DOCSLIB.ORG

Implications for Anti-CD200 Therapy Absence of Antibody Effector Function

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Implications for Anti-CD200 Therapy Absence of Antibody Effector Function Blockade of CD200 in the Presence or Absence of Antibody Effector Function: Implications for Anti-CD200 Therapy This information is current as Anke Kretz-Rommel, Fenghua Qin, Naveen Dakappagari, of September 29, 2021. Roxanne Cofiell, Susan J. Faas and Katherine S. Bowdish J Immunol 2008; 180:699-705; ; doi: 10.4049/jimmunol.180.2.699 http://www.jimmunol.org/content/180/2/699 Downloaded from References This article cites 37 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/180/2/699.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 29, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Blockade of CD200 in the Presence or Absence of Antibody Effector Function: Implications for Anti-CD200 Therapy Anke Kretz-Rommel,* Fenghua Qin,* Naveen Dakappagari,* Roxanne Cofiell,† Susan J. Faas,† and Katherine S. Bowdish1* CD200 is an immunosuppressive molecule overexpressed in multiple hematologic malignancies such as B cell chronic lymphocytic leukemia, multiple myeloma, and acute myeloid leukemia. We previously demonstrated that up-regulation of CD200 on tumor cells suppresses antitumor immune responses and that antagonistic anti-human CD200 mAbs enabled human PBMC-mediated tumor growth inhibition in xenograft NOD/SCID human (hu)-mouse models. Ab variants with effector function (IgG1 constant region (G1)) or without effector function (IgG2/G4 fusion constant region (G2G4)) exhibited high antitumor activity in a human tumor xenograft model in which CD200 was expressed. In this report, we seek to select the best candidate to move forward into the clinic and begin to decipher the mechanisms of tumor cell killing by comparing anti-CD200-G1 vs anti-CD200-G2G4 in two Downloaded from related animal models. In a CD200-expressing xenograft NOD/SCID hu-mouse model where CD200 ligand/receptor interactions are already established before initiating treatment, we find that anti-CD200-G1 is a less effective Ab compared with anti-CD200- G2G4. Separately, in a model that evaluates the effect of the Abs on the immune cell component of the xenograft NOD/SCID hu-mouse model distinctly from the effects of binding to CD200 on tumor cells, we find that the administration of anti-CD200-G1 Abs completely abolished human PBMC-mediated tumor growth inhibition. Along with supporting in vitro studies, our data indicate that anti-CD200-G1 Abs efficiently mediate Ab-dependent cellular cytotoxicity of activated T cells, critical cells involved http://www.jimmunol.org/ in immune-mediated killing. These studies suggest important implications regarding the selection of the constant region in anti- CD200 immunotherapy of cancer patients. The Journal of Immunology, 2008, 180: 699–705. he immunosuppressive molecule CD200 (1, 2) is up-reg- (9), alemtuzumab (anti-CD52) in chronic B cell leukemia (10), ulated on B cells in all patients with B cell chronic lym- trastuzumab (anti-human epidermal growth factor receptor 2) in T phocytic leukemia (3), and the up-regulation of CD200 breast cancer (11), and bevacizumab (anti-vascular endothelial correlates with a negative prognosis in multiple myeloma (4) and growth factor) (12) and cetuximab (anti-epidermal growth factor acute myeloid leukemia (5). CD200 is a type 1a membrane protein receptor) (13) in colorectal cancer. Although these Abs have by guest on September 29, 2021 related to the B7 family of costimulatory receptors with two ex- shown clinical effectiveness for many years, the critical in vivo tracellular Ig superfamily domains, a single transmembrane region, mechanisms of action are still in question (14, 15) but most likely and a short cytoplasmic tail with no known signaling motifs (6). It include direct effects on target cells via signaling mechanisms and is normally expressed on thymocytes, T and B lymphocytes, some by blocking growth factors, as well as mediating effector function dendritic cells, neurons, kidney glomeruli, syncytiotrophoblasts, through the constant region (C region) of the Ab. and endothelial cells (7). We previously demonstrated that the Because CD200 does not have a signaling domain it appears that presence of human CD200 on Namalwa tumor cells (Burkitt’s anti-CD200 Abs do not directly affect targeted tumor cells, and we 2 lymphoma cell line) coinjected with human PBMCs (hPBMCs) demonstrated that the anti-CD200 mAbs, as expected, did not in- into NOD/SCID mice prevents hPBMCs from eradicating the can- hibit tumor cell proliferation or directly induce cell death (8). cer cells. Administration of antagonistic anti-CD200 Abs blocked Therefore, antagonistic anti-CD200 Abs are expected to exert their tumor growth in this model (8), suggesting possible use in cancer effect by blocking immune suppression and, depending on the therapy. C region of the Ab, by potentially mediating Ab-dependent A number of mAbs have been successfully used in cancer ther- cellular cytotoxicity (ADCC) or complement-dependent cyto- apy such as rituximab (anti-CD20) in non-Hodgkin’s lymphoma toxicity (CDC). Our previous studies demonstrated that in an im- mediate treatment model where the first dose of Ab was injected at the same time as the tumor cells and effector hPBMCs, the effects *Alexion Antibody Technologies, San Diego, CA 92121; and †Alexion Pharmaceu- ticals, Cheshire, CT 06410 of anti-CD200-IgG1 constant region (G1) compared with those of Received for publication July 26, 2007. Accepted for publication October 27, 2007. anti-CD200-IgG2/G4 fusion constant region (G2G4) were nearly indistinguishable and confirm that a blockade of CD200 immune The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance suppression by an effectorless anti-CD200 mAb is sufficient to with 18 U.S.C. Section 1734 solely to indicate this fact. achieve high antitumor activity. 1 Address correspondence and reprint requests to Dr. Katherine S. Bowdish, Alexion To gain more insight into the mechanism of action of our anti- Antibody Technologies, 3985 Sorrento Valley Boulevard, Suite A, San Diego, CA 92121. E-mail address: [email protected] CD200 Abs in the xenograft NOD/SCID human (hu)-mouse model and to provide more information in support of one Ab C region 2 Abbreviations used in this paper: hPBMC, human PBMC; ADCC, Ab-dependent cellular cytotoxicity; CDC, complement dependent cytotoxicity; C region, constant over the other in the context of anti-CD200 therapy in the clinic, region; G1, IgG1 C region; G2G4, IgG2/G4 fusion C region; hu, human; mOKT3, we evaluated both anti-CD200-G1 and anti-CD200-G2G4 Abs in mouse OKT3. two additional tumor models, a delayed treatment model and a Copyright © 2008 by The American Association of Immunologists, Inc. 0022-1767/08/$2.00 model where the tumor xenograft lacks CD200 expression. These www.jimmunol.org 700 ANTI-CD200-G1 ANTIBODY MEDIATES KILLING OF ACTIVATED T CELLS studies, along with supporting in vitro data, indicate that mecha- Isolation of human CD3ϩ T cells nisms of cell killing provide clues that may be critical to the suc- hPBMCs were obtained from normal healthy donors as described above cess of CD200 blockade in cancer. We demonstrate that a blockade and passaged over a human T cell enrichment column (HTCC-5; R&D of CD200 in the presence or absence of effector function has Systems) according to the manufacturer’s instructions to obtain purified ϩ implications for cancer therapy and support the use of anti- CD3 T cells. Eluted cells were washed, counted, and resuspended in CD200-G2G4 in the clinic. complete RPMI 1640 medium (Mediatech) containing 5% heat-inactivated single donor serum, 2 mM L-glutamine (Mediatech), 10 mM HEPES (Mediatech) and 1% penicillin/streptomycin (Mediatech). Materials and Methods Mice PHA activation of hPBMCs hPBMCs from healthy human donors were cultured in 6-well plates (Fal- Four- to six-week-old NOD.CB17-Prkdcscid/J (NOD/SCID) mice were con) at a concentration of 2 ϫ 106 cells/ml in complete medium in the obtained from The Jackson Laboratory. Animals were housed at Perry absence or presence of 5–10 ␮g/ml PHA (Sigma-Aldrich). Cells were Scientific. All procedures and protocols were approved by the Perry Sci- maintained for 72 h at 37°C in a humidified incubator containing 5% CO . entific Institutional Animal Care and Use Committee. 2 At the end of the culture period, the cells were either chromated for use as Anti-CD200 mAbs targets in ADCC or for flow cytometric analysis. ϩ Engineering of chimeric and humanized anti-CD200 mAbs with either an Activation of CD3 T cells with plate-bound mouse IgG1 (G1) or an IgG2/G4 fusion C region (G2G4) have been described OKT3 (mOKT3) previously (8) and designated chB7-G1 and chB7-G2G4 to denote chi- meric anti-CD200 Abs derived from anti-CD200 Fab B7 or hB7V2-G1 and The wells of 12-well plates were coated by overnight incubation at 4°C ␮ Downloaded from hB7V2-G2G4 to denote humanized variants derived from anti-CD200 Fab with 10 g/ml mOKT3 (Ortho Pharmaceuticals) diluted in PBS. Residual Ab was removed and the plates were gently rinsed with PBS.
Load more

Recommended publications
  • CD226 T Cells Expressing the Receptors TIGIT and Divergent Phenotypes of Human Regulatory
    The Journal of Immunology Divergent Phenotypes of Human Regulatory T Cells Expressing the Receptors TIGIT and CD226 Christopher A. Fuhrman,*,1 Wen-I Yeh,*,1 Howard R. Seay,* Priya Saikumar Lakshmi,* Gaurav Chopra,† Lin Zhang,* Daniel J. Perry,* Stephanie A. McClymont,† Mahesh Yadav,† Maria-Cecilia Lopez,‡ Henry V. Baker,‡ Ying Zhang,x Yizheng Li,{ Maryann Whitley,{ David von Schack,x Mark A. Atkinson,* Jeffrey A. Bluestone,‡ and Todd M. Brusko* Regulatory T cells (Tregs) play a central role in counteracting inflammation and autoimmunity. A more complete understanding of cellular heterogeneity and the potential for lineage plasticity in human Treg subsets may identify markers of disease pathogenesis and facilitate the development of optimized cellular therapeutics. To better elucidate human Treg subsets, we conducted direct transcriptional profiling of CD4+FOXP3+Helios+ thymic-derived Tregs and CD4+FOXP3+Helios2 T cells, followed by comparison with CD4+FOXP32Helios2 T conventional cells. These analyses revealed that the coinhibitory receptor T cell Ig and ITIM domain (TIGIT) was highly expressed on thymic-derived Tregs. TIGIT and the costimulatory factor CD226 bind the common ligand CD155. Thus, we analyzed the cellular distribution and suppressive activity of isolated subsets of CD4+CD25+CD127lo/2 T cells expressing CD226 and/or TIGIT. We observed TIGIT is highly expressed and upregulated on Tregs after activation and in vitro expansion, and is associated with lineage stability and suppressive capacity. Conversely, the CD226+TIGIT2 population was associated with reduced Treg purity and suppressive capacity after expansion, along with a marked increase in IL-10 and effector cytokine production. These studies provide additional markers to delineate functionally distinct Treg subsets that may help direct cellular therapies and provide important phenotypic markers for assessing the role of Tregs in health and disease.
    [Show full text]
  • Associations Between FCGR3A Polymorphisms and Susceptibility to Rheumatoid Arthritis: a Metaanalysis YOUNG HO LEE, JONG DAE JI, and GWAN GYU SONG
    Associations Between FCGR3A Polymorphisms and Susceptibility to Rheumatoid Arthritis: A Metaanalysis YOUNG HO LEE, JONG DAE JI, and GWAN GYU SONG ABSTRACT. Objective. To investigate whether the Fcγ receptor (FCGR) polymorphism confers susceptibility to rheumatoid arthritis (RA). Methods. We conducted metaanalyses on the associations between FCGR polymorphisms and RA susceptibility as determined using (1) allele contrast, (2) recessive models, (3) dominant models, and (4) contrast of homozygotes, using fixed or random effects models. Results. A total of 10 separate comparisons were considered, which comprised 6 European and 4 Asian population samples. Metaanalysis of FCGR3A polymorphism revealed a significant associa- tion between the VV genotype and the risk of developing RA relative to the VF+FF genotype (OR 1.256, 95% CI 1.045–1.510, p = 0.015), with no evidence of between-study heterogeneity (p = 0.167). In subjects of European descent, a stronger association was observed between the VV geno- type and RA than for the FF genotype (OR 1.374, 95% CI 1.101–1.714, p = 0.005). In Asians, no such association was found. Metaanalysis of the VV vs FF genotype revealed a significantly increased OR in Europeans (OR 1.399, 95% CI 1.107–1.769, p = 0.005), but not in Asians. No asso- ciation was found between RA and the FCGR2A and FCGR3B polymorphisms in all subjects and in European and Asian populations, except for the NA22 vs NA11 of FCGR3B in Europeans. Conclusion. No relation was found between the FCGR2A polymorphism and susceptibility to RA in Europeans or Asians. The FCGR3A polymorphism was found to be associated with RA in Europeans but not in Asians.
    [Show full text]
  • Single-Cell RNA Sequencing Demonstrates the Molecular and Cellular Reprogramming of Metastatic Lung Adenocarcinoma
    ARTICLE https://doi.org/10.1038/s41467-020-16164-1 OPEN Single-cell RNA sequencing demonstrates the molecular and cellular reprogramming of metastatic lung adenocarcinoma Nayoung Kim 1,2,3,13, Hong Kwan Kim4,13, Kyungjong Lee 5,13, Yourae Hong 1,6, Jong Ho Cho4, Jung Won Choi7, Jung-Il Lee7, Yeon-Lim Suh8,BoMiKu9, Hye Hyeon Eum 1,2,3, Soyean Choi 1, Yoon-La Choi6,10,11, Je-Gun Joung1, Woong-Yang Park 1,2,6, Hyun Ae Jung12, Jong-Mu Sun12, Se-Hoon Lee12, ✉ ✉ Jin Seok Ahn12, Keunchil Park12, Myung-Ju Ahn 12 & Hae-Ock Lee 1,2,3,6 1234567890():,; Advanced metastatic cancer poses utmost clinical challenges and may present molecular and cellular features distinct from an early-stage cancer. Herein, we present single-cell tran- scriptome profiling of metastatic lung adenocarcinoma, the most prevalent histological lung cancer type diagnosed at stage IV in over 40% of all cases. From 208,506 cells populating the normal tissues or early to metastatic stage cancer in 44 patients, we identify a cancer cell subtype deviating from the normal differentiation trajectory and dominating the metastatic stage. In all stages, the stromal and immune cell dynamics reveal ontological and functional changes that create a pro-tumoral and immunosuppressive microenvironment. Normal resident myeloid cell populations are gradually replaced with monocyte-derived macrophages and dendritic cells, along with T-cell exhaustion. This extensive single-cell analysis enhances our understanding of molecular and cellular dynamics in metastatic lung cancer and reveals potential diagnostic and therapeutic targets in cancer-microenvironment interactions. 1 Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
    [Show full text]
  • Associations Between FCGR3A Polymorphisms and Susceptibility to Rheumatoid Arthritis: a Metaanalysis YOUNG HO LEE, JONG DAE JI, and GWAN GYU SONG
    Associations Between FCGR3A Polymorphisms and Susceptibility to Rheumatoid Arthritis: A Metaanalysis YOUNG HO LEE, JONG DAE JI, and GWAN GYU SONG ABSTRACT. Objective. To investigate whether the Fcγ receptor (FCGR) polymorphism confers susceptibility to rheumatoid arthritis (RA). Methods. We conducted metaanalyses on the associations between FCGR polymorphisms and RA susceptibility as determined using (1) allele contrast, (2) recessive models, (3) dominant models, and (4) contrast of homozygotes, using fixed or random effects models. Results. A total of 10 separate comparisons were considered, which comprised 6 European and 4 Asian population samples. Metaanalysis of FCGR3A polymorphism revealed a significant associa- tion between the VV genotype and the risk of developing RA relative to the VF+FF genotype (OR 1.256, 95% CI 1.045–1.510, p = 0.015), with no evidence of between-study heterogeneity (p = 0.167). In subjects of European descent, a stronger association was observed between the VV geno- type and RA than for the FF genotype (OR 1.374, 95% CI 1.101–1.714, p = 0.005). In Asians, no such association was found. Metaanalysis of the VV vs FF genotype revealed a significantly increased OR in Europeans (OR 1.399, 95% CI 1.107–1.769, p = 0.005), but not in Asians. No asso- ciation was found between RA and the FCGR2A and FCGR3B polymorphisms in all subjects and in European and Asian populations, except for the NA22 vs NA11 of FCGR3B in Europeans. Conclusion. No relation was found between the FCGR2A polymorphism and susceptibility to RA in Europeans or Asians. The FCGR3A polymorphism was found to be associated with RA in Europeans but not in Asians.
    [Show full text]
  • Single-Cell RNA Sequencing of Tocilizumab-Treated Peripheral Blood Mononuclear Cells As an in Vitro Model of Inflammation
    UCSF UC San Francisco Previously Published Works Title Single-Cell RNA Sequencing of Tocilizumab-Treated Peripheral Blood Mononuclear Cells as an in vitro Model of Inflammation. Permalink https://escholarship.org/uc/item/8q84n3jb Authors Zarinsefat, Arya Hartoularos, George Rychkov, Dmitry et al. Publication Date 2020 DOI 10.3389/fgene.2020.610682 Peer reviewed eScholarship.org Powered by the California Digital Library University of California fgene-11-610682 December 19, 2020 Time: 19:44 # 1 BRIEF RESEARCH REPORT published: 05 January 2021 doi: 10.3389/fgene.2020.610682 Single-Cell RNA Sequencing of Tocilizumab-Treated Peripheral Blood Mononuclear Cells as an in vitro Model of Inflammation Arya Zarinsefat1*, George Hartoularos2, Dmitry Rychkov1, Priyanka Rashmi1, Sindhu Chandran3, Flavio Vincenti1, Chun J. Yee2 and Minnie M. Sarwal1 1 Department of Surgery, University of California, San Francisco, San Francisco, CA, United States, 2 Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, United States, 3 Department of Medicine, University of California, San Francisco, San Francisco, CA, United States COVID-19 has posed a significant threat to global health. Early data has revealed that IL-6, a key regulatory cytokine, plays an important role in the cytokine storm of COVID-19. Multiple trials are therefore looking at the effects of Tocilizumab, an IL-6 receptor antibody that inhibits IL-6 activity, on treatment of COVID-19, with promising findings. As part of a clinical trial looking at the effects of Tocilizumab treatment on Edited by: kidney transplant recipients with subclinical rejection, we performed single-cell RNA Gabriele Bucci, sequencing of comparing stimulated PBMCs before and after Tocilizumab treatment.
    [Show full text]
  • CD29 Identifies IFN-Γ–Producing Human CD8+ T Cells With
    + CD29 identifies IFN-γ–producing human CD8 T cells with an increased cytotoxic potential Benoît P. Nicoleta,b, Aurélie Guislaina,b, Floris P. J. van Alphenc, Raquel Gomez-Eerlandd, Ton N. M. Schumacherd, Maartje van den Biggelaarc,e, and Monika C. Wolkersa,b,1 aDepartment of Hematopoiesis, Sanquin Research, 1066 CX Amsterdam, The Netherlands; bLandsteiner Laboratory, Oncode Institute, Amsterdam University Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands; cDepartment of Research Facilities, Sanquin Research, 1066 CX Amsterdam, The Netherlands; dDivision of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands; and eDepartment of Molecular and Cellular Haemostasis, Sanquin Research, 1066 CX Amsterdam, The Netherlands Edited by Anjana Rao, La Jolla Institute for Allergy and Immunology, La Jolla, CA, and approved February 12, 2020 (received for review August 12, 2019) Cytotoxic CD8+ T cells can effectively kill target cells by producing therefore developed a protocol that allowed for efficient iso- cytokines, chemokines, and granzymes. Expression of these effector lation of RNA and protein from fluorescence-activated cell molecules is however highly divergent, and tools that identify and sorting (FACS)-sorted fixed T cells after intracellular cytokine + preselect CD8 T cells with a cytotoxic expression profile are lacking. staining. With this top-down approach, we performed an un- + Human CD8 T cells can be divided into IFN-γ– and IL-2–producing biased RNA-sequencing (RNA-seq) and mass spectrometry cells. Unbiased transcriptomics and proteomics analysis on cytokine- γ– – + + (MS) analyses on IFN- and IL-2 producing primary human producing fixed CD8 T cells revealed that IL-2 cells produce helper + + + CD8 Tcells.
    [Show full text]
  • FCGR Polymorphisms Influence Response to IL2 in Metastatic Renal Cell Carcinoma
    Published OnlineFirst October 14, 2016; DOI: 10.1158/1078-0432.CCR-16-1874 Cancer Therapy: Clinical Clinical Cancer Research FCGR Polymorphisms Influence Response to IL2 in Metastatic Renal Cell Carcinoma Amy K. Erbe1,WeiWang1, Jacob Goldberg1, Mikayla Gallenberger1, KyungMann Kim2, Lakeesha Carmichael2,DustinHess1,EneidaA.Mendonca2,3, Yiqiang Song2, Jacquelyn A. Hank1,Su-ChunCheng4, Sabina Signoretti5, Michael Atkins6,7, Alexander Carlson8,JamesW.Mier6,8, David J. Panka8, David F. McDermott6,8,and Paul M. Sondel1,3 Abstract Purpose: Fc-gamma receptors (FCGRs) are expressed on Results: When higher-affinity genotypes for FCGR2A, FCGR3A, immune cells, bind to antibodies, and trigger antibody-induced and FCGR2C were considered together, they were associated with cell-mediated antitumor responses when tumor-reactive antibo- significantly increased tumor shrinkage and prolonged survival in dies are present. The affinity of the FCGR/antibody interaction is response to HD-IL2. variable and dependent upon FCGR polymorphisms. Prior stud- Conclusions: Although associations of higher-affinity FCGR ies of patients with cancer treated with immunotherapy indicate genotype with clinical outcome have been demonstrated that FCGR polymorphisms can influence antitumor response for with mAb therapy and with idiotype vaccines, to our knowl- certain immunotherapies that act via therapeutically administered edge,thisisthefirst study to show associations of FCGR mAbs or via endogenous tumor-reactive antibodies induced from genotypes with outcome following HD-IL2 treatment. We tumor antigen vaccines. The previously published "SELECT" trial hypothesize that endogenous antitumor antibodies may of high-dose aldesleukin (HD-IL2) for metastatic renal cell carci- engage immune cells through their FCGRs, and HD-IL2 may noma resulted in an objective response rate of 25%.
    [Show full text]
  • First Infusion Reactions Are Mediated by Fcγriiib and Neutrophils
    Pharm Res (2018) 35: 169 https://doi.org/10.1007/s11095-018-2448-8 RESEARCH PAPER First Infusion Reactions are Mediated by FcγRIIIb and Neutrophils Felix Weber 1 & Daniel Breustedt 1,2 & Sonja Schlicht 3 & Claas A. Meyer 3 & Jens Niewoehner 4 & Martin Ebeling 1 & Per-Ola Freskgard5 & Peter Bruenker6 & Thomas Singer1 & Michael Reth7 & Antonio Iglesias1 Received: 29 March 2018 /Accepted: 15 June 2018 /Published online: 27 June 2018 # The Author(s) 2018 ABSTRACT FIR. FcγRIIIb-mediated FIR was abolished by depleting neu- Purpose Administration of therapeutic monoclonal antibod- trophils or blocking FcγRIIIb with CD11b antibodies. ies (mAbs) is frequently accompanied by severe first infusion Conclusions Human FcγRIIIb and neutrophils are primarily reactions (FIR). The mechanism driving FIR is still unclear. responsible for triggering FIR. Clinical strategies to prevent This study aimed to investigate the cellular and molecular FIR in patients should focus on this pathway and may include mechanisms causing FIR in humanized mouse models and transient depletion of neutrophils or blocking FcγRIIIb with their potential for evaluating FIR risk in patients. specific mAbs. Methods Mice humanized for Fc gamma receptors (FcγRs) were generated by recombination-mediated genomic replace- KEY WORDS human FcγRIIIb . humanized mouse model . ment. Body temperature, cytokine release and reactive oxygen immunotoxicology . infusion reactions . neutrophils species (ROS) were measured to assess FIR to mAbs. Results Infusion of human mAb specific for mouse transferrin receptor (HamTfR) into FcγR-humanized mice, produced marked transient hypothermia accompanied by an increase ABBREVIATIONS in inflammatory cytokines KC and MIP-2, and ROS. FIR ADA Anti-drug antibodies were dependent on administration route and Fc-triggered ef- ADCC Antibody-dependent cellular cytotoxicity fector functions mediated by neutrophils.
    [Show full text]
  • Identi Cation of Key Survival Genes of the Tumor Microenvironment And
    Identication of key survival genes of the tumor microenvironment and immune inltration in head and neck squamous cell carcinoma Min Wang Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Tao Lu Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Yanshi Li Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Min Pan Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Zhihai Wang Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Guohua Hu ( [email protected] ) Department of Otorhinolaryngology, The First Aliated Hospital of Chongqing Medical University Research Article Keywords: Head and neck cancer, Tumor microenvironment, Immune scores, Bioinformatics analysis, Survival Posted Date: March 13th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-283947/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/23 Abstract Background: Head and neck cancer (HNC) are highly aggressive solid tumors with poor prognoses. The tumor microenvironment (TME) plays a critical role in angiogenesis, invasion, and metastasis of HNC. In the TME, immune and stromal cells inuence tumor initiation, response, and therapy. Our study aimed to evaluate the progression and prognosis of HNC by analyzing the key genes involved in immunization and stromal cells. Methods: Gene expression proles, demographics, and survival data were downloaded from the TCGA database. Patients with HNC were divided into high immune/stromal score groupss or low immune/stromal score groups based on the ESTIMATE algorithm. Differentially expressed genes (DEGs) were identied via functional enrichment analysis and protein-protein interaction networks, and survival analysis based on DEGs was also performed.
    [Show full text]
  • High-Resolution Melting Analysis As an Alternative Method for Human Neutrophil Antigen Genotyping
    O RIGINAL R EP O RT High-resolution melting analysis as an alternative method for human neutrophil antigen genotyping K. Yasui, M. Tanaka, T. Hayashi, N. Matsuyama, A. Kuroishi, R.A. Furuta, Y. Tani, and F. Hirayama Human neutrophil antigen (HNA)-typed granulocyte panels are and Japanese (88%) populations.19,20 Polymorphism of this widely used to screen for the presence of HNA antibodies and to antigen has not been reported. Antibodies against this antigen determine antibody specificity. Many laboratories screen donors are associated with ANN, autoimmune neutropenia, febrile for HNA genotypes using low-throughput methods such as 12,21–23 allele-specific polymerase chain reaction (PCR), PCR–restriction transfusion reactions, and TRALI. HNA-3, comprising fragment–length polymorphism, and multiplex PCR. In the HNA-3a and HNA-3b, is expressed on granulocytes, present study, we used a high-resolution melting (HRM) analysis lymphocytes, platelets, endothelial cells, kidney, spleen, and to determine HNA genotypes. For the HRM analysis, purified 24 genomic DNA samples were amplified via PCR with HNA-specific placental cells. Alloantibodies to HNA-3a are associated with primers. Nucleotide substitutions in genes encoding HNAs were occasional cases of febrile transfusion reactions,25 ANN,26 and differentiated on the basis of the HRM curves, and the results of HRM serious cases of TRALI.14,27,28 The HNA-4 and HNA-5 antigens and DNA sequencing analyses were determined to be in complete reside on the subunits of the β-2 integrin family (CD11a and agreement. The gene frequency of HNA-1a, -1b, -1c, -3a, -3b, -4a, -4b, -5a, and -5b in the Japanese population was consistent CD11b, respectively).
    [Show full text]
  • ID AKI Vs Control Fold Change P Value Symbol Entrez Gene Name *In
    ID AKI vs control P value Symbol Entrez Gene Name *In case of multiple probesets per gene, one with the highest fold change was selected. Fold Change 208083_s_at 7.88 0.000932 ITGB6 integrin, beta 6 202376_at 6.12 0.000518 SERPINA3 serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 3 1553575_at 5.62 0.0033 MT-ND6 NADH dehydrogenase, subunit 6 (complex I) 212768_s_at 5.50 0.000896 OLFM4 olfactomedin 4 206157_at 5.26 0.00177 PTX3 pentraxin 3, long 212531_at 4.26 0.00405 LCN2 lipocalin 2 215646_s_at 4.13 0.00408 VCAN versican 202018_s_at 4.12 0.0318 LTF lactotransferrin 203021_at 4.05 0.0129 SLPI secretory leukocyte peptidase inhibitor 222486_s_at 4.03 0.000329 ADAMTS1 ADAM metallopeptidase with thrombospondin type 1 motif, 1 1552439_s_at 3.82 0.000714 MEGF11 multiple EGF-like-domains 11 210602_s_at 3.74 0.000408 CDH6 cadherin 6, type 2, K-cadherin (fetal kidney) 229947_at 3.62 0.00843 PI15 peptidase inhibitor 15 204006_s_at 3.39 0.00241 FCGR3A Fc fragment of IgG, low affinity IIIa, receptor (CD16a) 202238_s_at 3.29 0.00492 NNMT nicotinamide N-methyltransferase 202917_s_at 3.20 0.00369 S100A8 S100 calcium binding protein A8 215223_s_at 3.17 0.000516 SOD2 superoxide dismutase 2, mitochondrial 204627_s_at 3.04 0.00619 ITGB3 integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61) 223217_s_at 2.99 0.00397 NFKBIZ nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, zeta 231067_s_at 2.97 0.00681 AKAP12 A kinase (PRKA) anchor protein 12 224917_at 2.94 0.00256 VMP1/ mir-21likely ortholog
    [Show full text]
  • Current Understanding of an Emerging Role of HLA-DRB1 Gene in Rheumatoid Arthritis–From Research to Clinical Practice
    cells Review Current Understanding of an Emerging Role of HLA-DRB1 Gene in Rheumatoid Arthritis–From Research to Clinical Practice Tomasz Wysocki 1,*, Marzena Olesi ´nska 1 and Agnieszka Paradowska-Gorycka 2 1 Department of Systemic Connective Tissue Diseases, National Institute of Geriatrics, Rheumatology and Rehabilitation, Sparta´nska1, 02-637 Warsaw, Poland; [email protected] or [email protected] 2 Department of Molecular Biology, National Institute of Geriatrics, Rheumatology and Rehabilitation, Sparta´nska1, 02-637 Warsaw, Poland; [email protected] or [email protected] * Correspondence: [email protected] Received: 27 March 2020; Accepted: 30 April 2020; Published: 2 May 2020 Abstract: Rheumatoid arthritis (RA) is an autoimmune disease with an unclear pathogenic mechanism. However, it has been proven that the key underlying risk factor is a genetic predisposition. Association studies of the HLA-DRB1 gene clearly indicate its importance in RA morbidity. This review presents the current state of knowledge on the impact of HLA-DRB1 gene, functioning both as a component of the patient’s genome and as an environmental risk factor. The impact of known HLA-DRB1 risk variants on the specific structure of the polymorphic HLA-DR molecule, and epitope binding affinity, is presented. The issues of the potential influence of HLA-DRB1 on the occurrence of non-articular disease manifestations and response to treatment are also discussed. A deeper understanding of the role of the HLA-DRB1 gene is essential to explore the complex nature of RA, which is a result of multiple contributing factors, including genetic, epigenetic and environmental factors.
    [Show full text]