bluuebunny/crossref_metadata_2025_split huggingface dataset:
ADAM9 drives immunosuppressive environment for lung tumor progression
Authors: Sher Yuh-Pyng , Liu Jing-Pei , Liu Shih-Jen 
Published: August 2023

Abstract
Cancer cells develop the immune suppressive tumor environment, but the underlying molecular mechanism for it is still elusive. Here, we demonstrated that ADAM9 contributed to lung tumor progression and promoted IFN-related gene expression in lung cancer cells that participated in immune regulation in the tumor environment. ADAM9 knockout (KO) lung cancer cells reduced lung cancer metastasis with a decrease in neutrophil infiltration and an increase in CD8+ T cell infiltration. By performing the genome-wide approach to explore ADAM9-regulated genes in lung cancer cells, we revealed that ADAM9 regulated cytokine production and lymphocyte activation. It enhanced the master transcription factor of IFN signaling, STAT1 activation in lung cancer cells. Moreover, ADAM9 promoted IL6/IL6 receptor-STAT3 axis signaling that lead to suppression of IL12p40 secretion and MHC class II expression in dendritic cells and macrophages. Notably, we found that type I IFN related-proteins and steroid biosynthesis-related proteins were related to ADAM9-mediated STAT signaling activation by LC-MS/MS analysis. We demonstrated that ADAM9 created an immune suppressive environment by regulating IFN signaling for tumor progression.
Supported by grants from NHRI (NHRI-EX112-11219BI)
CAR Tregs protect alloimmune islets against autoimmune destruction
Authors: Wardell Christine M , Fung Vivian , Sanchez Isaac Rosado , Mojibian Majid , Levings Megan K 
Published: August 2023

Abstract
Regulatory T cell (Treg) therapy is a promising approach to induce tolerance of transplanted tissues and/or self antigens that drive autoimmunity. Early clinical trials with polyclonal Tregs have proven feasible, safe and tolerable. The next wave of Treg therapies is genetically enhanced with antigen specificity. We have previously shown that Tregs engineered to express a chimeric antigen receptor specific for HLA-A2 (A2-CAR) delay rejection of HLA-A2+ grafts; however, it was unknown whether they would limit T cell responses targeting antigens other than HLA-A2 via bystander suppression. To investigate this possibility, we used a model of HLA-A2-mismatched islet transplantation and asked if A2-CAR Tregs could inhibit rejection mediated by diabetogenic T cells. HLA-A2+ islets (150–200) isolated from NOD-HLA-A2+ mice were transplanted into immunodeficient NSG mice. After 3.5 weeks, diabetogenic BDC2.5 CD4+ T cells were adoptively transferred with or without polyclonal or A2-CAR Tregs (1 CD4+:5 Tregs). Mice that received BDC2.5 CD4+ T cells without Tregs or polyclonal Tregs rapidly developed hyperglycemia. In contrast, mice that received A2-CAR Tregs were completely protected from hyperglycemia. Longitudinal monitoring of injected Tregs showed that A2-CAR Tregs were more numerous and activated relative to polyclonal Tregs. Removal of the HLA-A2+ islet graft from a protected A2-CAR Treg-treated mouse did not induce hyperglycemia, suggesting that the ability of BDC2.5 CD4+ T cells to mediate autoimmune attack of the endogenous (HLA-A2-) pancreas was suppressed. These data suggest that A2-CAR Tregs mediate bystander suppression in vivo and may have the ability to induce long-term tolerance to both allo- and autoantigens.
Supported by a CIHR Canada Graduate Scholarship doctoral research award and a CIHR foundation grant (FDN-154304).
Inflammasome activation occurs in CD4+ and CD8+ T cells during graft-versus-host disease
Authors: Talley Sarah , Campbell Edward 
Published: August 2023

Abstract
A severe complication of hematopoietic stem cell transplantation is graft-versus-host disease (GvHD), a reaction that occurs following the transfer of donor immune cells (the graft) into an allogeneic host. Transplanted cells recognize host alloantigens as foreign, resulting in the activation of donor T cells and migration of these pathological cells into host tissues. In this study, we found that caspase-1 is active in alloreactive murine and human CD4+ and CD8+ T cells early during acute GvHD (aGvHD) development. The presence of inflammasome-bound, active caspase-1 (p33) and ASC speck formation confirmed inflammasome activation in these cells. Isolated T cells with high levels of active caspase-1 had a strong inflammatory transcriptional signature and a metabolic phenotype similar to inflammatory myeloid cells, including the upregulation of proinflammatory cytokines and metabolic switch from oxidative phosphorylation to aerobic glycolysis. We also observed oxidative stress and mitochondrial dysfunction, in alloreactive T cells. For the first time, this study characterizes caspase-1 activation in transplanted T cells during aGvHD, using mouse and human models, adding to a body of literature supporting inflammasome function in cells of the adaptive immune system.
NIH
CD45.1/CD45.2 congenic markers induce a selective bias for CD8+ T cells during adoptive lymphocyte proliferation in lymphocytopenia mice
Authors: Kandy Rakhee Rathnam Kalari , Fan Xiaoxuan , Cao Xuefang 
Published: August 2023

Abstract
CD45.1/CD45.2 congenic markers have been frequently used to track hematopoietic lineage differentiation following hematopoietic stem and progenitor cell (HPSC) transplantation. However, several studies suggest that a bias exists in CD45.1 versus CD45.2 hematopoietic cell reconstitution from the transplanted HPSCs. Meanwhile, no definitive comparison has been reported for mature immune cells as to whether the CD45.1/CD45.2 disparity can skew immune cell response. In this study using lymphocytopenia Rag1−/− CD45.2 mice as hosts, we assessed the proliferative potential of CD45.1 versus CD45.2 lymphocytes following adoptive transfer of mature T and B cells harvested from the spleens of CD45.1 and CD45.2 mice. We have found that a selective bias for CD8+ T cells in that CD45.1 CD8+ T cells showed significantly higher proliferation and higher expression of PD-1 compared CD45.2 CD8+ T cells in the Rag1−/− CD45.2 hosts. This bias is likely due to MHC-I restricted allogeneic stimulation as CD45.1 versus CD45.2 CD4+ T cells and CD19+ B cells contained the same graft as CD8+ T cells showed equivalent proliferation. These results suggest that CD45.1/CD45.2 markers induce an alloreactive response specific to CD8+ T cells, and therefore call for caution for using them as congenic markers in immunologic models.
This study was supported by funds through the National Cancer Institute (R01CA184728), the Maryland Department of Health’s Cigarette Restitution Fund (CRF) Program and used shared core facilities supported by University of Maryland Greenebaum Comprehensive Cancer Center (UMGCC) Support Grant (P30CA134274).
Identification and function of a C3hiIL-33+ fibroblast population in the lungs
Authors: Atakkatan Aishwarya , Gour Naina , Magesh Aishwarya , Yong Hwan Mee , Lajoie Stephane 
Published: August 2023

Abstract
Complement component C3 is known to be predominantly expressed by epithelial and immune cells, however, we have found that fibroblasts are a primary source of C3 in murine and human lungs. Specifically, C3 is majorly expressed in adventitial fibroblasts. Moreover, we found that C3hi fibroblasts are characterized by the expression of IL-33. Further, C3 and IL-33 are not only markers of a novel subset of pulmonary adventitial fibroblasts but form a functional axis where C3 drives the expression of IL-33. As compared to other adventitial fibroblasts, this C3hiIL-33+ subset is enriched for matrix genes, especially type I collagen (COL1A1). These cells require a functional C3− IL-33 axis to maintain their matrix identity. Impairing C3 not only reduces their ability to make collagen but guides them towards a lipofibroblast program characterized by lower collagen content and higher lipid accumulation. As we and others have published that C3 is a key driver of allergic airway inflammation, we wanted to understand how exposure to allergen affected C3hiIL-33+ fibroblasts. Using a murine model of airway allergy, we observed a significant increase in the C3hiIL33+ fibroblast population in lung samples from house dust mite (HDM)-treated mice compared to the PBS control group. Recently, IL-33+ fibroblasts were found to support ILC2 recruitment during helminth infection. As we have found C3 is critical to maintain IL-33 in these cells, we aim to understand if C3hiIL-33+ fibroblasts participate in allergy by mediating cellular crosstalk with ILC2s and T cells during type 2 immune responses.
Supported by grants from NIH R01AI27644 and the Johns Hopkins Catalyst Award to Dr. Stephane Lajoie.
Manipulation of Leucocytic NADPH Oxidase Maturation May Drive Macrophage Differentiation and Affect Atherogenesis
Authors: Cheng Wan-Ting , Shieh Chi-Chang 
Published: August 2023

Abstract
Atherosclerosis is a process of arterial intima thickening caused by lipid deposition as a consequence of insufficient efferocytosis of apoptotic cells by M2 macrophages and excessive inflammatory cytokine secretion from M1 macrophages. NADPH oxidase 2 (NOX2) is recognized as a major source of ROS for enhancing macrophage efferocytosis and functional polarization. We previously found that human macrophages contain significant amount of endoplasmic reticulum (ER)-retained gp91phox, a major component of NOX2, which may hinder the formation of functional NOX2. To address the role of macrophage NOX2 in atherosclerosis, we established a mouse model bearing Apoe−/− and the CYBB point mutation (C1024T), which leads to ER-retained gp91phox in immune cells. We also developed a method for targeted delivery of a SERCA inhibitor with poly (lacticcoglycolic acid) (PLGA) nanoparticles (NPs), which may promote the glycosylation and maturation of ER-retained gp91phox. We first found that oxLDL-administrated M1 macrophages expressed increased M2 marker CD163 and increased efferocytotic ability after being treated with NPs. We also found that there was an increased glycosylation of gp91phox in those macrophages from human and mice with CYBB point mutation (C1024T) after the treatment of NPs. Macrophage ATP-binding cassette transporters, which could decrease cholesterol overloading in macrophages, were upregulated in macrophages after macrophages were treated with NPs. Our results demonstrated that NPs not only can recover the maturation of ER-retained gp91phox in macrophages but also may facilitate the differentiation of M1 to M2-like macrophages with enhanced efferocytotic activity and cholesterol efflux.
Microanatomical lipid niche controls intestinal memory B cell migration and effector function
Authors: Raso Fiona Ann , Howley Kelsey , Reboldi Andrea 
Published: August 2023

Abstract
Enteric infections are a leading global burden, yet few oral vaccines elicit long-lasting immunity and prevent disease, likely due to our lack of understanding of intestinal immunological memory. Long-lived memory B cells can be rapidly reactivated to differentiate into plasma cells, thus further increasing antibody titers and mediating host protection. Such long-term protection requires memory B cells to migrate and persist in sub-anatomical niches within secondary lymphoid organs. While survival niches have been described for memory B cells in lymph nodes and spleen, no niche have been identified in mucosal lymphoid organs, such as Peyer’s patches (PPs).
Here, we show that memory B cells require positioning in the PP subepithelial dome (SED) for maintenance and effector function. In PPs, memory B cells express the migratory receptors EBI2, which binds the oxysterol 7alpha, 25-hydroxycholesterol, and CCR6, which recognizes the chemokine CCL20. Both ligands are present in PPs and drive memory B cell migration in vitro. Conditional removal of EBI2 from memory B cells, either in a constitutive or acute fashion, alters their zonation in PPs, reduces their persistency, and impairs their antigen-specific response. While most memory B cells in PPs are IgA+, IgA B cell receptor expression is dispensable for memory B cell positioning in the SED.
Using conditional mice for the enzymes generating the oxysterol gradient and cell specific Cre lines, we are currently mapping the cellular and metabolic requirements for EBI2 dependent memory B cell maintenance and effector function in PPs.
Understanding the mechanisms for memory B cell persistence is fundamental to our understanding of the intestinal B cell memory response against pathogens.
Previously supported by training grants from NIH (5 T32 AI 132152-2, 5 T32 AI 007349-29)
Does MHC Heterozygosity Promote IgA Plasmablast Diversity in the Gut?
Authors: Peacock Tori , Kubinak Jason 
Published: August 2023

Abstract
The diversity of protein antigens the adaptive immune system can respond to is governed by polymorphic major histocompatibility complex (MHC) molecules. Because MHC alleles are co-dominantly expressed, MHC heterozygosity should allow for a wider diversity of antigens to activate T cells. B cells present antigen via MHC molecules to T cells, and through the ensuing T cell interaction, differentiate into plasma cells. Because of this, MHC heterozygosity should result in the generation of a plasma cell pool able to generate antibodies that target a wider array of antigens. To investigate this claim, we compared B cell selection dynamics between homozygote and heterozygote MHC congenic mice within the germinal centers (GCs) of Peyer’s patches; the major secondary lymphoid tissues of the gut where plasma cells develop in response to the microbiota. I have observed differences in the distribution of GC B cells between the dark zone (DZ) and light zone (LZ) of GCs. Specifically, GC B cells in MHC heterozygotes are found in greater abundance in the LZ and lower abundance in the DZ compared the GC B cells from MHC homozygote mice, which suggests that B cells are more capable of interacting with T cells in MHC heterozygotes. MHC heterozygotes also have significantly higher abundance of GC-TFH cells in their Peyer’s patches. I have observed that MHC heterozygote mice show enhanced binding of IgA to SI-resident bacteria. Future experiments seek to sort-purify IgA-bound SI-resident bacteria from MHC heterozygotes and homozygotes for 16S analysis to determine if MHC heterozygotes bind a more diverse array of bacterial species. We will also perform IgH-sequencing to define the effect of MHC heterozygosity on repertoire diversity of plasma cell populations.
Supported by grants from the NIH (R21AI142409, R01AII55887)
Bruton’s tyrosine kinase impacts IgA repertoire and microbiome composition in non-obese diabetic mice
Authors: Olin Ilana Rae , Cohen Lucy , Brown Alana C , Verma Sonam , Kendall Peggy L 
Published: August 2023

Abstract
The microbiome has emerged as a contributor to type 1 diabetes (T1D), but how interactions between the microbiome and the mucosal immune system lead to or prevent T1D is not clear. Previously, we found that deletion of the B cell signaling protein Bruton’s Tyrosine Kinase (BTK) selectively eliminates autoreactive B cells and autoantibodies, preventing T1D development. However, we discovered that this disease protection is microbiome-dependent, as it disappears in cleaner facilities. We hypothesize that disruption of BTK-mediated B cell receptor (BCR) signaling promotes the growth of particular microbes that are protective against T1D by altering IgA production. Thus, the objective of this study is to determine how BTK-deficiency affects the IgA repertoire, shapes the gut microbiome, and synergistically prevents T1D. First, we performed IgA-seq to assess how BTK-deficiency alters IgA-coating of the microbiota. Then, we characterized the microbiomes of non-obese diabetic (NOD) mouse colonies living in distinct environments using 16S sequencing and conducted disease studies to track T1D development. We found that BTK-deficiency altered the ability of IgA to coat microbiota in the small intestine and colon. We also found that BTK-deficiency significantly impacts the composition of the microbiome in NOD mice living in distinct facilities. BTK-deficient mice have increased commensal diversity and enrichment of specific microbes. Based on these findings, we conclude that disruption of BTK-mediated BCR signaling alters IgA production and microbiome composition, with downstream effects on T1D pathogenesis.
Supported by grants from NIH (R01-DK-084246I) and Veteran’s Affairs (101-BX-002882)
Segmented filamentous bacteria (SFB) drives enhanced T cell-dependent IgA and IgG2b responses in Peyer’s patches
Authors: Kang Byunghyun , Kim Eun-Do , Kim Bong-Hyun , Tomachi Tomohiro , He Jianping , Kelsall Brian L 
Published: August 2023

Abstract
Peyer’s patches (PPs) are the major site for T-cell dependent IgA B-cell differentiation. Segmented filamentous bacteria (SFB) are commensal bacteria that preferentially colonize the ileum, mainly over PPs. SFB colonization of germ-free mice induces polyclonal IgA responses by unclear mechanisms. Using scmRNAseq and flow cytometry we found SFB colonization of SPF mice induced an increase in the size and overall cell number in PPs that includes enhanced numbers of naïve, follicular, and germinal center B-cells as well as Th 17 and Tfh cells. SFB colonization increased the numbers of IgA+ and IgG2b+, but not IgG1+ B-cells in PPs. Enhanced class-switching to IgA and IgG2b was dependent on T-cells, CCR6, CD40-CD40L and DC-expression of MHCII, as well as to some degree on IL-6. An increased number of activated CD11c+ SIRPa+ ST2− CD11b+ DCs with CCR6+ Th17 of SFB+ compared to SFB− mice suggests increased initiation of IgA class switching in the PP subepithelial dome (SED). SFB colonization increased the conversion of Th17 into Tfh cells, however unlike previous reports, SFB-colonized IL-17R−/− and CD4CRE-RORgtfl/fl mice indicated that neither IL-17, nor the enhanced Tfh conversion from Th17 cells is required for increased IgA and IgG2b responses. Finally, B cell repertoire analysis from IgA+, IgG1+ and IgG2b+ PP B cells showed SFB colonization affected the B-cell repertoire qualitatively as well as quantitatively. Together, these data indicate that SFB colonization enhances the recruitment of naïve B cells, as well as drives IgA and IgG2b antibody responses through enhanced T cell-dependent class switching in PPs, and contribute to our understanding of how commensal microbes affect humoral immune responses to infection and vaccination.
This work was supported by the Division of Intramural Research, NIAID, NIH
Small intestine and colon tissue-resident memory CD8+ T cells exhibit molecular heterogeneity and differential dependence on Eomesodermin
Authors: Lin Yun Hsuan Elena , Duong Han G , Limary Abigail E , Kim Eleanor S , Hsu Paul , Patel Shefali A , Wong William H , Indralingam Cynthia S , Liu Yi Chia , Yao Priscilla , Chiang Natalie R , Vandenburgh Sara A , Ferry Amir , Takehara Kennidy K , Tsai Matthew S , Yeo Gene W , Goldrath Ananda W , Chang John T 
Published: August 2023

Abstract
Tissue-resident memory CD8+T (TRM) cells are a subset of memory T cells positioned at barrier sites, critical for host defense against infection. Recent studies have begun to reveal substantial heterogeneity in CD8+ TRM populations across different tissues. The gut consists of four anatomically distinct compartments, the epithelia and lamina propria in the small intestine (siIEL, siLPL) and colon (cIEL, cLPL). While CD8+ TRM in the small intestine epithelia are well-described, findings are often generalized to the whole gut, thus obscuring potential differences in CD8+ TRM in the much-less studied small intestine lamina propria and colon compartments. We thereby characterized CD8+ TRM in the four intestinal layers using single cell RNA-sequencing and flow cytometry. We show that CD8+ TRM across the intestinal compartments exhibit differences in the expression of surface molecules, cytokines, granzymes, and transcription factors. For example, small intestine intraepithelial CD8+ TRM produce the highest levels of granzymes, whereas colon intraepithelial CD8+ TRM have the greatest capacity for cytokine production. We also uncover an unexpected, tissue-specific role for the T-box transcription factor Eomesodermin (Eomes), known to suppress TRM formation in the skin, liver, and kidney. Our data demonstrate that Eomes is dispensable for CD8+ TRM formation in both the small intestine and colon. By contrast, following CD8+ TRM formation, Eomes plays a critical role in promoting the maintenance of established CD8+ TRM in the small intestine, but not in the colon. Together, our study provides new insights into intestinal CD8+ TRM heterogeneity and differential transcriptional regulation of intestinal TRM formation vs. maintenance.
Presenting author Y.H.L. is supported by the Canadian Institutes of Health Research (CIHR) Doctoral Foreign Study Award. This study is supported by grants from NIH (P01 AI132122).
The genomic landscape of infant CD4 T cells and monocytes after BCG vaccination
Authors: Kandiyil Vidya Vijayan Karuvan , Kamthunzi Portia , Tegha Gerald , Hosseinipour Mina , De Paris Kristina 
Published: August 2023

Abstract
The live-attenuated Bacille Calmette-Guérin (BCG) vaccine has been introduced a century ago to prevent tuberculosis and is still administered to &gt;80% of newborns worldwide. Independent of its effect on TB, BCG introduction markedly reduced childhood mortality. Although BCG vaccine is routinely administered to infants, there is a gap in understanding the influence of BCG on the neonatal immune system. In adults, BCG induces trained or heterologous immunity. Here, to better understand the molecular changes in the infant immune system after BCG immunization, we performed RNA sequencing and ATAC sequencing in circulating CD4 T cells and monocytes of 28 infants prior to and 6–12 weeks after administering BCG.
Consistent with data from BCG-vaccinated adults, the expression profile of 168 differentially expressed genes (DEG) supports the activation of monocytes after BCG vaccination, and ATAC sequencing data identified epigenetic modification that impact accessibility of genes in Nod1 and Nod 2 signaling pathways. In CD4 T cells, we identified a total of 322 DEG, including 95 upregulated and 226 downregulated genes. The functional enrichment and transcription factor analysis showed the enrichment of ErbB signaling pathways which regulate cell proliferation and differentiation mediated through MAPK signaling pathways. These results combined with peaks identified by ATAC sequencing were indicative of the expansion of Th1 CD4 T cells in BCG vaccinated infants. This study provides an understanding of molecular changes in monocytes and CD4 T cells after BCG vaccination that may explain the improved capacity of the infant immune system to respond to intracellular pathogens.