bluuebunny/crossref_metadata_2025 huggingface dataset:
MAIT cell-mediated inhibition of colorectal tumor growth is associated with increase in IL-5 and accumulation of eosinophils in the tumor microenvironment
Authors: Cheng Olivia J , Lebish Eric J , Jensen Owen , Jacenik Damian , Trivedi Shubhanshi , Beswick Ellen J , Leung Daniel T
Published: August 2023

Mucosal-associated invariant T (MAIT) cells are innate-like lymphocytes with cytotoxic and pro-inflammatory function and are enriched in mucosal tissues such as the colon. With studies demonstrating both pro- and anti-tumor capability in various cancer models, the role of MAIT cells in colorectal cancer remains unclear. Our study uses a mouse model of colorectal cancer to show that adoptive transfer of MAIT cells from C57BL/6 donors into MC38-bearing RAG1−/− mice leads to a decrease in tumor growth compared to sham-transfer control. Using qRT-PCR and multiplex cytokine analysis, we show that extracted tumors from MAIT adoptive transfer group have a higher expression of eosinophil markers, Siglecf and Ccr3, as well as higher levels of eosinophil-activating cytokines, such as EOTAXIN-1, IL-5, GM-CSF, suggesting an association between tumor inhibition and eosinophil prevalence and activation. Consistent with this, flow cytometry analysis of human peripheral leukocytes shows increases in IL-5 expression in MAIT cells and activation of eosinophils upon stimulation with MAIT ligand, 5-A-RU, suggesting that MAIT cell activation could modulate eosinophil function. Together, these data suggest that the anti-tumor effect of MAIT cells may involve IL-5- and eosinophil-dependent mechanisms. This study provides evidence of the ability of MAIT cells to modulate the tumor microenvironment and immune cell profile, specifically the potential to recruit and activate eosinophils.
BAFF and APRIL signaling through BCMA regulates microglial cells in the central nervous system
Authors: Kumar Gaurav , Massey Kaylea , Ko Rose M , Axtell Robert C
Published: August 2023

B cell Maturation Antigen (BCMA) is a receptor that engages the cytokines BAFF and APRIL. BCMA is known to play a critical role in regulating B cell proliferation, survival, and differentiation into plasma cells. However, the effect of BCMA on non-B cells has been understudied. We have previously shown that BCMA regulates EAE, where BCMA−/− mice have exacerbated disease compared to BCMA+/+ mice. The increased disease in the BCMA−/− mice was associated with increased infiltration of B cells, T cells and inflammatory macrophages in the central nervous system (CNS). In this study, we assessed the expression patterns of BCMA on different immune cell types infiltrating the CNS of mice with EAE by flow cytometry. We found BCMA is expressed on plasma cells, microglia and inflammatory macrophages that are in the CNS, but not T cells or neutrophils. Next, we compared the numbers and phenotype of different myeloid cells in the CNS and lymph nodes in healthy BCMA−/− and BCMA+/+ mice. We found that BCMA−/− mice have an increased number of resident microglial cells with low CD86 expression in the brain compared to BCMA+/+ mice. Also, we found elevated numbers of macrophages with M1 (measured by the expression of iNOS) phenotype in the brain of BCMA−/− mice compared to BCMA+/+ mice. In lymph nodes, we found an increased number of M1 type macrophages in the BCMA−/− in comparison to BCMA+/+ mice with low CD86 expression. These data suggest a novel role for BCMA in regulating the phenotype and function of microglial and macrophages in CNS and in the peripheral immune tissues. Our data points towards a new pathway regulated by BAFF and APRIL signaling through BCMA and provides clues behind the immunopathogenesis of neuroautoimmune diseases.

This study was supported by grants from the NMSS (RG-1602-07722), the NIH (R01AI137047 and R01EY027346) awarded to R.C. Axtell.
C57BL/6 mice cohoused with pet shop mice phenocopy immune-related adverse events associated with checkpoint inhibition
Authors: Hapak Sophie , Vezys Vaiva
Published: August 2023

Checkpoint inhibition has been successful in the treatment of a number of malignancies. These medications block inhibitory pathways in T cells and allow for their improved function in anti-tumor immune responses. An important factor limiting checkpoint inhibitor therapy is immune-related adverse events (irAE), which can involve any organ system. It is currently not possible to predict which patients will develop irAE or which tissues will be affected. Our understanding of the cellular and molecular factors driving irAE development remains limited, as immune sufficient specific pathogen free (SPF) C57BL/6 mice do not develop irAE when treated with checkpoint inhibition. We have previously reported that C57BL/6 mice cohoused with pet shop mice recapitulate aspects of the human adult immune system, while SPF C57BL/6 mice have immune systems resembling that of human neonates. We hypothesized that checkpoint inhibition in C57BL/6 mice cohoused with pet shop mice will phenocopy irAE development in humans. Here, we report that treating “dirty” C57BL/6 mice with anti-CTLA4 and anti-PD-L1 resulted in the development of irAE, including nephritis, dermatitis, and hepatitis, as evaluated by histology and serum lab values. Ongoing experiments include depletion of CD4 and CD8 T cells to determine whether irAE are mediated by these cells. To our knowledge, this represents the first model of irAE in immune sufficient C57BL/6 mice, enabling further investigation into the mechanistic understanding of irAE and development of treatments targeting irAE while leaving anti-tumor responses intact.

DIABETES T32 RESEARCH TRAINING PROGRAM
Abrogating Regulatory T Cells Overcomes Tumor-Specific T cell Exhaustion and Prevents Metastatic Pancreatic Cancer
Authors: Schmiechen Zoe C , Burrack Adam L , Miller Ebony , Butler Jonah , Cruz Eduardo , Rollins Meagan , Patterson Michael , Wang Iris , Stromnes Ingunn
Published: August 2023

Pancreatic ductal adenocarcinoma (PDA) has a dismal 5-year survival rate of 10%. Lethality is attributed to late diagnosis, early metastasis, and therapeutic resistance. We identified pancreatic tumor cells derived from mice which resist immunotherapy (e.g., escape variants, EV) rapidly metastasize upon re-implantation into the pancreas of syngeneic and immunocompetent mice. EVs retain the dominant tumor antigen, and despite a defect in IFNg-inducible MHC class I upregulation, EVs remain sensitive to tumor antigen specific T cell-mediated lysis in vitro, suggesting EVs may confer unique qualities in vivo to resist T cell killing. Using a peptide:MHC tetramer based approach, we identified that tumor-specific CD8 T cells in EV tumors have increased Granzyme B production and a reduction in prototypical exhaustion markers Pd1, Lag3, and Tox. Further, restoring MHC class I upregulation reversed this phenotype. Notably, primary tumors from EVs were significantly enriched with Foxp3+ Tregs compared to parental tumors. Using a genetic model, we identified that Treg depletion resulted in a drastic reduction in tumor burden and metastasis and improved tumor-specific T cell function in EV tumors, expanding a putative anti-metastatic CD8 T cells subset. Investigation of tumor-cell intrinsic changes identified elevated expression of the receptor tyrosine kinase Ddr2in multiple EVs compared to immunotherapy naïve cells. Our data supports the novel hypothesis that Ddr2 overexpression drives intratumoral Treg accumulation and promotes metastasis. In summary, Tregs are key drivers of T cell exhaustion and immunosuppression in pancreatic cancer and may prove a valuable clinical target for tumors that evade immune checkpoint blockade.

Supported by NIH T32 grant – AG029796 – Functional Proteomics of Aging Training Program
T cell-dependent regulatory roles of 18-kDa Translocator protein (TSPO)
Authors: Sendai Yuka , Takeda Kazuyoshi , Ichinohe Tatsuya , Koshika Kyotaro , Okumura Ko , Azuma Toshifumi , Ohno Tatsukuni
Published: August 2023

Objective

TSPO (also called PBR) is a translocator protein that is localized on the outer mitochondrial membrane. TSPO is broadly and constitutively expressed in various types of immune cells including macrophages, monocytes, T cells, and B cells. Although many studies have shown that TSPO negatively regulates innate immune cell, such as macrophage and monocyte, activity, the roles of TSPO in T cell-mediated immune responses remain unclear. In this study, we examine the effects of Ro5-4864, a TSPO agonistic ligand, in an DNFB-induced contact hypersensitivity murine model.

Results and Discussion

TSPO expression in draining lymph nodes (dLNs) was elevated after sensitization. Ro5-4864 treatment at sensitization, but not at challenge, significantly suppressed ear swelling. The total cell numbers in the dLNCs were decreased by Ro5-4864 treatment. In the Ro5-4864-treated mice, the ratios of IFN-γ expressing CD4+ T cells and CD8+ T cells in dLNs were decreased. Adoptive transfer of Ro5-4864-treated sensitized T cells into naive mice inhibited ear swelling after ear challenge. Ro5-4864-treated sensitized T cells showed lower proliferative responses against DNFB-pulsed antigen presenting cells. Our results demonstrated that treatment with Ro5-4864 inhibited DNFB-induced contact hypersensitivity, suggesting the regulatory roles of TSPO in T cell-depending immune responses.
Assays for profiling changes in T cell metabolism
Authors: Bach Maggie L , Sylvester Kayla , Leippe Donna , Valley Mike , Cali Jim , Vidugiriene Jolanta
Published: August 2023

Immune cell therapy, including engineered T cells, engineered NK cells, and treatments to enhance native T cell response, is a rapidly advancing field of research with great promise to treat multiple diseases. However, multiple challenges, from isolating the correct cell type, determining its mode of action, manufacturing and establishing appropriate quality control criteria, must be overcome to reach its potential. In recent years, cellular energy metabolism has emerged as an important consideration for efficacy and persistence when developing cell therapies.

Here we evaluate a set of bioluminescent cell-based assays for measuring rate of metabolite uptake, monitoring activity for key metabolic pathways and determining changes in intracellular metabolite levels during T cell transition from naïve to activated to memory phenotypes. Metabolite levels measured include ATP, glucose uptake, malate, branched chain amino acids, and glycogen levels. Depending on the activation state of the cells, the cells respond differently to glycolysis and oxidative phosphorylation inhibitors, as demonstrated by changes to metabolite levels. Energy storage as measured by intracellular glycogen levels also differs depending on the T cell phenotype (naïve, activated, or memory).

Understanding the processes governing dynamic metabolic changes during cell modification and expansion is required for next-generation cell therapy development. The developed bioluminescent cell-based assays provide the sensitivity, throughput and robustness required for rapid evaluation of such changes.
Mitochondrial cristae disruption enables optimal T cell activation and expansion
Authors: Carvalho Edmund , Su Irene , Ecker Christopher , Srivatsa Sriram , Riley James
Published: August 2023

T cell activation is an anabolic process requiring both large amounts of energy and biomaterials to construct daughter T cells. We examined structural changes to the mitochondria during T cell activation, given its predominant role in generating ATP and harboring key building blocks such as NADH. Upon T cell activation, the mitochondria massively disturb their cristae and swell in a process that peaks after 48 hours. As the T cells begin to rest down, we observed that mitochondria begin to revert back to their pre-activation phenotype. Intriguingly environments that limit T cell activation have less cristae disruption, suggesting that this process is beneficial to the immune response. We saw rapid downregulation of electron transport chain subunits required for complexes 1,2 and 4. Moreover, NADH/NADPH, which is largely housed in the mitochondria of a resting T cell leaves the organelle to the cytoplasm, coincident with mitochondria cristae disruption, suggesting that the mitochondrial cristae disturbance causes a shift in the equilibrium towards of these molecules for energy production so that it can be used as a building block for nucleotides. This data suggest that the mitochondria alter their structure and function to promote optimal T cell activation.
Hypoxia promotes inflammatory fibroblast formation in pancreatic cancer
Authors: Mello Ashley M , Ngodup Tenzin , Donahue Katelyn L , Di Magliano Marina Pasca , Lee Kyoung Eun
Published: August 2023

Pancreatic ductal adenocarcinoma (PDAC) is a deadly malignancy characterized by poor response to all existing therapies. Although immunotherapy has shown great promise against multiple deadly cancers, it has been largely ineffective in PDAC. This lack of response is in part attributed to its extensive, fibroinflammatory stroma and hypoxic microenvironment. Cancer-associated fibroblasts (CAFs) are the predominant stromal cell type in PDAC, and single-cell transcriptomics of human and mouse PDAC has recently revealed a distinct CAF subpopulation, inflammatory CAFs (iCAFs). These inflammatory fibroblasts produce high levels of cytokines and chemokines in PDAC, and have the potential to contribute to its immunosuppressive microenvironment and tumorigenesis. By injecting a hypoxia probe into PDAC mouse models, we recently found that iCAFs predominantly reside in hypoxic tumor regions. We also observed that the hypoxia-related gene signature is positively enriched in iCAFs in human PDAC samples. Importantly, by exposing three-dimensional (3D) co-cultures of pancreatic cancer cells and fibroblasts to either hypoxia or normoxia, we showed that hypoxia induces IL1α from cancer cells and that IL1α is required for hypoxia-mediated iCAF formation. Our data implicate hypoxia as a critical regulator of tumor stroma. Efforts are ongoing to understand the role of hypoxia in the crosstalk between cancer cells, fibroblasts, and immune cells in driving an immunosuppressive tumor microenvironment.

Supported by a grant from the NIH (T32 AI007413) and the University of Michigan Rackham Merit Fellowship.
Melanocyte PDL1 promotes early tumor progression in a novel autochthonous NRAS-mutant melanoma model
Authors: Ontiveros Carlos Oscar , Chen Tiffany , Murray Clare , Kornepati Anand , Deng Yilun , Blinkiewicz Paige , Bai Haiyan , Crossland Grace , Souto-Padron Alvaro , Gupta Harshita B , Turk Mary Jo , Burd Christin , Curiel Tyler J
Published: August 2023

Tumor-intrinsic PDL1 drives melanoma treatment resistance. Melanocytes (melanoma cell-of-origin) do not express PDL1, but PDL1 is expressed in malignant melanomas, suggesting melanocyte PDL1 promotes melanomagenesis. We developed a novel autochthonous mouse model that develops NRASQ61R-mutant melanomas lacking PDL1 only in melanocytes (PDL1KOTNQ61R) and littermate controls (PDL1+/+ TNQ61R). We induced mice with 4-OH tamoxifen ± UV exposure to accelerate melanoma development. Tumor latency significantly increased in PDL1KO vs. PDL1+/+ TNQ61R mice at 0 kJ/m2 and 2 kJ/m2UV(p<0.02), suggesting melanocyte/tumor PDL1 promotes melanomagenesis. Latencies were similar at 4.5 kJ/m2, suggesting immune contributions, e.g., from local immunosuppression. We derived transplantable cell lines from autochthonous tumors. PDL1+/+ TNQ61R cell lines had increased mTORC1 and stemness, and reduced STING and reduced Chk1 inhibitor sensitivity in vitro and in vivo vs. PDL1KOTNQ61R, phenocopied by PDL1KO B16. Tumor PDL1 suppressed ERK signals (NRAS target) in TNQ61R vs. PDL1KO cell lines and promoted resistance to NRAS-targeting small molecule drugs, suggesting tumor PDL1 promotes NRAS signals. Transplanted 4.5 kJ/m2UV-inducedPDL1KO TNQ61R tumors were resistant to αPD1, αPDL1, αPDL2, and CD122-biased IL2 in WT mice vs. B16 melanomas, which are sensitive to biased IL2. Our model allows studies of PDL1 signals in melanomagenesis and progression and distinguishes bona fide cell-intrinsic PDL1 signals.

This research was funded by the NIH T32GM113896 (STX MSTP) Award (C.O.) and the NCI (CA204965, CA054515) supported Curiel.
ST8Sia6 sialylates CD44 in cancer cells to generate ligands for inhibitory Siglecs
Authors: Chen Yin , Friedman David J , Saraswat Mayank , Chen Alex Shuaibo , Shapiro Michael , Wilfahrt Drew , Pandey Akhilesh , Shapiro Virginia Smith
Published: August 2023

Glycans decorate most proteins and lipids on the cell surface, and glycoproteins serve a critical role in cell-cell communication. The terminal modification of many glycans is a sialic acid. There are twenty different mammalian sialyltransferases, each with a unique specificity. Recently, we demonstrated that the sialyltransferase ST8Sia6, which adds a terminal alpha-2,8 disialic acid to a preexisting sialic acid on cell surface proteins, generates ligands for the inhibitory receptor Siglec-E. In patients, ST8Sia6 overexpression is associated with worse outcome. We have previously shown that ST8sia6 overexpression in a mouse tumor model accelerates tumor growth, dependent upon Siglec-E expression by immune cells. To understand the basis for ST8Sia6 function in tumors, proteomic analysis of murine tumor lines was performed to identify proteins modified by ST8Sia6. Here, we demonstrate that ST8Sia6 sialylates CD44. Siglecs vary significantly between species, with 5 non-conserved Siglecs in mice compared to 10 non-conserved Siglecs in humans. The closest ortholog to Siglec-E in mice is Siglec-7 in humans. We also demonstrate that ST8Sia6 sialylates CD44, generating ligands for Siglec-7. Thus, ST8sia6 has a conserved role in both mice and humans in modifying glycans on CD44 to generate ligands for Siglecs. As CD44 expression on tumors is associated with increased metastasis, we predict that ST8Sia6 may also have a role in this process which is under investigation.

Supported by grants from 2T32AI007425-23 to DJF and NIH Glycobiology Common Fund supplement to R01CA243545-01A1 to VSS.
A shift in the hepatic immunological pattern from predominant adaptive immunity to innate immunity compromises tissue-specific immunity and leads to HCC
Authors: Koelsch Nicholas James , Mirshahi Faridoddin , Olex Amy , Sanyal Arun , Manjili Masoud H
Published: August 2023

The incidence of hepatocellular carcinoma (HCC) associated with non-alcoholic fatty liver disease (NAFLD) is growing because of the global consumption of high fat, high sugar or Western diet (WD). Current data suggests a correlation between the hepatic inflammation and progression of NAFLD to HCC. However, contradictory reports on the identification of specific inflammatory cells or cytokines hinder further progress in the field. Such controversial reports are due to highly reductionist approaches and getting lost in the details. By using flow cytometry, we have demonstrated that the hepatic immunological patterns were associated with the progression or inhibition of HCC, which can be discovered through the detection of the proportion of immune cell types functioning as a network. However, flow cytometry utilizes a limited number of markers hindering a comprehensive detection of the patterns. Here, we perform scRNAseq of the liver collected from the mouse model of NAFLD (DIAMOND) being on a WD prior to or after the development of HCC, as well as control mice being on a chow diet (CD). We demonstrate that a WD alters the hepatic immunological pattern by dominating innate immunity, M1 macrophages, over adaptive immunity, B and T cells, associated with the activation of the hepatic fibroblasts. Each immunological pattern created a distinct collective function beyond the function of its cellular constituents. Also, shifts in the immunological pattern compromised the hepatic immune system, transforming its anti-tumor function to carcinogenesis-promoting process. Our data suggest that the discovery and modulation of the hepatic immunological pattern could offer a comprehensive understanding of and curative treatment for NAFLD and HCC.

This work was supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program under Award No. W81XWH2210793. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the U.S. Department of Defense.This work was also supported, in part, by CTSA award No. UL1TR002649 from the National Center for Advancing Translational Sciences.
Exploring the effects of tissue structural cell and tumor cell heparan sulfate on immune system trafficking, activation, and intercommunication via amphiregulin
Authors: Loffredo Lucas F , Li Fangda , Ringham Olivia R , Surpur Anmol , Cavero Renzo , Arpaia Nicholas
Published: August 2023

Heparan sulfate (HS), a glycan subtype, is ubiquitously present on the surface of structural tissue cells and tumor cells during homeostasis, disease, and cancer, but how it affects immune cell trafficking, activation, and signal transmission in these tissue contexts is poorly understood. Furthermore, amphiregulin (Areg), a growth factor used by immune cells for intercommunication with structural tissue and tumor cells, contains a HS binding domain. We investigated the downstream signaling modalities of Areg-mediated cell activation in the context of HS inhibition. Areg signaling is qualitatively altered, but still present, upon exogenous inhibition or genetic ablation of HS. We further probed the consequences of HS-dependent vs. -independent Areg signaling at the transcriptional level. Next, we used a panel of CRISPR-Cas9-mediated knockout Lewis Lung Carcinoma (LLC) cell lines to interrogate how various aspects of HS (core proteoglycan, HS chain extension, sulfation status) contribute to Areg signaling. We found that the presence of the glypican family of HS-harboring proteoglycans on the surface of cells is critical for Areg signaling. Lastly, we explored the effects of cancer cell HS on immune system trafficking to and activation within tumors. HS-deficient tumors experience similar levels of lymphoid cell trafficking and activation, but show a significant increase in the presence of certain myeloid cell type (neutrophils, monocytes, and macrophages). Additionally, tumor-associated macrophages within HS-deficient tumors show an altered phenotype, with increased maturation markers. Overall, our work has illuminated previously understudied aspects of the relationships between HS, Areg, and the immune system.

Supported by a grant from NIH (R21-AI149657)
Myocyte enhancer factor-2C is critical for the maintenance of ATLL cells by regulating the activity of retroviral antisense promoter and bZIP
Authors: Jain Pooja , Madugula Kiran , Joseph Julie , DeMarino Catherine , Ginwala Rashida , Teixeira Vanessa , Khan Zafar , Sales Dominic , Wilson Sydney , Kashanchi Fatah , Rushing Amanda , Lemasson Isabelle , Harhaj Edward , Janakiram Murali , Ye B Hilda , Jain Pooja
Published: August 2023

Adult T-cell leukemia and lymphoma (ATLL) is an intractable T-cell neoplasia caused by a retrovirus, namely human T-cell leukemia virus type 1 (HTLV-1). Patients suffering from ATLL present a poor prognosis and have a dearth of treatment options. In contrast to the sporadic expression of viral transactivator protein Tax present at the 5′ promoter region long terminal repeats (LTR), HTLV-1 bZIP gene (HBZ) is encoded by 3′LTR (the antisense promoter) and maintains its constant expression in ATLL cells and patients. The antisense promoter is associated with selective retroviral gene expression and has been an understudied phenomenon. Herein, we delineate the activity of transcription factor MEF (myocyte enhancer factor)-2 family members, which were found to be enriched at the 3′LTR and play an important role in the pathogenesis of ATLL. Of the four MEF isoforms (A to D), MEF-2A and 2C were highly overexpressed in a wide array of ATLL cell lines and in acute ATLL patients. The activity of MEF-2 isoforms were determined by knockdown experiments that led to decreased cell proliferation and regulated cell cycle progression. High enrichment of MEF-2C was observed at the 3′LTR along with cofactors Menin and JunD resulting in binding of MEF-2C to HBZ at this region. Chemical inhibition of MEF-2 proteins resulted in the cytotoxicity of ATLL cells in vitro and reduction of proviral load in a humanized mouse model. Taken together, this study provides a novel mechanism of 3′LTR regulation and establishes MEF-2 signaling a potential target for therapeutic intervention for ATLL.

NIH/NINDS: R01 NS097147 to PJ.
Antagonism of regulatory ISGs enhances the anti-melanoma efficacy of STING agonists
Authors: Filderman Jessica , Storkus Walter J
Published: August 2023

STimulator of INterferon Genes (STING) is a dsDNA sensor that triggers type I inflammatory responses. Recently this pathway has been leveraged as a cancer treatment with the use of STING agonists. Our lab has previously demonstrated that treating mice bearing established tumors with STING agonists leads to an inhibition in tumor growth in association with tumor vascular normalization and immune cell recruitment within the tumor microenvironment. However, STING agonism also results in the upregulated expression of regulatory interferon-stimulated genes (ISGs), including ARG2, enzymes involved in the production of immunosuppressive prostaglandins (i.e., PTGES and PTGS2/COX2), PD-L1, and ISG15, which limit the overall efficacy of this immunotherapeutic paradigm. We hypothesized that combined treatment of melanoma-bearing mice with STING agonists along with targeted inhibitors of ARG2, PTGES/COX2, PD-L1, and ISG15 would result in improved control of tumor growth and more robust anti-tumor immune responses in vivo. In the B16 (BRAFWTPTENWT) melanoma model, we observed improvement in tumor growth control after treatment with STING agonists + blocking anti-PD-L1 or anti-ISG15 antibodies vs. monotherapy. However, in the BPR (BRAFV600EPTEN−/−) melanoma model, tumor growth control over that observed for STING agonist alone was only noted upon combined addition of pharmacologic inhibitors of ARG2, COX2, and NOS2. The underlying therapy-associated changes in the TIME that support improved tumor growth control are currently being evaluated by single cell RNAseq, flow cytometry, and IFM to discern mechanism(s) of action.

Supported by grants from NIH (P01 CA234212) and the AAI Careers in Immunology Fellowship.
Regulation of immune-recognition molecules following delivery of plasmid backbone
Authors: Shi Guilan
Published: August 2023

Local intratumor delivery with electroporation of low levels of plasmids encoding molecules, such as IL-12, IL-15, induces an antitumor effect without causing systemic toxicity. However, previous studies have predominately focused on the function of the delivered molecule encoded within the plasmid, and ignored the plasmid backbone. In this study, we found backbones pUMVC3 and pVax1 induced up-regulation of MHC class I (MHC-I), PD-L1, CD95 (Fas) and CD155 on tumor cell surface. These molecules participate in a considerable number of immunoregulatory functions through their interactions with activating and inhibitory immune cell receptors. MHC molecules are well-known for their role in antigen (cross-) presentation, thereby functioning as key players in the communication between immune cells and tumor cells. Increased PD-L1 expression on tumor cells is an important monitor of tumor growth and effectiveness of immune inhibitor therapy. During cancer progression CD95 is frequently downregulated, raising the possibility that loss of CD95 is part of a mechanism for tumor evasion. Due to its prominent endogenous and immune functions, CD155 has been gaining interest as a therapeutic target in the field of tumor immunology. Results from flow cytometry confirmed increased expression of MHC-I and PDL-1 on B16F10, 4T1 and KPC tumor cell lines. Preliminary animal data from tumor-bearing models, B16F10 melanoma, 4T1 breast cancer and KPC pancreatic cancer mouse models showed that tumor growth was attenuated after pUMVC3 intratumoral electroporation. Our data also documented pSTAT1-NF-κB signaling pathway might be associated with plasmid backbones’ function of up-regulating MHC-I, PD-L1, CD95 (Fas) and CD155 on tumor cells.

R01 CA186730
Profiling cGAS-STING pathway in T-Acute Lymphoblastic Leukemia
Authors: Rocha Maria Theresa Araújo Balby , Tomchuck Suzanne , Askew David , Marti Luciana , Huang Alex
Published: August 2023

Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy, accounting for over one-quarter of all childhood malignancies. Approximately 15% of ALL derive from the T-cell lineage (T-ALL). Patients with newly diagnosed T-ALL undergo intensive multidrug combination chemotherapy resulting in many side effects. Despite chemotherapy regimens including steroids, and allogeneic transplantation, relapse remains the main challenge in children with T-ALL. Moreover, genetic alterations have been extensively examined in recent years; however, there are still currently few prognostic markers for therapy response. Therefore, alternative treatments are needed to treat this disease and improve overall survival.

The cyclic GMP-AMP synthase - Stimulator of Interferon Genes (cGAS-STING) pathway has emerged as a possible target in treatment of cancers, aiming to increase anti-tumor immunity. Most clinical trials to-date have focused on STING agonists used for solid tumors. Interestingly, the high activation of this pathway in normal T cells can lead to ER stress and cell death via the activation of the BH3-family. The status of cGAS-STING pathway in T-ALL is largely unknown. We hypothesize that the cGAS-STING pathway is defective in T-ALL, thus protecting these malignant cells from apoptosis, and that increasing cGAS-STING activity may be a way of enhancing the treatment of this disease. Our preliminary data shows that some T-ALL cell lines do not produce IFN-β after stimulation to STING agonist. Also, some of these cell lines have a decreased cGAS expression, and have a decreased rate of apoptosis compared to healthy T-cells. Our project will hopefully lay the foundation for future targeted immunotherapies for treating T-ALL.

Center for Pediatric Immunotherapy at Rainbow
Antigen-Specific CD4 T Cell and B Cell Responses to Borrelia burgdorferi
Authors: Hammond Elizabeth M , Olsen Kimberly J , Ram Shivneel , Tran Giang Vu Vi , Hall Laura S , Bradley John E , Lund Frances E , Samuels D Scott , Baumgarth Nicole
Published: August 2023

Long-lived T-dependent B cell responses fail to develop during persistent infection of mice with Borrelia burgdorferi, the causative agent of Lyme disease, raising questions about the induction and/or functionality of anti–B. burgdorferi adaptive immune responses. Yet, a lack of reagents has limited investigations into B. burgdorferi–specific T and B cells. We attempted two approaches to track B. burgdorferi–induced CD4 T cells. First, a B. burgdorferi mutant was generated with an influenza hemagglutinin (HA) peptide, HA111–119, inserted into the B. burgdorferi arthritis-related protein (Arp) locus. Although this B. burgdorferi arp::HA strain remained infectious, peptide-specific TCR transgenic CD4 T cells in vitro, or adoptively transferred into B. burgdorferi arp::HA–infected BALB/c mice, did not clonally expand above those of recipients infected with the parental B. burgdorferi strain or a B. burgdorferi mutant containing an irrelevant peptide. Some expansion, however, occurred in B. burgdorferi arp::HA–infected BALB/c SCID mice. Second, a (to our knowledge) newly identified I-Ab–restricted CD4 T cell epitope, Arp152–166, was used to generate Arp MHC class II tetramers. Flow cytometry showed small numbers of Arp-specific CD4 T cells emerging in mice infected with B. burgdorferi but not with Arp-deficient Borrelia afzelii. Although up to 30% of Arp-specific CD4 T cells were ICOS+PD-1+CXCR5+BCL6+ T follicular helper cells, their numbers declined after day 12, before germinal centers (GCs) are prominent. Although some Arp-specific B cells, identified using fluorochrome-labeled rArp proteins, had the phenotype of GC B cells, their frequencies did not correlate with anti-Arp serum IgG. The data suggest a failure not in the induction, but in the maintenance of GC T follicular helper and/or B cells to B. burgdorferi.
Expression and clinical significance of HER2/neu, aromatase P450 and adhesion molecule CD24 in endometrial cancer
Authors: Guan Liyun , Wang Ying , Cheng Jianxin , Zhang Jun , Kang Shan
Published: August 2023

This study aimed at exploring the expression and clinical significance of aromatase P450, adhesion molecule CD24 and HER2/neu in endometrial cancer. The expression of aromatase P450, adhesion molecule CD24 and HER2/neu was detected by immunohistochemistry in 15 cases of endometrial hyperplasia group, 50 cases of endometrial adenocarcinoma and 3 cases of uterine papillary adenocarcinoma, with 15 cases of normal endometrium as control group. We detected no expression of aromatase P450, adhesion molecule CD24 or HER2/neu in control group. Aromatase P450 positive expression rate was 66.7% in endometrial hyperplasia group and 70.3% in endometrial carcinoma group, without significant difference (p>0.05). There was no significant difference (p>0.05) in the positive expression rate of aromatase P450 between different myometrial invasion groups of endometrial adenocarcinomas. CD24 positive expression rate was 40.0% in endometrial hyperplasia group and 79.6% in endometrial carcinoma group, with significant difference (p<0.05). HER2/neu positive expression rate was 26.7% in the endometrial hyperplasia group and 57% in endometrial carcinoma group, with significant difference (p<0.05). In conclusion, aromatase P450 may be one factor associated with endometrial cancer cell proliferation, while CD24 and HER2/neu may be important factors associated with the invasion and metastasis of endometrial cancer.
The limits of political development and constitutionalism in South Africa
Authors: Van Riet Gideon
Published: August 2023

The article investigates political development and constitutionalism in post- apartheid South Africa by focusing on two general features of this society. These are firstly the enduring prevalence of violence defined broadly. The second feature is the particular democratic deficit manifested in the politics of professionalism associated with the New Public Management (NPM) informed developmental state. The article interprets these two trends as characteristic of ruptures and continuities with the apartheid state. It scrutinizes the underlying assumptions of political development and constitutionalism and critiques both as ideals for the post-apartheid state. It is concluded that political development and constitutionalism, as they have manifested in post-apartheid South Africa, are insufficient in alleviating the structural violence which characterizes the everyday for millions of South Africans. Ordinary citizens must obtain greater access to the decision-making processes in which they are currently not meaningfully included through contemporary developmental practices. Such inclusion would serve both as an end in itself and as a means towards greater two-directional integration between marginalized citizens and dominant processes of material and symbolic production and consumption. At the same time, constitutionalism, by enshrining a relatively inflexible approach to property rights, is impotent in the face of persistent and increasing material inequalities.
Duodenal Tuberculosis
Authors: Palaniappan S. , Munirathinam Manoj , Murali A.
Published: August 2023

Tuberculosis (TB) of the gastrointestinal tract is common in India. The most common site is the ileocecal region. TB involving the duodenum is rare. It can present as duodenal ulcer, gastric outlet obstruction, and rarely with periduodenal lymph nodal involvement and bile duct erosion. Here, we report a patient who presented with pain abdomen and significant weight loss. Evaluation revealed a duodenal ulcer with paraduodenal nodal mass with probable contained perforation. Endoscopic ultrasound with fine-needle aspiration cytology of the nodal mass revealed the diagnosis of TB. She was started on anti-TB therapy and responded well with complete resolution. This case report emphasizes the need for having TB as a differential diagnosis for duodenal ulcer in an endemic country like India.
Clinico-Epidemiological Study of Papulosquamous Disorders in Childhood at a Tertiary Care Hospital in South India
Authors: Gurudev Shradha , Pise Gajanan , Manohar Naveen , Sanagoudar Vinitha , Rao Kiran , Patil Malteshgauda N
Published: August 2023

Introduction:

Papulosquamous disorders in children differ from those in adults in terms of presentations, treatments, and prognosis. The aim of this study was to add to the existing data regarding the presentation of pediatric papulosquamous disorders and analyze the metabolic parameters in children with psoriasis.

Materials and Methods:

This observational study included 83 patients (age, 0–18 years) with clinically diagnosed papulosquamous disorders. Demographic data and detailed clinical history were collected followed by a detailed clinical examination. In children with psoriasis, we assessed the parameters of metabolic health, such as anthropometry and blood chemistry. The outcomes are expressed as percentages and proportions.

Results:

Papulosquamous disorders constituted 2.3% of all the pediatric dermatoses with a male-to-female ratio of 1.7:1. They were commonly noted in the age group of 7–12 years. Psoriasis vulgaris (26.5%) was the most common disease followed by lichen planus (19.2%), seborrheic dermatitis (16.8%), pityriasis rosea (13.2%), lichen striatus (12%), lichen nitidus (7.2%), pityriasis lichenoides chronica (3.6%), and pityriasis rubra pilaris (1.20%). None of the 22 patients with psoriasis fulfilled the criteria for metabolic syndrome. However, aberrant serum levels included elevated fasting glucose (n = 5, 22.7%), elevated fasting triglycerides (n = 3, 13.6%), and decreased high-density lipoproteins (n = 2, 9.1%). Anthropometric abnormalities were also not noted.

Conclusions:

Understanding the morphological and epidemiological features of pediatric papulosquamous disorders can aid in early diagnosis, treatment, and counseling the patients and parents, which can alleviate their anxiety and improve the psychological distress.
Comparison of the Efficacy of Intralesional Measles, Mumps, and Rubella Vaccine with Intralesional Vitamin D3 for the Treatment of Extragenital Warts in Pediatric Age Group (5–18 Years)
Authors: Ahmed Jafir , Kushwaha Ramesh Kumar , Sharma Avinash , Agarwal Morish , Sharma Neha , Jain Suresh Kumar
Published: August 2023

Introduction:

Warts are benign skin growths caused by human papillomavirus. Traditional methods of treatment usually include destructive modalities which are painful and scar-forming. Such modalities have disadvantage of recurrence. Consequently, immunotherapy is introduced to overcome these problems.

Aims and Objectives:

To study the comparative efficacy of intralesional measles, mumps, and rubella (MMR) vaccine with intralesional Vitamin D3 for the treatment of extragenital warts in the pediatric age group (5–18 years).

Materials and Methods:

Seventy-two immunocompetent patients aged between 5 and 18 years with multiple extragenital warts (2 or more) were enrolled and divided randomly into two groups (Group A and Group B). Group A and Group B received intralesional MMR vaccine and Vitamin D3, respectively, in the largest wart present. The injections were repeated every 2 weeks for a maximum of four injections.

Results:

Complete response (CR) in injected warts in Group A is seen in 25 (83.3%) patients, partial response (PR) in 3 (10%), and no or inadequate response (NR) in 2 (6.67%) patients. CR is seen in 23 (76.66%) patients of Group B, PR in 4 (13.33%) and no or NR in 3 (10%) patients. CR in distant warts in 22 (73.33%) patients in Group A and 20 (66.66) patients in Group B.

Conclusion:

Both MMR vaccine and Vitamin D3 are safe, efficacious, and less traumatic to the patients for multiple warts in comparison with destructive modalities.
Lymphocutaneous Sporotrichosis
Authors: Kutty Vijayaragavan , Soundaian Balasankar , Jeyaraman Balasubramanian , Manisekaran Sathishkumar
Published: August 2023

Sporotrichosis is a chronic subcutaneous mycosis caused by a fungus, Sporothrix schenckii complex. The first case in India was reported by Ghosh in 1932. The disease is prevalent primarily in the sub-Himalayan region, North-eastern states, and parts of Karnataka. Mostly seen in adults, it uncommonly presents in children over the face and limbs either as fixed cutaneous or lymphocutaneous form. Here, we present a case of 3-years-old boy hailing from Southern Tamil Nadu with multiple nodular lesions in linear fashion over the right upper limb. It was diagnosed as a case of lymphocutaneous sporotrichosis that developed following cut injury over the right index finger. We publish this case as it is rare to occur in this geographic location.
Sturge–Weber Syndrome in Childhood Lupus Complicated by Macrophage Activation Syndrome
Authors: Jethwa Manisha , Viswanath Vishalakshi , Dhakne Mayuri , Joshi Pradnya R
Published: August 2023

Sturge–Weber syndrome (SWS) is an uncommon, nonhereditary developmental condition characterized by nevus flammeus and ocular and neurological manifestations. Childhood-onset systemic lupus erythematosus (cSLE) is a rare disease with a female preponderance and is associated with multiple complications and a poor prognosis. The coexistence of cSLE and SWS is uncommon and can be associated with an increased risk of thromboembolic and neuropsychiatric complications. Macrophage activation syndrome (MAS) is one of the fatal complications of cSLE and an early diagnosis based on clinical findings and laboratory parameters is vital. We report a rare and unique case of SWS with cSLE in an 11-year-old boy, which was complicated by MAS.
Oligometastatic carcinoma prostate – An overview of the last decade
Authors: Shah Milap J , Sharma Aditya Prakash , Hameed B. M. Zeeshan , Jain Ravi , Patil Abhijit , Karthickeyan Naganathan , Singh Abhishek
Published: August 2023

Introduction:

Oligometastatic prostate cancer (OMPC) has gained profound interest lately due to its different tumor biology and our ability to use multimodality therapy for cure or prolonged survival. Selecting the appropriate patient for treatment has become the aim of treating urologists, medical oncologists, and radiation oncologists. Through this review, we try to highlight the management of OMPC in light of recent literature.

Methods:

Literature search was performed on Pubmed, Scopus and Embase using keywords “Oligometastatic”, “ Prostate Cancer” using operators such as “And” & “Or”. Relevant articles were screened and all the latest articles on this emerging entity were included in this review.

Results:

All trials relevant to oligometastatic prostate cancer defining the role of surgery, radiotherapy and systemic therapy were included and appropriate inferences were drawn. Relevant studies were compiled in tabular form for this article.

Conclusion:

The current standard of care of management for OMPC remains systemic therapy on the lines of hormone-sensitive metastatic prostate cancer. The evolving role of surgery, and radiotherapy along with systemic therapy is highlighted in this article.
Knowledge, Awareness and Practice of Hand Hygiene among HealthCare Workers for the Prevention of Infection: A Cross-Sectional Study
Authors: Gaur Namrata , Gaur Dushyant Singh
Published: August 2023

Background:

Hand hygiene protocols, if followed stringently reduce the possibility of the spread of infection from healthcare workers (HCWs) to patients, to almost 99%. However, in actual practice, compliance of these protocols is poor.

Aims:

(1) To observe the practice of hand hygiene among HCWs and (2) assess knowledge and awareness regarding hand hygiene protocol and identify the factors hampering its effective practice. Prospective observational study at an multispeciality hospital.

Patients and Methods:

Consultants, junior doctors, nursing in-charges and nurses and ancillary staff like ward attendants, ward secretaries, housekeeping staff and lift operators were (1) Observed during their hand hygiene practice and (2) 200 of these HCWs were provided a written questionnaire, in order to assess their depth of hand hygiene knowledge. Data were studied and analysed.

Results:

Female HCWs, right from consultants to ward attendants and ancillary workers showed better knowledge and compliance with hand hygiene practices. The junior nurses, interns and junior residents were more compliant with hand hygiene procedures in comparison to consultants and in-charge nurses. Hand hygiene compliance was poor in outpatient department, particularly on days with heavy workloads. Lack of knowledge regarding hand hygiene was observed amongst ancillary staff.

Conclusions:

The study emphasised stringent compliance and constant re-enforcement of protocol of correct hand hygiene practice amongst HCWs including ancillary staff.
Comparative Analysis of Conventional Cytology and Liquid-Based Cytology in the Detection of Carcinoma Cervix and its Precursor Lesions
Authors: Maheshwari Yashika , Handa Uma , Aggarwal Phiza , Goel Bharti
Published: August 2023

Context:

The conventional smears (CS) and Liquid based cytology (LBC) are important tools to detect carcinoma cervix and its precursor lesions.

Aims:

The present study was done to compare the cytomorphological features of cervical lesions using both techniques and compare with the histopathological diagnosis.

Settings and Design:

This was a prospective observational study over a period of 1.5 years at a tertiary care hospital.

Methods and Material:

A total of 969 women in the age group of 21–65 years presenting with either routine screening or complaints of vaginal bleeding, discharge, or pelvic pain were enrolled for the study. Both the CS and LBC smears were analyzed and compared with the corresponding histopathology diagnosis. The data was analyzed using Statistical Package for the Social Sciences (SPSS) software and P values <0.05 were considered significant.

Results:

There were 8.57% unsatisfactory smears in CS as compared to 0.5% in LBC smears. Liquid-based cytology was superior to conventional preparations in terms of smear adequacy, lesser hemorrhagic and inflammatory background, and presence of more endocervical cells. Liquid-based cytology showed a better yield in detecting all the types of epithelial cell lesions with a concordance rate of 73.9% between the two techniques. On histopathology correlation of these lesions, LBC had a higher sensitivity (96.67%) and diagnostic accuracy (99.08%) as compared to CS (73.33% and 92.66%, respectively).

Conclusions:

Liquid-based cytology is superior to conventional cytology for the detection of epithelial cell lesions. Reduction in the unsatisfactory smears, a cleaner background, and better representation of the sample are more significantly appreciated on LBC in contrast to CS.
Cytological Diagnosis of Castleman Disease by Endoscopic Ultrasound Guided Fine Needle Aspiration
Authors: Bhatia Vikram , Singh Satender Pal , Mishra Ajay Kumar , Rath Indira , Prabhu Meghana , Bihari Chhagan
Published: August 2023

Castleman disease is a nonmalignant, lymphoproliferative disorder. Unicentric disease type involves a single enlarged lymph node or nodal regions, and multicentric disease involves multiple lymph node site involvement. We present a case of 26-year-old young female presented to outpatient with complaint of generalized weakness and abdominal pain for last 2 months. Her imaging included positron emission computed tomography which showed soft-tissue mass along the body of pancreas. Endoscopic ultrasound done showed nodal mass with extensive calcification and doppler endoscopic ultrasound showing arborising vessels. Fine needle aspiration cytology (FNAC) and biopsy taken showed lymphoid cells and diffuse hyaline material. After suboptimal response to chemotherapy regimen, she underwent open laparotomy and excision of the retroperitoneal mass.