Pub Date : 2023-05-01DOI: 10.4049/jimmunol.210.supp.59.15
I. Košík, Jefferson J.J. Santos, Mathew Angel, Zhe Hu, J. Holly, J. Gibbs, Tanner U. Gill, Sarah F. Andrews, Rebecca A. Gillespie, M. Kanekiyo, A. McDermott, J. Yewdell
The broadly neutralizing hemagglutinin (BN-HA) stem antibodies represent an universal vaccine prospect. However, our understanding of how these antibodies interact with the influenza virus is limited by excluding other immune elements, such as complement, in our experimental systems. Here, we show that the Fc binding protein of the complement system, C1q, grants BN-HA stem Abs a novel attachment inhibition (AI) and boosts virus neutralization (VN) activity. C1q promotes AI even if added to preformed BN-HA Ab-virion-RBC complexes, pointing at the steric nature of the phenomenon. Most probably by the same mechanism of steric hindrance, C1q also contributes to fusion inhibition. By employing a panel of engineered avidity-reverted anti-BNHA Abs we show that the C1q-mediated VN enhancement is independent of Ab avidity. We also show that C1q’s presence expands the escape viral repertoire beyond the HA stem domain. Among selected viral variants, near-RBS mutations (E156K-Sb, R224I-Ca, S145N-Ca) modulate HA avidity while causing antigenic drift. Expanding our discoveries to SARS-CoV-2, we identified non-RBD anti-SARS-CoV-2 spike Abs which exhibit C1q-VN enhancement. With an increased SARS-CoV-2 spike per viral particle amount, C1q-VN increased concomitantly. Altogether, our results emphasize the general role of C1q in Ab-mediated antiviral activity and viral evolution. NIH/NIAID intramural funding
广泛中和的血凝素(BN-HA)干细胞抗体代表了通用疫苗的前景。然而，我们对这些抗体如何与流感病毒相互作用的理解是有限的，因为在我们的实验系统中排除了其他免疫元素，如补体。在这里，我们发现补体系统的Fc结合蛋白C1q赋予BN-HA干细胞抗体一种新的附着抑制(AI)并增强病毒中和(VN)活性。即使将C1q添加到预形成的BN-HA ab -病毒粒子- rbc复合物中，也能促进AI，这表明了该现象的立体性质。很可能通过相同的位阻机制，C1q也有助于融合抑制。通过使用一组工程化的抗bnha抗体，我们发现c1q介导的VN增强与抗体的亲和力无关。我们还表明，C1q的存在将逃逸病毒库扩展到HA茎域之外。在选定的病毒变体中，接近rbs的突变(E156K-Sb, R224I-Ca, S145N-Ca)在引起抗原漂移的同时调节HA的亲和力。将我们的发现扩展到SARS-CoV-2，我们发现了非rbd抗SARS-CoV-2刺突抗体，其表现出C1q-VN增强。随着每病毒颗粒数量的SARS-CoV-2尖峰增加，C1q-VN也随之增加。总之，我们的研究结果强调了C1q在抗体介导的抗病毒活性和病毒进化中的一般作用。NIH/NIAID内部资助
{"title":"The complement protein C1q modulates the activities of anti-influenza HA-stem and anti-SARS-Cov2-nonRBD antibodies and alters viral evolution","authors":"I. Košík, Jefferson J.J. Santos, Mathew Angel, Zhe Hu, J. Holly, J. Gibbs, Tanner U. Gill, Sarah F. Andrews, Rebecca A. Gillespie, M. Kanekiyo, A. McDermott, J. Yewdell","doi":"10.4049/jimmunol.210.supp.59.15","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.59.15","url":null,"abstract":"\u0000 The broadly neutralizing hemagglutinin (BN-HA) stem antibodies represent an universal vaccine prospect. However, our understanding of how these antibodies interact with the influenza virus is limited by excluding other immune elements, such as complement, in our experimental systems.\u0000 Here, we show that the Fc binding protein of the complement system, C1q, grants BN-HA stem Abs a novel attachment inhibition (AI) and boosts virus neutralization (VN) activity. C1q promotes AI even if added to preformed BN-HA Ab-virion-RBC complexes, pointing at the steric nature of the phenomenon. Most probably by the same mechanism of steric hindrance, C1q also contributes to fusion inhibition. By employing a panel of engineered avidity-reverted anti-BNHA Abs we show that the C1q-mediated VN enhancement is independent of Ab avidity. We also show that C1q’s presence expands the escape viral repertoire beyond the HA stem domain. Among selected viral variants, near-RBS mutations (E156K-Sb, R224I-Ca, S145N-Ca) modulate HA avidity while causing antigenic drift. Expanding our discoveries to SARS-CoV-2, we identified non-RBD anti-SARS-CoV-2 spike Abs which exhibit C1q-VN enhancement. With an increased SARS-CoV-2 spike per viral particle amount, C1q-VN increased concomitantly. Altogether, our results emphasize the general role of C1q in Ab-mediated antiviral activity and viral evolution.\u0000 NIH/NIAID intramural funding","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"2011 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73509022","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.160.12
Jennifer Tran, Pamela Wong, Changxu Fan, David A. Russler-Germain, Celia C. Cubitt, Nancy D. Marin, Timothy Schappe, M. Berrien-Elliott, Ting Wang, J. Foltz, T. Fehniger
NK cells exhibit memory properties after activation with haptens, viral infection, or cytokines with each stimulus having distinct biology. Memory-like (ML) NK cells differentiate over one week after brief activation with combined IL-12/15/18, and have augmented responses upon restimulation by cytokines, activating receptor, or tumor targets. While much is known about their functional characteristics, there are large gaps in our understanding of the molecular mechanisms involved in programming conventional (c)NK cells into ML NK cells. As ML NK cells have enhanced ability to produce IFN-γ after multiple cell divisions, this suggests that the IFNG gene, and others, may be epigenetically poised. Therefore, we hypothesize that activation with IL-12/15/18 triggers epigenetic remodeling of cNK cells to ML NK cells that drives enhanced functionality. To test this, we performed ATAC-seq on baseline cNK cells, IL-12/15/18 activated NK cells, and in vitro differentiated cNK and ML NK cells after one week. Activation with IL-12/15/18 induces dramatic early epigenetic remodeling with >10,000 differentially accessible regions (DARs) compared to cNK cells. Moreover, ML differentiation generates epigenetically distinct ML NK cells (>1,000 DARs) compared to in vitro differentiated cNK cells, including increased IFNG locus accessibility, suggesting epigenetic mechanisms regulate their enhanced functionality. Also, ML NK cells are markedly different from IL-12/15/18 activated NK cells (>15,000 DARs) suggesting not all epigenetic changes are immediately induced, and time is required for full implementation of the memory-like state. Together, this supports that ML NK cells rely on epigenetic programming for enhanced function. Supported by grants from NIH (P50CA171963, R01CA205239, P30CA91842, 1F31GM146361-01).
{"title":"Combined IL-12, IL-15 and IL-18 activation induces epigenetic changes in human NK cells during the memory-like state transition","authors":"Jennifer Tran, Pamela Wong, Changxu Fan, David A. Russler-Germain, Celia C. Cubitt, Nancy D. Marin, Timothy Schappe, M. Berrien-Elliott, Ting Wang, J. Foltz, T. Fehniger","doi":"10.4049/jimmunol.210.supp.160.12","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.160.12","url":null,"abstract":"\u0000 NK cells exhibit memory properties after activation with haptens, viral infection, or cytokines with each stimulus having distinct biology. Memory-like (ML) NK cells differentiate over one week after brief activation with combined IL-12/15/18, and have augmented responses upon restimulation by cytokines, activating receptor, or tumor targets. While much is known about their functional characteristics, there are large gaps in our understanding of the molecular mechanisms involved in programming conventional (c)NK cells into ML NK cells. As ML NK cells have enhanced ability to produce IFN-γ after multiple cell divisions, this suggests that the IFNG gene, and others, may be epigenetically poised. Therefore, we hypothesize that activation with IL-12/15/18 triggers epigenetic remodeling of cNK cells to ML NK cells that drives enhanced functionality. To test this, we performed ATAC-seq on baseline cNK cells, IL-12/15/18 activated NK cells, and in vitro differentiated cNK and ML NK cells after one week. Activation with IL-12/15/18 induces dramatic early epigenetic remodeling with >10,000 differentially accessible regions (DARs) compared to cNK cells. Moreover, ML differentiation generates epigenetically distinct ML NK cells (>1,000 DARs) compared to in vitro differentiated cNK cells, including increased IFNG locus accessibility, suggesting epigenetic mechanisms regulate their enhanced functionality. Also, ML NK cells are markedly different from IL-12/15/18 activated NK cells (>15,000 DARs) suggesting not all epigenetic changes are immediately induced, and time is required for full implementation of the memory-like state. Together, this supports that ML NK cells rely on epigenetic programming for enhanced function.\u0000 Supported by grants from NIH (P50CA171963, R01CA205239, P30CA91842, 1F31GM146361-01).","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73446805","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.150.06
Deepika Sharma, Ankit Malik, Shaina McGrath, Sarah Zabala, Daping Yang, Isaac M. Chiu, B. Jabri
Epithelial response to injury is coordinated through an intricate interaction with neuronal and myeloid cells, however the signaling modules involved are not well understood. In humans, somatic mutations in Tet methylcytosine dioxygenase 2 (TET2), a DNA demethylase, are commonly observed during ageing in myeloid cells and known to modulate inflammatory responses. Using a mouse model that lacks TET2 in myeloid cells (Tet2 ΔLysM), we show that myeloid cells and sympathetic neurons form a signaling nexus that controls differentiation of enterochromaffin cells and serotonin production during colonic inflammation. Under physiological conditions, TET2 restricts IL-1β production by myeloid cells which in turn controls the intestinal sympathetic architecture. During inflammation, IL1R signaling limits sympathetic cues that drive differentiation of enterochromaffin cells through α1-adrenergic signaling. As a result, enterochromaffin differentiation and colitis progression in response to mucosal injury is attenuated in Tet2 ΔLysMmice. Further, protection from colitis in Tet2 ΔLysMmice is mediated by its catalytic activity, and dependent on sympathetic neurons and IL1R signaling. Adrenergic control of epithelial response and pro-colitic serotonin production is also evident under conditions of physiological stress that leads to increased colitis susceptibility. Overall, our study reveals a sympathetic-epithelial axis that controls the severity of colitis and is modulated by myeloid-derived IL-1β and physiological stress. Our findings may also explain why inflammatory bowel disease in the elderly, where TET2 mutations in myeloid cells are common, is less severe and suggests TET2 activity as an attractive target for IBD. Supported by the US National Institutes of Health (DK067180) to B.J, the University of Chicago's Center for Interdisciplinary Study of Inflammatory Intestinal Disorders (C-IID) Pilot & Feasibility Award (NIDDK P30 DK042086) to A.M. and D.S, Crohn’s and Colitis Foundation Career Development Award #964209 to A.M. and G.I. Research Foundation Associates Board Award to A.M. and D.S.
{"title":"Neuronal cells co-ordinate myeloid-epithelial cell interaction to regulate intestinal inflammation","authors":"Deepika Sharma, Ankit Malik, Shaina McGrath, Sarah Zabala, Daping Yang, Isaac M. Chiu, B. Jabri","doi":"10.4049/jimmunol.210.supp.150.06","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.150.06","url":null,"abstract":"\u0000 Epithelial response to injury is coordinated through an intricate interaction with neuronal and myeloid cells, however the signaling modules involved are not well understood. In humans, somatic mutations in Tet methylcytosine dioxygenase 2 (TET2), a DNA demethylase, are commonly observed during ageing in myeloid cells and known to modulate inflammatory responses. Using a mouse model that lacks TET2 in myeloid cells (Tet2 ΔLysM), we show that myeloid cells and sympathetic neurons form a signaling nexus that controls differentiation of enterochromaffin cells and serotonin production during colonic inflammation. Under physiological conditions, TET2 restricts IL-1β production by myeloid cells which in turn controls the intestinal sympathetic architecture. During inflammation, IL1R signaling limits sympathetic cues that drive differentiation of enterochromaffin cells through α1-adrenergic signaling. As a result, enterochromaffin differentiation and colitis progression in response to mucosal injury is attenuated in Tet2 ΔLysMmice. Further, protection from colitis in Tet2 ΔLysMmice is mediated by its catalytic activity, and dependent on sympathetic neurons and IL1R signaling. Adrenergic control of epithelial response and pro-colitic serotonin production is also evident under conditions of physiological stress that leads to increased colitis susceptibility. Overall, our study reveals a sympathetic-epithelial axis that controls the severity of colitis and is modulated by myeloid-derived IL-1β and physiological stress. Our findings may also explain why inflammatory bowel disease in the elderly, where TET2 mutations in myeloid cells are common, is less severe and suggests TET2 activity as an attractive target for IBD.\u0000 Supported by the US National Institutes of Health (DK067180) to B.J, the University of Chicago's Center for Interdisciplinary Study of Inflammatory Intestinal Disorders (C-IID) Pilot & Feasibility Award (NIDDK P30 DK042086) to A.M. and D.S, Crohn’s and Colitis Foundation Career Development Award #964209 to A.M. and G.I. Research Foundation Associates Board Award to A.M. and D.S.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"102 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73465499","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.247.08
Samantha L. Coss, R. Aziz, Danlei Zhou, Bi Zhou, K. Miller, Yee-Ling Wu, S. Ardoin, E. Oberle, K. Driest, O. Al Ahmed, A. Patwardhan, S. Akoghlanian, V. Sivaraman, F. Barbar‐Smiley, Joanne Drew, C. Spencer, L. Pachman, Gabrielle Morgan, Gulnara Mamyrova, R. Curiel, O. Jones, Terry O’Hanlon, L. Rider, F. Miller, L. Padyukov, I. Lundberg, A. Notarnicola, J. Vencovský, B. Stibůrková, O. Kryštůfková, Chack-Yung Yu
Juvenile dermatomyositis (JDM) is an autoimmune myopathy characterized by rash and muscle weakness. Complement C4 gene copy number (GCN) variation is a known risk factor for JDM. JDM patients often develop autoantibodies. We investigated the relationship between C4 GCN, clinical features, and antibody development. Subjects were recruited (n=255) from the US, Sweden, and the Czech Republic. C4 GCN was determined by real time PCR. Comparative analyses were performed via student’s t test or Mann Whitney U test. Correlation was assessed via linear regression or Fisher’s exact test. All studies were IRB approved, and informed consent was obtained. C4 GCN correlated with muscle pathology and extra-muscular disease at diagnosis. Lower C4A and C4L GCN were associated with a higher number of abnormal muscle enzymes (p=0.0062 and p=0.0029), while the opposite was true for C4S (p=0.024). Higher C4S GCN correlated with lower muscle strength scores (p=0.039). Lower C4L correlated with abnormal MRI (p= 0.042). C4B GCN correlated with dysphagia (p=0.035). Higher GCN of C4S was a risk factor for arthritis (p=0.032), systemic symptoms (p=0.020), and dysphagia (p=0.0078). C4 GCN was associated with myositis-specific (MSA) and myositis-associated (MAA) antibodies. Patients with homozygous C4A deficiency were more likely to test positive for anti-NXP2 (OR 20.0, p=0.011), and C4A GCN was lower in anti-NXP2 positive subjects (p=0.030). A higher C4B GCN correlated with positive MAA (p=0.043). Here, we show that low C4A/C4L GCN and high C4B/C4S GCN correlate with worse muscle disease, extra-muscular pathology, and autoantibodies. Our data suggest that complement C4 may play a key role in the ongoing pathogenesis of JDM. Supported by grants from NIH (R21 AR070509) and the CureJM Foundation
{"title":"Complement C4 gene copy number variations and polymorphisms, autoantibodies, and clinical manifestations of juvenile dermatomyositis– a multi-center study","authors":"Samantha L. Coss, R. Aziz, Danlei Zhou, Bi Zhou, K. Miller, Yee-Ling Wu, S. Ardoin, E. Oberle, K. Driest, O. Al Ahmed, A. Patwardhan, S. Akoghlanian, V. Sivaraman, F. Barbar‐Smiley, Joanne Drew, C. Spencer, L. Pachman, Gabrielle Morgan, Gulnara Mamyrova, R. Curiel, O. Jones, Terry O’Hanlon, L. Rider, F. Miller, L. Padyukov, I. Lundberg, A. Notarnicola, J. Vencovský, B. Stibůrková, O. Kryštůfková, Chack-Yung Yu","doi":"10.4049/jimmunol.210.supp.247.08","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.247.08","url":null,"abstract":"\u0000 Juvenile dermatomyositis (JDM) is an autoimmune myopathy characterized by rash and muscle weakness. Complement C4 gene copy number (GCN) variation is a known risk factor for JDM. JDM patients often develop autoantibodies. We investigated the relationship between C4 GCN, clinical features, and antibody development.\u0000 \u0000 \u0000 Subjects were recruited (n=255) from the US, Sweden, and the Czech Republic. C4 GCN was determined by real time PCR. Comparative analyses were performed via student’s t test or Mann Whitney U test. Correlation was assessed via linear regression or Fisher’s exact test. All studies were IRB approved, and informed consent was obtained.\u0000 \u0000 \u0000 \u0000 C4 GCN correlated with muscle pathology and extra-muscular disease at diagnosis. Lower C4A and C4L GCN were associated with a higher number of abnormal muscle enzymes (p=0.0062 and p=0.0029), while the opposite was true for C4S (p=0.024). Higher C4S GCN correlated with lower muscle strength scores (p=0.039). Lower C4L correlated with abnormal MRI (p= 0.042). C4B GCN correlated with dysphagia (p=0.035). Higher GCN of C4S was a risk factor for arthritis (p=0.032), systemic symptoms (p=0.020), and dysphagia (p=0.0078). C4 GCN was associated with myositis-specific (MSA) and myositis-associated (MAA) antibodies. Patients with homozygous C4A deficiency were more likely to test positive for anti-NXP2 (OR 20.0, p=0.011), and C4A GCN was lower in anti-NXP2 positive subjects (p=0.030). A higher C4B GCN correlated with positive MAA (p=0.043).\u0000 \u0000 \u0000 \u0000 Here, we show that low C4A/C4L GCN and high C4B/C4S GCN correlate with worse muscle disease, extra-muscular pathology, and autoantibodies. Our data suggest that complement C4 may play a key role in the ongoing pathogenesis of JDM.\u0000 \u0000 \u0000 \u0000 Supported by grants from NIH (R21 AR070509) and the CureJM Foundation\u0000","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"135 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73744354","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.145.01
Kirsty A McBain, J. Lovchik, N. Bevan
Complement dependent cytotoxicity (CDC) is a key Fc mediated function of monoclonal antibody (mAb) therapeutics. mAb binding to antigen triggers a complex molecular cascade with sequential recruitment of serum proteins, eventuating in target cell lysis. Here we demonstrate CDC quantification in a streamlined, in vitro, advanced flow cytometry assay. Anti-CD20 mAbs were incubated with target cell lines in 96- or 384-well plates. Human serum (15%) was then added to induce CDC. Cells were labeled with iQue® Cell Membrane Integrity (R/Red) Dye to enable assessment of cell death using the iQue® Advanced Flow Cytometry Platform. Induction of CDC by anti-CD20 mAbs correlated with target cell CD20 expression, with most cell death observed with high-CD20 expressing Ramos cells. Maximal Ramos cell death induced by Rituximab and Truxima® was 68% and 70%, respectively. Comparatively, cell death of mid-CD20 expressing Raji cells was lower at 34% and 24%. No CDC was induced with CD20 negative Jurkat cells. Another anti-CD20-IgG1 mAb was compared against two isotype mutants: IgG1fut (non-fucosylated) and IgG1NQ (non-glycosylated). Their activity was profiled towards three previously described effector functions: antibody-dependent cellular cytotoxicity (ADCC); antibody-dependent cellular phagocytosis (ADCP) and CDC. CDC induction was comparable between the three mAbs, however their ADCC and ADCP activity differed. The IgG1NQ did not exert ADCP or ADCC activity, whilst the IgG1fut showed more ADCC and less ADCP than the native. These data exemplify the use of advanced flow cytometry to quantify Fc function of mAbs towards a range of effector mechanisms, highlighting the potential to profile libraries of novel therapeutics in minimal time.
{"title":"Quantifying in vitrocomplement-dependent cytotoxicity (CDC) using advanced flow cytometry","authors":"Kirsty A McBain, J. Lovchik, N. Bevan","doi":"10.4049/jimmunol.210.supp.145.01","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.145.01","url":null,"abstract":"\u0000 Complement dependent cytotoxicity (CDC) is a key Fc mediated function of monoclonal antibody (mAb) therapeutics. mAb binding to antigen triggers a complex molecular cascade with sequential recruitment of serum proteins, eventuating in target cell lysis. Here we demonstrate CDC quantification in a streamlined, in vitro, advanced flow cytometry assay.\u0000 Anti-CD20 mAbs were incubated with target cell lines in 96- or 384-well plates. Human serum (15%) was then added to induce CDC. Cells were labeled with iQue® Cell Membrane Integrity (R/Red) Dye to enable assessment of cell death using the iQue® Advanced Flow Cytometry Platform.\u0000 Induction of CDC by anti-CD20 mAbs correlated with target cell CD20 expression, with most cell death observed with high-CD20 expressing Ramos cells. Maximal Ramos cell death induced by Rituximab and Truxima® was 68% and 70%, respectively. Comparatively, cell death of mid-CD20 expressing Raji cells was lower at 34% and 24%. No CDC was induced with CD20 negative Jurkat cells. Another anti-CD20-IgG1 mAb was compared against two isotype mutants: IgG1fut (non-fucosylated) and IgG1NQ (non-glycosylated). Their activity was profiled towards three previously described effector functions: antibody-dependent cellular cytotoxicity (ADCC); antibody-dependent cellular phagocytosis (ADCP) and CDC. CDC induction was comparable between the three mAbs, however their ADCC and ADCP activity differed. The IgG1NQ did not exert ADCP or ADCC activity, whilst the IgG1fut showed more ADCC and less ADCP than the native.\u0000 These data exemplify the use of advanced flow cytometry to quantify Fc function of mAbs towards a range of effector mechanisms, highlighting the potential to profile libraries of novel therapeutics in minimal time.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"31 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74133283","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.65.14
L. Siransy, A. Adou, Jokebed Kira, N. Moussa, Jocelyne Sery, Doris Oura
The immune system is a particularly sophisticated protection system. As a true guardian of our health, the immune system defends our body against external or internal aggressions. Sometimes, the immune system is not able to fight because it lacks or does not function properly causing an ineffective or absent immune response. Although more than 400 diseases exist, there is no data available in our country. We analyze through a retrospective study carried out in the pediatric department at Hospital University Center, in Abidjan, between February and April 2022, involving 93 PID suspect children, the epidemiological, clinical, paraclinical and therapeutic aspects. The average age of our patients was 2 years with extremes of 22 days to 15 years, with a male predominance: sex ratio of 1.45. The predominant geographical origin was Abidjan (85.32%). Consanguinity was found in 12.9%, death in siblings was found in 3.49% of patients. The most common clinical manifestations were repeated respiratory infections (60.71%), fever (16%), severe infections at least twice a year (13.04%), persistent fungal infections (12.90), thrive retardation in 48.7%. Abnormalities such as hyperleukocytosis (42.4%), neutropenia (29.23%), lymphopenia (16.44%), thrombocytopenia (38.36%) and mostly anemia (62.36%) were noted. Phagocytosis deficiency combined immune deficiencies and humoral deficiencies were respectively found at 95.16%, 89.78%.79.03%. This work draws attention to the importance of PID which are often underdiagnosed, with absent or delayed treatment and poor prognosis. It is important to build a cohort with confirmed data to better support children with PID in our country. no support
{"title":"Primary immunodeficiency disorders (PID) or inborn errors of immunity : preliminary work West Africa Côte d’Ivoire","authors":"L. Siransy, A. Adou, Jokebed Kira, N. Moussa, Jocelyne Sery, Doris Oura","doi":"10.4049/jimmunol.210.supp.65.14","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.65.14","url":null,"abstract":"\u0000 The immune system is a particularly sophisticated protection system. As a true guardian of our health, the immune system defends our body against external or internal aggressions. Sometimes, the immune system is not able to fight because it lacks or does not function properly causing an ineffective or absent immune response. Although more than 400 diseases exist, there is no data available in our country.\u0000 We analyze through a retrospective study carried out in the pediatric department at Hospital University Center, in Abidjan, between February and April 2022, involving 93 PID suspect children, the epidemiological, clinical, paraclinical and therapeutic aspects.\u0000 The average age of our patients was 2 years with extremes of 22 days to 15 years, with a male predominance: sex ratio of 1.45. The predominant geographical origin was Abidjan (85.32%). Consanguinity was found in 12.9%, death in siblings was found in 3.49% of patients. The most common clinical manifestations were repeated respiratory infections (60.71%), fever (16%), severe infections at least twice a year (13.04%), persistent fungal infections (12.90), thrive retardation in 48.7%. Abnormalities such as hyperleukocytosis (42.4%), neutropenia (29.23%), lymphopenia (16.44%), thrombocytopenia (38.36%) and mostly anemia (62.36%) were noted. Phagocytosis deficiency combined immune deficiencies and humoral deficiencies were respectively found at 95.16%, 89.78%.79.03%.\u0000 This work draws attention to the importance of PID which are often underdiagnosed, with absent or delayed treatment and poor prognosis. It is important to build a cohort with confirmed data to better support children with PID in our country.\u0000 no support","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74161630","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.60.02
A. Bouteau, S. Zurawski, G. Zurawski, B. Igyártó
Langerhans cells (LCs) can support the induction of antibody responses against foreign antigens without a danger signal. These findings contradict the danger model proposed to define how the adaptive immune responses are mounted. Deciphering how LCs, and some other dendritic cell subsets, support adaptive immune responses in steady-state could lead to a better understanding of how immune responses are induced and regulated and, ultimately, to developing more efficient immunotherapeutics for autoimmune- and infectious diseases. Here we used a well-established steady-state antigen targeting model to dissect the mechanism by which LCs support T follicular helper (Tfh) cells and antibody responses. Using bone marrow chimeras, Cre-lox system, and blocking antibodies, we found that IL-6, which is critical in the induction of Tfh cells and antibody responses in inflammatory conditions, played no role in LC-induced adaptive immune responses at steady-state. Type I interferon signaling was also dispensable in this regard. However, our preliminary data support that induction of humoral immune responses by steady-state LCs depends on membrane-bound co-stimulatory molecules, such as ICOS/ICOS-L. Thus, these data suggest that adaptive immune responses against foreign antigens in the absence of inflammation are generated through a distinct mechanism that likely does not involve inflammatory cytokines. Supported by grants from NIH R01AI146420 and Thomas Jefferson University startup funds.
{"title":"Steady-state Langerhans cells induce Tfh and B cell responses through a type I interferon and IL-6 independent mechanism","authors":"A. Bouteau, S. Zurawski, G. Zurawski, B. Igyártó","doi":"10.4049/jimmunol.210.supp.60.02","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.60.02","url":null,"abstract":"\u0000 Langerhans cells (LCs) can support the induction of antibody responses against foreign antigens without a danger signal. These findings contradict the danger model proposed to define how the adaptive immune responses are mounted. Deciphering how LCs, and some other dendritic cell subsets, support adaptive immune responses in steady-state could lead to a better understanding of how immune responses are induced and regulated and, ultimately, to developing more efficient immunotherapeutics for autoimmune- and infectious diseases. Here we used a well-established steady-state antigen targeting model to dissect the mechanism by which LCs support T follicular helper (Tfh) cells and antibody responses. Using bone marrow chimeras, Cre-lox system, and blocking antibodies, we found that IL-6, which is critical in the induction of Tfh cells and antibody responses in inflammatory conditions, played no role in LC-induced adaptive immune responses at steady-state. Type I interferon signaling was also dispensable in this regard. However, our preliminary data support that induction of humoral immune responses by steady-state LCs depends on membrane-bound co-stimulatory molecules, such as ICOS/ICOS-L. Thus, these data suggest that adaptive immune responses against foreign antigens in the absence of inflammation are generated through a distinct mechanism that likely does not involve inflammatory cytokines.\u0000 Supported by grants from NIH R01AI146420 and Thomas Jefferson University startup funds.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74309008","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.72.08
Alexi Zastrow, D. J. Friedman, S. Crotts, Matthew J. Rajcula, Brady Hammer, Mai Elissa, V. Shapiro
At the terminal position of many glycan chains are unique sugars known as sialic acids. Sialic acids bind sialic acid immunoglobin-like lectins (Siglecs). Our lab is interested in the role of CD22 (Siglec-2), which binds to α2,6-linked sialic acid generated by ST6GalI. CD22 can associate with itself or other molecules on the same cells in a “cis” configuration or bind in “trans” with ligands on other cells. CD22 contains four ITIMs within its cytoplasmic tail and is primarily known to function as an inhibitory co-receptor of the B cell receptor (BCR). CD22-ligand interactions restrain CD22 and BCR association which dampens BCR signaling. While CD22 is usually characterized as a B cell specific protein, we found that CD22 is highly expressed in hybrid macrophages (CD11b+ CD11c+ F4/80+) and expression decreases with activation, implying that CD22 may modulate myeloid cell responses. Using novel macrophage and dendritic cell-specific CD22 cKO mouse models, we will explore how CD22 impacts myeloid cell activation. We propose that CD22 impacts myeloid cell function and modulating this interaction may improve immune responses.
{"title":"Understanding the role of CD22 on Macrophage and Dendritic Cell Function","authors":"Alexi Zastrow, D. J. Friedman, S. Crotts, Matthew J. Rajcula, Brady Hammer, Mai Elissa, V. Shapiro","doi":"10.4049/jimmunol.210.supp.72.08","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.72.08","url":null,"abstract":"\u0000 At the terminal position of many glycan chains are unique sugars known as sialic acids. Sialic acids bind sialic acid immunoglobin-like lectins (Siglecs). Our lab is interested in the role of CD22 (Siglec-2), which binds to α2,6-linked sialic acid generated by ST6GalI. CD22 can associate with itself or other molecules on the same cells in a “cis” configuration or bind in “trans” with ligands on other cells. CD22 contains four ITIMs within its cytoplasmic tail and is primarily known to function as an inhibitory co-receptor of the B cell receptor (BCR). CD22-ligand interactions restrain CD22 and BCR association which dampens BCR signaling. While CD22 is usually characterized as a B cell specific protein, we found that CD22 is highly expressed in hybrid macrophages (CD11b+ CD11c+ F4/80+) and expression decreases with activation, implying that CD22 may modulate myeloid cell responses. Using novel macrophage and dendritic cell-specific CD22 cKO mouse models, we will explore how CD22 impacts myeloid cell activation. We propose that CD22 impacts myeloid cell function and modulating this interaction may improve immune responses.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"78 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75152834","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.250.19
Alex Jin Woong Kim, Hyunseok Kim, Kubra Akyildiz
Immune cells have a variety of functions to defend against pathogens under certain circumstances. It is not until recently, However, that these miscellaneous responses can be observed in vivo. Herein, we optimized some solutions of researchers keeping track of the changes of the immune response by utilizing the IVM-C intravital imaging system and in vivo fluorescence labeling for target cells. It is possible to successfully acquire a crystal-clear cellular level images of immune cells in the different animal model including RNA adjuvant-injected mouse and sepsis-induced one by using intravital microscope, respectively. In case of the RNA adjuvant injection, it is obvious that retention of the antigen presenting cells in draining lymph node is on the rise to promote T/B cell activation. A dramatic increase in the number of the infiltrated dendritic cells into the organ was observed at the same time. Moreover, Pathogenesis of sepsis by monitoring of pulmonary capillary circulation was investigated. In order to visualize pulmonary microcirculation, we’ve established a pulmonary imaging window which makes it feasible to image the real-time dynamics of rapidly circulating neutrophils in live mouse. We found out that many clusters of neutrophils aggregates in the pulmonary vasculature or, even in the arteriole of the sepsis-induced mouse, thereby disturbing the microcirculation of that region and generating dead space in the pulmonary structure. Taking everything into account, we make certain it is appropriate to utilize the intravital imaging method for cellular level visualization of the immune system under various pathological conditions. we believe that it could be an invaluable tool for a comprehensive understanding of immunology.
{"title":"In vivo imaging system for cellular level visualization of immune responses in various animal models","authors":"Alex Jin Woong Kim, Hyunseok Kim, Kubra Akyildiz","doi":"10.4049/jimmunol.210.supp.250.19","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.250.19","url":null,"abstract":"\u0000 Immune cells have a variety of functions to defend against pathogens under certain circumstances. It is not until recently, However, that these miscellaneous responses can be observed in vivo. Herein, we optimized some solutions of researchers keeping track of the changes of the immune response by utilizing the IVM-C intravital imaging system and in vivo fluorescence labeling for target cells. It is possible to successfully acquire a crystal-clear cellular level images of immune cells in the different animal model including RNA adjuvant-injected mouse and sepsis-induced one by using intravital microscope, respectively. In case of the RNA adjuvant injection, it is obvious that retention of the antigen presenting cells in draining lymph node is on the rise to promote T/B cell activation. A dramatic increase in the number of the infiltrated dendritic cells into the organ was observed at the same time. Moreover, Pathogenesis of sepsis by monitoring of pulmonary capillary circulation was investigated. In order to visualize pulmonary microcirculation, we’ve established a pulmonary imaging window which makes it feasible to image the real-time dynamics of rapidly circulating neutrophils in live mouse. We found out that many clusters of neutrophils aggregates in the pulmonary vasculature or, even in the arteriole of the sepsis-induced mouse, thereby disturbing the microcirculation of that region and generating dead space in the pulmonary structure. Taking everything into account, we make certain it is appropriate to utilize the intravital imaging method for cellular level visualization of the immune system under various pathological conditions. we believe that it could be an invaluable tool for a comprehensive understanding of immunology.","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"40 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75164254","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.164.09
Manikandan Palrasu, Khadija Kakar, Ahmed K. Aladhami, Tayler Carter, K. Wilson, Y. Zhong, X. Yang, Narendra P. Singh, P. Busbee, P. Nagarkatti, M. Nagarkatti
Indole-3-carbinol (I3C) is a dietary compound that acts as a ligand for the aryl hydrocarbon receptor (AhR), an important sensor for environmental polyaromatic chemicals and a regulator of the immune response. The pathogenesis of colitis involves multiple interactions between microbial dysbiosis, dysregulation of the host immune response, environmental factors, and host genetic profile. Indeed, studies have shown defective expression of AhR and antimicrobial peptides (AMPs) particularly defensins in inflammatory bowel disease (IBD) patients. Here, we investigated how I3C influences the production of AMPs, particularly b-defensins, through AhR and colonic microbiota composition in Anti-CD40 or dextran sulfate sodium (DSS)-induced colitis. Our analysis found that I3C attenuates colitis through AhR activation leading to increased expression of murine b-defensins (mBDs) by Colonic Epithelial Cells, resulting in the restoration of healthy gut microbiota and prevention of colonic inflammation. We analyzed the colonic expression of AMPs and mucins. Our analysis found that I3C significantly increased the mRNA expression of AhR, AMPs such as mBDs, Reg4, and mucins, when compared to vehicle-treated mice with colitis. Similarly, the mRNA levels of AhR, AMPs, and mucins were significantly increased in DSS+I3C-treated colon adenocarcinoma cells MC-38 (murine) and Caco2 (human) when compared to DSS-treated cells. To conclude, our findings demonstrate that I3C inhibits colitis primarily through AhR-mediated induction of AMPs and protective mucins, resulting in the enrichment of healthy gut microbiota (This work was supported in part by NIH grants R01ES030144, P01AT003961, P20GM103641, and R01AI123947, R01AI160896). This work was supported in part by NIH grants R01ES030144, P01AT003961, P20GM103641, and R01AI123947, R01AI160896
{"title":"Indol-3-Carbinol ameliorates colitis in mice by AhR-mediated production of antimicrobial peptides and mucins","authors":"Manikandan Palrasu, Khadija Kakar, Ahmed K. Aladhami, Tayler Carter, K. Wilson, Y. Zhong, X. Yang, Narendra P. Singh, P. Busbee, P. Nagarkatti, M. Nagarkatti","doi":"10.4049/jimmunol.210.supp.164.09","DOIUrl":"https://doi.org/10.4049/jimmunol.210.supp.164.09","url":null,"abstract":"\u0000 Indole-3-carbinol (I3C) is a dietary compound that acts as a ligand for the aryl hydrocarbon receptor (AhR), an important sensor for environmental polyaromatic chemicals and a regulator of the immune response. The pathogenesis of colitis involves multiple interactions between microbial dysbiosis, dysregulation of the host immune response, environmental factors, and host genetic profile. Indeed, studies have shown defective expression of AhR and antimicrobial peptides (AMPs) particularly defensins in inflammatory bowel disease (IBD) patients. Here, we investigated how I3C influences the production of AMPs, particularly b-defensins, through AhR and colonic microbiota composition in Anti-CD40 or dextran sulfate sodium (DSS)-induced colitis. Our analysis found that I3C attenuates colitis through AhR activation leading to increased expression of murine b-defensins (mBDs) by Colonic Epithelial Cells, resulting in the restoration of healthy gut microbiota and prevention of colonic inflammation. We analyzed the colonic expression of AMPs and mucins. Our analysis found that I3C significantly increased the mRNA expression of AhR, AMPs such as mBDs, Reg4, and mucins, when compared to vehicle-treated mice with colitis. Similarly, the mRNA levels of AhR, AMPs, and mucins were significantly increased in DSS+I3C-treated colon adenocarcinoma cells MC-38 (murine) and Caco2 (human) when compared to DSS-treated cells. To conclude, our findings demonstrate that I3C inhibits colitis primarily through AhR-mediated induction of AMPs and protective mucins, resulting in the enrichment of healthy gut microbiota (This work was supported in part by NIH grants R01ES030144, P01AT003961, P20GM103641, and R01AI123947, R01AI160896).\u0000 This work was supported in part by NIH grants R01ES030144, P01AT003961, P20GM103641, and R01AI123947, R01AI160896","PeriodicalId":22698,"journal":{"name":"The Journal of Immunology","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75148665","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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