bluuebunny/crossref_metadata_2025_split huggingface dataset:
Levels of anti-Gal IgG2 negatively correlate with the survival of porcine islet graft in non-human primates under immunosuppression of anti-CD154
Authors: Kang Hee Jung , Lee Haneulnari , Park Eun Mi , Kim Jong Min , Park Chung-Gyu 
Published: January 2023

Abstract

Background
Anti-CD154, the most successful immunosupppression (IS) regimen in porcine islet transplantation (PITx) is prohibited for clinical use. Anti-CD40, an alternative to anti-CD154, is not as effective as anti-CD154. To understand the reason, the relationship between anti-Gal responses and the graft survival was investigated in rhesus monkeys of PITx.

Methods
Intraportal PITx was performed in 18 diabetic monkeys: 3 received various IS without CD40 pathway blockade (Tx-control); 12, IS with anti-CD154 (Tx-anti-CD154); and 3, IS with anti-CD40 (Tx-anti-CD40). Blood samples were weekly obtained from the monkeys after PITx until the graft ceased to function, and additionally from 3 monkeys without transplantation (non-Tx-control) for 6 months. Levels of anti-Gal IgG, IgG1, IgG2 and IgM in the samples were analyzed by in-house ELISA, and levels of D-dimer and high-sensitive CRP by turbidimetry.

Results
Intraportal PITx led to strong induction of anti-Gal IgG1 response, not IgG2, in Tx-control, which was suppressed in both Tx-anti-CD154 and Tx-anti-CD40. The level of anti-Gal IgG2 revealed negative association with the levels of CRP and D-dimer in Tx-anti-CD40 and non-Tx-control, not in Tx-anti-CD154. The time-weighted average levels of anti-Gal IgG2 in Tx-anti-CD154 were significantly lower than in non-Tx-control. Additionally, the levels of anti-Gal IgG2 and D-dimer negatively correlated with the graft survival in Tx-anti-CD154.

Conclusions
The strong correlation between the suppression of anti-Gal IgG2 response and the graft survival in Tx-anti-CD154 suggests that IgG2 response is implicated with the effector mechanism mediating rejection, possibly NK cell activation which is known to promote IgG2 response.
Biological role of T Memory Stem Cells as a Cellular Reservoir for Graft-versus-Host Disease
Authors: Tkachev Victor , Furlan Scott N , Watkins Benjamin K , Panoskaltsis-Mortari Angela , Blazar Bruce R , Kean Leslie S 
Published: January 2023

Abstract
T memory stem cells (Tscm) are a subset of antigen-experienced, long-lived, self-renewing and multipotent T cells, which reconstitute immunity following hematopoietic cell transplantation (HCT). Here we investigated the biological role of Tscm in acute Graft-versus-Host Disease (GVHD) using a translational non-human primate model.
Tscm cells were tracked longitudinally following autologous or allogeneic HCT under different immunoprophylaxis regimens, resulting either in fulminant Th1-dependent ‘Primary GVHD’, late-onset Th17-driven ‘Breakthrough GVHD’, or GVHD-free immune tolerance.
Following HCT, Tscm acquired an activated phenotype (Ki67+, CD69+, PD-1+ and CCR5+), but produced low amounts of IFNγ, IL-2, IL-17A, TNFα and Granzyme B. Tscm were a minor population in donor grafts but expanded in peripheral blood (with a reciprocal decline of naïve T cells) early after HCT and then contracted (with a concomitant increase of effector T cells) when recipients developed clinical GVHD. The kinetics of Tscm expansion/contraction correlated closely with clinical disease: Tscm expanded slower and contracted later in animals progressing to Breakthrough compared to Primary GVHD, and Tscm numbers remained stable under tolerizing conditions. At necropsy, animals with GVHD had increased numbers of Tscm in both lymphoid organs (blood, lymph nodes, spleen) and non-lymphoid GVHD-target organs (colon, lungs, liver) compared to GVHD-free animals.
Our data demonstrates that Tscm dynamics correlate with GVHD-free survival and suggest that pathogenic T cells infiltrating both lymphoid and non-lymphoid organs transition from the naïve into the effector state via Tscm. Thus Tscm may play a key role as a cellular reservoir for GVHD.
EOMESODERMIN-EXPRESSING INNATE-LIKE CD8+ T CELLS IN HUMANS ARE IMPAIRED IN CHRONIC MYELOID LEUKEMIA PATIENTS AT DIAGNOSIS
Authors: Herbelin Andre , Jacomet Florence , Cayssals Emilie , Desmier Deborah , Barbarin Alice , Basbous Sara , Levescot Anaïs , Robin Aurélie , Giraud Christine , Barra Anne , Guilhot François , Roy Lydia , Gombert Jean-Marc 
Published: January 2023

Abstract
Starting with our recent identification in healthy individuals of a new distinct CD8+ T cell subset harboring Eomes and exhibiting a marked “innate” (panKIR/NKG2A+) phenotype, we present here the first report on this unique T cell population in a physiopathological context, namely chronic myelogenous leukemia (CML), a clonal myeloproliferative disorder. The current first-line therapies for this hematopoietic malignancy are tyrosine kinase inhibitors such as Imatinib (IM). Knowing that the generation of innate Eomes-expressing CD8+ T cells in mice depends on PLZF-expressing NKT cells and that iNKT cells are functionally deficient in CML, we surmised that panKIR/NKG2A+Eomes+ CD8+ T cells could be altered during CML. Accordingly, in CML patients at diagnosis, the pool of blood KIR/NKG2A(+)Eomes(+) CD8(+) T cells was found to be severely reduced. Moreover, like for iNKT and NK cells, the functions of these innate-like CD8(+) T cells were impaired, as attested by their lost of potent antigen-independent cytotoxic activity and IFN-γ production in response to innate-like stimulation by IL-12+IL-18. Remarkably, both innate-like CD8(+) T cells and iNKT cells returned to normal after complete CML remission upon IM therapy, supporting evidence for an iNKT cell-dependent generation of circulating KIR/NKG2A+Eomes+ CD8+ T cells in CML patients, as in mice. Our study reveals a possible contribution of iNKT/innate-like CD8(+) T cells as a new innate immune axis to tumor control in CML.
The Hepatic Recruitment of Innate Natural Killer T cells and Immune-Microenvironment Modulation after Colorectal Cancer Liver Metastasis
Authors: Feng Hao , Lu Aiguo , Thasler Wolfgang 
Published: January 2023

Abstract

Aims
To investigate the modulation of the immune microenvironment by human NKT cells and the crosstalk between NKT cells and Kupffer cells (KC), dendritic cells (DC) in colorectal cancer liver metastasis.

Methods
The numbers of iNKT cells and other cells from tumor margin, tumor or adjacent normal tissues were analysed by flow cytometry and AMNIS system. Ex vivo co-culture, 3D culture, Elisa array were established for this study.

Results
In primary colorectal cancers, percentage of iNKT cells was higher in early stage patients (T0~2). In the metastatic CRC, CXCR6+ cells are recruited and enriched at the invasive margin, elevated CXCR6 density was negatively related with Rees’ score, which suggests better prognosis. iNKT cells were the major population (30~40%) of CXCR6+cells, and the proportion and counts of iNKT cells were higher at the invasive margin (&lt;2cm from liver- tumor interphase). CXCL16+ cells were found to be enriched at the invasive margin using immunofluorescence. 85±14% of CXCL16+ (the ligand of CXCR6) cells were CD68+ macrophage, of which 89% cells were Clec4f+Kupffer cells. And the Ex vivo chemotaxis experiments showed that rhCXCL16 and Clec4f+Kupffer cell conditional medium promote iNKT cells recruitment. The ex vivo and in vitro experiments showed that iNKT cells could lead KC to M2 differentiation, however, antigen presented by KC restricted the CRC elimination capability of iNKT cells. In contrast, antigen presenting by DC enhanced clearance of CRC but at meanwhile also induce hepatocytes cell death. Furthermore, we found by in situ staining and FACS that the majority of the KC was M1 KC at intra-tumor area and M2 KC in the adjacent normal tissues. The iNKT cells as well as KC showed Th2 polarization at the invasive margin.
VGX-3100 Drives Regression of HPV16/18 CIN2/3 and Robust Cellular Immune Responses in Blood and Cervical Tissue in a Blinded, Randomized, Placebo-controlled Phase 2b Study
Authors: Morrow Matthew P , Trimble Connie , Shen Xuefei , Dallas Michael , Weiner David , Boyer Jean , Yan Jian , Kraynyak Kimberly A , Sylvester Albert J , Giffear Mary , Marcozzi-Pierce Kathleen , Shah Divya , Broderick Kate , Khan Amir S , Lee Jessica , Humeau Laurent , Sardesai Niranjan , Bagarazzi Mark 
Published: January 2023

Abstract

Objectives
Assessment of the safety and efficacy and immunogenicity of VGX-3100 in women with biopsy-proven CIN2/3 with concurrent HPV16 and/or HPV18 infection.

Methods
The randomized, placebo-controlled, double-blind study, which was stratified by age and severity of CIN, evaluated cervical tissue changes after three 6 mg intramuscular doses of VGX-3100 followed by electroporation with Inovio’s CELLECTRA2000 device at weeks 0, 4, and 12.

Results
Among 167 vaccinated women, the study met its primary efficacy endpoint; the percentage of patients who had regression of CIN2/3 to CIN1 or no disease at 6 months post third dose was significantly higher in the VGX-3100 group compared to placebo (p=0.034). In addition, the trial demonstrated the ability of VGX-3100 to clear HPV infection concurrent with regression of CIN2/3 (p=0.003). Post-hoc immune analysis also revealed significantly elevated immune responses in treated patients who had CIN2/3 regression concurrent with HPV clearance when compared to those who did not. This included the presence of CD8+ T cells in the blood exhibiting CD137 expression concurrent with perforin (p=0.032) as well as perforin in addition to granzyme A (p=0.036) as well as an influx of CD8+ T cells into cervical tissue (p=0.008).

Conclusion
The successful phase 2b results represent a significant milestone in the development of active immunotherapies to treat HPV-related dysplasia and cancer. The data generated from the trial reveal statistically significant correlations between antigen specific T cell activity and clinical benefits afforded by VGX-3100. Thus VGX-3100 has the potential to provide an important alternative or adjunct to surgery in treating CIN 2/3 based on HPV specific T cell activity.
Salmonella promote T cell-mediated anti-tumor immuno-therapy
Authors: Xiao Yichuan , Wang Yan , Liu Chenli 
Published: January 2023

Abstract
B16 melanoma is widely used as an animal model to investigate the efficacy, mechanism and immune response of anti-tumor immuno-therapy. In this study, the anti-tumor efficacy was examined by using a Salmonella strain 7207, which cannot survive in the circulation system and aerobic tissues, but can thrived in the anaerobic environment of solid tumors. We found that Salmonella injection could significantly inhibited tumor growth in B16 tumor-bearing mice. Interestingly, injection of heat-killed Salmonella also showed comparable anti-tumor efficacy as alive Salmonella, with no bacteria detectable in the tumor and circulation system. In addition, both alive and heat-killed Salmonella could promote robust pro-inflammatory gene induction in bone marrow-derived macrophages and dendritic cells, and increased tumor-infiltrating IFN-g-producing cytotoxic T cells (CTL). Moreover, deletion of T cells abolished the anti-tumor effect by injecting of alive or heat-killed Salmonella in Rag1-deficient tumor-bearing mice, suggesting Salmonella -mediated anti-tumor effect is due to the cytotoxic function of CTL. Taken together, our study provides a new insight of the development of anti-tumor immuno-therapy, and a potential therapeutic strategy for B16 melanoma.
Nrf2 expression is upregulated in tumor infiltrating T cells and induces T cell anergy
Authors: JO Yuna , Lee Byunghyuk , Joo Myungsoo , Hong Changwan 
Published: January 2023

Abstract
The transcription factor nuclear factor-erythroid 2-related-2 (Nrf2) controls cellular redox homeostasis and displays immunomodulatory properties. Hypoxic microenvironment in solid tumors generated by their outgrowth activates Nrf2 which induces the expression of anti-oxidant genes and supports continuous growth and proliferation via metabolic reprogramming. It is easy to imagine that hypoxic situation may affect function of immune cells infiltrated into tumor mass. We have thus questioned how hypoxic condition affect T cell immune response against tumor and the role of Nrf2 is involved. First, tumor infiltrating T cells (TILs) were sorted from tumor mass to investigate the alteration of Nrf2 expression. Interestingly, we found that mRNA levels of Nrf2 are significantly increased in TILs compared to control T cells. In order to test whether T cell responses are affected by Nrf2 levels, we stimulated T cells and measured Nrf2 expression. Nrf2 expression and their target genes are significantly downregulated by TCR activation. We further confirmed that IFN-γ production in Nrf2 deficient T cells was dramatically enhanced. Our findings offer novel insights into how immune response and oxidative stress is integrated in tumor cells, and we highlight Nrf2 as candidate of molecular target to control tumor in hypoxic condition.
Low signal strength enhances the antitumor activity of human CAR Th17 cells
Authors: Neal Lilli R , Huff Logan W. , Nelson Michelle , Meek Megan E. , Bailey Stefanie R , Bowers Jacob S. , Kundimi Sreenath , Mehrotra Shikhar , Varela Juan C. , Paulos Chrystal M 
Published: January 2023

Abstract
We reported the antitumor activity of human Th17 cells (engineered to express a chimeric antigen receptor (CAR) that recognizes mesothelin) regresses large human mesothelioma tumors when infused into mice, but it remains unknown what type and what level of co-signal strength impacts their functional fate and antitumor activity. To address this question, human Th17 cells were activated with a high (3 beads: 1 T cell), medium (1:10) or low (1:100) signal via CD3-beads coated with either CD28 or ICOS agonists. We discovered Th17 cells activated with low ICOS signal were highly multifunctional (IL-17A, IFN-γ, IL-2 and IL-22) and possessed a less differentiated phenotype (high CD62L and CD45RA expression) than cells activated with a high ICOS signal. In comparison to low ICOS signal strength, Th17 cells activated with low CD28 signal strength were poorly functional and terminally differentiated. Of clinical significance, in two distinct tumor models (human mesothelioma and murine melanoma), we found that Th17 cells regressed large tumors to a better extent when primed with a low or medium ICOS signal strength compared to if they were activated with a high ICOS signal strength, or with any level of CD28 signal strength. Our data is compelling given that most ACT clinical trials use high CD3/CD28 activation to expand patients’ cells. Specifically, our work suggests the use of a low bead to T cell ratio will result in a more simple and clinically feasible method to expand highly therapeutic T cells for the clinic.
Tonic 4-1BB signaling from chimeric antigen receptors (CARs) impairs expansion of T cells due to Fas-mediated apoptosis
Authors: Mamonkin Maksim , da Silva Diogo Gomes , Mukherjee Malini , Sharma Sandhya , Srinivasan Madhuwanti , Orange Jordan S , Brenner Malcolm K 
Published: January 2023

Abstract
High expression of CARs in T cells or the presence of CAR ligands in cell culture results in tonic CAR signaling that enhances differentiation and exhaustion of T cells and compromises anti-tumor activity. A recent study demonstrated tonic signaling negatively affected T cells expressing a CD28.zeta GD2-specific CAR, indicating 4-1BB costimulation may improve their function. However, whether 4-1BB is a uniformly optimal costimulatory signal for CAR-transduced T cells is unclear.
We found that retroviral expression of several 2nd generation CARs with a 4-1BB domain (4-1BB.zeta) unexpectedly impaired T cell expansion. High expression of 4-1BB.zeta CD19 and kappa CARs led to upregulation of Fas and FasL and enhanced caspase 8-dependent apoptosis. Decreasing CAR expression in retro- or lentiviral systems normalized Fas levels and restored T cell expansion. However, a similar approach did not rescue expansion of 4-1BB.zeta CD5 CAR T cells, likely because the presence of the target antigen CD5 on T cells sustains high tonic signaling. 4-1BB.zeta CD5 CAR T cells recruit Fas and FasL to form stable fratricidal immunologic synapses. Disrupting TRAF2 binding site in the 4-1BB domain prevented Fas upregulation and restored T cell function but compromised costimulation. Therefore, we developed a regulated retroviral expression system that uses a small molecule to reversibly inhibit CAR expression, preventing tonic signaling and the resulting fratricide and differentiation. Withdrawing the inhibitor restores CAR expression and the anti-tumor function of T cells. Hence, we have described a mechanism by which chronic 4-1BB signaling may undermine CAR T cell function and provide a solution that curtails detrimental tonic CAR signaling.
Expression of CLL Upregulated Gene1(CLLU) ,Telomere Reverse Transcriptase (hTERT) ,Zeta -Associated Protein Kinase -70 (ZAP-70) in patients with Chronic Lymphocytic Leukemia
Authors: Abdelrahman Zakia Ahmed , shamaa lobna abou , Sultan Magda , Abdelrahman Mohmed , Almorish Mohmed A 
Published: January 2023

Abstract

Background
Defining prognostic tool may help in B-CLL patient subgroups to benefit from early therapeutic intervention. ZAP-70 expression is one marker for IgVH mutational status. hTERT expression discriminated the Ig-unmutated from Ig-mutated B-CLL and may serve as a molecular prognostic marker. CLLU1 expression holds some prognostic information not offered by other biologic markers in B-CLL. This findings drove us forcibly to investigate their expression in B-CLL.

Methods
The current study was carried out on newly diagnosed 30 B-CLL patients before treatment. In Binet classification ,23% in stage A, 33.3% in stage B and 43.7% in stage C. lymphoadenopathy was in 76.7% , splenomegaly 66.7% ,hepatomegaly 20% of patients. PBMCs were isolated from B-CLL patients and tested by immunocytochemical staining technique for ZAP-70 expression and QRT-PCR for hTERT and CLLU1 expression using cDNA1 for CLLU1

Results
ZAP-70 in mononuclear cells was positive in 30% of patients . ZAP-70 was significantly increased with Binet stage progression and . It was positively correlated with TLC and lymphocytic count and negatively correlated with the hemoglobin level and platelets count. The expression levels of hTERT mRNA and CLLU1 mRNA were +ive in all BCLL and -ive in normal cells. They were not correlated with Binet stage, hematological parameters ,ZAP-70 . The expression levels of hTERT and CLLU1 were higher in ZAP-70 positive but statistically non significant

Conclusion
ZAP-70 expression by Immunocytochemistry is a standardized, simple and inexpensive cost effective as a prognostic. hTERT and CLLU1 expression were novel prognostic markers in B-CLL that can be measured using a rapid and more sensitive method (QRT-PCR) accessible to routine laboratories.
Leucine metabolism is a promising new approach to study the interplay between T cells and cancer cells
Authors: Ananieva-Stoyanova Elitsa A , Bostic Josh , Torres Ashley , Hutson Susan 
Published: January 2023

Abstract
Cancer and T cells undergo similar metabolic reprogramming during proliferation to support increased biosynthetic and energy demands. They both rely on high rates of glycolysis while retaining mitochondrial respiration, a phenomenon known as the Warburg effect. In the tumor microenvironment, tumor cells can escape immune surveillance by competing with T cells for common metabolites and redirecting nutrients for their own advantage. To gain insight into the relationship between T cells and cancer cells, we studied the effect of leucine metabolism on T cells and their cancerous counterpart, EL-4 lymphoma cells. Our previous results showed that a loss of expression of leucine degrading enzymes (cytosolic or mitochondrial branched chain aminotransferases, BCATc and BCATm, respectively) increased intracellular leucine concentrations and stimulated glycolytic rate in activated T cells. These results correlated with activation of complex 1 of the mammalian target of rapamycin (mTORC1). Continuation of this study with EL-4 lymphoma cells, grown in leucine free media or treated with the leucine antagonist, n-acetyl-leucine amide, revealed a decrease in glycolysis, glycolytic capacity, and glycolytic reserve while mTORC1 was only moderately impacted. The oxygen consumption rate was stimulated in T cells lacking BCATc and/or BCATm but was decreased in the absence of leucine in EL-4 cells. The results suggest that T cells and their cancerous counterpart use leucine in a similar manner and that leucine is essential for the regulation of glucose and oxygen metabolism in either cell type. Future studies targeting leucine metabolism in the tumor microenvironment will aim to improve the effectiveness of T cells to fight cancer cells.
A comprehensive system of congenic mouse melanoma models for evaluation of immune therapies
Authors: Bosenberg Marcus , Meeth Katrina , Damsky William 
Published: January 2023

Abstract
There has been longstanding interest in the relationship between the immune system and malignant melanoma, the deadliest form of skin cancer. Melanoma has been the tumor of choice for the evaluation of a wide variety of therapeutic approaches that rely on stimulation of the immune system to combat cancer. The striking responses to immune checkpoint inhibitors noted in some melanoma patients have now been observed in several other malignancies, including renal cell carcinoma, lung cancer, and prostate cancer. These findings are very encouraging, however the specific mechanisms of response and reasons why only a subset of patients respond to therapy are not known. The extraordinary potential of immune therapies in cancer suggests that human-relevant mouse models of cancer are required to understand mechanisms of response and to optimize therapy so that a larger proportion of patients respond.
We have generated and evaluated a series of genetically engineered mouse models driven by alterations in Braf, Pten, Cdkn2a, beta catenin, p53, and Nras that reflect the genetic diversity of the vast majority of human melanoma. We have backcrossed 7 of these models to C57Bl/6, generated tumors, and isolated a series of over 30 congenic mouse melanoma cell lines. Several independent lines have been isolated for each melanoma genotype generated. We are in the process of characterizing the tumor microenvironment in each of the models using flow cytometry and a novel quantitative imaging approach of histological sections. This series of congenic lines represents an ideal set of models for the study of cancer immunology and responses to immune therapies.