Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.234.19
Chad W. Euler, V. Fischetti
Acute rheumatic fever (ARF) is an autoimmune disease that occurs in a subset of patients as a delayed complication of an improperly treated throat infection with a rheumatogenic strain of Streptococcus pyogenes. ARF / Rheumatic heart disease is a leading cause of preventable cardiovascular morbidity and mortality in children worldwide, affecting >33 million people. A method of detecting children susceptible to ARF (3–5% of the total population) would have major global health and economic benefits. One potential ARF diagnostic is the IgM antibody, D8/17, which binds B cells from ARF patients of diverse ethnic groups and geographical locations at a higher % versus matched controls. While this antibody has had mixed results in the past, no group has identified the antigen that D8/17 binds. To aid in this endeavor, we used recombinant antibody engineering to produce new IgM and IgG1 versions of D8/17 that give more consistent results in our in vitrodiagnostic tests. These new derivatives were then utilized in a multi-omics approach to characterize the differences between immortalized B cell lines of ARF patients and controls. Western blot and MS/MS proteomic analyses of lymphocyte lysates identified cytoskeletal proteins from ARF B cells that cross-react more readily with our recombinant D8/17. RNA sequencing and microarray analysis of B cells from ARF patients versus controls confirmed our proteomic results. Further, we discovered differences in the gene expression of other cell surface proteins, kinases and signaling pathways in ARF B cells that bind these antibodies. We hope our analyses will help identify markers or genetic factors related to ARF development and pathogenesis, or aid in designing diagnostic assays for ARF susceptibility. Supported by a grant from the NIH /NIAD (SC2 1SC2AI134947-01)
{"title":"Using novel diagnostic antibodies to identify lymphocyte markers of rheumatic fever and heart disease.","authors":"Chad W. Euler, V. Fischetti","doi":"10.4049/jimmunol.210.supp.234.19","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.234.19","url":null,"abstract":"\u0000 Acute rheumatic fever (ARF) is an autoimmune disease that occurs in a subset of patients as a delayed complication of an improperly treated throat infection with a rheumatogenic strain of Streptococcus pyogenes. ARF / Rheumatic heart disease is a leading cause of preventable cardiovascular morbidity and mortality in children worldwide, affecting >33 million people. A method of detecting children susceptible to ARF (3–5% of the total population) would have major global health and economic benefits.\u0000 One potential ARF diagnostic is the IgM antibody, D8/17, which binds B cells from ARF patients of diverse ethnic groups and geographical locations at a higher % versus matched controls. While this antibody has had mixed results in the past, no group has identified the antigen that D8/17 binds. To aid in this endeavor, we used recombinant antibody engineering to produce new IgM and IgG1 versions of D8/17 that give more consistent results in our in vitrodiagnostic tests.\u0000 These new derivatives were then utilized in a multi-omics approach to characterize the differences between immortalized B cell lines of ARF patients and controls. Western blot and MS/MS proteomic analyses of lymphocyte lysates identified cytoskeletal proteins from ARF B cells that cross-react more readily with our recombinant D8/17. RNA sequencing and microarray analysis of B cells from ARF patients versus controls confirmed our proteomic results. Further, we discovered differences in the gene expression of other cell surface proteins, kinases and signaling pathways in ARF B cells that bind these antibodies. We hope our analyses will help identify markers or genetic factors related to ARF development and pathogenesis, or aid in designing diagnostic assays for ARF susceptibility.\u0000 Supported by a grant from the NIH /NIAD (SC2 1SC2AI134947-01)","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"52 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84999439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.77.05
Xin Zhang, J. Meng, Gitanjali Lobo, Ronak Patel, A. Ray, R. Quinet, W. Davis, J. Zakem, S. Hayat, Z. You
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that is more common in women than men. SLE has been reported with diverse clinical phenotypes of varying severity, which may be caused by sex hormones and environmental factors. Our recent studies showed that high-fat diet (HFD) exacerbated lupus development in MRL/lpr lupus-prone mice. Here we explored the gender difference in lupus progress and autoimmune response to HFD. Twenty male and twenty female MRL/lpr mice were evenly grouped and fed with a regular diet (RD, 10% fat) or HFD (60% fat) for 14 weeks. Body weights were recorded weekly. SLE progression was monitored by skin lesions, urine protein, titers of anti-dsDNA antibody in serum. Kidney and skin from the dorsum of the neck were embedded for H&E, PAS, and Masson’s staining quantified as kidney index and skin score. Immune cells in the spleens were identified by immunofluorescence staining and flow cytometry. HFD induced a greater weight gain in female mice than male mice (p<0.01). Skin rash showed up as early as week 6 in female HFD group with a greater histopathological skin score (p<0.01). Splenomegaly, proteinuria, and anti-dsDNA level were increased only in male HFD group. Kidney pathological changes were more severe in male HFD mice with significant increase of kidney index (p<0.05). Significant increases of germinal center B cells, plasma cells, and T follicular helper cells were observed in HFD mice (p<0.05). SLE progression is sexually dimorphic in lupus-prone mice in response to HFD. Our results parallel many known clinical lupus phenotypes and sexual dimorphism in which male patients are more likely to have severe disease (such as nephritis) than female lupus patients who may have a broader range of lupus symptoms. None
{"title":"Sex disparities in pathological features and autoimmunity in high fat diet associated lupus mouse model","authors":"Xin Zhang, J. Meng, Gitanjali Lobo, Ronak Patel, A. Ray, R. Quinet, W. Davis, J. Zakem, S. Hayat, Z. You","doi":"10.4049/jimmunol.210.supp.77.05","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.77.05","url":null,"abstract":"\u0000 Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that is more common in women than men. SLE has been reported with diverse clinical phenotypes of varying severity, which may be caused by sex hormones and environmental factors. Our recent studies showed that high-fat diet (HFD) exacerbated lupus development in MRL/lpr lupus-prone mice. Here we explored the gender difference in lupus progress and autoimmune response to HFD.\u0000 Twenty male and twenty female MRL/lpr mice were evenly grouped and fed with a regular diet (RD, 10% fat) or HFD (60% fat) for 14 weeks. Body weights were recorded weekly. SLE progression was monitored by skin lesions, urine protein, titers of anti-dsDNA antibody in serum. Kidney and skin from the dorsum of the neck were embedded for H&E, PAS, and Masson’s staining quantified as kidney index and skin score. Immune cells in the spleens were identified by immunofluorescence staining and flow cytometry.\u0000 HFD induced a greater weight gain in female mice than male mice (p<0.01). Skin rash showed up as early as week 6 in female HFD group with a greater histopathological skin score (p<0.01). Splenomegaly, proteinuria, and anti-dsDNA level were increased only in male HFD group. Kidney pathological changes were more severe in male HFD mice with significant increase of kidney index (p<0.05). Significant increases of germinal center B cells, plasma cells, and T follicular helper cells were observed in HFD mice (p<0.05).\u0000 SLE progression is sexually dimorphic in lupus-prone mice in response to HFD. Our results parallel many known clinical lupus phenotypes and sexual dimorphism in which male patients are more likely to have severe disease (such as nephritis) than female lupus patients who may have a broader range of lupus symptoms.\u0000 None","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85039315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.148.14
Lou-Ella George-Alexander, Anna K. Kania, Christopher D. Scharer, J. Boss
Plasma cell differentiation is a tightly regulated process coordinated by the timed expression of several transcription factors as well as histone modifying enzymes that modulate chromatin accessibility. During this process, the histone methyltransferase (HMT) G9a, dimethylates histone H3 lysine 9 (H3K9) at promoters and inhibits gene expression through the recruitment of proteins that impair chromatin accessibility. HMTs are expressed ubiquitously but display distinct enzymatic activities and patterns of chromosomal localization. During plasma cell differentiation, G9a was found to co-localize with Blimp-1, which is required to silence genes associated with a B cell fate and cellular proliferation. However, the processes that are modulated by G9a mediated dimethylation during plasma cell formation remain to be elucidated. To study the role of G9a in plasma cell differentiation, we crossed G9a fl/flmice onto the CD19 Cre/+background (G9aKO mice). Stimulation of CD19 Cre/+(CreCtrl) and G9aKO mice with the T cell independent antigen LPS resulted in a significant increase of activated B cells and plasmablast in G9aKO mice. Further characterization of this phenotype, identified a skewing of the mature B cell sub-populations in G9aKO mice, accompanied by altered proliferation rates when challenged. ATAC-Seq and RNA-Seq will be used to identify chromatin accessibility and expression changes in G9a deficient mice. The CUT&Tag assay will also be used to validate regions that are subject to direct modulation by G9a during plasma cell differentiation. Together, our data suggests that G9a contributes to regulating proliferation during plasma cell differentiation. his work is supported by grants from NIH/NIAID (RO1 AI123733 and P01 AI125180 to JMB)
{"title":"H3K9 dimethyltransferase G9a is an important epigenetic modulator of B cell differentiation","authors":"Lou-Ella George-Alexander, Anna K. Kania, Christopher D. Scharer, J. Boss","doi":"10.4049/jimmunol.210.supp.148.14","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.148.14","url":null,"abstract":"\u0000 Plasma cell differentiation is a tightly regulated process coordinated by the timed expression of several transcription factors as well as histone modifying enzymes that modulate chromatin accessibility. During this process, the histone methyltransferase (HMT) G9a, dimethylates histone H3 lysine 9 (H3K9) at promoters and inhibits gene expression through the recruitment of proteins that impair chromatin accessibility. HMTs are expressed ubiquitously but display distinct enzymatic activities and patterns of chromosomal localization. During plasma cell differentiation, G9a was found to co-localize with Blimp-1, which is required to silence genes associated with a B cell fate and cellular proliferation. However, the processes that are modulated by G9a mediated dimethylation during plasma cell formation remain to be elucidated. To study the role of G9a in plasma cell differentiation, we crossed G9a fl/flmice onto the CD19 Cre/+background (G9aKO mice). Stimulation of CD19 Cre/+(CreCtrl) and G9aKO mice with the T cell independent antigen LPS resulted in a significant increase of activated B cells and plasmablast in G9aKO mice. Further characterization of this phenotype, identified a skewing of the mature B cell sub-populations in G9aKO mice, accompanied by altered proliferation rates when challenged. ATAC-Seq and RNA-Seq will be used to identify chromatin accessibility and expression changes in G9a deficient mice. The CUT&Tag assay will also be used to validate regions that are subject to direct modulation by G9a during plasma cell differentiation. Together, our data suggests that G9a contributes to regulating proliferation during plasma cell differentiation.\u0000 his work is supported by grants from NIH/NIAID (RO1 AI123733 and P01 AI125180 to JMB)","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"64 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85485405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.63.16
Nina C. Nwade, N. James, Akanksha D. Nagarkar, S. Desse, Ummugulsum Yildiz-Altay, G. Okoye, A. Byrd, J. Richmond
Mycosis fungoides (MF) is the most common type of cutaneous T cell lymphoma.There are notable differences in MF presentation in Skin of Color (SOC) versus White skin, such as asymptomatic hyperpigmented and hypopigmented lesions resulting from malignant T-cell interaction from keratinocytes and melanocytes. Therefore, this work aims at elucidating MF immunopathogenesis and gene expression in SOC patients and determining whether clinical presentation differences are dependent on the cytokine type produced by the tumor. We used both hypopigmented and hyperpigmented MF biopsy samples from SOC patients with MF from Howard University Dermatology and compared their gene expression to that of biopsy samples from SOC healthy patients. Preliminary data showed an upregulation of genes such as PRDX1, HLA-DRA, CTNNB1, CSTB, and S100A4 in MF samples versus their healthy counterparts. The MF samples also exhibited downregulation of CCL3, CCRL2, PDGFB and HOXD4 genes when compared to their healthy counterparts. Lastly, the pathways ‘cell cycle and apoptosis’, ‘antigen presentation’, and ‘interferon signaling’ were increased in MF and decreased in the healthy samples. Overall, these data will elucidate MF gene expression in SOC patients as well as the immunopathogenesis that results in varying presentations. This may facilitate the development of more concise diagnostic criteria and personalized targeted immunotherapies to better health outcomes within a minority population. Supported by grants from Dermatology Foundation DRSA (SD), the Skin of Color Society Research Grant (JMR), and the American Skin Association Milstein Research Scholar Award for Melanoma/Non-Melanoma Skin Cancer (ASB).
{"title":"Elucidating Mycosis Fungoides Gene Expression in Skin of Color Patients from Howard University Dermatology","authors":"Nina C. Nwade, N. James, Akanksha D. Nagarkar, S. Desse, Ummugulsum Yildiz-Altay, G. Okoye, A. Byrd, J. Richmond","doi":"10.4049/jimmunol.210.supp.63.16","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.63.16","url":null,"abstract":"\u0000 Mycosis fungoides (MF) is the most common type of cutaneous T cell lymphoma.There are notable differences in MF presentation in Skin of Color (SOC) versus White skin, such as asymptomatic hyperpigmented and hypopigmented lesions resulting from malignant T-cell interaction from keratinocytes and melanocytes. Therefore, this work aims at elucidating MF immunopathogenesis and gene expression in SOC patients and determining whether clinical presentation differences are dependent on the cytokine type produced by the tumor. We used both hypopigmented and hyperpigmented MF biopsy samples from SOC patients with MF from Howard University Dermatology and compared their gene expression to that of biopsy samples from SOC healthy patients. Preliminary data showed an upregulation of genes such as PRDX1, HLA-DRA, CTNNB1, CSTB, and S100A4 in MF samples versus their healthy counterparts. The MF samples also exhibited downregulation of CCL3, CCRL2, PDGFB and HOXD4 genes when compared to their healthy counterparts. Lastly, the pathways ‘cell cycle and apoptosis’, ‘antigen presentation’, and ‘interferon signaling’ were increased in MF and decreased in the healthy samples. Overall, these data will elucidate MF gene expression in SOC patients as well as the immunopathogenesis that results in varying presentations. This may facilitate the development of more concise diagnostic criteria and personalized targeted immunotherapies to better health outcomes within a minority population.\u0000 Supported by grants from Dermatology Foundation DRSA (SD), the Skin of Color Society Research Grant (JMR), and the American Skin Association Milstein Research Scholar Award for Melanoma/Non-Melanoma Skin Cancer (ASB).","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"37 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85748669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.63.15
Ling Cao, James Withers, Elizabeth N. Bean
Previously we have shown that CD137–CD137 ligand (CD137L) mediates pro-nociceptive behaviors in the sciatic nerve crush (SNC) model of neuropathic pain. Global CD137L knockout (KO) mice and wildtype (WT) mice intrathecally treated with neutralizing antibody against either CD137L or CD137 displayed significantly reduced neuropathic pain-like behaviors and faster functional recovery compared to WT mice following SNC. To identify the cellular sources of CD137 and CD137L in the spinal cord, a series of immunohistochemistry analyses was performed. While we could not detect significant expression of CD137L (in part due to both technical and biological issues), we observed CD137 expression exclusively co-localized with astrocytic marker glial fibrillary acidic filament (GFAP). We then conducted a time course study of CD137 expression in the lumbar spinal cord dorsal horn region, the region relevant to nociceptive behaviors, in B6-CD137L KO and B6 WT mice. Significant increases in CD137 expression (represented by integrated density) were observed in WT mice from days 7 to 28 following either SNC or sham surgery. The expression of CD137 in KO mice was slightly (however not statistically significantly) higher than that in WT mice at baseline, and did not change overtime post-either surgery. When RNA expression of CD137 was evaluated, although SNC induced significant upregulation of GFAP RNA expression in WT mice, no changes in CD137 RNA expression were detected in any treatment groups or any genotype of mice. Altogether, we report a novel expression of CD137 on murine spinal cord astrocytes. The underlying mechanism through which astrocytic CD137 mediates SNC-induced nociceptive behavior requires further investigation. NIH/NINDS R01NS098426 (Cao) and Peter Caradonna from the COBRE Histology and Imaging Core funded by NIH/NIGMS P20GM103643 (Meng).
{"title":"Astrocytic expression of CD137 in the lumbar spinal cord following sciatic nerve crush model of neuropathic pain in mice","authors":"Ling Cao, James Withers, Elizabeth N. Bean","doi":"10.4049/jimmunol.210.supp.63.15","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.63.15","url":null,"abstract":"\u0000 Previously we have shown that CD137–CD137 ligand (CD137L) mediates pro-nociceptive behaviors in the sciatic nerve crush (SNC) model of neuropathic pain. Global CD137L knockout (KO) mice and wildtype (WT) mice intrathecally treated with neutralizing antibody against either CD137L or CD137 displayed significantly reduced neuropathic pain-like behaviors and faster functional recovery compared to WT mice following SNC. To identify the cellular sources of CD137 and CD137L in the spinal cord, a series of immunohistochemistry analyses was performed. While we could not detect significant expression of CD137L (in part due to both technical and biological issues), we observed CD137 expression exclusively co-localized with astrocytic marker glial fibrillary acidic filament (GFAP). We then conducted a time course study of CD137 expression in the lumbar spinal cord dorsal horn region, the region relevant to nociceptive behaviors, in B6-CD137L KO and B6 WT mice. Significant increases in CD137 expression (represented by integrated density) were observed in WT mice from days 7 to 28 following either SNC or sham surgery. The expression of CD137 in KO mice was slightly (however not statistically significantly) higher than that in WT mice at baseline, and did not change overtime post-either surgery. When RNA expression of CD137 was evaluated, although SNC induced significant upregulation of GFAP RNA expression in WT mice, no changes in CD137 RNA expression were detected in any treatment groups or any genotype of mice. Altogether, we report a novel expression of CD137 on murine spinal cord astrocytes. The underlying mechanism through which astrocytic CD137 mediates SNC-induced nociceptive behavior requires further investigation.\u0000 NIH/NINDS R01NS098426 (Cao) and Peter Caradonna from the COBRE Histology and Imaging Core funded by NIH/NIGMS P20GM103643 (Meng).","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85900636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.160.01
Joey H. Li, Vignesh Senthilkumar, Siya Shah, Eddie T. Padilla, Luke Riggan, Andréa B. Ball, Nedas Matulionis, Ajit S. Divakaruni, H. Christofk, Timothy E. O’Sullivan
Natural killer (NK) cells are critical for the first line of defense against viral infection and malignancy. While the transcriptional and epigenetic regulation of activated NK cells is well characterized in mice, these processes are not yet fully understood in human NK cells. We performed a targeted CRISPR/Cas9 ribonucleoprotein (RNP) transcription factor screen in mature primary human NK cells to identify putative transcription factors required for optimal effector function. The transcription factor myocyte enhancer factor 2C (MEF2C) emerged as a previously unidentified regulator of NK cell homeostasis, cytokine production, and cytotoxicity. MEF2C-deficient NK cells activated with IL-2 and IL-15 displayed impaired proliferation, degranulation, and production of granzyme B. CRISPR-mediated deletion of MEF2C resulted in disrupted glycolysis and oxidative phosphorylation using both SCENITH and Seahorse extracellular flux analysis. Liquid chromatography/mass spectrometry (LC/MS)-based metabolomics of MEF2C-deficient NK cells revealed reductions in the metabolites glyceraldehyde-3-phosphate and a-ketoglutarate/succinate, accompanied by increased accumulation of lipid metabolism products including phosphorylethanolamine and saturated long-chain fatty acids. In vivo, CRISPR/Cas9 RNP-mediated disruption of MEF2C expression in mouse NK cells resulted in decreased expansion during mouse cytomegalovirus infection. Together, these studies reveal MEF2C as a novel regulator of NK cell effector function through control of multiple critical cell-intrinsic metabolic pathways.
{"title":"MEF2C is a critical regulator of human NK cell metabolism.","authors":"Joey H. Li, Vignesh Senthilkumar, Siya Shah, Eddie T. Padilla, Luke Riggan, Andréa B. Ball, Nedas Matulionis, Ajit S. Divakaruni, H. Christofk, Timothy E. O’Sullivan","doi":"10.4049/jimmunol.210.supp.160.01","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.160.01","url":null,"abstract":"\u0000 Natural killer (NK) cells are critical for the first line of defense against viral infection and malignancy. While the transcriptional and epigenetic regulation of activated NK cells is well characterized in mice, these processes are not yet fully understood in human NK cells. We performed a targeted CRISPR/Cas9 ribonucleoprotein (RNP) transcription factor screen in mature primary human NK cells to identify putative transcription factors required for optimal effector function. The transcription factor myocyte enhancer factor 2C (MEF2C) emerged as a previously unidentified regulator of NK cell homeostasis, cytokine production, and cytotoxicity. MEF2C-deficient NK cells activated with IL-2 and IL-15 displayed impaired proliferation, degranulation, and production of granzyme B. CRISPR-mediated deletion of MEF2C resulted in disrupted glycolysis and oxidative phosphorylation using both SCENITH and Seahorse extracellular flux analysis. Liquid chromatography/mass spectrometry (LC/MS)-based metabolomics of MEF2C-deficient NK cells revealed reductions in the metabolites glyceraldehyde-3-phosphate and a-ketoglutarate/succinate, accompanied by increased accumulation of lipid metabolism products including phosphorylethanolamine and saturated long-chain fatty acids. In vivo, CRISPR/Cas9 RNP-mediated disruption of MEF2C expression in mouse NK cells resulted in decreased expansion during mouse cytomegalovirus infection. Together, these studies reveal MEF2C as a novel regulator of NK cell effector function through control of multiple critical cell-intrinsic metabolic pathways.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"224 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85967108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.252.10
B. Poole, D. Redd, Jessica D Altman, Jamie L. Jensen, Chantel Sloan-Aagard, Triston B Crook, Aaron E Asay, Bryce U Nielson, Ruth J Bodily, Dashiell S Miner
Safe and effective vaccines have been developed that protect against high-risk strains of HPV, but uptake is relatively low. We previously identified factors such as sexual attitudes and HPV knowledge that impact the intent of Christian parents to vaccinate their children against HPV. One of the major factors that decreased vaccine utilization was the belief that they did not need to vaccinate their children because of their moral beliefs. We hypothesized that culturally specific interventions in the form of short videos would be effective at improving HPV vaccine attitudes. We made three short educational videos, one with a Christian focus, one informational about HPV, and one control. Videos were distributed electronically with accompanying surveys, and attitudes were measured before and after watching a randomly selected video. The religious-focused and educational interventions significantly (P=0.001) improved attitudes about HPV vaccination. The religiously-focused video also significantly diminished the belief that the HPV vaccine is unnecessary because of a family’s values (p=0.023). Parents significantly credited both interventions with improving their intent to vaccinate their children against HPV (p<0.001 for both). These results suggest that culturally focused educational interventions are effective at influencing vaccine attitudes, even when those attitudes are based on religious or cultural feelings. Highly specific interventions are likely to be necessary for optimal improvement in vaccine hesitancy. Supported in part by a research grant from Investigator-Initiated Studies Program of Merck Sharp & Dohme LLC.
{"title":"Culturally-specific education can change perception of risk of Human Papillomavirus infection and need for vaccination","authors":"B. Poole, D. Redd, Jessica D Altman, Jamie L. Jensen, Chantel Sloan-Aagard, Triston B Crook, Aaron E Asay, Bryce U Nielson, Ruth J Bodily, Dashiell S Miner","doi":"10.4049/jimmunol.210.supp.252.10","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.252.10","url":null,"abstract":"\u0000 Safe and effective vaccines have been developed that protect against high-risk strains of HPV, but uptake is relatively low. We previously identified factors such as sexual attitudes and HPV knowledge that impact the intent of Christian parents to vaccinate their children against HPV. One of the major factors that decreased vaccine utilization was the belief that they did not need to vaccinate their children because of their moral beliefs. We hypothesized that culturally specific interventions in the form of short videos would be effective at improving HPV vaccine attitudes. We made three short educational videos, one with a Christian focus, one informational about HPV, and one control. Videos were distributed electronically with accompanying surveys, and attitudes were measured before and after watching a randomly selected video. The religious-focused and educational interventions significantly (P=0.001) improved attitudes about HPV vaccination. The religiously-focused video also significantly diminished the belief that the HPV vaccine is unnecessary because of a family’s values (p=0.023). Parents significantly credited both interventions with improving their intent to vaccinate their children against HPV (p<0.001 for both). These results suggest that culturally focused educational interventions are effective at influencing vaccine attitudes, even when those attitudes are based on religious or cultural feelings. Highly specific interventions are likely to be necessary for optimal improvement in vaccine hesitancy.\u0000 Supported in part by a research grant from Investigator-Initiated Studies Program of Merck Sharp & Dohme LLC.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"49 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76912857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.59.19
C. Jondle
Epstein-Barr virus (EBV) is a human specific gammaherpesvirus that establishes lifelong infection in >95% of all adults and is associated with multiple cancers, including B cell lymphomas. Murine gammaherpesvirus 68 (MHV68) is a natural rodent pathogen that is genetically and biologically related to EBV and importantly provides a tractable animal model to study viral and host factors that impact chronic infection. Both of these viruses have a natural tropism for B cells and usurp B cell differentiation to drive a robust polyclonal germinal center response, which is needed to establish the latent viral reservoir in memory B cells. The factors which these viruses use to usurp the host germinal center response to establish chronic infection are not clearly defined. Using MHV68 we discovered that global IL-17RA signaling as well as T-cell intrinsic IL-17RA signaling is proviral and supports the gammaherpesvirus-driven germinal center response needed to establish chronic infection. Loss of global and T-cell intrinsic IL-17RA signaling resulted in a significant attenuation in the germinal center response as well as viral latency and reactivation. Given the B cell tropism of MHV68 we generated a B cell specific model of IL-17RA deficiency to understand the significance of IL-17RA signaling in virally infected cells. Loss of IL-17RA signaling in B cells during MHV68 infection resulted in an attenuated germinal center response as well as reduced viral reactivation and latency, similar to what was observed in the loss of global and T-cell intrinsic IL-17RA signaling. This indicates that IL-17RA signaling in both virally infected and uninfected cells play an important proviral role in the establishment of chronic gammaherpesvirus infection. WMed Start up funds and an American Cancer Society Postdoctoral Award (134165-PF-19-176-01-MPC) which was used to generate the B cell specific IL-17RA deficient mice.
{"title":"The proviral role of B cell-intrinsic IL-17RA signaling in the establishment of chronic gammaherpesvirus infection","authors":"C. Jondle","doi":"10.4049/jimmunol.210.supp.59.19","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.59.19","url":null,"abstract":"\u0000 Epstein-Barr virus (EBV) is a human specific gammaherpesvirus that establishes lifelong infection in >95% of all adults and is associated with multiple cancers, including B cell lymphomas. Murine gammaherpesvirus 68 (MHV68) is a natural rodent pathogen that is genetically and biologically related to EBV and importantly provides a tractable animal model to study viral and host factors that impact chronic infection. Both of these viruses have a natural tropism for B cells and usurp B cell differentiation to drive a robust polyclonal germinal center response, which is needed to establish the latent viral reservoir in memory B cells. The factors which these viruses use to usurp the host germinal center response to establish chronic infection are not clearly defined.\u0000 Using MHV68 we discovered that global IL-17RA signaling as well as T-cell intrinsic IL-17RA signaling is proviral and supports the gammaherpesvirus-driven germinal center response needed to establish chronic infection. Loss of global and T-cell intrinsic IL-17RA signaling resulted in a significant attenuation in the germinal center response as well as viral latency and reactivation.\u0000 Given the B cell tropism of MHV68 we generated a B cell specific model of IL-17RA deficiency to understand the significance of IL-17RA signaling in virally infected cells. Loss of IL-17RA signaling in B cells during MHV68 infection resulted in an attenuated germinal center response as well as reduced viral reactivation and latency, similar to what was observed in the loss of global and T-cell intrinsic IL-17RA signaling. This indicates that IL-17RA signaling in both virally infected and uninfected cells play an important proviral role in the establishment of chronic gammaherpesvirus infection.\u0000 WMed Start up funds and an American Cancer Society Postdoctoral Award (134165-PF-19-176-01-MPC) which was used to generate the B cell specific IL-17RA deficient mice.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"164 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77247944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.64.11
Seungho Choi, S. Mehrotra
Uncoupling protein 2 (UCP2) is an anion transporter in the mitochondrial matrix that controls ROS production, calcium influx, and C4 metabolites. It is regarded as a metabolic regulator. A lack of UCP2 has led to an extreme clinical score with higher oxidative stress and inflammation in autoimmune illness, even though elevated UCP2 has been found in many tumors and is known to be a potential target for tumor therapy. Uncertainty still exists regarding UCP2’s function in T cells. A significant rise in the Th2/Th1 ratio was seen in this work using CD4 cells from UCP2−/− mice. These cells exhibited an improved Th2 phenotype with decreased calcium influx, elevated iNOS expression, and elevated urea metabolism. CD4 cells were treated with NOC-18, a source of nitric oxide, or Genipin to demonstrate nitric oxide-mediated Th2 polarization (inhibitor for UCP2). There were more cytotoxic markers and Th2/Th1 double positive cells. The B16/F10 tumor cells were controlled in vitro in a tumor killing assay utilizing genipin-treated CD4 cells, according to the findings. However, in UCP2−/− mice, the severity of EAE worsened, and heterogenic Th2 cells were infiltrated into central nerves system. Our findings imply that the loss of UCP2 in CD4 increases Th2 polarization with a cytotoxic phenotype and may be relevant to various illnesses. R01 CA250458, R01 CA236379
{"title":"UCP2 Deficiency Renders Th1/2 Pathogenic Phenotype to CD4+ T cells","authors":"Seungho Choi, S. Mehrotra","doi":"10.4049/jimmunol.210.supp.64.11","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.64.11","url":null,"abstract":"\u0000 Uncoupling protein 2 (UCP2) is an anion transporter in the mitochondrial matrix that controls ROS production, calcium influx, and C4 metabolites. It is regarded as a metabolic regulator. A lack of UCP2 has led to an extreme clinical score with higher oxidative stress and inflammation in autoimmune illness, even though elevated UCP2 has been found in many tumors and is known to be a potential target for tumor therapy. Uncertainty still exists regarding UCP2’s function in T cells. A significant rise in the Th2/Th1 ratio was seen in this work using CD4 cells from UCP2−/− mice. These cells exhibited an improved Th2 phenotype with decreased calcium influx, elevated iNOS expression, and elevated urea metabolism. CD4 cells were treated with NOC-18, a source of nitric oxide, or Genipin to demonstrate nitric oxide-mediated Th2 polarization (inhibitor for UCP2). There were more cytotoxic markers and Th2/Th1 double positive cells. The B16/F10 tumor cells were controlled in vitro in a tumor killing assay utilizing genipin-treated CD4 cells, according to the findings. However, in UCP2−/− mice, the severity of EAE worsened, and heterogenic Th2 cells were infiltrated into central nerves system. Our findings imply that the loss of UCP2 in CD4 increases Th2 polarization with a cytotoxic phenotype and may be relevant to various illnesses.\u0000 R01 CA250458, R01 CA236379","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"50 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80876428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.88.08
A. Putelo, Tzu-Yu Feng, Sree H Kolli, Mitchell T McGinty, Melanie R Rutkowski
Hormone receptor-positive (HR +), HER2 −is the most prevalent metastatic breast cancer subtype, constituting 73.1% of the metastatic disease population in the US. Despite targeted therapies that have increased long-term survival, many patients develop and eventually succumb to metastatic disease. Tumor dissemination occurs early during disease progression and is driven by cellular and molecular changes in the tissue. Though host-intrinsic factors that preferentially predispose certain individuals to metastatic disease are poorly defined, gut microbial health has become increasingly recognized as a determinant of the metastatic potential of breast tumors. We demonstrated that commensal dysbiosis, an inflammatory gut microbiome with low biodiversity, drives HR +tumor metastasis. Our work identified that a dysbiosis-induced CCL2/mast cell axis in normal (non-tumor-bearing) mammary tissues acts as a master regulator of early metastasis. Our present goal is to define how gut dysbiosis triggers tissue inflammation. We present evidence that dysbiosis-induced metabolic dysregulation increases early dissemination of HR +tumors by initiating cellular and molecular changes in normal mammary tissue. Dysbiosis results in systemic insulin-glucose dynamics that resemble an insulin resistant phenotype. Additionally, we find that acylcarnitine species, byproducts of incomplete fatty acid β-oxidation which we hypothesize promote local tissue inflammation, accumulate in normal mammary tissues of dysbiotic mice. Altogether, we propose that commensal dysbiosis triggers a systemic metabolic shift that enhances the metastatic potential of HR +tumors by shaping the immune landscape of the pre-cancerous mammary tissues. Supported by grants from NIH (1RO1 CA253285, 2T32AI007496-26A1)
{"title":"Distal inflammatory priming of normal mammary tissue by the gut microbiome drives breast tumor metastasis via metabolic dysregulation","authors":"A. Putelo, Tzu-Yu Feng, Sree H Kolli, Mitchell T McGinty, Melanie R Rutkowski","doi":"10.4049/jimmunol.210.supp.88.08","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.88.08","url":null,"abstract":"\u0000 Hormone receptor-positive (HR +), HER2 −is the most prevalent metastatic breast cancer subtype, constituting 73.1% of the metastatic disease population in the US. Despite targeted therapies that have increased long-term survival, many patients develop and eventually succumb to metastatic disease. Tumor dissemination occurs early during disease progression and is driven by cellular and molecular changes in the tissue. Though host-intrinsic factors that preferentially predispose certain individuals to metastatic disease are poorly defined, gut microbial health has become increasingly recognized as a determinant of the metastatic potential of breast tumors. We demonstrated that commensal dysbiosis, an inflammatory gut microbiome with low biodiversity, drives HR +tumor metastasis. Our work identified that a dysbiosis-induced CCL2/mast cell axis in normal (non-tumor-bearing) mammary tissues acts as a master regulator of early metastasis. Our present goal is to define how gut dysbiosis triggers tissue inflammation. We present evidence that dysbiosis-induced metabolic dysregulation increases early dissemination of HR +tumors by initiating cellular and molecular changes in normal mammary tissue. Dysbiosis results in systemic insulin-glucose dynamics that resemble an insulin resistant phenotype. Additionally, we find that acylcarnitine species, byproducts of incomplete fatty acid β-oxidation which we hypothesize promote local tissue inflammation, accumulate in normal mammary tissues of dysbiotic mice. Altogether, we propose that commensal dysbiosis triggers a systemic metabolic shift that enhances the metastatic potential of HR +tumors by shaping the immune landscape of the pre-cancerous mammary tissues.\u0000 Supported by grants from NIH (1RO1 CA253285, 2T32AI007496-26A1)","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80888658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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