inflammation.
Pulmonary Dendritic Cells Trans-present IL-15 to Directly Promote the Survival of Effector Influenza Virus-Specific CD8 T Cells in the Lungs (39.6)
Authors: McGill Jodi , Legge Kevin L. 
Published: January 2023

Abstract
We have previously shown that aDC depletion at 48 hours post influenza virus infection results in increased mortality and decreased virus-specific CD8 T cell responses. Reconstitution with pulmonary DC rescues the CD8 T cell response in a manner that requires direct cell-cell contact, MHC I and viral antigen. In aDC depleted mice, influenza-specific CD8 T cells undergo increased apoptosis in the lungs and express decreased levels of the anti-apoptotic factor Bcl-2. Reconstitution with pulmonary DC results in decreased levels of T cell apoptosis, suggesting that pulmonary DC provide a survival signal to effector CD8 T cells in the lungs during infection. Here, we demonstrate that pulmonary DC express IL-15 and IL-15Rα, and blocking these molecules on their surface ablates the rescue of pulmonary CD8 T cell immunity. Together, our results suggest that virus-specific CD8 T cells require interactions with viral antigen-bearing, IL-15 trans-presenting DC in the lungs to up-regulate anti-apoptotic signals such as Bcl-2, promoting T cell survival and accumulation. Our results suggest a "two-hit" model of CD8 T cell activation during influenza infection where T cells require a primary interaction with DC in the lymph node to initiate programmed activation; and a novel, secondary interaction with viral antigen-bearing, IL-15 trans-presenting DC in the lungs to survive, accumulate and mediate viral clearance and recovery.
A pivotal role of stromal NFkappaB-inducing kinase in sphingosine 1-phosphate-medaited peritoneal B cell trafficking for the intestinal IgA production (39.9)
Authors: Kunisawa Jun , Gohda Masashi , Kurashima Yosuke , Ishikawa Izumi , Higuchi Morio , Kiyono Hiroshi 
Published: January 2023

Abstract
Our previous study showed that sphingosine 1-phosphate (S1P) regulates peritoneal B cell trafficking for the subsequent intestinal IgA production. In this study, we demonstrate that stromal NFkappaB-inducing kinase (NIK) is specifically involved in this pathway. Although peritoneal B cells from NIK-mutated alymphoplasia (aly) mice expressed substantial levels of S1P receptor and showed normal migration toward S1P, aly peritoneal B cells showed decreased sensitivity to FTY720, an S1P modulator. NIK-mutated stromal cells showed aberrant expression of VCAM-1, ICAM-1, and CXCL13, leading to the impaired ability to support S1P-mediated peritoneal B cell emigration. Therefore, aly peritoneal B cells exhibited normal S1P-mediated peritoneal B cell trafficking from peritoneum to intestine for IgA production when they were adoptively transferred into SCID mice. Further, transfer of wild-type stromal cells into the peritoneum restored S1P-mediated trafficking of aly peritoneal B cells. These findings suggest that NIK in stromal cells has a specific role in the regulation of S1P-mediated peritoneal B cell-derived intestinal IgA production.
This work was supported by grants from the Ministry of Education, Science, Sports, and Culture of Japan and the Ministry of Health and Welfare of Japan.
CCL1: a novel therapeutic target for the modulation of Treg function: Implications for immmunotherapy of cancer (40.10)
Authors: Hoelzinger Dominique B , Dominguez Ana Lucia , Smith Shannon E , Zorro Manrique Soraya , Lustgarten Joseph 
Published: January 2023

Abstract
Effective anti-tumor immunotherapy must overcome the immunosuppressive networks created by the tumor. Previous studies from our group indicate that intratumoral injection of CpG-ODN reduces Treg levels within the tumor by 80%. This decrease in Tregs is mainly mediated by the activation of antigen presenting cells secreting IL-6. To explore the effect of IL6 on Tregs, an expression microarray analysis of genes involved in TGFÎ2 dependent Treg conversion, which are also sensitive to IL6, was done. It yielded gene candidates which can be exploited to modulate the function of Tregs. In vitro analysis of two such candidates: chemokine ligand 1 (CCL1) and integrinÎ±E (ITGÎ±E), shows that blocking these proteins reduces de novo conversion of Tregs (CCL1 and ITGÎ±E) and inhibits the suppressive function of Tregs (CCL1 only). In vivo assays using the neu tolerant breast cancer mouse model demonstrates that injection of Î±CCL1 or Î±ITGÎ±E blocking antibodies significantly reduced the level of Tregs within the tumor and the tumor draining lymph nodes. Furthermore, the combination of intratumoral injections of CpG-ODN and Î±CCL1 induce complete tumor rejection in neu mice. These results demonstrate that blockage of CCL1 signaling modulated the function of Tregs and subsequently enhances immunotherapeutic approaches.
Sex-related differences in regulatory T cell (Treg) function and resistance to B16 melanoma in the absence of B7-H1 co-signaling (40.18)
Authors: Lin Pei Yi , Livi Carolina , Thibodeaux Suzanne R , Wierman Margaret E , Kious Mark , Jeansonne Duane P , Shin Tahiro , Brumlik Michael J , Zhang Bin , Daniel Benjamin J , Curiel Tyler J 
Published: January 2023

Abstract
We tested Treg function in B7-H1-/- (KO) mice. 6-10 wk male (M) or female (F) KO and M or F wildtype (WT) had equal numbers of CD4+CD25hiFoxP3+ T cells (Tregs) with similar CD25 and Foxp3 MFI. CTLA-4 trended lower in F vs M KO. M and F WT and M KO Tregs suppressed T cell proliferation equally in vitro but F KO Tregs were ~30% less suppressive vs the other 3 groups. Transfer of M KO Tregs fully protected RAG1-/- mice from CD4+RBhi T cell colitis but F KO Tregs did not, with up to 30% weight loss demonstrating poor in vivo Treg function. In M or F WT or KO with subcutaneous B16 melanoma, tumor was different at day 15 in F (159 mm3) vs M (458 mm3, p = 0.04) and F KO vs F WT (593 mm3, p = 0.01). We detected OVA-specific immunity in 0/3 M KO bearing OVA+ B16 vs 3/3 in F KO based on in vivo OT-I cell proliferation. Tregs in pre-pubertal 3-wk F KO had similar phenotype and function vs post-pubertal WT and M KO Tregs. Culture with TGF-β +IL-2 converted 6-wk CD4+CD25- F KO T cells into CD4+CD25hiFoxP3+ T cells with suppression similar to WT, but lower CD25. Thus, F KO T cells can become functional Tregs ex vivo. This previously unknown sex/T cell co-signaling interaction could explain sex-related differences in cancer, autoimmune disease, infection susceptibility and response to hormone antagonists. We hypothesize Treg dysfunction to improve immunity and tumor resistance in KO and estrogen as a B7-H1 co-factor in Treg development. CTLA-4 defects remain undefined.
Supported by CA105207, FD003118, and Voelcker and Greehey Trusts
Interferon-alpha improves immunologic and clinical response to regulatory T cell (Treg) depletion in ovarian cancer (41.15)
Authors: Wall Shawna , Thibodeaux Suzanne R , Lin Pei Yi , Kious Mark , Jeansonne Duane , Brumlik Michael J , Livi Carolina , Daniel Benjamin J , Curiel Tyler J 
Published: January 2023

Abstract
Treg depletion improves anti-tumor immunity but is unlikely to be curative alone. Denileukin diftitox (DT) depletes Tregs in BL6 mice and prolongs survival 11% at 5 µg/mouse weekly starting 2 weeks after challenge with ID8 ovarian cancer. Weekly human interferon (hIFN)-α 20,000 U/day on 4 consecutive days of 7 plus weekly DT survival improved 26% vs DT alone. Neither DT nor DT+hIFN-α improved survival in RAG1-/- mice with ID8, showing a requirement for adaptive immunity. hIFN-α alone reduced in vitro Treg suppression ~15% vs PBS, but did not clearly enhance DT-mediated Treg depletion, or increase T cell activation by CD69 expression. A patient in our clinical trial of DT for ovarian cancer failed, with CA-125 increasing 4.5-fold after 4 monthly cycles. Addition of weekly pegylated IFN-α2a to failed monthly DT improved blood Treg depletion 50% over DT alone and reduced CA-125 by 9-fold, demonstrating immunologic and clinical efficacy. To provide additional mechanistic insight we studied dendritic cell (DC) activation in hIFN-α treatment. In mice with ID8 tumor, hIFN-α significantly increased CD86 expression on myeloid and plasmacytoid DC. IFN-α may thus have an expanded role in treatment of epithelial carcinomas when used with Treg depletion, through enhanced Treg depletion or DC activation among other mechanisms.
Supported by FD003118, the Fanny Rippel Foundation, the Voelcker Trust and the Greehey Fund
Diversification of TCR repertoire in melanoma patients treated with chemotherapy before peptides-vaccination (41.19)
Authors: Nistico Paola , Palermo Belinda , Del Bello Duilia , Gualtieri Novella , Ferraresi Virginia , Belardelli Filippo , Proietti Enrico , Natali Pier Giorgio , Sottini Alessandra , Ghidini Claudia , Serana Federico , Imberti Luisa 
Published: January 2023

Abstract
Combining chemotherapy and immunotherapy is an attractive antitumor strategy. Recently we have reported that the administration of dacarbazine (DTIC) one day before peptide-vaccination in disease-free melanoma patients induced an increase of peptide-specific effector-memory CD8+ lymphocytes. To investigate the functional differences between anti-Melan-A CD8+ T cell response elicited by vaccination alone (Arm 1) or by chemotherapy plus vaccination (Arm 2), we generated T cell clones from 5 patients of both Arms, before and after the treatments. A significant enhancement of tumor lytic activity of Melan-A+ clones generated by patients of Arm 2 after the treatment was observed, whereas no significant differences of tumor recognition efficiency were evident among the two arms before treatment. Ninety-six sequences were obtained and 47 clonotypes with different CDR3 regions were observed. A progressive enhancement of TCR repertoire diversity was found only in patients treated with DTIC before vaccination, whereas patients treated with vaccine alone showed a reduction of TCR repertoire diversity accompanied by a reduction of tumor lytic activity in one patient. These results suggest that a diverse Melan-A-specific T cell repertoire occurs in melanoma patients treated with DTIC plus vaccination, which may offer a new clinically advantageous anti-tumor strategy.
Peripheral NK Cell Activation and Expansion by Artificial Antigen Presenting Cells, 2D11 (41.25)
Authors: Zhang Hua , Levine Bruce , June Carl , Mackall Crystal 
Published: January 2023

Abstract
Natural killer (NK) cells play an important role in early immune defenses against both nonself cells and autologous cells infected with viruses, bacteria, or parasites or in malignant transformation. NK cells mediate direct cytotoxic attack without prior sensitization on their targets through activating receptors, NKG2D, NCR, TRAIL, and inhibitory receptors ("absence of self"), and/or produce cytokines and chemokines that contribute to cytotoxicity and immunoregulation. A number of studies have reported that activation/expansion enhances NK cytotoxicity, but optimal methods for activating and expanding NK cells have not been well defined. Here we demonstrate that artificial APCs (2D11), previously reported and used to expand antigen-specific T cells, also efficiently expand NK cells via NKG2D and 4-1BB signaling mediated by MICA/B and 4-1BBL expression respectively on 2D11 aAPCs. In the presence of exogenous IL15, which is trans-presented by IL-15Rα on 2D11, robust expansion of highly lytic activated NK cells occurs. Compared to resting NK cells, IL15/2D11-activated NK cells upregulated TRAIL and NKp30, 44, 46 expression, and exerted enhanced cytotoxicity against K562, Ewing's tumor cell lines, including autologous tumors, ALL, as well as NK-resistant Daudi cells. In previous studies, expression of KIR ligands on tumor cells has been put forth as a mechanism of tumor immune escape from NK cell mediated killing. Our preliminary results demonstrate that expression of KIR ligands on tumor targets no longer inhibits killing by NK cells activated with 2D11. Ongoing studies are underway to test the hypothesis that NK cells activated by IL15/2D11 will prove more effective therapeutics due to breaking up of KIR-mediated inhibition of tumor cell killings.
Immunomodulatory activity of Resveratrol and its potential for combination with anti-tumor immunotherapy (41.26)
Authors: Soto Brenda Liz , Hank Jackie A , Polans Arthur S , Sondel Paul M 
Published: January 2023

Abstract
Resveratrol (RV) is a natural compound which is currently being investigated for many possible health promoting activities. It has anti-tumor activity in vitro on cancer cells and in vivo in some animal models of cancer. RV at a dose of 50mg/kg (i.g.) showed antitumor activity when treating nude mice bearing human neuroblastoma (Nb) xenografts. Prior studies on the effect of RV on the immune system found RV to be immunosuppressive in vitro and ex vivo in a dose-dependent manner, but not immunosuppressive in vivo. We are currently examining the ability of RV to be combined with immunotherapy to treat cancer. Using proliferative and cytotoxicity assays we are defining a dose in vitro and a regimen in vivo that does not suppress the immune system and is additive or synergistic in mediating anti-tumor effects when combined with immunotherapy. In our in vitro studies, 50uM or higher of RV inhibited the ability of PHA to stimulate proliferation of human PBMCs and inhibited IL2-stimulated murine splenocytes from dividing as measured by the ability of the cells to pass the dye CFSE to the daughter cells. These cells were found to be arrested in the G1 phase of their cycle, at those same concentrations. We are currently examining the effects of various concentrations of RV on target and effector cells in ADCC and NK killing assays. Also, studies on the antitumor and immunomodulatory effect of RV in vivo in Nb tumor-bearing animals are underway.
Synergy of radiation and immune therapy in tumor eradication (41.27)
Authors: Ruocco Maria Grazia , Kawashima Noriko , Huang Julie , Formenti Silvia C , Dustin Michael L , Demaria Sandra 
Published: January 2023

Abstract
We have investigated the mechanism of synergy between CTLA-4 blockade and local radiation therapy (RT) in treatment of breast cancer using the 4T1 mouse model. We have utilized CXCR6 GFP+ mice and CFP+ 4T1 tumor cells and intravital 2-photon laser scanning microscopy to follow the interaction of T cells with tumor cells. In mice treated with anti-CTLA-4 antibody 9H10 T cell speed was increased preventing stable interactions with tumor cells or stromal elements, critical for efficient tumor killing. RT also increased T cell velocity, but not to the same extent as 9H10 treatment. When RT was combined with 9H10 we observed increased T cell arrest in contact with tumor cells followed by tumor elimination. It has been shown that CTLA-4 ligation can increase T cell motility and this can overcome a TCR mediated stop signal. We confirmed that 9H10 treatment increased T cell motility on ICAM-1 coated surfaces and 9H10 was dominant over stop signals provided by soluble anti-CD3 antibodies. However, when a 9H10 go signal was subordinated to a stronger stop signal provided by solid phase anti-CD3 T cells did stop. These data suggest that anti-CTLA-4 therapy may impair killing of weak immunogenic tumors by suppressing IS formation. However, this effect can be overcome by RT-mediated enhancement of tumor antigen presentation. We have found that RT increases MHC class I and ICAM-1 expression on 4T1 cells, and are currently testing this in vivo.
WT1 peptide mix used for in vitro generation of WT1 CTL (41.29)
Authors: Stamer Mindy M , Herse Zachary , Dunham Kimberly , Bao Lei , Lucas Kenneth G 
Published: January 2023

Abstract
We developed a method to in vitro prime and expand WT1-specific CD8+ T cells using a WT1 peptide mix which could facilitate the expansion of WT1 specific T cells from donors of different HLA backgrounds. Monocyte-derived dendritic cells or PBMC isolated from 4 normal donors were pulsed with WT1 overlapping peptides prior to T cell stimulation. CD8 selected T cells were primed using WT1 peptide pulsed dendritic cells and then re-stimulated 11 days later using WT1 peptide pulsed autologous PBMC. IL-7 and IL-2 were added to the T cell cultures 4 days after the first stimulation to promote CTL differentiation. 7 days after the second CD8 stimulation, WT1 specificity was measured by tetramer staining. All donors exhibited tetramer positive T cells (donor 1=3.11%, donor 2=6.32%, donor 3=2.17%, donor 4=1.08%). WT1-specific cytotoxicity measured by 51Cr-release assays was also assayed 7 days post the second stimulation; however, WT1-specific cytotoxicity was not observed. WT1-specific CD8 T cells from 3 of the 4 donors were cloned and enriched by limited dilution. Clones were removed from OKT3 and WT1-specific cytotoxicity was measured in 51Cr-release assays. At least 1 clone from each of the 3 donors exhibited WT1 specific cytotoxicty (donor 1=54%, donor 2=20%, donor 3=26%). In conclusion, CD8 selected PBMC stimulated by antigen presenting cells pulsed with a WT1 peptide mix stained positive for HLA-A2 WT1-tetramers and exhibited WT1-specific cytotoxicity after cloning and subsequent expansion.
Phase 1 study of infusion of HER-2/neu specific T cells in patients with advanced stage HER-2/neu overexpressing cancers who have received a HER-2/neu vaccine (41.3)
Authors: Dang Yushe , Salazar Lupe G , Coveler Andrew , Waisman James , Higgins Doreen , Childs Jennifer , Bates Nicole , Slota Meredith , Disis Mary L 
Published: January 2023

Abstract
Adoptive T cell therapy has great potential for the treatment of patients with advanced-stage cancers. We have previously reported that HER-2/neu (HER2) specific T cell expansion from peripheral blood mononuclear cells (PBMN) can be greatly facilitated by in vivo antigen specific MHC class II epitope-priming. To test whether vaccine primed T cells could be administered to patients, we assessed the safety and feasibility of infusion of ex vivo expanded HER2 specific T cells in patients who have advanced HER2 overexpressing cancers. Three escalating doses of T cells were given at 10 day intervals with cyclophosphamide used for host lymphodepletion prior to the first dose of T cells. To date, 5 of 10 subjects have been enrolled. The maximal T cell doses infused were 1x109- 41x109 cells (median 10x109) with and average 35% of CD4+ and 60% of CD8+ T cells. IFN-gamma ELISPOT was used to monitor antigen specific T cells in both the infused product and to track the cells in vivo. HER2 specific T cells could be generated at high levels in vivo after infusion. Moreover, the magnitude of HER2 specific CD4+ and CD8+ T cells post-infusion was maintained in responding patients, with precursor frequencies 1:253 and 1:286 in total PBMN after 130 and 450 days respectively. Epitope spreading, which was induced with immunization, was also maintained and in some cases augmented after adoptive T cell therapy. TCR Vβ analysis performed showed the outgrowth of several HER2 specific clones in the CD4+ population post infusion. Our preliminary data suggests that adoptive transfer of autologous HER2 specific polyclonal T cells is safe and results in persistent high levels of tumor specific T cell immunity in vivo.
Generation of WT1-specific CTL from CMV stimulated CD8 cells (41.32)
Authors: Stamer Mindy M , Herse Zachary , Dunham Kimberly , Bao Lei , Lucas Kenneth G 
Published: January 2023

Abstract
WT1 responsive CTL were generated from CMVpp65 stimulated CD8 cells isolated from three CMV-seropositive HLA-A2 normal donors. CD8 selected T cells were stimulated with dendritic cells pulsed with CMVpp65 overlapping peptides. 10 days after the primary stimulation, CD8 T cells were restimulated with CMV pp65 peptide pulsed PBMC. The CMV-stimulated CTL exhibited CMV-specific but not WT1-specific cytotoxicity after the second stimulation as measured by 51CR release assay (Donor 1=54% and 7%, Donor 2=77% and 14%). However, all CMV-stimulated CTL stained with PE-RMFPNAPYL, a WT1A2 tetramer (Donor 1=2.51%, Donor 2=3.85%, Donor 3=0.49%). EBV-CTL generated from an HLA-A2 donor, and WT1 CTL generated from a non-HLA-A2 donor did not stain with the WT1A2 tetramer (0.02% and 0.01% respectively). Therefore, the WT1A2 tetramer was cross-reactive with pp65-CTL and exhibited HLA-A2 specificity. To enrich the CMV-CTL WT1A2 tetramer staining population, we PE-selected donor 1 PE-WT1 tetramer binding pp65 stimulated CTL. The WT1-selected cells were cloned by limiting dilution. 7 of 8 WT1-selected clones stained with the WT1A2 tetramer ranging from 0.89%-3.56%. WT1-selected clones showed three types of specific cytotoxicity: 1) against pp65-only (3 clones=22-34%), 2) against WT1-only (2 clones=43-44%), or 3) against both WT1 and CMV (1 clone=55% and 24% respectively). 2 of the 8 clones did not exhibit either WT1 or CMV specific cytotoxicity. Only 1 of the 2 clones that exhibit WT1 specific cytotoxicity exhibited WT1-specifc IFN-γ production, as measured by ELISPOT. All clones that displayed CMV-specific cytotoxicity also exhibited CMV-specific IFN-γ production. To our knowledge this is the first description of WT1 cross-reactive, CMV-specific CTL.
Intratumoral (IT) injection of hu14.18-IL2 immunocytokine (IC) enhances antitumor activity and augments NK infiltration into tumors: analyses of cellular internalization of hu14.18-IL2 by human NK cells (41.44)
Authors: Buhtoiarov Ilia N. , Neal Zane C. , Gan Jacek , Buhtoiarova Tatiana N. , Hank Jacquelyn A. , Yamane Brett H. , Rakhmilevich Alexander L. , Patankar Manish S. , Gubbels Jennifer A. A. , Gillies Stephen D , Sondel Paul M. 
Published: January 2023

Abstract
Hu14.18-IL2 (EMD 273063) IC, consisting of GD2-specific mAb and IL2, is in trials for treatment of melanoma and neuroblastoma. Preclinical studies showed augmented antitumor activity when IC is injected IT vs. IV. Human IL2R+CD16+NK cells mediate ex vivo ADCC in the presence of IC that binds them to GD2+ tumor cells. We studied the kinetics of IC presence on the surface of human M21 melanoma and CD25+CD16-NK cell lines, NKL and RL12, after IC binding to the cells via GD2 and IL2R, respectively. For NK cells, ~50% of IC was internalized by 6 h, and ~90% by 24 h of cell culture. Excess free IL2 in the culture did not slow internalization kinetics for these NK cells. IL2-deprivation for 24 h prior to IC arming of NK cells decreased IL2Rα expression and overall IC binding but did not affect internalization kinetics of the bound IC. For NK cells that were prelabeled with IC, the decrease of surface IC levels over 24 h correlated with reduction of ADCC in vitro. Unlike NK cells, M21 cells retained ~70% of IC on the surface following 24 h of culture.
When CFSE-labeled NKL cells were injected IV into M21-bearing SCID mice, they migrated better into the tumor when IC was delivered IT than when they were pre-armed with IC before implantation. These preliminary studies suggest that once IC gets to tumor sites, it may stay on the tumor surface for a prolonged time, and that IT injections of IC enhance the recruitment of NK cells to the tumor site.