bluuebunny/crossref_metadata_2025_split huggingface dataset:
Potent interaction between CMV reactivation and GVHD: Immunologic evidence for the blunting of CMV-driven immune reconstitution in the setting of GVHD in transplant patients
Authors: Suessmuth Yvonne , Betz Kayla , Yu Alison , Bratrude Brandi , Watkins Benjamin , Qayed Muna , Horan John , Kean Leslie S , Langston Amelia 
Published: January 2023

Abstract
Allogeneic hematopoietic cell transplantation (HCT) may be curative for patients with marrow and immune disorders, but acute graft-vs-host-disease (aGVHD) and infections cause significant morbidity and non-relapse mortality. We conducted a multicenter, double blind, placebo-controlled phase II trial of costimulation blockade with abatacept (Aba) combined with standard GVHD prophylaxis with a calcineurin inhibitor and methotrexate (CNI + MTX) following HLA matched unrelated donor transplant (n=140).
In order to assess whether Aba produces clinically relevant compromise of immune reconstitution, and whether this reconstitution is influenced during CMV reactivation, we longitudinally evaluated post-transplant whole blood samples with multiparameter flow cytometry using markers for CD3, CD4, CD8, CD197 and CD45RA to measure reconstitution of CD4 and CD8 T cell populations and their respective memory subsets over time.
We show here that post-transplant CMV reactivation induces a marked expansion of CD8 effector memory (EM) cells, which is similar in Aba vs placebo pts. We found that development of moderate (gr 2–4) or severe (gr 3–4) GVHD does not cause more CMV reactivation in patients, but patients with moderate GVHD show a blunted expansion of CD8 EM cells compared to those without GVHD. Remarkably, CD8 EM expansion was essentially absent among CMV reactivating patients with severe aGVHD.
Our results suggest that adding abatacept to CNI/MTX does not materially affect reconstitution of T cell immunity in the presence or absence of CMV reactivation, but aGVHD remains a major driver of compromised immune recovery after HCT.
Plasma cells are not necessary for regulatory B cells and preventing their differentiation markedly prolongs allograft survival
Authors: Ding Qing , Rothstein David 
Published: January 2023

Abstract
Plasma Cells produce Abs. Recently, PCs were reported to be the major source of B-cell IL-10, and absence of PCs worsened EAE, implying that PCs underlie “Breg” activity. To test the hypothesis in transplantation, we transplanted Bc islets into chemically diabetic B6 Blimp-1fl/flCD19-Cre+/− (PCKO) mice. Surprisingly, allograft survival (GS) was prolonged by ~42% in untreated PCKO vs. Cre-neg Ctl recipients (Mean GS: 27 vs. 19d; p&lt;0.05). This prompted us to re-examine the EAE model where the regulatory role of PCs was first identified. Surprisingly, in our hands, the severity of EAE (MOG35–55 induced) was dramatically reduced in PCKO vs. Ctl mice (p&lt;0.05). These unexpected results suggested that PCs are not essential for Breg activity, but might actually promote EAE and allograft rejection. These improved results do not appear to be mediated simply by loss of Ab production, since EAE is worse in the complete absence of B cells. To investigate the differences between our own and published results, we examined B cells from PCKO mice and found a 1.4-fold ↑ in IL-10+and &gt;2-fold ↓ in IL-17+ B cells vs. Ctl mice (intracellular staining; flow cytometry). Moreover, CD4+T cells from PCKO mice exhibited a ~2.5× ↑ in IL-10 and ~2× ↓ in IL-17.
Therefore, we examined cytokine expression by splenic PCs in alloimmunized Blimp-1-YFP reporter mice. While PCs are enriched for IL-10 expression (42±3%), they are also highly enriched for IL-17 (50 ±9%).
Thus, PCs are not essential for Breg activity in either auto- or allo- immunity. We hypothesize preventing newly induced PCs may reduce immune responsiveness through the loss of potentially proinflammatory PCs. To further address these questions, we are now developing novel mice with PC-specific deletion of IL-10 or IL-17.
Recovery of innate lymphoid cells after allogeneic stem cell transplant: A single institution pilot study
Authors: Srinivasan Anand , Crawford David , Bajana Sandra , Yuen Carrie , Sun Xiao-Hong , Shah Rikin 
Published: January 2023

Abstract
The response of Innate Lymphoid Cells (ILCs) to conditioning regimens and subsequent recovery in the setting of allogeneic hematopoietic stem cell transplant (aHSCT) in pediatric patients is not known. The impact of this recovery pattern on graft versus host disease (GVHD) has not been prospectively studied. We aim to study the recovery of ILCs after aHSCT in children and discover its role in GVHD. Pediatric patients undergoing aHSCT were enrolled in a prospective translational study since May 2019. Patients blood were analysed by flow cytometry at following time points: prior to the conditioning regimen, weekly starting on day 0 until +30 (+/− 3 days), then +60, +90 and +180 days. Ten patients have been enrolled to date - demographics: median age 9.5 years; 4 malignant, 6 non-malignant; haploidentical donor (n=1), matched sibling (n=3), unrelated (n= 6). Graft source was bone marrow for all patients. Most common conditioning regimen was fludarabine, busulfan and rabbit antithymocyte globulin (40%). GVHD prophylaxis for most patients was tacrolimus with mini-methotrexate (90%). We saw substantial drop in number of ILCs in response to conditioning regimen in all patients, with nadir on +14 that plateaued through +30, followed by a spike on +60 and return to normal levels by +90. Similar trend was seen in type 1 ILCs (Lin− CD127+ CRTH2− CD117−) but showed a slower drop, reaching nadir on +21 with slow recovery by +60. Type 2 ILCs (Lin− CD127+ CRTH2+) and Type 3 ILCs (Lin− CD127+ CRTH2-ve CD117+) however showed a small increase on +7 compared to day 0 with both cell types recovering by +60. We showed that it is feasible to prospectively study the recovery of ILCs after aHSCT. Continued enrollment will allow us to evaluate the correlation between ILCs and GVHD.
Pancreatic and colon cancer stem cells escape NK cell effector function via PCNA–NKp44 interaction
Authors: Malaer Joseph D. , Mathew Porunelloor A. 
Published: January 2023

Abstract
Natural killer (NK) cells play an important role in the innate immune response against cancer. Unlike B or T lymphocytes, NK cells do not require prior sensitization and can immediately respond to target cells. NK cell function depends on a balance of signals transmitted from activating and inhibitory receptors interacting with ligands on the surface of cells. Cancer cells may escape NK-mediated responses by expressing inhibitory ligands. Proliferating cell nuclear antigen (PCNA), in association MHC I, forms the inhibitory ligand for NKp44. Cancer stem cells (CSC), a unique subset of tumor cells, possess a stem-cell-like phenotype and are thought to facilitate metastasis by escaping NK cell effector function. CSC can be identified by a variety of markers such as surface co-expression of CD44 and CD133. In both Panc-1 and HCT 116 cell lines, surface PCNA is associated with co-expression of CD44 and CD133. Triple positive (PCNA+CD44+CD133+) cells have increased CSC transcription factor (NANOG, SOX2, and Oct-4) expression compared to double positive (PCNA−CD44+CD133+) or negative cells. Additionally, blocking the PCNA–NKp44 interaction alters IFN-g secretion and increases specific lysis of cancer cells by NK cells. Taken together, these data suggest that surface co-expression of PCNA, CD44, and CD133 are markers of pancreatic and colon CSC and blocking the PCNA–NKp44 interaction may provide an immunotherapeutic approach to target pancreatic and colon CSC and prevent metastasis.
Post-translational modification of cytoskeleton regulates anti-tumor cytotoxicity in γδ T cells
Authors: Hu Yi 
Published: January 2023

Abstract
γδ T cells based immunotherapy has emerged as a new treatment strategy for malignant tumors. The underlying cytotoxic mechanism against cancer cells involves surface receptor recognition molecules such as LFA-1, NKG2D, TRAIL, CD16, CD107a, and FasL, as well as cytokines such TNFα, IFNγ, perforin, and granzyme B. Previously, it has been shown that the cytoskeleton plays pivotal roles in T cell immunity. Nevertheless, the role of cytoskeleton in γδ T cell anti-tumor immunity remains largely unknown.
In our work, we attempted to elucidate the functions of cytoskeleton in regulating the antitumor cytotoxicity of γδ T cells. We discovered that the acetylation of tubulin-α is positively correlated with γδ T cells cytotoxicity. While the down-regulation of tubulin-α acetylation in γδ T cells leads to decreased cancer cells killing efficacy, up-regulation could potentiate it through the expression of cytotoxic related molecules such as NKG2D, IFN-γ, CD16, and suppression of PD-1 molecule. Moreover, we observed that the acetylation of tubulin-α in γδ T cells drastically differs after challenged by MDA-MB-231, SkBr3, and MCF7 breast cancer cells, respectively, implying diverse γδ T cell immune responses towards these cell types. This result was further supported by the discrepancies in γδ T cell in vitro killing efficacy.
Together, our work will provide important scientific references to further understand the cytoskeletal function in γδ T cell anti-tumor immunity.
Two-color flow cytometric detection of serial cell-mediated cytotoxicity by natural killer (NK) lymphocytes
Authors: Navarrete-Galvan Lydia T , Guglielmo Michael , Smith-Gagen Julie , Merica Rebecca , Lombardi Vincent C , Hudig Dorothy 
Published: January 2023

Abstract
Serial killers are much more productive than one-hit wonders, especially when it comes to killing cancer. Natural killer lymphocytes are significant cells in many anti-cancer immunotherapies, such as monoclonal antibody based- and adoptive cell transfer therapies. However, current data show that less than 30% of peripheral blood NK cells are capable of sequentially killing more than one cell, i.e., serial killing. These serial killers are not only capable of killing up to 6 target cancer cells each, but are also reported to deliver lytic hits at a faster rate and with more killing power than their counterpart NK cells. Current methods to identify serial killer NK cells, such as time-lapse cinematography, are laborious, expensive, and require highly skilled expertise and equipment. In order to enrich and grow serial NK cells, an inexpensive, simple assay is needed to detect and isolate them from blood for subsequent in vitro growth. Here we developed a flow cytometric method to identify serial killers via 2-color detection of the cell surface marker CD107a, expressed on the NK cell surface following killing. The NK-92 cell line was incubated with target Raji B leukemia cells for 2-hours, then labeled with one color, AF647, anti-CD107a mAb. Cells were then re-incubated for an additional 4-hours and subsequently labeled with a second color, PE, anti-CD107a mAb. Serial killers labeled double positive for both colors at 6 hours. We found that NK-92s are capable of killing at least 14 target Raji cells each and about 18% of the NK-92s can be identified as serial killers via two-color CD107a mAb labeling.
Role of the KLRG1 pathway during the immune response to cancer
Authors: Tata Angela , Vivier Eric , Brossay Laurent 
Published: January 2023

Abstract
Immunotherapy drugs targeting T cells have been successfully developed for a variety of different cancers. In contrast, comparably less work has been done on targeting natural killer (NK) cells for immunotherapy. NK cells are intriguing targets for a number of different reasons, including their cytotoxic abilities and relatively lower autoimmunity risks. Thus, treatments that target NK cells, either alone or in conjunction with T cells, should be explored for the development of new treatments. One of the most well-known immunotherapy targets, programmed death-1 (PD-1) has recently been implicated in modulating NK cell function in cancer and demonstrates that targeting NK cells is an important aspect of cancer treatment. In addition to PD-1, there are many checkpoint molecules that may be capable of impacting NK cell function. One such target is KLRG1, a well conserved C-type lectin inhibitory receptor expressed on NK cells, which contains an ITIM in its cytoplasmic domain. To determine the role of KLRG1, and expand on the role of PD-1, as potential NK cell checkpoints, we developed a series of mice deficient for KLRG1 and PD-1, and took advantage of the NK sensitive cancer model B16. Following administration of B16-E-cadherin, we found that KLRG1 deficient animals had significantly fewer tumors in the lung tissue, compared to wild type controls. In addition, B16-E-cadherin resulted in a higher number of tumors in wild-type mice, compared to the parental B16 cell line. Altogether, these data suggest that KLRG1 is a checkpoint target candidate.
Therapeutic potential of follicular helper T cells for melanoma treatment
Authors: Nurieva Roza I , Liu Zhongshan , Zhao Yang-Zhi , Divenko Margarita , Alekseev Andrei , Sahoo Anupama 
Published: January 2023

Abstract
Clinical relapse and metastases are the major causes of death in melanoma. Currently, adoptive T cell therapy using tumor-infiltrating lymphocytes rich in cytotoxic CD8+ T cells (CTLs) is one of the promising approaches that helps overcome this major challenge in melanoma; however, CTL cell transfer alone does not improve clinical response, suggesting a supportive role of helper CD4+ T cells. There is increasing evidence on the potential contribution of tumor-infiltrated CD4+ T follicular helper (Tfh) cells and intratumoral lymphoid structure formation in tumor eradication; however to date, the role of Tfh cells in anti-tumor immunity has not been clearly elucidated. Remarkably, using a murine melanoma model, we found that the percentage of intratumoral Tfh cells negatively correlates with melanoma growth. In addition, we observed enhanced tumor growth in Bcl6 T-cell conditional knockout mice, which lack Tfh cells as compared to wild-type mice, suggesting that Tfh cells might play a protective role in tumor growth and could serve as a marker for tumor regression. Importantly, in support of mouse studies, in melanoma patients the number of Tfh cells correlate with the rate of patient survival. Importantly, adoptive transfer of antigen specific Tfh cells into tumor bearing mice results in increased number of intratumoral CTLs, as well as in an efficient tumor eradication, indicating the essential role of Tfh cells in promoting CTL expansion and anti-tumor reactivity. Our results thus, for the first time, indicate the therapeutic potential of Tfh cells in promoting CTL-mediated anti-tumor immunity against melanoma and provide the basis for potential usage of these cells to improve current immunotherapy approaches.
STINGing the tumor microenvironment to promote therapeutic tertiary lymphoid organogenesis
Authors: Chelvanambi Manoj , Taylor Jennifer , Storkus Walter 
Published: January 2023

Abstract
Classical tertiary lymphoid structures (TLS) are non-encapsulated aggregates of B cells, T cells and dendritic cells (DCs) that form around high endothelial venules (HEVs) in tissue sites affected by chronic inflammation. Homeostatic chemokines/cytokines CCL19, CCL21, lymphotoxin α, lymphotoxin β, IL-36γ, type I interferons, CXCL9 and CXCL10 have collectively been implicated in secondary lymphoid organogenesis as well as in formation of TLSs. As these factors can be produced by appropriately activated dendritic cells (DC), we posit that conditioning of intratumoral DCs/stromal cells can lead to TLS formation, improved immune cell infiltration and slowed tumor growth. We report that STING activation in bone marrow derived CD11c+ DCs leads to the production of various TLS forming factors through a STING-TBK1-IRF3 signaling cascade. In vivo, B16.F10 bearing mice treated intratumorally with a human/murine STING agonist, ML-RR-S2-CDA, showed slowed tumor growth dependent on CD8+ T cells. Immunofluorescent imaging of treated tumors showed increased aggregation of CD11c+ DCs with CD3+ T cells around PNAd+/MAdCAM1+ HEVs as early as 5 days post-treatment. These changes occurred in association with evidence for normalization of the tumor vasculature based on enhanced PDGFRβ+ pericyte coverage of CD31+ VECs. Future studies will analyze how the presence of TLS shapes the evolving local (tumor) vs. systemic therapeutic anti-tumor T cell repertoire.
Therapeutic vascular normalization to promote tumor-associated tertiary lymphoid structures
Authors: Filderman Jessica , Storkus Walter 
Published: January 2023

Abstract
Tertiary lymphoid structures (TLS) are non-encapsulated immune cell aggregates that form at sites of chronic inflammation. Recent studies have shown that the presence of TLS in human tumors indicates positive clinical outcome. However, many tumors have poorly organized and leaky vasculature that impedes entry of immune effector cells into tumors and consequently TLS formation. Recently, studies have shown that low doses of antiangiogenic agents normalize tumor vasculature, leading us to hypothesize that treating tumors with low doses of vascular normalizing (VN) therapies will improve immune cell infiltration and TLS formation within the tumor microenvironment (TME). To test this hypothesis, tumor-bearing mice were treated intratumorally with VN agents. RNA was isolated from digested tumors and transcript levels of TLS-promoting factors and markers of inflammation/immune cell infiltration were measured and compared to PBS treated controls. Changes in tumor vasculature were evaluated using immunofluorescence microscopy. Additionally, primary cultures of murine T cells and DCs and the BPR melanoma cell line were treated in vitro with VN agents. We observed that the VN agents dasatinib, STING agonist, bevacizumab, and agonist anti-TNFR1 antibody each induced global changes in the TME that are potentially supportive of immune cell infiltration and TLS formation. In vitro, dasatinib induced DCs and BPR melanoma cells to express higher levels of co-stimulatory receptors, MHC class I and II, and TLS-promoting chemokines. Treatment with a STING agonist was able to enhance T cell activation, while treatment with dasatinib inhibited T cell activation.
Exosomal PD-L1 expression as non-invasive biomarker for immune checkpoint inhibitors in non-small cell lung cancer
Authors: Gunasekaran Muthukumar , Russo Alessandro , Cardona Andrés F , de Miguel Perez Diego , Lapidus Rena , Cooper Brandon , Kaushal Sunjay , Adamo Vincenzo , Rolfo Christian 
Published: January 2023

Abstract
The development of immune checkpoint inhibitors (ICIs) significantly improved survival outcomes of non-small cell lung cancer (NSCLC) patients compared to chemotherapy. However, identification of predictive exosomal biomarker to reduce the invasive biopsy as well as to understand the efficacy of ICIs in NSCLC patients are unexplored. In this investigation, we demonstrate for the first time exosomal PD-L1 expression as predictive biomarker (liquid biopsy) for immune checkpoint blockage with PD-1/PD-L1 inhibitors in advanced/metastatic NSCLC.
Plasma samples from NSCLC patients (n=25) with matched paired samples at baseline and after 8 weeks of PD-1 blockade (Nivolumab, pembrolizumab and atezolizumab) were utilized for exosomal PD-L1 analysis by immunoblot. No statistically significant differences were reported for PD-L1 changes between pretreatment samples and at 8 weeks in both progressive patients (p=0.3783) and non-progressing patients (p=0.8066). Interestingly, when considering PD-L1 changes at 8 weeks between patients with progressive disease and partial response/stable disease (non-progressors), a trend towards PD-L1 reduction was observed in the non-progressors group, albeit this difference was not statistically significant (p=0.1941) due to the small sample size. Interestingly, dynamic changes during treatment of exosomal PD-L1 expression, assessed as exosomal PD-L1 ratio between pretreated samples and after 8 weeks of treatment, predicted the outcome of ICI treated patients in terms of both PFS and OS.
In conclusion, reduced exosomal PD-L1 expression at 8 weeks observed in non-progressors suggesting that exosomal PD-L1 might be used as non-invasive serum marker for ICIs treatment in NSCLC patients.
Mechanism of Heterodimeric IL-15 in tumor reduction: Induction of myeloid-lymphoid cell interactions leads to IFN-γ dependent lymphocyte recruitment by dendritic cells into tumors
Authors: Pavlakis George N , Bergamaschi Cristina , Stellas Dimitri , Karaliota Sevasti , Nagy Bethany , Stravokefalou Vasiliki , Dimas Konstantinos , Jensen Shawn , Fox Bernard A. , Felber Barbara K. 
Published: January 2023

Abstract

Background
hetIL-15 is evaluated in on-going immunotherapy clinical trials. We studied the mechanism of hetIL-15 antitumor function in several mouse models (MC38, TC-1, 4T1 and EO771).

Methods
We evaluated tumor immune infiltrate by flow cytometry, multi-color immunohistochemistry (IHC), transcriptomics and proteomics.

Results
hetIL-15 treatment delayed primary tumor growth, induced tumor regression and reduced metastases. Flow cytometry and IHC showed increased intratumoral NK and CD8+ T cell infiltration. Tumor-resident NK and CD8+ T cells were activated with enhanced IFN-γ production, increased proliferation (Ki67+), expression of survival factors (Bcl-2) and cytotoxic potential (Granzyme B+). hetIL-15 therapy resulted in increased IFN-γ and XCL1 levels and enhanced tumor accumulation of conventional type 1 dendritic cells (cDC1) expressing XCR1, IRF-8 and CD103. Production of CXCL9 and CXCL10 chemokines by tumor infiltrating cDC1 was boosted in an IFN-γ-dependent manner. Increased frequency of circulating NK and CD8+ T cells expressing CXCR3 was found, suggesting their migration toward tumors following the CXCL9/10 chemokine gradient.

Conclusions
hetIL-15 inhibits tumor growth by triggering lymphocyte-myeloid cell interactions modifying the tumor chemokine and cytokine milieu. hetIL-15 causes induction of XCL1 by lymphocytes, inducing DC intratumoral number, subsequent IFN-γ dependent CXCL9/10 production by DC and increased attraction of effector lymphocytes.
Investigating microbiome-based anti-tumor immune modulation using patient-derived mouse models
Authors: Fessler Jessica , Matson Vyara , Gajewski Thomas 
Published: January 2023

Abstract
Growing data supports that the gut microbiota can impact cancer immunotherapy. We previously found certain bacterial taxa associated with response to immune checkpoint blockade. These correlations warrant deeper mechanistic studies using tractable mouse models to establish the role of the microbiota in determining immunotherapy efficacy.
To understand how the microbiota affects anti-tumor immunity we generated patient-based microbiota models. Germ-free mice were colonized using patient feces, bred, and subsequent generations studied using transplantable tumor models. We focused on two representative patient-based models: one from a responder (R) to immunotherapy and one from a non-responder (NR).
Mice established from the R demonstrated greater benefit from anti-PD-L1 therapy compared to NR colonized mice and had superior CD8+ T cell responses with greater accumulation of tumor-specific CD8+ T cells and increased proliferation. In addition, compared to NR-derived mice, R-derived mice had increased intra-tumoral CD103+ dendritic cells (DCs), as well as increased BATF3-lineage DCs in the mesenteric lymph nodes and spleen. In agreement with a stronger anti-tumor immune response, R mice had higher circulating levels of IFN-γ, IL-12p70, and CXCL5 compared to NR mice.
To identify potential systemic signals originating from the gut microbiome and capable of altering global immune populations we profiled serum using an unbiased metabolomics-based approach. We found distinct metabolite profiles in the R- and NR-derived mice, suggesting potential bacteria-derived or -modified metabolite modulating immune function. These data support a role for the gut microbiome determining host sensitivity to cancer immunotherapy.