bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of Cdk5/p35 kinase activity in natural killer cell cytotoxicity
Authors: Wong Derek P , Parameswaran Reshmi 
Published: January 2023

Abstract
Natural killer (NK) cells and T cells are immune cells with cytotoxic function, but while the distinct role of Cyclin-dependent kinase 5 (Cdk5) in the activation and effector function of T cells has been explored, its role in NK cells is unknown. Here, we show that both Cdk5 and its obligatory co-activator p35 protein are expressed in NK cells. We also detect Cdk5 kinase activity in NK cells, and kinase inhibition via p35 shRNA results in increased NK cell cytotoxicity. Overexpression of p35, which increases Cdk5 kinase activity, results in decreased NK cell cytotoxicity. Exogenous expression of a kinase-dead Cdk5 mutant that is still capable of binding p35 also increases NK cell cytotoxicity. Meanwhile, p35 knockout mouse NK cells exhibit significantly higher cytotoxicity against murine melanoma cells compared to wild-type NK cells. Based on our preliminary data, we conclude that Cdk5/p35 kinase activity negatively regulates NK cell cytotoxicity, with implications for the potential improvement of NK cell-based cancer immunotherapies.
Supported by grants from the NIH (R21 CA246194, T32 GM007250) and the St. Baldrick’s Scholar Award.
TL1A promotes a multi-cytokine Th9 cell phenotype
Authors: Chu Michelle , Kaplan Mark H 
Published: January 2023

Abstract
The TNF superfamily member TL1A is a costimulatory molecule that signals through its receptor DR3 on T lymphocytes. The Th9 subset of T lymphocytes secretes the pleiotropic cytokine IL-9 which has functions in allergic airway disease, helminth infections, and tumor immunity. TL1A signaling has been shown to increase IL-9 production by Th9 cells. However, its role in regulating other functions of Th9 cells is unknown. Here we demonstrate that TL1A increases expression of IL-9 and IL-13 as well as the frequency of IL-9 and IL-13 co-expressing cells in Th9 cell cultures through flow cytometric analyses. We also show that the Il9 and Il13 promoter and enhancer regions are differentially accessible in response to TL1A over a five-day culture period through chromatin accessibility assays. At the Il9 locus, TL1A enhances binding of IRF4, BATF, and PPARg. Mechanistically, this is linked to decreased H3K9 tri-methylation and increased H3K4 tri-methylation at Il9 enhancer region CNS2. At the Il13 locus, TL1A enhances binding of BATF at the Il13 promoter alongside decreased H3K9 tri-methylation and increased H3K4 tri-methylation. Ongoing experiments define the function of TL1A-induced multi-cytokine producing cells. Together, these data indicate that TL1A contributes to heterogeneity of IL-9-secreting T cell populations.
Supported by NIH Combined Adult and Pediatrics Pulmonary Research Training Program T32
Dietary iron deficiency impairs the functionality but not generation or establishment of memory T cells following influenza infection
Authors: Bradley Marissa C , Idzikowski Emma , La Carpia Francesca , Gray Joshua I , Rybkina Ksenia , Guyer Rebecca S , Pethe Kalpana , Hod Eldad A , Connors Thomas J 
Published: January 2023

Abstract
Protection from viral respiratory tract infections (VRTIs) is dependent on the activation, differentiation, and maintenance of antigen specific memory T cells. Iron is essential for fundamental metabolic processes in the human body, with iron deficiency, the most prevalent micronutrient deficiency worldwide, resulting in increased morbidity and mortality to VRTIs. The role of iron in the establishment and maintenance of memory T cells is not fully defined. Using a murine model of dietary iron modulation, we define the impact of iron deficiency on anti-viral T cell memory formation, functionality, and resultant protection during primary and heterosubtypic influenza infections. Multi-parameter flow cytometry and stimulation assays were performed on T cells isolated from lung and spleen at effector and memory time points following primary (X31) infection, as well as 5 days post heterosubtypic (PR8) challenge. Strikingly, there was no significant difference in the numbers or proportions of CD4+ and CD8+ influenza specific memory T-cells at any time point when comparing iron deficient to iron replete mice. However, memory T cells generated by iron deficient mice displayed impaired production of key effector cytokines, most notably IFN-γ and IL-17a. Importantly, following heterosubtypic challenge, iron deficient mice exhibited increased morbidity, as defined by increased weight loss, and significantly diminished IFN-γ production by T cells across all sites. This work reveals that distinct iron profiles did not interfere with the generation or maintenance of influenza specific T cells but iron deficiency significantly impacts memory T cell anti-viral functionality resulting in impaired protective responses.
Supported by -K23 AI141686 -UL1 TR001873
Helpless CD8 T cell memory explained: prolonged antigen presentation drives a temporal rather than terminal defect
Authors: van der Heide Verena , Davenport Bennett , Jhun Kevin , Homann Dirk 
Published: January 2023

Abstract
CD4 T cell help is critical for long-term antiviral CD8 T cell immunity. Despite seemingly normal effector properties, “helpless” CD8 memory T cells (TM) generated in the absence of CD4 T cells during initial priming exhibit a substantially blunted recall response. Yet, how lack of CD4 help specifically impairs CD8 TM formation in the context of acute viral infections remains elusive. Here, we demonstrate that in transiently and constitutively CD4 T cell-deficient mice “helpless” antiviral CD8 TM to acute lymphocytic choriomeningitis virus (LCMV) exhibit a wide array of homeostatic, phenotypic, and functional defects that are broadly consistent with delayed CD8 TM maturation. Mechanistically, “helpless” CD8 T cell memory is not grounded in altered CD8 effector T (TE) cell differentiation in the absence of CD4 T cells but in exposure of post-effector CD8 T cells to protracted viral antigen presentation. Conversely, efficient generation of “helped” CD8 T cell memory is contingent on CD40L-mediated activities by LCMV-specific CD4 T cells that promote timely clearance of viral antigen. Importantly, we show that “helpless” memory-precursor CD8 TE display comparable memory potential to their “helped” counterparts, and that, once viral antigen is cleared, complete resolution of “helpless” CD8 memory is achieved over a period of ~1.5 years during which “helpless” CD8 TM eventually acquire mature TM properties including fully restored recall capacities. Collectively, our data redefine “helpless” CD8 T cell memory as a temporal rather than terminal defect, with implications for our general conceptualization of CD8 T cell memory as well as for the development of preventive and therapeutic strategies to improve protective CD8 TM function.
Supported by grant NIH R01AI093637.
TRM and mucosal tissue sites are protected from age-associated phenotypic changes in secondary lymphoid organs
Authors: Lam Nora , Caron Daniel P , Davis-Porada Julia , Jensen Isaac J , Lee YoonSeung , Morrison-Colvin Rory , Poon Maya M L , Szabo Peter A , Ural Basak B , Wells Steven B , Kubota Masaru , Matsumoto Rei , Farber Donna L 
Published: January 2023

Abstract
The persistence and maintenance of immunological memory is critical for long-term immunity generated in response to infection and vaccination. Aging of memory T cells in humans has predominantly been studied in the peripheral blood, and considerably less is known about the aging of memory T cells within tissues. Using tissues obtained from human organ donors, we investigated how tissue localization, expression of the CD103 integrin, and age influence the phenotype of tissue resident memory T cells (TRM). Using multi-parameter flow cytometry, we characterized diverse tissue sites from donors ages 9 to 93 years old and demonstrate that age-associated changes in the T cell compartment exhibit tissue- and subset- specific dynamics. CD8 T cells in blood-rich tissues spleen and lung have elevated expression of replicative senescence markers CD57 and KLRG1 relative to lung-associated lymph nodes (LLN), jejunum, and mesenteric lymph nodes (MLN). CD103+ TRM have the lowest expression of CD57 and KLRG1 across all tissue sites relative to both CD103− TRM and circulating TEM. CD8 T cells in lymphoid tissues spleen, LLN, and MLN exhibit the largest age-associated increases in expression of CD57, KLRG1, CD244, and PD-1. Together, these results demonstrate that age-associated phenotypic changes are most prominent in secondary lymphoid organs, while TRM and mucosal sites exhibit few senescent changes.
Supported by the National Science Foundation (NSF) Graduate Research Fellowship Program (GRFP) and grants from NIH (P01 AI106697, U19 AI057266)
The Rv2623-Rv1747 interaction influences phosphatidyl-myo-inositol levels on the cell envelope of Mycobacterium tuberculosis
Authors: Schami Alyssa , Ke Wei , Allué-Guardia Anna , Olmo-Fontánez Angélica M , Chan John , Torrelles Jordi B 
Published: January 2023

Abstract
Mycobacterium tuberculosis (M.tb), the causative agent of tuberculosis (TB), has a complex cell envelope that provides a barrier of protection to various environments. Peripheral lipids on the M.tb cell envelope act as virulent factors influencing the host immune response to infection. Of these, some phosphatidyl-myo-inositol mannosides (PIMs) and their associated lipoglycans (lipomannan; mannose-capped lipoarabinomannan) influence M.tb-host interactions by increasing phagocytosis, blocking the maturation of M.tb-containing phagosomes, and increasing the anti-inflammatory response of infected macrophages. However, there is limited knowledge regarding how M.tb regulates PIMs levels, and how this regulation influences infection outcomes. In our previous studies, we hypothesized that Rv2623 and Rv1747 work together to regulate PIMs on the M.tb cell surface. Indeed, deleting the universal stress protein Rv2623 increases PIMs levels, growth, and virulence in vivo; while deleting the ABC transporter Rv1747 decreases PIMs levels. Here we investigate the mechanistic regulation of M.tb PIMs levels and its impact on growth and pathogenesis in vitro by assessing multiple M.tb strains with Rv2623 or Rv1747 mutated at different amino acids. Depending on the mutation, Rv2623 interacts with Rv1747 to modulate M.tb cell envelope PIMs levels at different degrees. Mutants with increased PIMs levels have increased growth in macrophages, and PIMs levels on the M.tb cell envelope may also correlate to resistance to certain anti-TB drugs. Overall, our data indicate that Rv2623 negatively regulates Rv1747 to modulate PIMs levels on the M.tb cell envelope, ultimately influencing growth and M.tb-host interactions in vitro and in vivo.
Supported by grants from NIH/NIAID (1R01AI146340-01A1)
Gut microbiome regulates serotonin production in the neonatal intestine to promote immune tolerance in early life
Authors: Sanidad Katherine Z , Rager Stephanie L , Ananthanarayanan Aparna , Callaghan Ryann , Li Tingting , Jin Jenny C , Amir Mohammed , Luo Ryan , Silver Randi , Artis David , Guo Chun-Jun , Krumsiek Jan , Inohara Naohiro , Zeng Melody Y 
Published: January 2023

Abstract
The gut microbiome in neonates is vastly different from that in adults due to dietary changes and gut maturation. However, the neonatal gut metabolome, largely shaped by gut bacteria, remains largely undefined. Using high-throughput metabolomics, we compared &gt;500 metabolites in the ileum of specific pathogen-free (SPF) wild-type neonatal and adult mice. We found that most metabolites at significantly higher levels in neonates were neurotransmitters, including serotonin (5-HT), a neurotransmitter critical in the regulation of gut motility and mood stabilization. SPF neonates had significantly higher 5-HT levels in the ileum compared to both SPF adults and germ-free (GF) neonates, suggesting the gut microbiome as a key driver of generation in the neonatal intestine. Gut bacteria in neonates modulate the enzymes TPH-1 and MAO-A involved in the synthesis and breakdown of 5-HT. In addition, unlike in the adult intestine where 5-HT is produced primarily by enterochromaffin cells, gut bacteria are major producers of 5-HT in the neonatal intestine. The immunomodulatory role of 5-HT in the neonatal intestine has not been explored. We demonstrate that 5-HT directly promotes the differentiation regulatory T cells while diminishing interferon gamma and interleukin-17 produced in CD4 T cells, ex vivo and in vivo, in the neonatal intestine. Oral gavage of 5-HT into neonatal mice enhanced immune tolerance toward both commensal bacteria and dietary antigens. Our study has elucidated unique mechanisms by gut microbiota to regulate 5-HT generation in the neonatal intestine, and a novel immunomodulatory role for intestinal 5-HT to promote immune tolerance towards both dietary antigens and commensal bacteria during the early developmental stage.
Age associated differential innate immune response to strains of Escherichia coli in neonatal sepsis model
Authors: Muske Josey , Greenfield Kara G , Harlow Olivia , Knoop Kathryn 
Published: January 2023

Abstract
Bloodstream infections, causing late-onset neonatal sepsis (LOS), have an incidence rate of 10% in very low birth weight (&lt;1500 grams) infants. Despite the prevalence in neonates, the current understanding of bloodstream infections and the resulting immune response is predominately based on research in adults, which is often a complication of surgery. In contrast, sepsis in infants is often a result of the dissemination of bacteria from the gut. Additionally, observations have shown differences in the neonatal immune response, such as a decrease in gene expression associated with TLR signaling pathways, compared to adults. One question remaining in the field is the age dependent differences in pathogen detection by innate cells, including dendritic cells and macrophages. Use RAW-Blue reporter cells, and splenocytes cultured from mice at different ages, we have found disparate responses to gut-resident E. coli dependent on pathogen status of the bacteria. Specifically, we have observed differential increases in IL-6, TNFα, and IL-1β, along with NFκB activation that is dependent on live bacteria. Additionally, changes in this response were associated with age, suggesting differences in the innate immune response throughout early life. Future directions include measuring RNA expression of dendritic cells and macrophages between the age groups and exploring how the differences contribute to infection outcomes in a model of sepsis.
Supported by grants from NIH (DK122187).
Assessing mucosal antibody responses to SARS-CoV-2 in humans and mice.
Authors: Isho Baweleta , Abe Kento , Zuo Michelle , Jamal Alainna J , Cao Eric , Chao Gary , Li Zhijie , Rini James M , Tan Darrell , McGeer Allison J , Gingras Anne-Claude , Gommerman Jennifer L 
Published: January 2023

Abstract
COVID-19 is a respiratory disease caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). We previously developed an enzyme-linked immunosorbent assay (ELISA) protocol to profile antibody responses to the SARS-CoV-2 Spike protein and its receptor-binding domain (RBD) in the saliva of patients with COVID-19. Anti-SARS-CoV-2 antibody responses were readily detected in saliva, peaking at 16–30 days post-symptom onset. While anti-SARS-CoV-2 IgM/IgA were found to decay, IgG responses were more long-lived, persisting up to 105 days. Now we wish to know (1) if anti-SARS-CoV-2 antibodies are generated locally in the oral cavity, (2) the relative kinetics of IgA versus IgG appearance in the oral cavity, and (3) whether salivary antibodies have the capacity to neutralize SARS-CoV-2. To answer these, I have adapted our ELISA to detect secretory component-associated SARS-CoV-2-specific antibodies. I found that 33.3% and 26.9% of COVID-19 patients were positive for secretory component-associated antibodies to Spike and RBD, respectively. Secretory component-associated antibodies correlated with antigen-specific IgA levels, particularly for the Spike antigen. Moreover, using saliva from contact-traced subjects, I found that anti-Spike/RBD IgM/IgA are detected in the saliva 7 days post-exposure, prior to IgG. Lastly, I detected Spike-specific antibody-secreting cells in lymphoid tissues draining the upper respiratory tract upon intranasal infection of K18-hACE2 mice with a non-lethal dose of SARS-CoV-2. In summary, I have provided evidence in humans and mice that a local antibody response that quickly class switches to IgA occurs in the oral cavity and draining lymph nodes upon infection with SARS-CoV-2.
Decoding atopic dermatitis: patient heterogeneity and genetic variation contributing to immune-mediated dermatologic disease in the UK Biobank
Authors: Truong Dongnhu T , Molineros Julio , Chen Yanqing , Guan Meijian , Hart Amy , Keyes Bryce , Seiber Karsten B , Li Shuwei , Waterworth Dawn , Black Mary Helen 
Published: January 2023

Abstract

Background
Dupilumab is the only approved biologic used to treat moderate to severe atopic dermatitis (AD) but has a non-response rate of 55–60%. This suggests underlying heterogeneity in AD etiology, requiring alternate therapies for patients.

Study Objective
Identify biomarkers and disease conditions in the UK Biobank that stratify AD patients into homogenous subgroups for genetic characterization.

Methods
We identified 13,137 AD cases and 13,136 age and sex-matched controls using hospital ICD9/10 and primary care codes in the UK Biobank. A total of 94 biomarkers and 1,596 disease PheCodes were used to isolate traits that vary in AD patients relative to controls; K-means (quantitative) and K-modes (binary) clustering was performed to identify subgroups within AD patients. Genetic analysis was performed for subgroups vs. controls using REGENIE.

Results
We observed enrichment of dermatologic, gastrointestinal, and rheumatologic conditions in AD patients, as well as differences in eosinophil percentage, Cystatin C, C-reactive protein, and erythrocyte distribution width relative to controls. There were eight subgroups of AD patients including groups with co-occurring psoriasis, asthma and gastrointestinal problems, penicillin allergy, and AD with no co-morbidities. Genetic analyses revealed novel HLA and non-HLA associations with subgroups, while also replicating known associations with FLG.

Conclusion
There were eight distinct AD subgroups based on co-occurring disease states. Genetic analyses show subgroup-specific factors can contribute to disease pathology—signaling potential utility for differential therapies and highlight mechanisms for targeted therapeutic development in the treatment of AD.
The effect of induced lymphatic circulation on lymphangiogenesis and inflammatory mediators in rats with adjuvant induced arthritis
Authors: Volin Michael V , Blucher Kelly , Zanotti Brian , Incrocci Ryan , Del Toro Rosalinda Monroy , Jain Shreya , Weber Daniel , Gober Calley , Swanson-Mungerson Michelle 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a chronic inflammatory disease involving dysfunctional lymphatic circulation leading to edema. The adjuvant-induced arthritis (AIA) rat model was used to study the effect of lymphatic pump treatment (LPT) on the expression levels of inflammatory cytokines and lymphangiogenesis within the draining popliteal lymph nodes (PLN). PLN from both control and LPT treatment groups were harvested, and their cDNA were analyzed by qPCR to determine the levels of cytokines, VEGF-C, and VEGFR-3. Additionally, immunohistochemistry was used to visualize the expression of VEGF-C and VEGFR-3 and flow cytometry was used to quantify leukocyte cell types in the PLN. The LPT group showed a decrease in ankle circumference and arthritis grade a few days after the initiation of treatment. Flow cytometry of popliteal lymph nodes showed an increase in CD8, CD4, and CD3 cells in arthritic rats, however cell populations were not significantly different between treatment groups. The qPCR data indicated that the LPT group had a slight reduction in levels of IL-17a, IL-4, IL-6, and IL-1β. IHC showed that the PLN from the LPT group had the highest expression area of VEGF-C and the lowest expression area of VEGFR-3 compared to control. In conclusion, The LPT group showed a reduction in ankle circumference and arthritis score, suggesting that LPT helped alleviate edema and inflammation. When compared to control, the LPT group had lower expression of pro-inflammatory cytokines and elevated levels of VEGF-C, an important lymphatic vessel growth factor, in PLN. Thus, the data suggests that LPT may elevate the swelling and inflammation in rat AIA through the reduction of inflammatory cytokines and the elevation of a lymphatic growth factor.
Deficiency in B Cell Maturation Antigen reveals sex differences in Experimental Autoimmune Encephalomyelitis.
Authors: Kumar Gaurav , Ko Rose M , Bhatt Neal , Axtell Robert C 
Published: January 2023

Abstract
Multiple sclerosis (MS) is immune mediated demyelinating disease of the central nervous system (CNS) leading to neuronal damage. The disease is heterogeneous with respect to onset and clinical course, being more prevalent in women then in men with a ratio of 3:1. Further sex differences have been observed with males having a more severe and debilitating disease course compared to females. The complex interplay of immune cells, sex hormones or genetic differences could be contributing to these clinical phenomena, which remains to be elucidated. In experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, several reports have demonstrated no sex differences in disease onset or severity in C57BL/6 mice. Herein we report that deficiency in B Cell Maturation Antigen (BCMA) in C57BL/6 mice reveals a sex difference in EAE. BCMA plays a prominent role in B cell function, however the role BCMA plays in MS and EAE is unknown. Our initial goal of this project was to determine the impact BCMA deficiency has on EAE. In doing so we confirmed that EAE disease was not different in males and females from BCMA+/+ mice. Strikingly in the BCMA−/− mice, there was a significant increase in EAE disease severity in males compared to females. The increased EAE in the BCMA−/− male mice was associated with increased demyelination and infiltration of inflammatory T cells and macrophages into CNS. Also, we observed decreased splenic transitional B cells in male BCMA−/− mice compared to females. Castration of mice significantly reduces EAE in BCMA−/− mice as compared to sham controls. Altogether, our data demonstrate that male hormones exacerbates EAE in BCMA−/− mice by augmenting inflammatory responses compared to females leading to increased neuro-inflammation.
Supported by NIH (R01AI137047 and R01EY027346)