Publications by authors named "Denise Hinz"

13 Publications

  • Page 1 of 1

Molecular Signatures of Dengue Virus-Specific IL-10/IFN-γ Co-producing CD4 T Cells and Their Association with Dengue Disease.

Cell Rep 2019 12;29(13):4482-4495.e4

Division of Vaccine Discovery, La Jolla Institute for Immunology, La Jolla, CA 92037, USA.

Dengue virus (DENV) can cause diseases ranging from dengue fever (DF) to more severe dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). Whether antiviral T cells contribute to the protection against or pathogenesis of severe disease is not well defined. Here, we identified antigen-specific IL-10IFN-γ double-positive (DP) CD4 T cells during acute DENV infection. While the transcriptomic signatures of DP cells partially overlapped with those of cytotoxic and type 1 regulatory CD4 T cells, the majority of them were non-cytotoxic/Tr1 and included IL21, IL22, CD109, and CCR1. Although we observed a higher frequency of DP cells in DHF, the transcriptomic profile of DP cells was similar in DF and DHF, suggesting that DHF is not associated with the altered phenotypic or functional attributes of DP cells. Overall, this study revealed a DENV-specific DP cell subset in patients with acute dengue disease and argues against altered DP cells as a determinant of DHF.
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http://dx.doi.org/10.1016/j.celrep.2019.11.098DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6942518PMC
December 2019

A Modified Injector and Sample Acquisition Protocol Can Improve Data Quality and Reduce Inter-Instrument Variability of the Helios Mass Cytometer.

Cytometry A 2019 09 30;95(9):1019-1030. Epub 2019 Jul 30.

Human Immune Monitoring Center, Icahn School of Medicine at Mt. Sinai, New York, New York.

Mass cytometry is a powerful tool for high-dimensional single cell characterization. Since the introduction of the first commercial CyTOF mass cytometer by DVS Sciences in 2009, mass cytometry technology has matured and become more widely utilized, with sequential platform upgrades designed to address specific limitations and to expand the capabilities of the platform. Fluidigm's third-generation Helios mass cytometer introduced a number of upgrades over the previous CyTOF2. One of these new features is a modified narrow bore sample injector that generates smaller ion clouds, which is expected to improve sensitivity and throughput. However, following rigorous testing, we find that the narrow-bore sample injector may have unintended negative consequences on data quality and result in lower median and higher coefficients of variation in many antibody-associated signal intensities. We describe an alternative Helios acquisition protocol using a wider bore injector, which largely mitigates these data quality issues. We directly compare these two protocols in a multisite study of 10 Helios instruments across 7 institutions and show that the modified protocol improves data quality and reduces interinstrument variability. These findings highlight and address an important source of technical variability in mass cytometry experiments that is of particular relevance in the setting of multicenter studies. © 2019 International Society for Advancement of Cytometry.
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http://dx.doi.org/10.1002/cyto.a.23866DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6750971PMC
September 2019

Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells.

J Vis Exp 2019 06 19(148). Epub 2019 Jun 19.

Division of Inflammation Biology, La Jolla Institute for Immunology;

In this article, we present a protocol that is optimized to preserve neutrophil-lineage cells in fresh BM for whole BM CyTOF analysis. We utilized a myeloid-biased 39-antibody CyTOF panel to evaluate the hematopoietic system with a focus on the neutrophil-lineage cells by using this protocol. The CyTOF result was analyzed with an open-resource dimensional reduction algorithm, viSNE, and the data was presented to demonstrate the outcome of this protocol. We have discovered new neutrophil-lineage cell populations based on this protocol. This protocol of fresh whole BM preparation may be used for 1), CyTOF analysis to discover unidentified cell populations from whole BM, 2), investigating whole BM defects for patients with blood disorders such as leukemia, 3), assisting optimization of fluorescence-activated flow cytometry protocols that utilize fresh whole BM.
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http://dx.doi.org/10.3791/59617DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6726111PMC
June 2019

The benzene metabolite 1,4-benzoquinone reduces regulatory T-cell function: A potential mechanism for tobacco smoke-associated atopic dermatitis.

J Allergy Clin Immunol 2017 08 6;140(2):603-605. Epub 2017 Mar 6.

Department of Environmental Immunology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany. Electronic address:

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http://dx.doi.org/10.1016/j.jaci.2017.01.034DOI Listing
August 2017

Immunodominance in allergic T-cell reactivity to Japanese cedar in different geographic cohorts.

Ann Allergy Asthma Immunol 2016 12;117(6):680-689.e1

La Jolla Institute for Allergy and Immunology, La Jolla, California.

Background: Japanese cedar (JC) pollen is a common trigger for allergic rhinitis in Japan. Pollen proteins targeted by IgE, including Cry j 1 and Cry j 2, and isoflavone reductase (IFR) have been identified.

Objective: To compare antigen-specific IgE titers and T-cell responses to JC pollen-derived extract and peptides in cohorts with high and low pollen exposure.

Methods: Peripheral blood mononuclear cells from JC pollen allergic or nonallergic patients who have lived in Japan for at least 1 year and JC pollen allergic patients who have never been to Japan were tested for T-cell responses against JC pollen extract and peptide pools derived from Cry j 1, Cry j 2, or IFR. T-cell reactivity was assessed by interleukin 5 and interferon γ production by ELISPOT.

Results: JC pollen-specific T-cell reactivity and IgE titers were significantly higher in the allergic compared with the nonallergic Japanese cohort, which was also associated with different patterns of polysensitization. Interestingly, a significant overlap was observed in the hierarchy of the T-cell epitopes in the allergic Japanese cohort compared with the allergic non-Japanese cohort. In all 3 cohorts, T-cell reactivity was dominantly directed against peptides from the major allergens Cry j 1 and 2, with few T-cell responses detected against IFR.

Conclusion: Our studies identify common denominators of T-cell reactivity in patient populations with different sensitization patterns, suggesting that generally applicable immunotherapeutic approaches might be developed irrespective of exposure modality.
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http://dx.doi.org/10.1016/j.anai.2016.10.014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5172395PMC
December 2016

Prenatal phthalate exposure associates with low regulatory T-cell numbers and atopic dermatitis in early childhood: Results from the LINA mother-child study.

J Allergy Clin Immunol 2017 04 5;139(4):1376-1379.e8. Epub 2016 Nov 5.

Department of Environmental Immunology, UFZ - Helmholtz Centre for Environmental Research Leipzig, Leipzig, Germany. Electronic address:

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http://dx.doi.org/10.1016/j.jaci.2016.09.034DOI Listing
April 2017

Critical role for syndecan-4 in dendritic cell migration during development of allergic airway inflammation.

Nat Commun 2015 Jul 13;6:7554. Epub 2015 Jul 13.

Department of Dermatology, Venerology and Allergology, Leipzig University Medical Center, Leipzig 04103, Germany.

Syndecan-4 (SDC4), expressed on dendritic cells (DCs) and activated T cells, plays a crucial role in DC motility and has been shown as a potential target for activated T-cell-driven diseases. In the present study, we investigate the role of SDC4 in the development of T-helper 2 cell-mediated allergic asthma. Using SDC4-deficient mice or an anti-SDC4 antibody we show that the absence or blocking of SDC4 signalling in ovalbumin-sensitized mice results in a reduced asthma phenotype compared with control animals. Most importantly, even established asthma is significantly decreased using the anti-SDC4 antibody. The disturbed SDC4 signalling leads to an impaired motility and directional migration of antigen-presenting DCs and therefore, to a modified sensitization leading to diminished airway inflammation. Our results demonstrate that SDC4 plays an important role in asthma induction and indicate SDC4 as possible target for therapeutic intervention in this disease.
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http://dx.doi.org/10.1038/ncomms8554DOI Listing
July 2015

Development and validation of a broad scheme for prediction of HLA class II restricted T cell epitopes.

J Immunol Methods 2015 Jul 7;422:28-34. Epub 2015 Apr 7.

La Jolla Institute for Allergy and Immunology, 9420 Athena Circle, La Jolla, CA 92037, USA.

Computational prediction of HLA class II restricted T cell epitopes has great significance in many immunological studies including vaccine discovery. In recent years, prediction of HLA class II binding has improved significantly but a strategy to globally predict the most dominant epitopes has not been rigorously defined. Using human immunogenicity data associated with sets of 15-mer peptides overlapping by 10 residues spanning over 30 different allergens and bacterial antigens, and HLA class II binding prediction tools from the Immune Epitope Database and Analysis Resource (IEDB), we optimized a strategy to predict the top epitopes recognized by human populations. The most effective strategy was to select peptides based on predicted median binding percentiles for a set of seven DRB1 and DRB3/4/5 alleles. These results were validated with predictions on a blind set of 15 new allergens and bacterial antigens. We found that the top 21% predicted peptides (based on the predicted binding to seven DRB1 and DRB3/4/5 alleles) were required to capture 50% of the immune response. This corresponded to an IEDB consensus percentile rank of 20.0, which could be used as a universal prediction threshold. Utilizing actual binding data (as opposed to predicted binding data) did not appreciably change the efficacy of global predictions, suggesting that the imperfect predictive capacity is not due to poor algorithm performance, but intrinsic limitations of HLA class II epitope prediction schema based on HLA binding in genetically diverse human populations.
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http://dx.doi.org/10.1016/j.jim.2015.03.022DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4458426PMC
July 2015

Allergy-associated T cell epitope repertoires are surprisingly diverse and include non-IgE reactive antigens.

World Allergy Organ J 2014 22;7(1):26. Epub 2014 Oct 22.

La Jolla Institute for Allergy & Immunology, 9420 Athena Circle, La Jolla, CA 92037 USA.

We recently identified T cell epitopes associated with human allergic responses. In a majority of cases, responses focused on a few immunodominant epitopes which can be predicted on the basis of MHC binding characteristics. Several observations from our studies challenged the assumption that T cell epitopes are derived from the same allergen proteins that bind IgE. Transcriptomic and proteomics analysis identified pollen proteins, not bound by IgE. These novel Timothy Grass proteins elicited vigorous Th2 responses, suggesting that unlinked T cell help is operational in pollen-specific responses. Thus, the repertoire of antigens recognized by T cells is much broader than IgE-binding allergens. Additionally, we evaluated the use of epitopes from these novel antigens to assess immunological changes associated with Specific Immunotherapy (SIT). We found that a marked decrease in IL5 production is associated with clinically efficacious SIT, suggesting that these novel antigens are potential immunomarkers for SIT efficacy.
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http://dx.doi.org/10.1186/1939-4551-7-26DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4210551PMC
October 2014

Generation of IL-8 and IL-9 producing CD4⁺ T cells is affected by Th17 polarizing conditions and AHR ligands.

Mediators Inflamm 2014 20;2014:182549. Epub 2014 Feb 20.

UFZ-Helmholtz Centre for Environmental Research Leipzig, Department of Environmental Immunology, Permoserstrasse, 04318 Leipzig, Germany.

The T helper cell subsets Th1, Th2, Th17, and Treg play an important role in immune cell homeostasis, in host defense, and in immunological disorders. Recently, much attention has been paid to Th17 cells which seem to play an important role in the early phase of the adoptive immune response and autoimmune disease. When generating Th17 cells under in vitro conditions the amount of IL-17A producing cells hardly exceeds 20% while the nature of the remaining T cells is poorly characterized. As engagement of the aryl hydrocarbon receptor (AHR) has also been postulated to modulate the differentiation of T helper cells into Th17 cells with regard to the IL-17A expression we ask how far do Th17 polarizing conditions in combination with ligand induced AHR activation have an effect on the production of other T helper cell cytokines. We found that a high proportion of T helper cells cultured under Th17 polarizing conditions are IL-8 and IL-9 single producing cells and that AHR activation results in an upregulation of IL-8 and a downregulation of IL-9 production. Thus, we have identified IL-8 and IL-9 producing T helper cells which are subject to regulation by the engagement of the AHR.
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http://dx.doi.org/10.1155/2014/182549DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3945483PMC
December 2014

Maternal and cord blood miR-223 expression associates with prenatal tobacco smoke exposure and low regulatory T-cell numbers.

J Allergy Clin Immunol 2014 Feb 24;133(2):543-50. Epub 2013 Aug 24.

Department of Environmental Immunology, UFZ-Helmholtz Centre for Environmental Research Leipzig, Leipzig, Germany. Electronic address:

Background: There is evidence that microRNAs (miRNAs) are sensitive to environmental stressors, including tobacco smoke. On the other hand, miRNAs are involved in immune regulation, such as regulatory T (Treg) cell differentiation. The aim of the present study was to investigate the association between prenatal tobacco smoke exposure, miRNAs, and Treg cell numbers.

Methods: Within a prospective mother-child study (Lifestyle and Environmental Factors and Their Influence on Newborns Allergy Risk), we analyzed the expression of miR-155 and miR-223 together with Treg cell numbers in maternal blood during pregnancy, as well as in cord blood (n = 441). Tobacco smoke exposure was assessed based on questionnaire answers and maternal urine cotinine levels. Additionally, the concentration of smoking-related volatile organic compounds was measured in dwellings of study participants.

Results: Both maternal and cord blood miR-223 expressions were positively correlated with maternal urine cotinine levels. An association was also found between maternal miR-223 expression and indoor concentrations of benzene and toluene. High miR-223 expression was associated with lower Treg cell numbers in maternal and cord blood. Furthermore, children with lower Treg cell numbers at birth had a higher risk of atopic dermatitis during the first 3 years of life. The concentration of the toluene metabolite S-benzylmercapturic acid in maternal urine was associated with decreased cord blood, but not maternal blood, miR-155 expression. A relationship between miR-155 expression and Treg cell numbers was not found.

Conclusions: For the first time, we show that maternal tobacco smoke exposure during pregnancy correlates with the level of miRNA-223 expression in blood, with an effect on children's cord blood Treg cell numbers and subsequent allergy risk.
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http://dx.doi.org/10.1016/j.jaci.2013.06.036DOI Listing
February 2014

A strategy to determine HLA class II restriction broadly covering the DR, DP, and DQ allelic variants most commonly expressed in the general population.

Immunogenetics 2013 May 8;65(5):357-70. Epub 2013 Feb 8.

La Jolla Institute for Allergy and Immunology, La Jolla, San Diego, CA 92037, USA.

Classic ways to determine MHC restriction involve inhibition with locus-specific antibodies and antigen presentation assays with panels of cell lines matched or mismatched at the various loci of interest. However, these determinations are often complicated by T cell epitope degeneracy and promiscuity. We describe a selection of 46 HLA DR, DQ, and DP specificities that provide worldwide population (phenotypic) coverage of almost 90 % at each locus, and account for over 66 % of all genes at each locus. This panel afforded coverage of at least four HLA class II alleles in over 95 % of the individuals in four study populations of diverse ethnicity from the USA and South Africa. Next, a panel of single HLA class II-transfected cell lines, corresponding to these 46 allelic variants was assembled, consisting of lines previously developed and 15 novel lines generated for the present study. The novel lines were validated by assessing their HLA class II expression by FACS analysis, the in vitro peptide binding activity of HLA molecules purified from the cell lines, and their antigen presenting capacity to T cell lines of known restriction. We also show that these HLA class II-transfected cell lines can be used to rapidly and unambiguously determine HLA restriction of epitopes recognized by an individual donor in a single experiment. This panel of lines will enable high throughput determination of HLA restriction, enabling better characterization of HLA class II-restricted T cell responses and facilitating the development of HLA tetrameric staining reagents.
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http://dx.doi.org/10.1007/s00251-013-0684-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3633633PMC
May 2013

Renovation activities during pregnancy induce a Th2 shift in fetal but not in maternal immune system.

Int J Hyg Environ Health 2013 Jun 2;216(3):309-16. Epub 2012 Aug 2.

UFZ - Helmholtz Centre for Environmental Research Leipzig, Department of Environmental Immunology, Leipzig, Germany.

The aim of the present study was to investigate to which extent environmental influences like indoor renovation activities affect the immune system of mother and child during the gestation period. Within the LINA (Lifestyle and Environmental Factors and their Influence on Newborn Allergy risk) birth cohort study blood samples of mothers during pregnancy and cord blood samples were analyzed for concentrations of the Th1/Th2 cytokines IL-4, IL-5, IL-13, IFN-γ and IgE. Data on indoor renovation activities (painting, flooring and new furniture) were assessed with questionnaires. Data on cytokine blood concentrations and exposure variables were available for 422 mother/child pairs. Neonates, who were strongly affected by renovation activities (especially floor covering and new furniture) during pregnancy, had significantly higher concentrations of IL-4 and IL-5 in cord blood. Among the single activities, new furniture, particularly flake board, were associated with increased IL-4 levels. Elevated IL-4 levels were also observed in the cord blood of children whose mothers reported wall-to-wall carpeting. Among flooring, polyvinylchloride (PVC) showed the strongest effect with increased IL-5 concentrations. The Th1/Th2 imbalance towards Th2 at birth was related to allergic sensitization in children at the age of one. There were only few and negative associations between renovation activities and Th1/Th2 cytokine concentration in maternal blood. Our study shows that under similar exposure situations the fetal immune system is more susceptible to the influence of environmental factors, in particular renovation products (flake board, wall-to-wall carpets and PVC) compared to the maternal.
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http://dx.doi.org/10.1016/j.ijheh.2012.06.002DOI Listing
June 2013