bluuebunny/crossref_metadata_2025_split huggingface dataset:
Altered Trained immunity in AID−/− versus wild type mice following infection with Pneumocystis murina
Authors: Curran Shelly , Kovacs Joseph 
Published: January 2023

Abstract
Pneumocystis pneumonia continues to be a life-threatening infection in immunocompromised individuals. Dendritic cells are key antigen presenters to CD4+ T cells, which are critical to controlling Pneumocystis infection along with B cells. β-1,3 glucans are found in the cell wall of the cyst form of Pneumocystis. Glucans derived from other fungi have been shown to induce trained immunity. Therefore, we explored if Pneumocystis infection can induce a trained immunity response in C57Bl/6 (wild type) mice following exposure via a natural route of infection. We also tested activation-induced cytidine deaminase knockout mice (AID−/−), a deficiency that affects B cells, in the same infection model to determine whether a mutation known to affect the adaptive immune response can also alter the manifestation of trained immunity. We found that bone marrow derived dendritic cells (BMDCs) from C57BL/6 Pneumocystis exposed mice respond differently to homologous and heterologous antigen stimulation compared to BMDCs from naïve mice. Furthermore, although the differential response between cells from AID−/− exposed and naïve mice exists, specifically the TNFα response is opposite to that from C57BL/6 mice. In addition, when trained immunity was induced in BMDCs derived from naïve C57Bl/6 mice and AID−/− mice, the cytokine response was distinctive between the cells derived from C57BL/6 mice and AID−/− mice. These results suggest that not only can Pneumocystis infection induce trained immunity in BMDCs, this response is alterable by gene mutations traditionally thought to affect the adaptive immune response.
Intramural Research Program of the NIH Clinical Center
Mice infected with Mycobacterium tuberculosis are resistant to secondary infection with SARS-CoV-2
Authors: Mejia Oscar Rosas , Gloag Erin S , Li Jianying , Ruane-Foster Marisa , Claeys Tiffany A , Farkas Daniela , Farkas Laszlo , Xin Gang , Robinson Richard T 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb)and SARS-CoV-2 (CoV2) are the leading causes of death due to infectious disease; however, little is known regarding the immunological environment in the Mtb/CoV2 coinfected lung. The goal of this study was to use a mouse model of Mtb/CoV2 coinfection to determine if CoV2 affects Mtb bacterial burden and/or alters the lung immune profile. Using lung collected from human ACE2 transgenic (K18-hACE2) mice infected with both pathogens (Mtb only, SARS-CoV-2 only, and Mtb/SARS-CoV-2 co-infected), we evaluated immune gene expression, cytokine production, and bacterial burden. Surprisingly, these data show that Mtb suppresses CoV2 -related weight loss and lung viral burden in the human ACE2 transgenic mouse model. We also report a reduction in lung interferon gamma production and expression in coinfected mice compared to the Mtb only infected group, suggesting a possible altered T cell profile in the coinfected group. To determine whether Mtb-induced resistance to CoV2 was specific to the ACE2 transgenic model of COVID19, we performed the same set of experiments using a second mouse model of COVID19: Mouse Adapted SARS-CoV-2 (MACoV2) infection of C57BL/6 (B6) mice.
In both model systems, Mtb-infected mice were resistant to secondary CoV2 infection and its pathological consequences, and CoV2 infection did not affect Mtb burdens. Single cell RNA sequencing of coinfected and monoinfected lungs demonstrated the resistance of Mtb-infected mice is associated with expansion of T and B cell subsets upon viral challenge. Collectively, these data demonstrate that Mtb infection conditions the lung environment in a manner that is not conducive to CoV2 survival.
Supported by OSU Advancing Research in Infection and Immunity Fellowship Award
HIV compromises Th17 and Th22 immunity in a humanized mouse model of Tuberculosis and HIV co-infection
Authors: Martinez-Martinez Yazmin Berenice , Huante Matthew B , Files Megan , Gelman Benjamin B , Endsley Mark , Endsley Janice J 
Published: January 2023

Abstract
Tuberculosis (TB) kills an estimated 1–2 million people per year and is the leading cause of death in people with HIV (PWH). The CD4+ T helper (Th) populations play significant roles in protective immunity to Mycobacterium tuberculosis (Mtb), and are also important host cells for HIV proliferation and persistence. Emerging evidence in PWH suggests that Th17 and Th22 cells may be preferentially depleted during HIV infection, an outcome that could pose a risk for Mtb containment during co-infection. We employed our humanized mouse model of co-infection to assess pulmonary and peripheral changes in Th17 and Th22 frequency and function due to infection with HIV, Mtb, or both. As determined with multi-plex ELISA, co-infection with HIV suppressed the cytokine response to Mtb including IL-17A, IL-22, IL-6, and IL-1β. Multi-parameter flow cytometric analysis revealed that Mtb and HIV/Mtb infection led to increased Th17+ cells in lung and decreased Th17+ cells in spleen. Following HIV or HIV/Mtb infection, Th22+ cells were significantly decreased in both lung and spleen. Importantly, HIV preferentially infected Th17 cells compared to Th22 and other Th subpopulations. Greater HIV viral load was also observed in the lung, but not spleen, of animals with Mtb/HIV co-infection compared to those with HIV mono-infection. Overall, these results suggest that HIV may compromise Th22 immunity, and exploit Th17+ cells to promote viral pathogenesis, in the setting of Mtb and HIV co-infection.
Y.B. Martinez-Martinez is supported by Conacyt-I2T2 in Mexico, Contex, and the James W. McLaughlin Fellowship Fund. Grant support was provided by NIH R01AI147948 award to J. Endsley
Scrub typhus weakens humoral immune responses through suppression of germinal center and B cell activation
Authors: Gonzales Casey A , Liang Yuejin , Card Galen , Fisher James , Walker David H , Soong Lynn , Sun Jiaren 
Published: January 2023

Abstract
Scrub typhus is a poorly studied, but life-threatening, disease due to uncontrolled growth of Orientia tsutsugamushi bacteria within endothelial cells and phagocytes, in addition to dysregulated Th1 immune responses and inflammation. Cellular and humoral immunity in infected patients and experimental animals are short-lived, and wane within 1–2 years; however, the underlying mechanism of this transient immunity is unclear. No reports have examined splenic germinal center (GC) or B cell activation during this infection. The study objective is to fill this knowledge gap by using endothelium-targeted C57BL/6 mouse models that mimic pathological features of human scrub typhus. Through NanoString, qRT-PCR, and flow cytometry, we found significant repression of key markers crucial for B cell development (CCR7, BAFFR, PAX5) and GC formation (SLAMF1, ICOS-L, CXCR5, BCL-6) in both mild and severe disease outcomes. The widespread impairments in B and T follicular helper (Tfh) cell activation correlated with excessive Th1 cell activation during disease progression. Immunofluorescence staining (B220, CD3, GL7) revealed evidence of GC formation with sublethal infection, but the near absence of organized GCs at late stages of lethal infection, which was consistent with results from our splenic multi-color flow cytometry and serum antigen-specific ELISA assays for IgG isotypes. Ongoing RNA-seq studies for splenic B cells isolated from sub-lethally or lethally infected mice will provide in-depth, B cell-specific insight during infection. In conclusion, this is the first study to show profound alterations in B cell activation and GC formation in murine models, which helps understand “poor” humoral immunity seen with human scrub typhus.
Supported by grants from NIH (R01 EY028773 to JS, R01 AI132674 to LS, R21 AI156536 to LS)
CD206+MHCII− macrophages present in the neonatal but not adult intestine do not derive from LysM monocytes and are decreased in experimental Necrotizing Enterocolitis
Authors: Managlia Elizabeth , Yan Xiaocai , De Plaen Isabelle G 
Published: January 2023

Abstract
Necrotizing enterocolitis (NEC) is an intestinal disease targeting premature infants. While not well understood, its pathogenesis involves an underdeveloped intestinal immune system. In the neonate, this system is primarily innate, with Cx3cr1+ intestinal macrophages (Mφ) being the predominant population. In the murine neonatal intestine, the Cx3cr1+ compartment contains embryonic self-renewing Mφ (Ly6C− CD64+ MHCII+ Tim4+) that are largely replaced by monocyte-derived-Mφ at weaning, while only a small population of Tim4+ cells persist in adults. However, how inflammation affects the neonatal intestinal Mφ compartment is not known. Using flow cytometry, we found the neonatal but not adult murine intestine to contain a population of Cx3cr1Hi CD64Hi MHCII− cells (30.62% ± 1.6 of Cx3cr1+ cells). 28.7% ± 1.8 of these cells express CD206, a known marker of embryonic origin in other tissues. While 34.4% ± 1.2 of total CD206+ cells expressed Tim4 in dam fed pups, it increased to 49.3% ± 0.51 p≤ 0.01 in pups exposed to experimental NEC for 24 hours. This CD206+ Tim4+ subset was mainly MHCII+. When Cx3cr1-expressing cells were depleted (Cx3cr1-Cre CSFR1-DTR mice), all Cx3cr1-expressing cells were depleted, including CD206+ MHCII−and+, but not Ly6C+ monocytes. In contrast, when LysM-Cre was used (LysM-Cre CSFR1-DTR mice), pups had decreased Ly6C+ monocytes but the CD206+ MHCII−and+ populations were not depleted. These results were confirmed by immunofluorescent staining for CD206 and IBA-1. Thus, we conclude that neonatal intestinal CD206+ MHCII−or+ cells do not derive from LysM+ monocytes. We speculate that CD206+ MHCII− cells acquire MHCII during NEC thus decreasing the CD206+ MHCII− pool.
Supported by NIH DK116568 (IDP)
Neonatal Antibiotic Exposure Alters Intestinal Macrophage Frequency and Polarization
Authors: Schill Ellen Merrick , Udayan Sreeram , Gaddipatti Shreya , John Vini , Barrios Bibiana E , Floyd Alexandria N , McDonald Keely G , Newberry Rodney D 
Published: January 2023

Abstract
More than 75% of premature infants are given antibiotics in the first week of life. This is lifesaving for neonates with serious bacterial infections but is associated with increased risks of necrotizing enterocolitis (NEC). Given the association of intestinal macrophages (Mϕ) in the pathogenesis in NEC, we characterized the effect of antibiotics on this cell type. Pups were exposed to ampicillin and neomycin via the dam from postnatal day 1 (P1) to P7/8. Mϕ were identified as CD45+ CX3CR1+ CD11b+ F480+ using flow cytometry. In the small intestine (SI) there was an increase in the percentage of Mϕ in antibiotic-exposed lamina propria, but no difference in the proportion of F480-myeloid cells to Mϕ. The percentage of Mϕ in exposed colonic lamina propria cells was unchanged, but the proportion of Mϕ to F480-myeloid cells was increased. This suggests that antibiotic exposure expands myeloid cells in both the SI and colon, but likely via different mechanisms. To further characterize the phenotype of the intestinal Mϕ, we assessed polarity by immunoassay of 13 Mϕ associated cytokines. There were no significant differences in the proximal small intestine cytokine expression between control and antibiotic-exposed tissue. In the distal SI, antibiotic exposed mice had higher levels of TGFβ and CCL2, M2 Mϕ markers. In the colon, control tissue had elevated M1 Mϕ markers compared to antibiotic-exposed mice. Antibiotic-exposed colon had significantly increased levels of TGFβ. These data demonstrate that antibiotic exposure alters Mϕ proportion and polarization. Further work will be necessary to determine how this initial shift away from pro-inflammatory polarization leaves antibiotic-exposed infants at risk for intestinal inflammatory diseases.
Supported by NIH T32DK077653
A dietary fiber limits autoimmune neuroinflammation by restricting Th1 activation, polarization, and migration
Authors: Fettig Naomi M , Robinson Hannah G , Allanach Jessica R , Simister Rachel L , Wang Elsie J , Jiayu Ye , Davis Katherine M , Seo Jung Hee , Gibson Deanna L , Crowe Sean A , Horwitz Marc S , Osborne Lisa C 
Published: January 2023

Abstract
Dietary fibers and their breakdown products have been shown to limit inflammation in multiple disease contexts. However, the term “dietary fiber” encompasses a biochemically diverse family of carbohydrates, and it remains unknown how different fiber sources influence immune cell function. To address this question, we assessed the immunomodulatory capacity of different fiber types in a mouse model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). C57BL/6 mice were fed a control diet (5% cellulose) or diets enriched (30%) with resistant starch, inulin, pectin, or guar gum for 2 weeks prior to EAE induction. A diet rich in guar gum uniquely ameliorated the clinical, pathological, and immunological features of EAE. Limited central nervous system (CNS) infiltration of IFNγ+ CD4+ T cells (Th1s) in guar gum-fed mice could be attributed to T cell-intrinsic impairment Th1 activation and polarization. Nanostring nCounter transcriptional analyses revealed downregulation of migration-associated markers in CD4+ T cells isolated from guar gum/EAE mice, which was supported by reduced cell-surface expression of integrins involved in migration into the CNS. Functionally, this translated to limited migration of guar gum-derived encephalitogenic Th1s into the CNS upon adoptive transfer into control-fed recipients, which reduced incidence and severity of EAE. However, the 16S rRNA microbiome signature and short chain fatty acid profiles were not unique in comparison to other fiber-supplemented diets, indicating a novel mechanism of action from those previously ascribed in high fiber diet studies. These data confirm the individuality of dietary fibers and identify a novel immunomodulatory function of guar gum.
This project was supported by the endMS doctoral studentship program (Multiple Sclerosis Society of Canada) (NMF, JRA), the Canadian Institutes for Health Research (PJT-148909), and the Canada Research Chair program (LCO).
Controllable regulation of CD3ζ and MyD88 signaling in T cells expands and enhances the neoantigen-reactive repertoire resulting in effective antitumor activity
Authors: Liu Jing , Davila Eduardo 
Published: January 2023

Abstract

Background
Adoptive cell therapy (ACT) of tumor-reactive T cells including tumor infiltrating lymphocytes (TIL) represents one of the most promising forms of therapies for treating patients with solid tumors. However, durable responses are observed in a small subset of patients and in certain cancer types. Major hurdles to achieving durable responses include: 1) suboptimal TCR responses due to sparse and/or weakly immunogenic tumor antigen presentation, 2) development of dysfunctional or exhausted T cells, 3) low TIL persistence, and 4) tumor infiltration. We and others have demonstrated that activating MyD88 signaling in T cells strongly amplifies weak TCR responses and imparts T cells novel anti-tumor properties.

Hypothesis
TILs engineered with a novel fusion protein consisting of CD3ζ:MyD88 will activate MyD88 signals in a TCR-dependent manner resulting in enhanced activity and against a broader panel of neoantigens.

Method & Results
CD4 and CD8 TILs from a variety of human tumors engineered CD3ζ:MyD88 demonstrated: a) dramatically improved ability to expand in vitro resulting (generating 10–100X TILs than conventional TILs within a few weeks), b) persist for months in tissue culture, c) more polyfunctional cells, d) destruction of established melanoma tumors in vivo, e) enhanced responses against a panel of antigens with no overt autoimmunity.

Conclusions
CD3ζ:MyD88 represents a unique and universal approach to safely enhance T cell response to tumor antigens.
Evaluation of dry DURAClone antibody panels for the characterization of human PBMCs, enriched T cell fractions and anti-BCMA CAR-T cells
Authors: Bowers Rita , Grazia Giulia , Kapinsky Michael , Larbi Anis , Lacaille Vashti 
Published: January 2023

Abstract
CAR-T cells are becoming a more common therapeutic approach and are setting forth to targeting of hematological disorders other than CD19+ entities and also solid tumors. However, the immunological determinants of CAR-T efficacy still need comprehensive investigation from starting material to final product, covering different CAR constructs, transfection methods and targets.
DURAClone dry antibody panels were designed to overcome standardization issues and operator-dependent impact on accuracy and precision in multicentric flow cytometry studies. The aim of this study was to evaluate the utility of DURAClone dry antibody panels for the characterization of PBMCs, derived T cell-enriched fractions as well as anti-BCMA CAR-T cells (ProMab Inc.), including MOCK-transfected controls.
DURAClone dry antibody panels were used as pre-configured or extended with CAR-specific probes and further antibodies. Viable CD45+ leukocyte and CD3+ T cell count, leukocyte composition (CD3/CD19/CD56/CD14/CD16) and the T cell (CD3/CD4/CD8) maturation profile (CD45RA, CD197/CCR7, CD27, CD57) including expression of checkpoint markers (LAG-3, PD-1) were assessed.
Regardless of sample type, absolute counts and cellular composition were assessed using streamlined procedures. Typical staining patterns for T cell maturation profile were observed, revealing unchanged proportions for the different stages between PBMCs and enriched T cell fractions and strong skewing toward effector cell phenotypes with elevated expression of a checkpoint markers in case of CAR-T cells. Our experiments demonstrate the flexible utility of DURAClone dry antibody panels for the phenotypic characterization from starting material to final CAR-T cell product.
Influenza virus-like particle-based hybrid vaccine containing RBD induces immunity against influenza and SARS-CoV-2 viruses
Authors: Bommireddy Ramireddy , Stone Shannon , Bhatnagar Noopur , Kumari Pratima , Munoz Luis E , Oh Judy , Berry Jameson L , Jacobson Kristen M , Jafaar Lahcen , Naing Swe-Htet , Blackerby Allison N , Van der Gaag Tori , Wright Chloe N , Kim Ki-Hye , Lai Lilin , Pack Christopher D , Ramachandiran Sampath , Suthar Mehul S , Kang Sang-Moo , Kumar Mukesh , Selvaraj Periasamy 
Published: January 2023

Abstract
Several approaches have produced an effective vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the influence of immunity against other vaccinations on the durability and efficacy of the immune response against SARS-CoV-2 is still unknown in settings where vaccines against other viruses need to be administered simultaneously. This will be an important factor in developing multivalent vaccines against seasonal viruses. We have developed a hybrid vaccine for SARS-CoV-2 and influenza viruses using influenza virus-like particles (VLP) incorporated with glycosylphosphatidylinositol (GPI)-anchored Spike RBD of SARS-CoV-2 fused to GM-CSF as an adjuvant. GPI-anchored fusion protein of GM-CSF and the SARS-CoV-2 S1 RBD was expressed in CHO-S cells, purified by immunoaffinity chromatography and incorporated onto influenza VLPs by protein transfer to make a hybrid VLP vaccine. The efficacy of the hybrid VLP vaccine was tested against both SARS-CoV-2 and influenza A/PR8 viruses in a mouse model. Our results show that the hybrid vaccine induced a strong antibody response and protected the mice from both influenza virus and mouse-adapted SARS-CoV-2 challenges, with vaccinated mice having less body weight loss and significantly lower lung viral titers compared to control mice. These results suggest that the hybrid vaccine is a promising candidate for preventing influenza A and SARS-CoV-2 infections.
This work was supported by NIH/NIAID (SBIR Contract# 75N93019C00017 Amendment to Pack/Ramachandiran), and Intel Corporation for the Intel COVID-19 Global Technology Response Initiative grant.
Comparison of the Immunogenicity of five COVID-19 vaccines in Sri Lanka
Authors: Jeewandara J M K Chandima , Aberathna Inoka Sepali , Danasekara Saubhagya , Gomes Laksiri , Fernando Suranga , Guruge Dinuka , Ranasinghe Thushali , Gunasekera Banuri , Kamaladasa Achala , Kuruppu Heshan , Somathilake Gayasha , Jayamali Jeewantha , Jayathilaka Deshni , Wijayatilake Helanka Dinesh Kumara , Pushpakumara Pradeep Darshana , Harvie Michael , Nimasha Thashmi , de Silva Shiromi Devika Grace , Wijayamuni Ruwan , Schimanski Lisa , Rijal Pramila , Tan Jack , Townsend Alain , Ogg Graham S , Malavige Gathsaurie Neelika 
Published: January 2023

Abstract
We assessed antibody responses 3 months post-vaccination in those who received mRNA-1273 (n=225), Sputnik V (n=128) or the first dose of Gam-COVID-Vac (n=184) and compared the results with previously reported data of Sinopharm and AZD1222 vaccinees. 99.5% of Moderna &gt;94% of AZD1222 or Sputnik V, 72% to 76% of Gam-COVID-Vac (first dose) and 38.1% to 68.3% of Sinopharm vaccinees had ACE2 blocking antibodies above the positive threshold. The ACE2 blocking antibody levels were highest to lowest was Moderna &gt; Sputnik V/AZD1222 (had equal levels)&gt; first dose of Gam-COVID-Vac &gt; Sinopharm. All Moderna recipients had antibodies above the positive threshold to the ancestral (WT), B.1.1.7, B.1.351.1 and 80% positivity rate for B.1.617.2. Positivity rates of Sputnik V vaccinees for WT and variants, were higher than AZD1222 vaccinees, while Sinopharm vaccinees had the lowest positivity rates (&lt;16.7%). These findings highlight the need for further studies to understand the effects on clinical outcomes.
Allergy, Immunology and Cell Biology Unit, University of Sri Jayewardenepura; the NIH, USA (grant number 5U01AI151788-02), UK Medical Research Council and the Foreign and Commonwealth Office for support. T.K.T. is funded by the Townsend-Jeantet Charitable Trust (charity number 1011770) and the EPA Cephalosporin Early Career Researcher Fund. A.T. are funded by the Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Science (CIFMS), China (grant no. 2018-I2M-2-002).