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Article ; Online: A machine learning approach toward automating spatial identification of LAG3+/CD3+ cells in ulcerative colitis.

Bonnevie, Edward D / Dobrzynski, Eric / Steiner, Dylan / Hildebrand, Deon / Monslow, James / Singh, Mohan / Decman, Vilma / Krull, David L

Scientific reports

2023 Volume 13, Issue 1, Page(s) 21759

Abstract: Over the past decade, automation of digital image analysis has become commonplace in both research and clinical settings. Spurred by recent advances in artificial intelligence and machine learning (AI/ML), tissue sub-compartments and cellular phenotypes ... ...

Abstract	Over the past decade, automation of digital image analysis has become commonplace in both research and clinical settings. Spurred by recent advances in artificial intelligence and machine learning (AI/ML), tissue sub-compartments and cellular phenotypes within those compartments can be identified with higher throughput and accuracy than ever before. Recently, immune checkpoints have emerged as potential targets for auto-immune diseases. As such, spatial identification of these proteins along with immune cell markers (e.g., CD3
MeSH term(s)	Humans ; Artificial Intelligence ; Colitis, Ulcerative ; Algorithms ; Fluorescent Antibody Technique ; Machine Learning ; Biomarkers
Chemical Substances	Biomarkers
Language	English
Publishing date	2023-12-08
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-49163-5
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: A machine learning approach toward automating spatial identification of LAG3+/CD3+ cells in ulcerative colitis

Edward D. Bonnevie / Eric Dobrzynski / Dylan Steiner / Deon Hildebrand / James Monslow / Mohan Singh / Vilma Decman / David L. Krull

Scientific Reports, Vol 13, Iss 1, Pp 1-

2023 Volume 12

Abstract: Abstract Over the past decade, automation of digital image analysis has become commonplace in both research and clinical settings. Spurred by recent advances in artificial intelligence and machine learning (AI/ML), tissue sub-compartments and cellular ... ...

Abstract	Abstract Over the past decade, automation of digital image analysis has become commonplace in both research and clinical settings. Spurred by recent advances in artificial intelligence and machine learning (AI/ML), tissue sub-compartments and cellular phenotypes within those compartments can be identified with higher throughput and accuracy than ever before. Recently, immune checkpoints have emerged as potential targets for auto-immune diseases. As such, spatial identification of these proteins along with immune cell markers (e.g., CD3+/LAG3+ T-cells) is a crucial step in understanding the potential and/or efficacy of such treatments. Here, we describe a semi-automated imaging and analysis pipeline that identifies CD3+/LAG3+ cells in colorectal tissue sub-compartments. While chromogenic staining has been a clinical mainstay and the resulting brightfield images have been utilized in AI/ML approaches in the past, there are associated drawbacks in phenotyping algorithms that can be overcome by fluorescence imaging. To address these tradeoffs, we developed an analysis pipeline combining the strengths of brightfield and fluorescence images. In this assay, immunofluorescence imaging was conducted to identify phenotypes followed by coverslip removal and hematoxylin and eosin staining of the same section to inform an AI/ML tissue segmentation algorithm. This assay proved to be robust in both tissue segmentation and phenotyping, was compatible with automated workflows, and revealed presence of LAG3+ T-cells in ulcerative colitis biopsies with spatial context preserved.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2023-12-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Modified skin-window technique in cynomolgus macaques (Macaca fasicularis) for assessing neutrophil extravasation.

Maguire, Sean / Dobrzynski, Eric / Alpaugh, R Katherine

Journal of the American Association for Laboratory Animal Science : JAALAS

2010 Volume 49, Issue 4, Page(s) 475–479

Abstract: Alterations in neutrophil extravasation are seen in disease states and in response to therapeutics. To investigate neutrophil extravasation during the acute inflammatory response, a skin-window technique used in humans was adapted for use in cynomolgus ... ...

Abstract	Alterations in neutrophil extravasation are seen in disease states and in response to therapeutics. To investigate neutrophil extravasation during the acute inflammatory response, a skin-window technique used in humans was adapted for use in cynomolgus macaques (Macaca fasicularis). Modulation of neutrophil extravasation was attempted with systemic methotrexate and local application of the anaphylatoxin recombinant C5a (rC5a). On day 1, skin windows were created in 4 ketamine-anesthetized monkeys on both forearms by mildly abrading the skin and then overlaying the abrasions with filter paper either saturated in saline or rC5a for 6 h. At 2.5 h prior to generation of new skin windows on day 2, the monkeys received 4.5 mg methotrexate IM, and skin windows and treatment with saline or rC5a were repeated on new forearm sites on each monkey. All papers were analyzed for albumin, neutrophil number, and the neutrophil chemoattractant IL8. Day 1 albumin levels did not differ between groups, indicating consistent abrasion. Methotrexate given prior to the day 2 abrasions reduced neutrophil extravasation and IL8 levels compared with those on day 1. rC5a partially abrogated the methotrexate-induced reduction in neutrophil extravasation and IL8 production. The skin-window technique was well tolerated by the monkeys and successfully accommodated measurement of changes in neutrophil extravasation in response to inflammatory modulators.
MeSH term(s)	Animals ; CD11a Antigen/metabolism ; Macaca fascicularis ; Methotrexate/pharmacology ; Neutrophils/drug effects ; Neutrophils/metabolism ; Neutrophils/pathology ; Skin Window Technique/adverse effects ; Skin Window Technique/methods ; Skin Window Technique/veterinary
Chemical Substances	CD11a Antigen ; Methotrexate (YL5FZ2Y5U1)
Language	English
Publishing date	2010-09-06
Publishing country	United States
Document type	Journal Article
ISSN	1559-6109
ISSN	1559-6109
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Article: Tolerance induction by viral in vivo gene transfer.

Dobrzynski, Eric / Herzog, Roland W

Clinical medicine & research

2005 Volume 3, Issue 4, Page(s) 234–240

Abstract: Treatment of genetic disease by protein or gene replacement therapy is hampered by immune responses to the therapeutic protein. An excellent example is formation of inhibitory antibodies to coagulation factors in treatment of the X-linked bleeding ... ...

Abstract	Treatment of genetic disease by protein or gene replacement therapy is hampered by immune responses to the therapeutic protein. An excellent example is formation of inhibitory antibodies to coagulation factors in treatment of the X-linked bleeding disorder hemophilia. Experiments in murine and canine models of hemophilia B (deficiency in factor IX) have demonstrated sustained therapeutic levels of factor IX transgene expression following hepatic adeno-associated viral gene transfer in animals with deletion and nonsense mutations in the factor IX gene. This article reviews experimental evidence for induction of immune tolerance to the factor IX transgene product by hepatic adeno-associated viral gene transfer, which has been shown to limit T helper cell responses and to substantially reduce the risk of antibody responses. Tolerance induction is associated with activation of regulatory CD4(+) T cells capable of suppressing antibody formation to factor IX protein. Hepatic administration of adeno-associated viral vector expressing ovalbumin in mice transgenic for a T cell receptor specific for this antigen provided direct evidence for induction of CD4(+) T cell tolerance, including T cell anergy and clonal deletion. Taken together, these data indicate the potential for viral in vivo gene transfer not only to provide sustained systemic expression, but moreover to induce immunological hypo-responsiveness to the therapeutic gene product.
MeSH term(s)	Animals ; Dependovirus/metabolism ; Dogs ; Factor IX/metabolism ; Gene Deletion ; Gene Transfer Techniques ; Genes, Viral ; Genetic Therapy/methods ; Hemophilia A/genetics ; Hemophilia A/metabolism ; Hemophilia B/genetics ; Hemophilia B/metabolism ; Immune System ; Immune Tolerance ; Mice ; Mice, Transgenic ; T-Lymphocytes/metabolism
Chemical Substances	Factor IX (9001-28-9)
Language	English
Publishing date	2005-09-08
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	2303793-3
ISSN	1539-4182
ISSN	1539-4182
DOI	10.3121/cmr.3.4.234
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Article: Immune implications of gene therapy for hemophilia.

Herzog, Roland W / Dobrzynski, Eric

Seminars in thrombosis and hemostasis

2004 Volume 30, Issue 2, Page(s) 215–226

Abstract: Similar to any novel treatment strategy for hemophilia, gene therapy faces the question of the risk of formation of inhibitory antibodies to the therapeutic factor VIII or factor IX protein. Activation of CD4 (+) or CD8 (+) T cells could lead to antibody ...

Abstract	Similar to any novel treatment strategy for hemophilia, gene therapy faces the question of the risk of formation of inhibitory antibodies to the therapeutic factor VIII or factor IX protein. Activation of CD4 (+) or CD8 (+) T cells could lead to antibody formation or cytotoxic T lymphocyte responses to transgene-expressing cells. Preclinical studies in animal models of hemophilia A and B with different mutations in the dysfunctional gene shed light on the risk for such immune responses and point toward strategies to avoid immune activation or even promote tolerance induction. The impacts of variables such as choice and design of gene transfer vector, underlying gene mutation, route of vector administration, and transient immune suppression are discussed. Maintenance of immunological hyporesponsiveness to the therapeutic gene product is critical for successful gene therapy. Recent studies provide evidence for tolerance induction to coagulation factor antigens by viral hepatic or neonatal in vivo gene transfer, by in utero gene delivery, and by oral or nasal administration of protein or peptides.
MeSH term(s)	Animals ; Factor IX/administration & dosage ; Factor IX/immunology ; Factor VIII/administration & dosage ; Factor VIII/immunology ; Female ; Genetic Therapy/methods ; Hemophilia A/immunology ; Hemophilia A/therapy ; Humans ; Immune Tolerance ; In Vitro Techniques ; Pregnancy
Chemical Substances	Factor VIII (9001-27-8) ; Factor IX (9001-28-9)
Language	English
Publishing date	2004-04
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	196901-8
ISSN	1098-9064 ; 0094-6176
ISSN (online)	1098-9064
ISSN	0094-6176
DOI	10.1055/s-2004-825635
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Article ; Online: Belantamab Mafodotin (GSK2857916) Drives Immunogenic Cell Death and Immune-mediated Antitumor Responses

Montes de Oca, Rocio / Alavi, Alireza S / Vitali, Nick / Bhattacharya, Sabyasachi / Blackwell, Christina / Patel, Krupa / Seestaller-Wehr, Laura / Kaczynski, Heather / Shi, Hong / Dobrzynski, Eric / Obert, Leslie / Tsvetkov, Lyuben / Cooper, David C / Jackson, Heather / Bojczuk, Paul / Forveille, Sabrina / Kepp, Oliver / Sauvat, Allan / Kroemer, Guido /

Creighton-Gutteridge, Mark / Yang, Jingsong / Hopson, Chris / Yanamandra, Niranjan / Shelton, Christopher / Mayes, Patrick / Opalinska, Joanna / Barnette, Mary / Srinivasan, Roopa / Smothers, James / Hoos, Axel

Molecular cancer therapeutics

2021 Volume 20, Issue 10, Page(s) 1941–1955

Abstract: B-cell maturation antigen (BCMA) is an attractive therapeutic target highly expressed on differentiated plasma cells in multiple myeloma and other B-cell malignancies. GSK2857916 (belantamab mafodotin, BLENREP) is a BCMA-targeting antibody-drug conjugate ...

Abstract	B-cell maturation antigen (BCMA) is an attractive therapeutic target highly expressed on differentiated plasma cells in multiple myeloma and other B-cell malignancies. GSK2857916 (belantamab mafodotin, BLENREP) is a BCMA-targeting antibody-drug conjugate approved for the treatment of relapsed/refractory multiple myeloma. We report that GSK2857916 induces immunogenic cell death in BCMA-expressing cancer cells and promotes dendritic cell activation
MeSH term(s)	Animals ; Antibodies, Monoclonal/chemistry ; Antibodies, Monoclonal, Humanized/pharmacology ; Apoptosis ; B-Cell Maturation Antigen/antagonists & inhibitors ; B-Cell Maturation Antigen/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cell Proliferation ; Female ; Humans ; Immunoconjugates/pharmacology ; Immunogenic Cell Death ; Lymphoma/drug therapy ; Lymphoma/immunology ; Lymphoma/metabolism ; Lymphoma/pathology ; Mice ; Mice, Inbred C57BL ; Multiple Myeloma/drug therapy ; Multiple Myeloma/immunology ; Multiple Myeloma/metabolism ; Multiple Myeloma/pathology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays
Chemical Substances	Antibodies, Monoclonal ; Antibodies, Monoclonal, Humanized ; B-Cell Maturation Antigen ; Immunoconjugates ; TNFRSF17 protein, human ; belantamab mafodotin (DB1041CXDG)
Language	English
Publishing date	2021-07-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2063563-1
ISSN	1538-8514 ; 1535-7163
ISSN (online)	1538-8514
ISSN	1535-7163
DOI	10.1158/1535-7163.MCT-21-0035
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Zs.A 5750: Show issues

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Article: Modified skin-window technique in cynomolgus macaques (Macaca fasicularis) for assessing neutrophil extravasation

Maguire, Sean / Dobrzynski, Eric / Alpaugh, R. Katherine

Journal of the American Association for Laboratory Animal Science : JAALAS. 2010 July, v. 49, no. 4

2010

Keywords	human diseases ; animal diseases ; inflammation ; blood ; immune system ; neutrophils ; animal models ; monkeys ; Macaca fascicularis ; acute course ; pathogenesis ; diagnostic techniques ; surgery ; new methods ; methotrexate ; intramuscular injection ; immunomodulators ; immune response ; validity
Language	English
Dates of publication	2010-07
Size	p. 475-479.
Document type	Article
ISSN	1559-6109
Database	NAL-Catalogue (AGRICOLA)

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Article: Immune Implications of Gene Therapy for Hemophilia

Herzog, Roland W / Dobrzynski, Eric

Seminars in Thrombosis and Hemostasis

2004 Volume 30, Issue 02, Page(s) 215–226

Keywords	Immune response ; inhibitor ; T cell ; B cell ; cytotoxic T lymphocyte
Language	English
Publishing date	2004-01-01
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	196901-8
ISSN	1098-9064 ; 0094-6176
ISSN (online)	1098-9064
ISSN	0094-6176
DOI	10.1055/s-2004-825635
Database	Thieme publisher's database

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Article: Prevention of cytotoxic T lymphocyte responses to factor IX-expressing hepatocytes by gene transfer-induced regulatory T cells.

Dobrzynski, Eric / Fitzgerald, Julie C / Cao, Ou / Mingozzi, Federico / Wang, Lixin / Herzog, Roland W

Proceedings of the National Academy of Sciences of the United States of America

2006 Volume 103, Issue 12, Page(s) 4592–4597

Abstract: Treatment of genetic disease such as the bleeding disorder hemophilia B [deficiency in blood coagulation factor IX (F.IX)] by gene replacement therapy is hampered by the risk of immune responses to the therapeutic gene product and to the gene transfer ... ...

Abstract	Treatment of genetic disease such as the bleeding disorder hemophilia B [deficiency in blood coagulation factor IX (F.IX)] by gene replacement therapy is hampered by the risk of immune responses to the therapeutic gene product and to the gene transfer vector. Immune competent mice of two different strains were tolerized to human F.IX by hepatic gene transfer mediated by adenoassociated viral vector. These animals were subsequently challenged by systemic administration of an E1/E3-deleted adenoviral vector, which is known to induce a cytotoxic T lymphocyte response to the transgene product. Immune tolerance prevented cytotoxic T lymphocyte activation to F.IX and CD8(+) cellular infiltrates in the liver. Moreover, a sustained and substantial increase in hepatic F.IX expression from the adenoviral vector was achieved despite in vitro T cell responses to adenoviral antigens. Cytolytic responses to therapeutic and to viral vector-derived antigens had been prevented in vivo by activation of regulatory CD4(+) T cells, which mediated suppression of inflammatory lymphocyte responses to the liver. This result suggests that augmentation of regulatory T cell activation should provide new means to avoid destructive immune responses in gene transfer.
MeSH term(s)	Adenoviridae/genetics ; Adoptive Transfer ; Animals ; Cytotoxicity, Immunologic/genetics ; Factor IX/genetics ; Factor IX/immunology ; Factor IX/metabolism ; Gene Transfer Techniques ; Genetic Therapy/methods ; Genetic Vectors/genetics ; Hepatocytes/immunology ; Hepatocytes/metabolism ; Immune Tolerance ; Immunosuppression Therapy/methods ; Male ; Mice ; Mice, Inbred Strains ; T-Lymphocytes, Cytotoxic/immunology ; T-Lymphocytes, Regulatory/immunology
Chemical Substances	Factor IX (9001-28-9)
Language	English
Publishing date	2006-03-10
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.0508685103
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Article: Systemic protein delivery by muscle-gene transfer is limited by a local immune response.

Wang, Lixin / Dobrzynski, Eric / Schlachterman, Alexander / Cao, Ou / Herzog, Roland W

Blood

2005 Volume 105, Issue 11, Page(s) 4226–4234

Abstract: Adeno-associated viral (AAV) vectors have been successfully used for therapeutic expression of systemic transgene products (such as factor IX or erythropoietin) following in vivo administration to skeletal muscle of animal models of inherited hematologic ...

Abstract	Adeno-associated viral (AAV) vectors have been successfully used for therapeutic expression of systemic transgene products (such as factor IX or erythropoietin) following in vivo administration to skeletal muscle of animal models of inherited hematologic disorders. However, an immune response may be initiated if the transgene product represents a neoantigen. Here, we use ovalbumin (OVA) as a model antigen and demonstrate immune-mediated elimination of expression on muscle-directed AAV-2 gene transfer. Administration to immune competent mice resulted in transient systemic OVA expression. Within 10 days, OVA-specific T-helper cells had been activated in draining lymph nodes, an inflammatory immune response ensued, and OVA-expressing muscle fibers were destroyed by a cytotoxic CD8(+) T-cell response. Use of a muscle-specific promoter did not prevent this immune response. Adoptively transferred CD4(+) cells transgenic for a T-cell receptor specific to OVA peptide-major histocompatibility complex class II showed antigen-specific, vector dose-dependent proliferation confined to the draining lymph nodes of AAV-OVA-transduced muscle within 5 days after gene transfer and subsequently participated in lymphocytic infiltration of transduced muscle. This study documents that a local immune response limits sustained expression of a secreted protein in muscle gene transfer, a finding that may have consequences for design of clinical protocols.
MeSH term(s)	Animals ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cytotoxicity, Immunologic ; Dependovirus/genetics ; Gene Transfer Techniques/adverse effects ; Genetic Therapy/adverse effects ; Genetic Therapy/methods ; Immunity, Cellular ; Inflammation/immunology ; Lymph Nodes/immunology ; Mice ; Mice, Inbred Strains ; Mice, Knockout ; Models, Animal ; Muscle, Skeletal/immunology ; Ovalbumin/administration & dosage ; Ovalbumin/immunology ; Proteins/administration & dosage ; Proteins/immunology ; Time Factors
Chemical Substances	Proteins ; Ovalbumin (9006-59-1)
Language	English
Publishing date	2005-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	80069-7
ISSN	1528-0020 ; 0006-4971
ISSN (online)	1528-0020
ISSN	0006-4971
DOI	10.1182/blood-2004-03-0848
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