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Article ; Online: Editorial: Innate Cells in the Pathogenesis of Food Allergy.

Olivera, Ana / Laky, Karen / Hogan, Simon Patrick / Frischmeyer-Guerrerio, Pamela

2021 Volume 12, Page(s) 709991

MeSH term(s)	Antigen-Presenting Cells/immunology ; Basophils/physiology ; Food Hypersensitivity/etiology ; Food Hypersensitivity/immunology ; Gastrointestinal Microbiome ; Humans ; Immunity, Innate ; Mast Cells/physiology ; T-Lymphocytes, Regulatory/immunology ; Th2 Cells/immunology
Language	English
Publishing date	2021-06-10
Publishing country	Switzerland
Document type	Editorial ; Introductory Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2021.709991
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Article ; Online: In Vivo Depletion of T Lymphocytes.

Laky, Karen / Kruisbeek, Ada M

Current protocols in immunology

2016 Volume 113, Page(s) 4.1.1–4.1.9

Abstract: In vivo depletion of T lymphocytes is a means of studying the role of specific T cell populations during defined phases of in vivo immune responses. In this unit, a protocol is provided for injecting monoclonal antibodies (mAbs) into wild-type adult mice. ...

Abstract	In vivo depletion of T lymphocytes is a means of studying the role of specific T cell populations during defined phases of in vivo immune responses. In this unit, a protocol is provided for injecting monoclonal antibodies (mAbs) into wild-type adult mice. Depletion of the appropriate subset of cells is verified by flow cytometry analysis of lymph node and spleen cell suspensions in pilot experiments. Once conditions have been established, depleted mice can be used to study the impact of T cell subsets on a variety of in vivo immune responses. The depleted condition may be maintained by repeated injections of the monoclonal antibody, or reversed by normal thymopoiesis following discontinuation of antibody administration.
MeSH term(s)	Animals ; Antibodies, Monoclonal/immunology ; Biomarkers ; Flow Cytometry ; Immunophenotyping ; Lymphocyte Depletion ; Mice ; Phenotype ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism
Chemical Substances	Antibodies, Monoclonal ; Biomarkers
Language	English
Publishing date	2016-04-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ISSN	1934-368X
ISSN (online)	1934-368X
DOI	10.1002/0471142735.im0401s113
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Article ; Online: TGFβ prevents IgE-mediated allergic disease by restraining T follicular helper 2 differentiation.

Haque, Tamara T / Weissler, Katherine A / Schmiechen, Zoe / Laky, Karen / Schwartz, Daniella M / Li, Jenny / Locci, Michela / Turfkruyer, Mathilde / Yao, Chen / Schaughency, Paul / Leak, Lashawna / Lack, Justin / Kanno, Yuka / O'Shea, John / Frischmeyer-Guerrerio, Pamela A

Science immunology

2024 Volume 9, Issue 91, Page(s) eadg8691

Abstract: Allergic diseases are common, affecting more than 20% of the population. Genetic variants in the TGFβ pathway are strongly associated with atopy. To interrogate the mechanisms underlying this association, we examined patients and mice with Loeys-Dietz ... ...

Abstract	Allergic diseases are common, affecting more than 20% of the population. Genetic variants in the TGFβ pathway are strongly associated with atopy. To interrogate the mechanisms underlying this association, we examined patients and mice with Loeys-Dietz syndrome (LDS) who harbor missense mutations in the kinase domain of
MeSH term(s)	Animals ; Humans ; Mice ; Hypersensitivity/metabolism ; Immunoglobulin E ; Th2 Cells ; TOR Serine-Threonine Kinases ; Transforming Growth Factor beta
Chemical Substances	Immunoglobulin E (37341-29-0) ; TOR Serine-Threonine Kinases (EC 2.7.11.1) ; Transforming Growth Factor beta
Language	English
Publishing date	2024-01-19
Publishing country	United States
Document type	Journal Article
ISSN	2470-9468
ISSN (online)	2470-9468
DOI	10.1126/sciimmunol.adg8691
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Article ; Online: Dendritic cells Trigger IFN-γ secretion by NK cells independent of IL-12 and IL-18.

Abdi, Kaveh / Laky, Karen / Abshari, Mehrnoosh / Hill, Elizabeth M / Lantz, Larry / Singh, Nevil J / Long, Eric O

European journal of immunology

2022 Volume 52, Issue 9, Page(s) 1431–1440

Abstract: It is commonly believed that IL-12 produced by DCs in response to pathogens is the first signal that stimulates the production of IFN-γ by NK cells. However, IL-12 production by DCs in response to bacterial LPS depends on either engagement of CD40 by ... ...

Abstract	It is commonly believed that IL-12 produced by DCs in response to pathogens is the first signal that stimulates the production of IFN-γ by NK cells. However, IL-12 production by DCs in response to bacterial LPS depends on either engagement of CD40 by CD40L on activated T cells or IFN-γ from NK cells. This suggests that during the primary immune response, NK cells produce IFN-γ before IL-12 production by DCs. Here, using single-cell measurements, cell sorting and mouse lines deficient in IL-12, IL-23, type I IFN receptor and the IL-18 receptor, we show that a subset of BM-derived DCs characterized by low expression of MHC class II (MHCII
MeSH term(s)	Animals ; Cells, Cultured ; Dendritic Cells ; Interferon-gamma/metabolism ; Interleukin-12/metabolism ; Interleukin-18/metabolism ; Interleukin-23/metabolism ; Lipopolysaccharides/pharmacology ; Mice
Chemical Substances	Interleukin-18 ; Interleukin-23 ; Lipopolysaccharides ; Interleukin-12 (187348-17-0) ; Interferon-gamma (82115-62-6)
Language	English
Publishing date	2022-07-28
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202149733
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Article ; Online: Hematopoietic reconstitution of neonatal immunocompetent mice to study conditions with a perinatal window of susceptibility.

Laky, Karen / Dugan, Philip / Frischmeyer-Guerrerio, Pamela A

Scientific reports

2018 Volume 8, Issue 1, Page(s) 12254

Abstract: Efficient hematopoietic reconstitution of wild type mice requires preconditioning. Established experimental protocols exist to transplant hematopoietic stem cells into lethally irradiated or chemically myeloablated adult mice or unirradiated ... ...

Abstract	Efficient hematopoietic reconstitution of wild type mice requires preconditioning. Established experimental protocols exist to transplant hematopoietic stem cells into lethally irradiated or chemically myeloablated adult mice or unirradiated immunodeficient mice. We sought to develop a protocol to reconstitute immuno-replete neonatal mice. We describe irradiation and injection procedures for two-day old mice that lead to efficient long-term reconstitution of primary and secondary lymphoid organs. We demonstrate that the frequencies of lymphoid and myeloid cells in primary and secondary lymphoid organs are indistinguishable from unirradiated uninjected sex- and age-matched control animals by 5 weeks post-reconstitution. Thus, this system will facilitate studies aimed at understanding the developmental and environmental mechanisms that contribute to conditions that have a window of susceptibility during the perinatal period.
MeSH term(s)	Animals ; Animals, Newborn ; Cell Proliferation ; Hematopoietic Stem Cell Transplantation/methods ; Hematopoietic Stem Cells/cytology ; Immunocompetence ; Mice ; Time Factors ; Whole-Body Irradiation
Language	English
Publishing date	2018-08-16
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-018-30767-1
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Article ; Online: Bone Marrow-Derived Dendritic Cell Cultures from RAG

Abdi, Kaveh / Thomas, L Michael / Laky, Karen / Abshari, Mehrnoosh / Matzinger, Polly / Long, Eric O

ImmunoHorizons

2020 Volume 4, Issue 7, Page(s) 415–419

Abstract: Dendritic cells (DCs) play a key role in the initiation of an immune response and are known as "professional" APCs because of their ability to activate naive T cells. A widely used method to generate DCs in vitro is to culture bone marrow (BM) cells or ... ...

Abstract	Dendritic cells (DCs) play a key role in the initiation of an immune response and are known as "professional" APCs because of their ability to activate naive T cells. A widely used method to generate DCs in vitro is to culture bone marrow (BM) cells or blood monocytes in the presence of GM-CSF and IL-4. In this study, we show that a small population of NK cells residing in the BM of RAG
MeSH term(s)	Animals ; Bone Marrow Cells/cytology ; Bone Marrow Cells/drug effects ; Bone Marrow Cells/immunology ; Cell Differentiation/drug effects ; Dendritic Cells/cytology ; Dendritic Cells/drug effects ; Dendritic Cells/immunology ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Interferon-gamma/metabolism ; Interleukin-4/pharmacology ; Killer Cells, Natural/immunology ; Lipopolysaccharides/toxicity ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes/immunology ; T-Lymphocytes/cytology ; T-Lymphocytes/drug effects ; T-Lymphocytes/immunology
Chemical Substances	Homeodomain Proteins ; Lipopolysaccharides ; RAG-1 protein (128559-51-3) ; Interleukin-4 (207137-56-2) ; Interferon-gamma (82115-62-6) ; Granulocyte-Macrophage Colony-Stimulating Factor (83869-56-1)
Language	English
Publishing date	2020-07-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ISSN	2573-7732
ISSN (online)	2573-7732
DOI	10.4049/immunohorizons.2000011
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Article ; Online: Hematopoietic reconstitution of neonatal immunocompetent mice to study conditions with a perinatal window of susceptibility

Karen Laky / Philip Dugan / Pamela A. Frischmeyer-Guerrerio

Scientific Reports, Vol 8, Iss 1, Pp 1-

2018 Volume 8

Abstract: Abstract Efficient hematopoietic reconstitution of wild type mice requires preconditioning. Established experimental protocols exist to transplant hematopoietic stem cells into lethally irradiated or chemically myeloablated adult mice or unirradiated ... ...

Abstract	Abstract Efficient hematopoietic reconstitution of wild type mice requires preconditioning. Established experimental protocols exist to transplant hematopoietic stem cells into lethally irradiated or chemically myeloablated adult mice or unirradiated immunodeficient mice. We sought to develop a protocol to reconstitute immuno-replete neonatal mice. We describe irradiation and injection procedures for two-day old mice that lead to efficient long-term reconstitution of primary and secondary lymphoid organs. We demonstrate that the frequencies of lymphoid and myeloid cells in primary and secondary lymphoid organs are indistinguishable from unirradiated uninjected sex- and age-matched control animals by 5 weeks post-reconstitution. Thus, this system will facilitate studies aimed at understanding the developmental and environmental mechanisms that contribute to conditions that have a window of susceptibility during the perinatal period.
Keywords	Medicine ; R ; Science ; Q
Subject code	572
Language	English
Publishing date	2018-08-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
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Article ; Online: Epithelial-intrinsic defects in TGFβR signaling drive local allergic inflammation manifesting as eosinophilic esophagitis.

Laky, Karen / Kinard, Jessica L / Li, Jenny Min / Moore, Ian N / Lack, Justin / Fischer, Elizabeth R / Kabat, Juraj / Latanich, Rachel / Zachos, Nicholas C / Limkar, Ajinkya R / Weissler, Katherine A / Thompson, Robert W / Wynn, Thomas A / Dietz, Harry C / Guerrerio, Anthony L / Frischmeyer-Guerrerio, Pamela A

Science immunology

2023 Volume 8, Issue 79, Page(s) eabp9940

Abstract: Allergic diseases are a global health challenge. Individuals harboring loss-of-function variants in transforming growth factor-β receptor (TGFβR) genes have an increased prevalence of allergic disorders, including eosinophilic esophagitis. Allergic ... ...

Abstract	Allergic diseases are a global health challenge. Individuals harboring loss-of-function variants in transforming growth factor-β receptor (TGFβR) genes have an increased prevalence of allergic disorders, including eosinophilic esophagitis. Allergic diseases typically localize to mucosal barriers, implicating epithelial dysfunction as a cardinal feature of allergic disease. Here, we describe an essential role for TGFβ in the control of tissue-specific immune homeostasis that provides mechanistic insight into these clinical associations. Mice expressing a TGFβR1 loss-of-function variant identified in atopic patients spontaneously develop disease that clinically, immunologically, histologically, and transcriptionally recapitulates eosinophilic esophagitis. In vivo and in vitro, TGFβR1 variant-expressing epithelial cells are hyperproliferative, fail to differentiate properly, and overexpress innate proinflammatory mediators, which persist in the absence of lymphocytes or external allergens. Together, our results support the concept that TGFβ plays a fundamental, nonredundant, epithelial cell-intrinsic role in controlling tissue-specific allergic inflammation that is independent of its role in adaptive immunity.
MeSH term(s)	Animals ; Mice ; Eosinophilic Esophagitis/genetics ; Receptors, Transforming Growth Factor beta/genetics ; Inflammation ; Hypersensitivity, Immediate
Chemical Substances	Receptors, Transforming Growth Factor beta
Language	English
Publishing date	2023-01-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Intramural
ISSN	2470-9468
ISSN (online)	2470-9468
DOI	10.1126/sciimmunol.abp9940
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Article ; Online: A randomized double-blinded trial to assess recurrence of systemic allergic reactions following COVID-19 mRNA vaccination.

Khalid, Muhammad B / Zektser, Ellen / Chu, Eric / Li, Min / Utoh, Joanna / Ryan, Patrick / Loving, Hanna S / Harb, Roa / Kattappuram, Robbie / Chatman, Lindsay / Hartono, Stella / Claudio-Etienne, Estefania / Sun, Guangping / Feener, Edward P / Li, Zhongbo / Lai, Samuel K / Le, Quang / Schwartz, Lawrence B / Lyons, Jonathan J /

Komarow, Hirsh / Zhou, Zhao-Hua / Raza, Haniya / Pao, Maryland / Laky, Karen / Holland, Steven M / Brittain, Erica / Frischmeyer-Guerrerio, Pamela A

The Journal of allergy and clinical immunology

2024

Abstract: Background: Systemic allergic reactions (sARs) following coronavirus disease 2019 (COVID-19) mRNA vaccines were initially reported at a higher rate than after traditional vaccines.: Objective: We aimed to evaluate the safety of revaccination in these ...

Abstract	Background: Systemic allergic reactions (sARs) following coronavirus disease 2019 (COVID-19) mRNA vaccines were initially reported at a higher rate than after traditional vaccines. Objective: We aimed to evaluate the safety of revaccination in these individuals and to interrogate mechanisms underlying these reactions. Methods: In this randomized, double-blinded, phase 2 trial, participants aged 16 to 69 years who previously reported a convincing sAR to their first dose of COVID-19 mRNA vaccine were randomly assigned to receive a second dose of BNT162b2 (Comirnaty) vaccine and placebo on consecutive days in a blinded, 1:1 crossover fashion at the National Institutes of Health. An open-label BNT162b2 booster was offered 5 months later if the second dose did not result in severe sAR. None of the participants received the mRNA-1273 (Spikevax) vaccine during the study. The primary end point was recurrence of sAR following second dose and booster vaccination; exploratory end points included biomarker measurements. Results: Of 111 screened participants, 18 were randomly assigned to receive study interventions. Eight received BNT162b2 second dose followed by placebo; 8 received placebo followed by BNT162b2 second dose; 2 withdrew before receiving any study intervention. All 16 participants received the booster dose. Following second dose and booster vaccination, sARs recurred in 2 participants (12.5%; 95% CI, 1.6 to 38.3). No sAR occurred after placebo. An anaphylaxis mimic, immunization stress-related response (ISRR), occurred more commonly than sARs following both vaccine and placebo and was associated with higher predose anxiety scores, paresthesias, and distinct vital sign and biomarker changes. Conclusions: Our findings support revaccination of individuals who report sARs to COVID-19 mRNA vaccines. Distinct clinical and laboratory features may distinguish sARs from ISRRs.
Language	English
Publishing date	2024-03-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	121011-7
ISSN	1097-6825 ; 1085-8725 ; 0091-6749
ISSN (online)	1097-6825 ; 1085-8725
ISSN	0091-6749
DOI	10.1016/j.jaci.2024.03.001
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Article ; Online: A genome catalog of the early-life human skin microbiome.

Genome biology

2023 Volume 24, Issue 1, Page(s) 252

Abstract: Background: Metagenome-assembled genomes have greatly expanded the reference genomes for skin microbiome. However, the current reference genomes are largely based on samples from adults in North America and lack representation from infants and ... ...

Abstract	Background: Metagenome-assembled genomes have greatly expanded the reference genomes for skin microbiome. However, the current reference genomes are largely based on samples from adults in North America and lack representation from infants and individuals from other continents. Results: Here we use deep shotgun metagenomic sequencing to profile the skin microbiota of 215 infants at age 2-3 months and 12 months who are part of the VITALITY trial in Australia as well as 67 maternally matched samples. Based on the infant samples, we present the Early-Life Skin Genomes (ELSG) catalog, comprising 9483 prokaryotic genomes from 1056 species, 206 fungal genomes from 13 species, and 39 eukaryotic viral sequences. This genome catalog substantially expands the diversity of species previously known to comprise human skin microbiome and improves the classification rate of sequenced data by 21%. The protein catalog derived from these genomes provides insights into the functional elements such as defense mechanisms that distinguish early-life skin microbiome. We also find evidence for microbial sharing at the community, bacterial species, and strain levels between mothers and infants. Conclusions: Overall, the ELSG catalog uncovers the skin microbiome of a previously underrepresented age group and population and provides a comprehensive view of human skin microbiome diversity, function, and development in early life.
MeSH term(s)	Humans ; Infant ; Microbiota/genetics ; Metagenome ; Bacteria/genetics ; Australia ; North America ; Metagenomics
Language	English
Publishing date	2023-11-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural
ZDB-ID	2040529-7
ISSN	1474-760X ; 1474-760X
ISSN (online)	1474-760X
ISSN	1474-760X
DOI	10.1186/s13059-023-03090-w
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