bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigating the potential of CD8α:MyD88 to restrain terminal exhaustion in tumor-specific T cells
Authors: Albert Gabriella , Cao Phoebe , Davila Eduardo 
Published: November 2024

Abstract
Despite the advancements in immunotherapies for solid tumors, many patients fail to develop durable responses. The immunosuppressive and antigen persistent environment drives T cell exhaustion, interfering with the efficacy of adoptive cell therapies. Terminally exhausted T cells (TTEX) co-express inhibitory receptors, lose effector function and fail to elicit tumor control. Our lab has reported that T cells engineered to express a CD8α:MyD88 fusion protein exhibit enhanced anti-tumor activity and appear less exhausted. We hypothesize that expression of CD8α:MyD88 in T cells restrains exhaustion by transcriptionally repressing inhibitory receptor expression thereby supporting effector function, persistence, and memory. Our RNAseq and RT-qPCR data revealed that expression of inhibitory receptor genes (Havcr2, Entpd1, CD244) was lower in CD8α:MyD88 compared to CD8α T cells. Moreover, collection of CD8α:MyD88 TIL from B16-F10 tumors (10 days post transfer) exhibited reduced frequencies of TTEX (PD1+, TIM3+, CD39+) cells, greater expression of TCF1 and co-stimulatory molecules, and remained functional upon ex vivo restimulation (IFNg). Additionally, a greater frequency of central memory T cells was present in secondary lymphoid tissues. Together, our results suggest that expression of CD8α:MyD88 in T cells may interfere with the expression of inhibitory receptors that dampen the anti-tumor response, thus enhancing the functional longevity and memory potential of T cells.
Novel tumor-specific Vd1/3 gd CAR T cells effectively control solid tumors
Authors: Lopez Bailon Luis , Anadon Carmen , Chaurio Ricardo , Mine Jessica , Plappert Noah , Antonia Scott , Conejo-Garcia Jose 
Published: November 2024

Abstract
Chimeric Antigen Receptor (CAR) T cell therapy has proven to be successful in treating hematological malignancies. However, its effectiveness against solid tumors has been suboptimal so far, due to the strong immunosuppressive conditions and metabolic restrictions present in the tumor microenvironment. Furthermore, the use of autologous T cells for CAR product manufacturing has several disadvantages. Here, we demonstrate the effectiveness of a new CAR T cell platform using cord blood-derived γδ T cell subsets, which can be used allogeneically (“off-the-shelf”) and recapitulate the populations primarily found in solid tumors. As targeting motifs, we used scFv sequences specifically targeting transmembrane molecules expressed in metastatic SCLC and other tumors, but not in vital tissues, which were identified through a comprehensive proteogenomic approach. To minimize PD-1-driven inhibition and enhance metabolic re-programming, we designed a new CD3z-less CAR architecture, which is more effective than conventional second-generation CARs. Our results demonstrate that γδ T cells are natural killers independent of MHC presentation, superior to αβ T cells, allowing the generation of multiples ‘off-the-shelf’ CAR T cell products. Moreover, the new architecture demonstrated better efficiency in killing tumor cells, thus supporting a rationale for future trials aimed at rendering CAR T cells effective against the most frequent and aggressive human solid tumors.
Enrichment using a selectable CD34t marker enhances TCR/CAR transduced T cell functionality and transgene expression in clinically relevant receptors
Authors: Quinn Suzanne , Scurti Gina , Dalheim Annika , Bochnacka Oliwia , Hossain Nasheed , Nishimura Michael 
Published: November 2024

Abstract
Gene-modified TCR transduced T cells and CAR T cells are used clinically and display a range of efficacy across multiple cancers. Many therapies have successfully controlled patient’s tumors; however, some have shown off-target adverse events which are harmful to patients’ overall outcomes. T cell-based therapies have high variation in patient outcome due to multiple factors, such as transduction efficiency and T cell phenotypes. To reduce product variability, our lab developed a selectable marker gene for use in cellular constructs. This selectable marker is a truncated, non-signaling CD34 molecule (CD34t), which can be used to remove the non-transduced T cells that function as bystanders. Our clinical trials have used two TCRs (TIL 1383I and HERV-E) and one CAR (CD19 CAR). Following transduction with these CD34t constructs, we determined transduction efficiency by expression of CD34t. CD34t expression correlates with transgene, allowing us to select high CD34t expressors, and thus high transgene expressors. The selected population produces higher levels of inflammatory cytokines and CD107a expression compared to the nonselected. This improved functionality is likely due to the selected population having higher levels of transgene across the population, whereas the nonselected has a broader range of transgene expressions with fewer expressors total. This selectable marker contributes to a more functional product, which may in turn improve patient response clinically.
Early exposure to exogenous IL-15 subverts ILC precursor priming and the generation of KIR+ NK cells
Authors: Ruesch Michael , Broughton Megan , Eisele Caprice , Kowalski Jessie , Nalin Ansel , Sara Kinda , Elmer Isaac , Anderson Stephen , Oakes Christopher , Mundy-Bosse Bethany , Freud Aharon 
Published: November 2024

Abstract
Killer immunoglobulin-like receptors (KIRs) allow natural killer (NK) cells to recognize and kill tumor cells. The mechanisms that control KIR acquisition during human NK cell development are not known. We previously observed that while human tonsil innate lymphoid cell precursors (ILCPs) could become KIR+ NK cells in immune deficient mice treated with interleukin (IL)-15, ILCPs cultured in vitro with IL-15 and OP9-DL1 stroma generated only rare KIR+ NK cells, suggesting that additional signals are required in vitro. To test this hypothesis, we cultured ILCPs with OP9-DL1 stroma and +/- allogeneic blood CD34+ hematopoietic progenitor cells (HPCs), which on their own can become KIR+ NK cells when cultured as follows: two weeks in Flt3 ligand (FL), c-Kit ligand (KL), and IL-3 followed by two weeks in FL, KL, and IL-15. Remarkably, among the ILCP-derived NK cells, significantly more were KIR+ when derived with HPCs (14.5+/-2.9% w/HPCs vs 1.9+/-0.8% w/out HPCs, p=0.004, n=7). Interestingly, we also observed that when IL-15 was added at day (d)0 instead of at d14, significantly fewer KIR+ NK cells were produced (IL-15 at d0: 3.3+/-0.7% vs IL-15 at d14: 14.5+/-2.9%, p=0.001, n=7). Lastly, we noted epigenetic and transcriptional alterations with IL-15 added at d0 vs d14. Collectively, these data suggest that ILCP differentiation into KIR+ NK cells requires epigenetic priming in a conducive microenvironment and that early exposure to exogenous IL-15 subverts this process.
The SRSF2P95H splicing factor mutation exerts distinct splicing effects across myelopoiesis that converge upon dysregulated G-CSFR signaling
Authors: Rich Abigail , Zinkel Sandra 
Published: November 2024

Abstract
SRSF2 P95H (P95H) splicing factor mutations are initiating genetic lesions in myeloid malignancies. They are associated with myeloid-biased hematopoiesis in mouse models and human disease. Mechanistically, P95H substitutions induce transcriptome-wide alterations in RNA splicing that vary by cell type and genetic background, complicating efforts to understand how P95H drives disease. Efforts to understand P95H mutations focus on splicing in progenitor cell populations, however, to date no studies have investigated this mutation in the mature myeloid cells that comprise &gt;40% of the marrow. We evaluated P95H-associated splicing across myelopoiesis. We subjected promyelocytes, myelocytes, and neutrophils from P95H mice to RNAseq and integrated these data with RNAseq from P95H hematopoietic stem cells and human MDS samples. A cluster-based splicing analysis approach identified cell-type specific splicing with overrepresentation of differentially spliced transcripts involved in myeloid effector function pathways in myelocytes and neutrophils. Missplicing of Csf3r/CSF3R, the granulocyte colony-stimulating factor receptor (GCSFR), was conserved. Notably, P95H mouse bone marrow exhibits impaired activation of GCSFR-dependent pathways (STAT3/5, ERK) that is unique to GCSFR. Overall, these results define P95H-driven alternative splicing as a differentiation-dependent phenomenon and implicate dysregulated GCSFR signaling as conserved feature of P95H-dependent myeloid hematopoiesis.
Ezrin drives adaptation of monocytes to the inflamed lung microenvironment.
Authors: Gudneppanavar Ravindra , Di Pietro Caterina , Oez Hasan , Zhang Ping-Xia , Huang Pamela , Braga Cassia , Tebaldi Toma , Biancon Giulia , Kim Catherine , Gonzalez Anjelica , Halene Stephanie , Krause Diane , Egan Marie , Gupta Neetu , Murray Thomas , Bruscia Emanuela 
Published: November 2024

Abstract
We have previously shown that the actin-binding protein ezrin is reduced and displaced from the plasma membrane in monocytes/macrophages (MΦs) from patients with cystic fibrosis (CF) and CF mice, impacting immune response to bacteria. However, its role in monocyte/MΦ is not fully understood. To investigate how the lack of ezrin affects monocyte/MΦ functions in response to lung infections, we developed a monocyte/MΦ-specific ezrin knock out mouse model (Ez-KOm) . We aerosolized WT and Ez-KOm mice with LPS and assessed the number of lung monocyte/MΦ by flow cytometry and their transcriptional profile via RNA-seq, followed by in vitro functional studies. In WT mice, ezrin is induced by LPS in all lung monocyte/MΦ populations, with the highest induction observed in the interstitial MΦs (IMs) (4-fold higher than monocytes). LPS treated Ez-KOm mice exhibit a significant reduction in IM numbers compared with WT and increased expression of pro-inflammatory markers (Il6, Tnfα, and Cxcl1). RNA-seq revealed differentially expressed genes in Ez-KOm monocytes/IMs related to adhesion, proliferation and cytoskeleton rearrangement. Consistently, Ez-KOm monocytes treated in vitro with LPS on collagen show stunted filopodia, altered F-actin distribution, reduced adhesion and increased cell death due to defective FAK and PI3K/AKT signaling. Our studies suggest that ezrin plays a critical role in the survival and adaptation of monocytes to the lung extracellular matrix during infections.
Non-neutralizing immune-mediated cross-protection against influenza viruses by chimeric M2e-H3 stalk protein or multi-subtype neuraminidase plus M2e virus-like particle vaccine in ferrets
Authors: Kang Sang-Moo , Kim Ki-Hye , Subbiah Jeeva , Bhatnagar Noopur , Liu Rong , Raha Jannatul , Pal Surya , Park Bo Ryoung 
Published: November 2024

Abstract
Current influenza vaccine based on hemagglutinin (HA) immunity is not effective in providing cross protection against circulating variants and new pandemic viruses. In prior studies, we developed universal influenza vaccine candidates of multi-subtype neuraminidase and M2 ectodomain virus-like particles (m-cNA-M2e VLP) and chimeric M2e-H3 stalk protein vaccine (M2e-H3stalk), which were effective in inducing cross-protective immunity in mice. Here, we evaluated the immunogenicity and efficacy of these recombinant universal influenza vaccines in ferrets. Our results showed that immunization of ferrets with recombinant universal vaccines induced high levels of IgG antibody responses (M2e, H3stalk, multi-subtype NA), NA inhibition (NAI), antibody-secreting plasma cells in spleens, and IFN-γ secreting blood mononucleate cells. Ferrets with either m-cNA-M2e VLP or M2e-H3stalk vaccine were moderately protected from H1N1 and H3N2 influenza viruses as evidenced by lower viral titers in nasal washes, trachea and lung after challenge. Tests of passive immunity of vaccinated ferret antisera indicate that multi-NA plus M2e vaccine conferred more effective and broader humoral immunity in naïve mice than M2e-H3stalk vaccine. Our findings support that immunity to M2e, HA-stalk, and multi subtypes NA will induce broader cross protection in ferrets, which is likely translational to humans.
Abnormal uterine bleeding after COVID-19 vaccination in South Korea
Authors: Park Dong Choon , Yeo Joon Hyung , Yeo Seung Geun 
Published: November 2024

Abstract
Background: Various adverse reactions have been reported after vaccination against COVID-19.
Objective: Few studies to date, however, have analyzed uterine bleeding after COVID-19 vaccination. The present study therefore evaluated the incidence of abnormal uterine bleeding after COVID-19 vaccination in South Korea.
Methods: Data on suspected adverse reactions to COVID-19 vaccination reported to the Korea Disease Control and Prevention Agency from February 26, 2021, to June 29, 2023 (week 121), were analyzed. Subjects who experienced abnormal uterine bleeding were identified, and the incidence of abnormal uterine bleeding was compared among the types of COVID-19 vaccines administered.
Results: During the study period, 135,894,788 doses of COVID-19 vaccine were administered, with suspected adverse reactions occurring after administration of 483,391 doses (3.56 adverse reactions per 1,000 vaccinations) and abnormal uterine bleeding observed after administration of 6,288 doses (4.9 per 100,000 vaccinations).
Conclusions: Abnormal uterine bleeding was the fifteenth most common adverse reaction after COVID-19 vaccination. Studies are needed to clarify the causal relationships of vaccination and type of vaccine with abnormal uterine bleeding, as well as to develop protocols for the diagnosis and treatment of abnormal uterine bleeding after COVID-19 vaccination.
Abnormal innate immune responses and associated genetic factors in Multisystem Inflammatory Syndrome in Children (MIS-C)
Authors: Khan Rehan , Ji Weizhen , Guzman-Rivera Jeisac , Mishra Abhilasha , Richlin Benjamin , Gaur Sunanda , Singh Aalok , Bukulmez Hulya , Kaelber David , Riordan Mary , Kimura Yukiko , Ganapathi Usha , Michailidis Ioannis , Ukey Rahul , Casseus Myriam , Andrews Tracy , Roy Jason 
Published: November 2024

Abstract
Multisystem inflammatory syndrome in children (MIS-C) is a rare, severe complication of COVID-19 characterized by multi-lineage immune activation and multi-organ system involvement. While baseline immunophenotypes have been characterized, innate immune cell functionality and genetic risk factors remain poorly understood. To address these knowledge gaps, we assessed inflammatory immune responses in children with MIS-C (n = 33) relative to healthy controls (n = 5). We found that Toll like receptor (TLR) signaling is severely dampened in peripheral blood cells in the MIS-C group, with weak cytokine responses to LPS stimulation and increased baseline gene expression of TLR negative regulators, compared to controls. We also found that the cytokine response to LPS was abnormally heterogeneous in the MIS-C group. In particular, cytokine response outliers were enriched for rare and potentially deleterious variants in the lysosomal trafficking regulator (LYST) locus. These LYST variants were associated with markers of lysosomal and mitochondrial dysfunction, altered energy metabolism, and low levels of TLR negative regulator expression. Improved knowledge of immune dysfunctions associated with MIS-C and risk factors may help explain aspects of MIS-C and other manifestations of long COVID and identify targets for therapy of MIS-C and other hyperinflammatory syndromes in children.
THE ONES AND TWOS: PATTERN RECOGNITION RECEPTOR NLRX1 IMPACTS VIRAL REPLICATION OF AND IMMUNE RESPONSE TO SARS-COV-2
Authors: Ivester Hannah , Woolls Mackenzie , Mott Madeline , Gregory Julia , Morrison Holly , Tupik Juselyn , Finkielstein Carla , Duggal Nisha , Weger James , Allen Irving 
Published: November 2024

Abstract
When pandemic virus SARS-CoV-2 (SCV-2) emerged in 2020, research was quickly generated to investigate the virus and host immune responses to it. Much research has focused on the cytokine storm- when overzealous immune responses in severe COVID-19 disease causes rampant production of cytokines following signaling increased by the activation of pattern recognition receptors (PRRs). A unique PRR known as NLRX1 has many diverse roles, including regulation of key antiviral pathways- NF-kB and interferon- autophagy, and understudied roles in cellular metabolism. We show that changes in NLRX1 expression impacts immune responses to and viral growth of SCV-2 in cell culture and in mice. While Nlrx1-/- mice challenged with SCV-2 showed similar overall morbidity to WT counterparts, there were temporary increases in weight loss at multiple time points post infection. SCV-2 infection of Nlrx1-/- mice resulted in more airway inflammation, seemingly contrary to their also having increased viral titer at 2dpi. Data in cell culture report an enhanced replication ability of SCV-2 when NLRX1 expression is lower. Both cell culture and animal studies report differences in immune activation, with differences in cytokine production when NLRX1 expression changes. Together, these experiments: (1) support the enigmatic function of NLRX1 (2) suggest a possible role of NLRX1 in the pathobiology of SCV-2 infection and (3) add to the growing field exploring NLRX1 and its many curious roles.
The role of macrophage phagocytosis on respiratory pneumovirus infection and host defense
Authors: Martinez-Espinoza Ivan , Guerrero Plata Antonieta 
Published: November 2024

Abstract
Lung macrophages play a pivotal role in the defense against viral and bacterial infections. These specialized cells are equipped with multiple mechanisms, such as phagocytosis, to detect the presence of pathogens and clear the infections. Human Metapneumovirus (HMPV) is a leading cause of acute lung infection, affecting particularly young children and the elderly. During and after the HMPV infection, individuals are at an increased risk of secondary bacterial infections due to a decreased immune response capacity, which increases the severity of the disease. However, the cellular contribution of macrophages in the HMPV infection remains understudied. In this work, we investigated the role of macrophage phagocytosis in HMPV infection and the effect of HMPV on macrophage phagocytic activity. For that, we used THP-1- and PBMC-derived macrophages as experimental models. We found that macrophages were susceptible to HMPV infection and that phagocytosis is one of the mechanisms used by the virus to infect the cells. However, macrophages do not replicate the virus efficiently. Furthermore, viral infection decreases the capacity of macrophages to phagocyte bacteria, as demonstrated by the activity of intracellular pH indicator dye. These data highlight the importance of phagocytosis as a mechanism to control respiratory viral infections and the subversion of the host immunity by HMPV that may contribute to subsequent opportunistic bacterial infections.
The role of CMKLR1 in lymphocyte activation and localization within the CNS during West Nile Virus Encephalitis
Authors: Cortes Alejandro , Durrant Douglas 
Published: November 2024

Abstract
Continued West Nile virus (WNV) outbreaks causing neuroinvasive disease in the US highlight the urgent need to decipher CNS viral control mechanisms. DCs are pivotal in recruiting and reactivating CNS-infiltrating lymphocytes for viral clearance. However, the molecular mechanisms that regulate DC recruitment and activation remain unknown. In CNS-related autoimmune disorders, CMKLR1, a chemokine receptor expressed by a DC subset, aids their recruitment and activation via its ligand, chemerin. To investigate the role of CMKLR1 during WNND, we infected WT and CMKLR1-/- mice with WNV. CMKLR1-/- mice exhibited a slight increase in mortality but elevated clinical scores accompanied by increased CNS viral loads compared to WT. We hypothesize that CMKLR1 restricts WNV neuropathogenesis by regulating the activation and localization of antiviral T cells within the CNS during WNV encephalitis. Determining the role of CMKLR1 in effector T cell activation and migration may provide insights into local molecular mechanisms controlling lymphocyte function and fate within the CNS during WNV.
Improving immunity against chronic virus infection using an autoimmunity associated Ptpn22 allele.
Authors: Shaikh Anam , Bevis Alec , Barnes Jenna , Schwarting Nancy , Cockerham Tammy , Orozco Robin 
Published: November 2024

Abstract
Dysfunctional dendritic cells (DC) lead to inadequate T cell activation causing chronic virus infection. Enhancing both DC and T cell function can prevent chronic virus infection. An approach to enhance immune function is to harness the immune-activating phenotype driven by the autoimmunity-associated allele of PTPN22, which is expressed in all immune cells. We previously found that mice expressing the Ptpn22 autoimmunity-associated allele (Ptpn22-R619W) successfully clear Lymphocytic choriomeningitis virus clone 13 (LCMV-cl13) unlike wildtype (WT) mice. This correlates with a more immunostimulatory DC phenotype and enhanced T cell function. However, the mechanism Ptpn22-R619W enhances DC and T cell function remains unclear. Using our CRISPR/cas9 generated Ptpn22-R619W mice, we addressed the hypothesis that Ptpn22-R619W increases DC immunostimulatory proteins which increases anti-viral T cell activation during virus infection. Post infection, Ptpn22-R619W DCs have reduced infection, increased CD86, and decreased PD-L1 expression both in vivo and ex vivo. Additionally, Ptpn22-R619W DCs have a differential gene expression profile. Also, Ptpn22-R619W mice have enhanced CD8 T cell function against the LCMV-cl13 minor epitope, GP276, but not the major epitope. Lastly, co-culture experiments demonstrate that Ptpn22-R619W in both DC and T cells is necessary to enhance T cell function. These results show how Ptpn22-R619W prevents DC and T cell dysfunction during virus infection.
NEK7 phosphorylation amplifies NLRP3 inflammasome activation downstream of gasdermin D and potassium efflux
Authors: Xu Jie , Zhang Lingzhi , Duan Yanhui , Sun Fangyuan , Odeh Nouha , He Yuan , Núñez Gabriel 
Published: November 2024

Abstract
The NLRP3 inflammasome plays a critical role in innate immunity and inflammatory disease. The interaction of NEK7, a member of the NIMA-related kinase family that is essential for inflammasome activation, with NLRP3 is enhanced by potassium efflux. However, the mechanism by which potassium efflux promotes the interaction between NEK7 and NLRP3 remains unknown. Here, we show that following NLRP3 activation NEK7 is rapidly phosphorylated at a specific threonine residue by JNK1 downstream of gasdermin D-mediated potassium efflux. NEK7 phosphorylation enhances the binding between NEK7 and NLRP3, which further promotes inflammasome assembly and activation. Mutant mice and macrophages in which the threonine residue of NEK7 was replaced by valine were impaired in NEK7 phosphorylation, NLRP3 inflammasome activation and IL-1β secretion. Thus, NEK7 phosphorylation is an important event that acts downstream of gasdermin D and potassium efflux to further enhance NLRP3 inflammasome activation.