646 results on '"Schulten V"'
Search Results
2. Distinct modulation of allergic T cell responses by subcutaneous vs. sublingual allergen-specific immunotherapy
- Author
-
Schulten, V., Tripple, V., Aasbjerg, K., Backer, V., Lund, G., Würtzen, P. A., Sette, A., and Peters, B.
- Published
- 2016
- Full Text
- View/download PDF
3. Different Bla-g T cell antigens dominate responses in asthma versus rhinitis subjects
- Author
-
Dillon, M. B. C., Schulten, V., Oseroff, C., Paul, S., Dullanty, L. M., Frazier, A., Belles, X., Piulachs, M.-D., Visness, C., Bacharier, L., Bloomberg, G. R., Busse, P., Sidney, J., Peters, B., and Sette, A.
- Published
- 2015
- Full Text
- View/download PDF
4. Pru p 3, the nonspecific lipid transfer protein from peach, dominates the immune response to its homolog in hazelnut
- Author
-
Schulten, V., Nagl, B., Scala, E., Bernardi, M. L., Mari, A., Ciardiello, M. A., Lauer, I., Scheurer, S., Briza, P., Jürets, A., Ferreira, F., Jahn-Schmid, B., Fischer, G. F., and Bohle, B.
- Published
- 2011
- Full Text
- View/download PDF
5. Sequence-based HLA-A, B, C, DP, DQ, and DR typing of 496 adults from San Diego, California, USA
- Author
-
Moore, E., Grifoni, A., Weiskopf, D., Schulten, V., Arlehamn, C.S.L., Angelo, M., Pham, J., Leary, S., Sidney, J., Broide, D., Frazier, A., Phillips, E., Mallal, S., Mack, S.J., Sette, A., Moore, E., Grifoni, A., Weiskopf, D., Schulten, V., Arlehamn, C.S.L., Angelo, M., Pham, J., Leary, S., Sidney, J., Broide, D., Frazier, A., Phillips, E., Mallal, S., Mack, S.J., and Sette, A.
- Abstract
DNA sequence-based typing at the HLA-A, -B, -C, -DPB1, -DQA1, -DQB1, and -DRB1 loci was performed on 496 healthy adult donors from San Diego, California, to characterize allele frequencies in support of studies of T cell responses to common allergens. Deviations from Hardy Weinberg proportions were detected at each locus except A and C. Several alleles were found in more than 15% of individuals, including the class II alleles DPB1∗02:01, DPB1∗04:01, DQA1∗01:02, DQA1∗05:01, DQB1∗03:01, and the class I allele A∗02:01. Genotype data will be available in the Allele Frequencies Net Database (AFND 3562).
- Published
- 2018
6. Ex vivo assays show human gamma-delta T cells specific for common allergens are Th1-polarized in allergic donors.
- Author
-
Yu ED, Wang E, Garrigan E, Sutherland A, Khalil N, Kearns K, Pham J, Schulten V, Peters B, Frazier A, Sette A, and da Silva Antunes R
- Subjects
- Humans, Animals, Mice, Allergens, Cytokines metabolism, Hypersensitivity, Intraepithelial Lymphocytes metabolism
- Abstract
Gamma-delta (γδ) T cells contribute to the pathology of many immune-related diseases; however, no ex vivo assays to study their activities are currently available. Here, we established a methodology to characterize human allergen-reactive γδ T cells in peripheral blood using an activation-induced marker assay targeting upregulated 4-1BB and CD69 expression. Broad and reproducible ex vivo allergen-reactive γδ T cell responses were detected in donors sensitized to mouse, cockroach, house dust mite, and timothy grass, but the response did not differ from that in non-allergic participants. The reactivity to 4 different allergen extracts was readily detected in 54.2%-100% of allergic subjects in a donor- and allergen-specific pattern and was abrogated by T cell receptor (TCR) blocking. Analysis of CD40L upregulation and intracellular cytokine staining revealed a T helper type 1 (Th1)-polarized response against mouse and cockroach extract stimulation. These results support the existence of allergen-reactive γδ T cells and their potential use in rebalancing dysregulated Th2 responses in allergic diseases., Competing Interests: The authors have declared no conflict of interest., (© 2022 The Author(s).)
- Published
- 2022
- Full Text
- View/download PDF
7. Characterisation of the allergic T cell response to Pru p 3, the nonspecific lipid transfer protein in peach: 99
- Author
-
Schulten, V, Radakovics, A, Hartz, C, Mari, A, Vázquez-Cortés, S, Fernández-Rivas, M, Lauer, I, Jahn-Schmid, B, Eiwegger, T, Scheurer, S, and Bohle, B
- Published
- 2009
8. Urinary peptides as a novel source of T Cell allergen epitopes
- Author
-
da Silva Antunes, R., Pham, J., McMurtrey, C., Hildebrand, W.H., Phillips, E., Mallal, S., Sidney, J., Busse, P., Peters, B., Schulten, V., Sette, A., da Silva Antunes, R., Pham, J., McMurtrey, C., Hildebrand, W.H., Phillips, E., Mallal, S., Sidney, J., Busse, P., Peters, B., Schulten, V., and Sette, A.
- Abstract
Mouse allergy in both laboratory workers and in inner-city children is associated with allergic rhinitis and asthma, posing a serious public health concern. Urine is a major source of mouse allergens, as mice spray urine onto their surroundings, where the proteins dry up and become airborne on dust particles. Here, we tested whether oligopeptides that are abundant in mouse urine may contribute to mouse allergic T cell response. Over 1,300 distinct oligopeptides were detected by mass spectrometry analysis of the low molecular weight filtrate fraction of mouse urine (LoMo). Posttranslationally modified peptides were common, accounting for almost half of total peptides. A pool consisting of 225 unique oligopeptides of 13 residues or more in size identified within was tested for its capacity to elicit T cell reactivity in mouse allergic donors. Following 14-day in vitro stimulation of PBMCs, we detected responses in about 95% of donors tested, directed against 116 distinct peptides, predominantly associated with Th2 cytokines (IL-5). Peptides from non-urine related proteins such as epidermal growth factor, collagen, and Beta-globin accounted for the highest response (15.9, 9.1, and 8.1% of the total response, respectively). Peptides derived from major urinary proteins (MUPs), kidney androgen-regulated protein (KAP), and uromodulin were the main T cell targets from kidney or urine related sources. Further ex vivo analysis of enrichment of 4-1BB expressing cells demonstrated that LoMo pool-specific T cell reactivity can be detected directly ex vivo in mouse allergic but not in non-allergic donors. Further cytometric analysis of responding cells revealed a bone fide memory T cell phenotype and confirmed their Th2 polarization. Overall, these data suggest that mouse urine-derived oligopeptides are a novel target for mouse allergy-associated T cell responses, which may contribute to immunopathological mechanisms in mouse allergy.
- Published
- 2018
9. Heterogeneity of magnitude, allergen immunodominance, and cytokine polarization of cockroach allergen-specific T cell responses in allergic sensitized children.
- Author
-
da Silva Antunes R, Sutherland A, Frazier A, Schulten V, Pomés A, Glesner J, Calatroni A, Altman MC, Wood RA, O'Connor GT, Pongracic JA, Khurana Hershey GK, Kercsmar CM, Gruchalla RS, Gill M, Liu AH, Zoratti E, Kattan M, Busse PJ, Bacharier LB, Teach SJ, Wheatley LM, Togias A, Busse WW, Jackson DJ, and Sette A
- Abstract
Background: Characterization of allergic responses to cockroach (CR), a common aeroallergen associated with asthma, has focused mainly on IgE reactivity, but little is known about T cell responses, particularly in children. We conducted a functional evaluation of CR allergen-specific T cell reactivity in a cohort of CR allergic children with asthma., Methods: Peripheral blood mononuclear cells (PBMCs) were obtained from 71 children, with mild-to-moderate asthma who were enrolled in a CR immunotherapy (IT) clinical trial, prior to treatment initiation. PBMC were stimulated with peptide pools derived from 11 CR allergens, and CD4+ T cell responses assessed by intracellular cytokine staining., Results: Highly heterogeneous responses in T cell reactivity were observed among participants, both in terms of the magnitude of cytokine response and allergen immunodominance. Reactivity against Bla g 9 and Bla g 5 was most frequent. The phenotype of the T cell response was dominated by IL-4 production and a Th2 polarized profile in 54.9% of participants, but IFNγ production and Th1 polarization was observed in 25.3% of the participants. The numbers of regulatory CD4+ T cells were also highly variable and the magnitude of effector responses and Th2 polarization were positively correlated with serum IgE levels specific to a clinical CR extract., Conclusions: Our results demonstrate that in children with mild-to-moderate asthma, CR-specific T cell responses display a wide range of magnitude, allergen dominance, and polarization. These results will enable examination of whether any of the variables measured are affected by IT and/or are predictive of clinical outcomes., Competing Interests: All authors declare no conflict of interest., (© 2021 The Authors. Clinical and Translational Allergy published by John Wiley & Sons Ltd on behalf of European Academy of Allergy and Clinical Immunology.)
- Published
- 2021
- Full Text
- View/download PDF
10. Development of nasal allergen challenge with cockroach in children with asthma.
- Author
-
Rudman Spergel AK, Sever ML, Johnson J, Gill MA, Schulten V, Frazier A, Kercsmar CM, Lovinsky-Desir S, Searing DA, Sette A, Shao B, Teach SJ, Gern JE, Busse WW, Togias A, Wood RA, and Liu AH
- Subjects
- Allergens, Animals, Child, Humans, Leukocytes, Mononuclear, Nasal Provocation Tests, Asthma drug therapy, Cockroaches
- Abstract
Background: Nasal allergen challenge (NAC) could be a means to assess indication and/or an outcome of allergen-specific therapies, particularly for perennial allergens. NACs are not commonly conducted in children with asthma, and cockroach NACs are not well established. This study's objective was to identify a range of German cockroach extract doses that induce nasal symptoms and to assess the safety of cockroach NAC in children with asthma., Methods: Ten adults (18-37 years) followed by 25 children (8-14 years) with well-controlled, persistent asthma and cockroach sensitization underwent NAC with diluent followed by up to 8 escalating doses of cockroach extract (0.00381-11.9 µg/mL Bla g 1). NAC outcome was determined by Total Nasal Symptom Score (TNSS) and/or sneeze score. Cockroach allergen-induced T-cell activation and IL-5 production were measured in peripheral blood mononuclear cells., Results: 67% (6/9) of adults and 68% (17/25) of children had a positive NAC at a median response dose of 0.120 µg/mL [IQR 0.0380-0.379 µg/mL] of Bla g 1. Additionally, three children responded to diluent alone and did not receive any cockroach extract. Overall, 32% (11/34) were positive with sneezes alone, 15% (5/34) with TNSS alone, and 21% (7/34) with both criteria. At baseline, NAC responders had higher cockroach-specific IgE (P = .03), lower cockroach-specific IgG/IgE ratios (children, P = .002), and increased cockroach-specific IL-5-producing T lymphocytes (P = .045). The NAC was well tolerated., Conclusion: We report the methodology of NAC development for children with persistent asthma and cockroach sensitization. This NAC could be considered a tool to confirm clinically relevant sensitization and to assess responses in therapeutic studies., (© 2021 European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd. This article has been contributed to by US Government employees and their work is in the public domain in the USA.)
- Published
- 2021
- Full Text
- View/download PDF
11. IgE and T Cell Reactivity to a Comprehensive Panel of Cockroach Allergens in Relation to Disease.
- Author
-
Pomés A, Schulten V, Glesner J, da Silva Antunes R, Sutherland A, Bacharier LB, Beigelman A, Busse P, Frazier A, and Sette A
- Subjects
- Adult, Animals, Female, Humans, Male, Middle Aged, Allergens immunology, Blattellidae, Hypersensitivity immunology, Immunoglobulin E immunology, Insect Proteins immunology, T-Lymphocytes immunology
- Abstract
IgE sensitization to cockroach allergens is associated with development of allergic diseases, such as asthma. To understand the relevance of different cockroach allergens for diagnosis and immunotherapy, a comprehensive analysis of IgE antibody levels and T cell reactivity to an expanded set of cockroach allergens and their relationship to disease was performed in a cohort of USA cockroach sensitized patients. IgE antibody levels to recombinant chitinase and hemocyanin were measured for 23 subjects by custom-made ImmunoCAPs and compared with IgE levels to eight cockroach allergens we previously reported for the same cohort. Ex vivo T cell activation (Ox40/PDL-1 expression) of PBMCs stimulated with peptide pools derived from 11 German cockroach proteins, including nine official cockroach allergens, plus chitinase and vitellogenin, was determined by flow cytometry. IgE prevalences to chitinase (17%) and hemocyanin (44%) were comparable to values for the other eight allergens that we previously reported (21-57%). Hemocyanin (Bla g 3), was a major allergen (one to which more than 50% of patients with an allergy to its source react) for a sub-group of 15 highly cockroach-sensitized subjects (IgE > 3.5 kU
A /L: 53%). Chitinase was officially named as new allergen Bla g 12. Cockroach-specific IgE levels in plasma showed excellent correlation with the sum of 10 allergen-specific IgE (r = 0.94, p < 0.001). T cell reactivity to 11 proteins was highly variable among subjects, the highest being for vitellogenin, followed by Bla g 3. The main finding was that cockroach allergen-specific IgE and T cell reactivity patterns were unique per subject, and lacked immunodominant allergens and correlation with clinical phenotype/disease severity in the studied cohort. Knowing the subject-specific B/T cell reactivity profiles to a comprehensive panel of cockroach allergens will contribute to diagnosis of cockroach allergy and will be important for planning and assessing allergen immunotherapy outcomes, according to the allergen content in therapeutic cockroach extracts., Competing Interests: AP is an employee of Indoor Biotechnologies, Inc. and the contact principal investigator of the NIH R01 Award that funded the study. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Pomés, Schulten, Glesner, da Silva Antunes, Sutherland, Bacharier, Beigelman, Busse, Frazier and Sette.)- Published
- 2021
- Full Text
- View/download PDF
12. Immunoproteomic analysis of house dust mite antigens reveals distinct classes of dominant T cell antigens according to function and serological reactivity.
- Author
-
Oseroff, C., Christensen, L. H., Westernberg, L., Pham, J., Lane, J., Paul, S., Greenbaum, J., Stranzl, T., Lund, G., Hoof, I., Holm, J., Würtzen, P. A., Meno, K. H., Frazier, A., Schulten, V., Andersen, P. S., Peters, B., and Sette, A.
- Subjects
HOUSE dust mites ,ALLERGENS ,ASTHMA ,T cells ,LIQUID chromatography-mass spectrometry - Abstract
Background House dust mite ( HDM) allergens are a common cause of allergy and allergic asthma. A comprehensive analysis of proteins targeted by T cells, which are implicated in the development and regulation of allergic disease independent of their antibody reactivity, is still lacking. Objective To comprehensively analyse the HDM-derived protein targets of T cell responses in HDM-allergic individuals, and investigate their correlation with IgE/IgG responses and protein function. Methods Proteomic analysis (liquid chromatography-tandem mass spectrometry) of HDM extracts identified 90 distinct protein clusters, corresponding to 29 known allergens and 61 novel proteins. Peripheral blood mononuclear cells ( PBMC) from 20 HDM-allergic individuals were stimulated with HDM extracts and assayed with a set of ~2500 peptides derived from these 90 protein clusters and predicted to bind the most common HLA class II types. 2D immunoblots were made in parallel to elucidate IgE and IgG reactivity, and putative function analyses were performed in silico according to Gene Ontology annotations. Results Analysis of T cell reactivity revealed a large number of T cell epitopes. Overall response magnitude and frequency was comparable for known and novel proteins, with 15 antigens (nine of which were novel) dominating the total T cell response. Most of the known allergens that were dominant at the T cell level were also IgE reactive, as expected, while few novel dominant T cell antigens were IgE reactive. Among known allergens, hydrolase activity and detectable IgE/IgG reactivity are strongly correlated, while no protein function correlates with immunogenicity of novel proteins. A total of 106 epitopes accounted for half of the total T cell response, underlining the heterogeneity of T cell responses to HDM allergens. Conclusions and Clinical Relevance Herein, we define the T cell targets for both known allergens and novel proteins, which may inform future diagnostics and immunotherapeutics for allergy to HDM. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
13. Immunoproteomic analysis of house dust mite antigens reveals distinct classes of dominant T cell antigens according to function and serological reactivity
- Author
-
Oseroff, C., primary, Christensen, L. H., additional, Westernberg, L., additional, Pham, J., additional, Lane, J., additional, Paul, S., additional, Greenbaum, J., additional, Stranzl, T., additional, Lund, G., additional, Hoof, I., additional, Holm, J., additional, Würtzen, P. A., additional, Meno, K. H., additional, Frazier, A., additional, Schulten, V., additional, Andersen, P. S., additional, Peters, B., additional, and Sette, A., additional
- Published
- 2016
- Full Text
- View/download PDF
14. Allergen-specific IgG + memory B cells are temporally linked to IgE memory responses.
- Author
-
Hoof I, Schulten V, Layhadi JA, Stranzl T, Christensen LH, Herrera de la Mata S, Seumois G, Vijayanand P, Lundegaard C, Niss K, Lund A, Ahrenfeldt J, Holm J, Steveling E, Sharif H, Durham SR, Peters B, Shamji MH, and Andersen PS
- Subjects
- Adult, B-Lymphocytes pathology, Double-Blind Method, Female, Humans, Male, Rhinitis, Allergic, Seasonal pathology, Allergens immunology, B-Lymphocytes immunology, Immunoglobulin E immunology, Immunologic Memory, Pollen immunology, Rhinitis, Allergic, Seasonal immunology
- Abstract
Background: IgE is the least abundant immunoglobulin and tightly regulated, and IgE-producing B cells are rare. The cellular origin and evolution of IgE responses are poorly understood., Objective: The cellular and clonal origin of IgE memory responses following mucosal allergen exposure by sublingual immunotherapy (SLIT) were investigated., Methods: In a randomized double-blind, placebo-controlled, time course SLIT study, PBMCs and nasal biopsy samples were collected from 40 adults with seasonal allergic rhinitis at baseline and at 4, 8, 16, 28, and 52 weeks. RNA was extracted from PBMCs, sorted B cells, and nasal biopsy samples for heavy chain variable gene repertoire sequencing. Moreover, mAbs were derived from single B-cell transcriptomes., Results: Combining heavy chain variable gene repertoire sequencing and single-cell transcriptomics yielded direct evidence of a parallel boost of 2 clonally and functionally related B-cell subsets of short-lived IgE
+ plasmablasts and IgG+ memory B cells. Mucosal grass pollen allergen exposure by SLIT resulted in highly diverse IgE and IgGE repertoires. These were extensively mutated and appeared relatively stable as per heavy chain isotype, somatic hypermutations, and clonal composition. Single IgGE + memory B-cell and IgE+ preplasmablast transcriptomes encoded antibodies that were specific for major grass pollen allergens and able to elicit basophil activation at very low allergen concentrations., Conclusion: For the first time, we have shown that on mucosal allergen exposure, human IgE memory resides in allergen-specific IgG+ memory B cells. These cells rapidly switch isotype, expand into short-lived IgE+ plasmablasts, and serve as a potential target for therapeutic intervention., (Crown Copyright © 2019. Published by Elsevier Inc. All rights reserved.)- Published
- 2020
- Full Text
- View/download PDF
15. Immune Therapy Targeting E6/E7 Oncogenes of Human Paillomavirus Type 6 (HPV-6) Reduces or Eliminates the Need for Surgical Intervention in the Treatment of HPV-6 Associated Recurrent Respiratory Papillomatosis.
- Author
-
Aggarwal C, Cohen RB, Morrow MP, Kraynyak KA, Sylvester AJ, Cheung J, Dickerson K, Schulten V, Knoblock D, Gillespie E, Bauml JM, Yan J, Diehl M, Boyer J, Dallas M, Kim JJ, Weiner DB, and Skolnik JM
- Abstract
: Background: Recurrent respiratory papillomatosis (RRP) is a rare disorder characterized by the generation of papillomas of the aerodigestive tract, usually associated with human papilloma virus (HPV) subtypes 6, 11. INO-3106 is a DNA plasmid-based immunotherapy targeting E6 and E7 proteins of HPV6, in order to create a robust immune T cell response., Methods: Testing of INO-3016 in animal models confirmed immunogenicity of the DNA-based therapy. A single-site open-label Phase 1 study was initiated for patients with HPV6-positive RRP. Patients were dosed with INO-3106 with or without INO-9012, a DNA plasmid immunotherapy that encodes IL-12, delivered intramuscularly (IM) in combination with electroporation (EP) with the CELLECTRA
® device. Patients received an escalating dose of INO-3106, 3 mg once and then 6 mg for three additional doses, each dose three weeks apart, with the third and fourth doses co-administered with INO-9012. The primary objective of the study was to evaluate the safety and tolerability of INO-3106 with and without INO-9012. The secondary objective was to determine cellular immune responses to INO-3106 with and without INO-9012. Exploratory objectives included preliminary clinical efficacy to the therapy., Results: Three patients were enrolled in this study, of which two had RRP. Study therapy was well-tolerated, with no related serious adverse events and all related adverse events (AEs) were low-grade. Injection site pain was the most common related AE reported. Immunogenicity was evidenced by multiple immune assays showing engagement and expansion of an HPV6-specific cellular response, including cytotoxic T cells. Preliminary efficacy was demonstrated in patients with RRP in the form of reduction in need for surgical intervention for papilloma growth. Prior to intervention, both patients required surgical intervention approximately every 180 days. One patient demonstrated a greater than three-fold increase in surgery avoidance (584 days) and the other patient remains completely surgery-free as of the last contact at 915 days, a greater than 5-fold increase in surgery interval., Conclusion: INO-3106 with and without INO-9012 was well tolerated, immunogenic and demonstrated preliminary efficacy in patients with HPV6-associated RRP aerodigestive lesions. Further clinical study is indicated., Competing Interests: Authors employed by Inovio Pharmaceuticals are supported financially by that entity. Non-Inovio authors have no conflicts to disclose.- Published
- 2020
- Full Text
- View/download PDF
16. Cross-reactivity in allergy: A double-edged sword.
- Author
-
Pomés A and Schulten V
- Subjects
- Cross Reactions, Humans, Adaptive Immunity immunology, Ambrosia immunology, Hypersensitivity, Immediate immunology
- Published
- 2020
- Full Text
- View/download PDF
17. The association of allergic sensitization patterns in early childhood with disease manifestations and immunological reactivity at 10 years of age.
- Author
-
Schulten V, Frazier A, Calatroni A, Kattan M, Bacharier LB, O'Connor GT, Sandel MT, Wood RA, Wheatley LM, Togias A, Visness CM, Dresen A, Gern JE, and Sette A
- Subjects
- Age of Onset, Animals, Asthma epidemiology, Asthma pathology, Child, Child, Preschool, Cytokines immunology, Female, Humans, Male, Allergens immunology, Asthma diagnosis, Asthma immunology, Blattellidae immunology, Epitopes, T-Lymphocyte immunology
- Abstract
Background: Allergy to German cockroach (CR) is common in urban environments and is an important allergen in children with asthma., Objective: We hypothesize that the evolution of allergic sensitization and clinical disease is associated with distinct patterns of allergen-specific T cell reactivity. To test this hypothesis, a subset of high-risk inner-city children participating in the URECA (Urban Environment and Childhood Asthma) birth cohort were selected to evaluate CR-specific T cell reactivity from three distinct groups based on acquisition of aeroallergen sensitivity from ages 2 to 10: low atopy with minimal to no sensitivity (n = 26), early-onset allergic sensitization (n = 25) and late-onset allergic sensitization (n = 25)., Methods: Using pools of previously identified CR-derived T cell epitopes, we characterized the allergen-specific T cell response in these 76 subjects from blood samples obtained at age 10. CR-specific production of IL-5, IFNγ and IL-10 was measured by ELISPOT following two-week in vitro culture with CR extract., Results: T cell responses were significantly higher in the early-onset atopy group compared to low atopy (P = 0.01), and a trend for higher cytokine production in the late onset compared to the low atopy cohort was also observed (P = 0.06). T cell responses were similar between early- and late-onset cohorts. Furthermore, a comparison of T cell reactivity between asthmatic and non-asthmatic individuals revealed significantly higher cytokine production in asthmatics compared to non-asthmatics (P = 0.02) within both the CR-allergic and non-allergic cohorts., Conclusions and Clinical Relevance: In conclusion, the present study reports that higher T cell reactivity is associated with allergen sensitization and asthma. Interestingly, no significant difference in T cell reactivity was observed in allergic children with early-onset versus late-onset atopy., (© 2019 John Wiley & Sons Ltd.)
- Published
- 2019
- Full Text
- View/download PDF
18. Analysis of Allergen-Specific T Cell and IgE Reactivity to Different Preparations of Cow's Milk-Containing Food Extracts.
- Author
-
Chen M, Sutherland A, Birrueta G, Laubach S, Leonard S, Peters B, and Schulten V
- Subjects
- Animals, Cells, Cultured, Child, Child, Preschool, Female, Humans, Infant, Interferon-gamma immunology, Interferon-gamma metabolism, Interleukin-5 immunology, Interleukin-5 metabolism, Male, Milk Hypersensitivity blood, Milk Hypersensitivity diagnosis, Primary Cell Culture, Skin Tests, T-Lymphocytes metabolism, Allergens immunology, Immunoglobulin E immunology, Milk immunology, Milk Hypersensitivity immunology, T-Lymphocytes immunology
- Abstract
Background: cow's milk allergy (CM) is among the most common food allergies in young children and is often outgrown by adulthood. Prior to developing a tolerance to CM, a majority of CM-allergic children may tolerate extensively-heated CM. This study aims to characterize the IgE- and T cell-reactivity to unheated CM and the progressively more heated CM-containing foods., Methods: CM-containing food extracts from muffin, baked cheese, custard and raw, pasteurized CM commercial extract were tested for skin prick test reactivity, IgE binding and T cell reactivity as assessed by IL-5 and IFNγ production., Results: the skin prick test (SPT) reactivity was significantly decreased to muffin extract compared to raw, pasteurized CM. Both IgE- and T-cell reactivity were readily detectable against food extracts from all forms of CM. Western blot analysis of IgE reactivity revealed variability between extracts that was protein-specific. T cell-reactivity was detected against all four extracts with no significant difference in IL-5 or IFNγ production between them., Conclusion: our data indicate that despite reduced clinical reactivity, extracts from heated CM-containing foods retain immunogenicity when tested in vitro, particularly at the T cell level.
- Published
- 2019
- Full Text
- View/download PDF
19. Circulating T cell-monocyte complexes are markers of immune perturbations.
- Author
-
Burel JG, Pomaznoy M, Lindestam Arlehamn CS, Weiskopf D, da Silva Antunes R, Jung Y, Babor M, Schulten V, Seumois G, Greenbaum JA, Premawansa S, Premawansa G, Wijewickrama A, Vidanagama D, Gunasena B, Tippalagama R, deSilva AD, Gilman RH, Saito M, Taplitz R, Ley K, Vijayanand P, Sette A, and Peters B
- Subjects
- Flow Cytometry, Humans, Microscopy, Blood Cells cytology, Cell Adhesion, Monocytes cytology, Monocytes immunology, T-Lymphocytes cytology, T-Lymphocytes immunology
- Abstract
Our results highlight for the first time that a significant proportion of cell doublets in flow cytometry, previously believed to be the result of technical artifacts and thus ignored in data acquisition and analysis, are the result of biological interaction between immune cells. In particular, we show that cell:cell doublets pairing a T cell and a monocyte can be directly isolated from human blood, and high resolution microscopy shows polarized distribution of LFA1/ICAM1 in many doublets, suggesting in vivo formation. Intriguingly, T cell-monocyte complex frequency and phenotype fluctuate with the onset of immune perturbations such as infection or immunization, reflecting expected polarization of immune responses. Overall these data suggest that cell doublets reflecting T cell-monocyte in vivo immune interactions can be detected in human blood and that the common approach in flow cytometry to avoid studying cell:cell complexes should be re-visited., Competing Interests: JB, MP, CL, DW, Rd, YJ, MB, VS, GS, JG, SP, GP, AW, DV, BG, RT, Ad, RG, MS, RT, KL, PV, AS, BP No competing interests declared, (© 2019, Burel et al.)
- Published
- 2019
- Full Text
- View/download PDF
20. Characterization and epitope identification of the T cell response in non-allergic individuals exposed to mouse allergen.
- Author
-
Grifoni A, da Silva Antunes R, Westernberg L, Pham J, Birrueta G, Peters B, Sette A, and Schulten V
- Abstract
Background: Exposure to airborne allergens is a frequent trigger of respiratory allergy and asthma in atopic individuals. While allergic patients suffer hypersensitivity reactions to these allergens, non-allergic individuals do not exhibit clinical symptoms despite environmental exposure to these ubiquitous allergen sources. The aim of this study was to characterize T cell responses in non-allergic laboratory workers, who are heavily exposed to mice allergens (Exposed Non-Allergics, ENA) and compare this data to previously published T cell responses measured in mouse (MO)-allergic patients. METHODS: Peripheral mononuclear cells (PBMC) from ENA subjects were expanded for 2 weeks in vitro with mouse urine extract and screened for IFNγ and IL-5 cytokine production in response to mouse antigen-derived peptides by ELISPOT. Ex vivo T cell reactivity in the ENA cohort was performed after 6hr stimulation with peptide pools by intracellular staining of CD154., Results: Vigorous responses were detected, associated with 147 epitopes derived from 16 mouse antigens. As expected, responses in ENA subjects were somewhat lower than those observed in MO-allergics for both responder frequency and overall response magnitude. While responses in allergics were polarized towards IL-5 production and associated with low IFNγ production, ENA responses were not polarized. The composition of targeted antigens and epitopes was overall similar between the two cohorts, with the majority of T cell reactivity directed against Mus m 1 and other major urinary proteins. However, kappa-casein precursor and odorant binding protein Ib were more abundantly recognized in MO-allergics compared to ENA subjects. Additionally, T cell responses against oligopeptides derived from the low molecular weight fraction of mouse urine were also assessed. Interestingly, no difference in the response frequency, magnitude or polarization between MO-allergic and ENA individuals was observed. Finally, assessment of ex vivo T cell activation also revealed T cell reactivity in the ENA cohort, with a non-significant trend for lower responses compared to MO-allergics., Conclusion: Exposure to mouse induces potent T cell responses in non-allergic individuals, targeting similar epitopes as seen in allergic patients.
- Published
- 2019
- Full Text
- View/download PDF
21. Allergen content in German cockroach extracts and sensitization profiles to a new expanded set of cockroach allergens determine in vitro extract potency for IgE reactivity.
- Author
-
Glesner J, Filep S, Vailes LD, Wünschmann S, Chapman MD, Birrueta G, Frazier A, Jeong KY, Schal C, Bacharier L, Beigelman A, Busse P, Schulten V, Sette A, and Pomés A
- Subjects
- Animals, Female, Humans, Hypersensitivity etiology, Male, Allergens immunology, Blattellidae immunology, Hypersensitivity immunology, Immunoglobulin E immunology, Insect Proteins immunology
- Abstract
Background: Cockroach allergens are an important cause of IgE-mediated sensitization in inner-city asthmatic patients. However, cockroach extracts used for diagnosis and immunotherapy are not standardized., Objective: We sought to determine the allergen content of nonstandardized German cockroach extracts and the levels of sensitization to an expanded set of cockroach allergens as determinants of in vitro extract potency for IgE reactivity., Methods: Twelve German cockroach extracts were compared for allergen content and potency of IgE reactivity. Bla g 1, Bla g 2, and Bla g 5 were measured by using immunoassays. IgE antibody levels to 8 purified recombinant allergens from groups 1, 2, 4, 5, 6, 7, 9, and 11 were measured by using ImmunoCAP. IgE antibody binding inhibition assays were performed to assess extract in vitro potencies (concentration inhibiting 30% of the total IgE antibody-binding inhibition) relative to an arbitrarily selected reference extract in 5 patients with cockroach allergy., Results: Allergen levels were highly variable. Three new major allergens (groups 6, 9, and 11), were identified among highly cockroach-sensitized subjects (CAP class ≥ 3). Sensitization profiles were unique per subject without immunodominant allergens. The sum of IgE to 8 allergen components showed a good correlation with cockroach-specific IgE levels (r = 0.88, P < .001). In vitro potencies varied among different extracts per subject and among subjects for each extract., Conclusions: The in vitro potency of German cockroach extracts for IgE reactivity depends on allergen content and allergen-specific IgE titers of patients with cockroach allergy. These factors are relevant for selection of potent extracts to be used for immunotherapy and for the design and interpretation of data from immunotherapy trials., (Copyright © 2018 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
22. Variability in German Cockroach Extract Composition Greatly Impacts T Cell Potency in Cockroach-Allergic Donors.
- Author
-
Birrueta G, Frazier A, Pomés A, Glesner J, Filep S, Schal C, Jeong KY, McMurtrey C, Vander Schans T, Hildebrand WH, Busse P, Beigelman A, Bacharier LB, Peters B, Sette A, and Schulten V
- Subjects
- Animals, Blattellidae chemistry, Cytokines biosynthesis, Humans, Hypersensitivity diagnosis, Hypersensitivity metabolism, Immunoglobulin E immunology, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear metabolism, Skin Tests, T-Lymphocytes metabolism, Tissue Donors, Tissue Extracts chemistry, Allergens immunology, Blattellidae immunology, Hypersensitivity immunology, Lymphocyte Activation immunology, T-Lymphocytes immunology, Tissue Extracts immunology
- Abstract
German cockroach extract is used clinically to evaluate allergen-specific sensitization and for subcutaneous allergen-specific immunotherapy, though there are no guidelines for standardization in its manufacture. We performed an immunological evaluation of 12 different cockroach extracts prepared from different sources and their potency to induce allergen-specific T cell reactivity. PBMC from 13 cockroach allergic donors were expanded in vitro with 12 different German cockroach extracts. After culture expansion, cells were re-stimulated with the different extracts and T cell responses were assessed by FluoroSpot (IL-5, IFNγ and IL-10 production). In parallel to the extracts, single allergen peptide pools for allergens from groups 1, 2, 4, 5, and 11 were tested to determine allergen immunodominance. Furthermore, to assess allergy specificity, PBMC from 13 non-allergic donors were also tested with the most potent extract and T cell responses were compared to the allergic cohort. Dramatic variations in T cell reactivity were observed to the different cockroach extract batches. Response magnitudes varied over 3 logs within a single donor. IL-5 production in the allergic cohort was significantly higher compared to the non-allergic cohort (p=0.004). Allergen content determination by ELISA detected much lower concentrations of Bla g 5 compared to Bla g 1 and 2. Mass spectrometric analysis revealed that Bla g 5 was present in similar amounts to Bla g 1 and 2 in extracts made from whole body, whereas it was not detected in extracts made from fecal matter, suggesting that Bla g 5 is not excreted into feces. Different donors exhibit different response patterns to different extracts, potentially dependent on the donor-specific T cell allergen immunodominance pattern and the allergen content of the extract tested. These findings have dramatic implications for the selection of potent extracts used for diagnostic purposes or allergen-specific immunotherapy.
- Published
- 2019
- Full Text
- View/download PDF
23. Sequence-based HLA-A, B, C, DP, DQ, and DR typing of 496 adults from San Diego, California, USA.
- Author
-
Moore E, Grifoni A, Weiskopf D, Schulten V, Arlehamn CSL, Angelo M, Pham J, Leary S, Sidney J, Broide D, Frazier A, Phillips E, Mallal S, Mack SJ, and Sette A
- Subjects
- Adolescent, Adult, Alleles, California, Female, Gene Frequency, Genotype, Genotyping Techniques methods, Histocompatibility Testing methods, Humans, Linkage Disequilibrium, Male, Middle Aged, Sequence Analysis, DNA methods, T-Lymphocytes immunology, T-Lymphocytes metabolism, Young Adult, HLA-A Antigens genetics, HLA-B Antigens genetics, HLA-C Antigens genetics, HLA-DP Antigens genetics, HLA-DQ Antigens genetics, HLA-DR Antigens genetics
- Abstract
DNA sequence-based typing at the HLA-A, -B, -C, -DPB1, -DQA1, -DQB1, and -DRB1 loci was performed on 496 healthy adult donors from San Diego, California, to characterize allele frequencies in support of studies of T cell responses to common allergens. Deviations from Hardy Weinberg proportions were detected at each locus except A and C. Several alleles were found in more than 15% of individuals, including the class II alleles DPB1∗02:01, DPB1∗04:01, DQA1∗01:02, DQA1∗05:01, DQB1∗03:01, and the class I allele A∗02:01. Genotype data will be available in the Allele Frequencies Net Database (AFND 3562)., (Copyright © 2018. Published by Elsevier Inc.)
- Published
- 2018
- Full Text
- View/download PDF
24. Epitope Specific Antibodies and T Cell Receptors in the Immune Epitope Database.
- Author
-
Mahajan S, Vita R, Shackelford D, Lane J, Schulten V, Zarebski L, Jespersen MC, Marcatili P, Nielsen M, Sette A, and Peters B
- Subjects
- Animals, Epitopes, T-Lymphocyte genetics, Humans, Mice, Receptors, Antigen, T-Cell genetics, Antibodies immunology, Antibody Specificity, Databases, Protein, Epitopes, T-Lymphocyte immunology, Receptors, Antigen, T-Cell immunology
- Abstract
The Immune Epitope Database (IEDB) is a free public resource which catalogs experiments characterizing immune epitopes. To accommodate data from next generation repertoire sequencing experiments, we recently updated how we capture and query epitope specific antibodies and T cell receptors. Specifically, we are now storing partial receptor sequences sufficient to determine CDRs and VDJ gene usage which are commonly identified by repertoire sequencing. For previously captured full length receptor sequencing data, we have calculated the corresponding CDR sequences and gene usage information using IMGT numbering and VDJ gene nomenclature format. To integrate information from receptors defined at different levels of resolution, we grouped receptors based on their host species, receptor type and CDR3 sequence. As of August 2018, we have cataloged sequence information for more than 22,510 receptors in 18,292 receptor groups, shown to bind to more than 2,241 distinct epitopes. These data are accessible as full exports and through a new dedicated query interface. The later combines the new ability to search by receptor characteristics with previously existing capability to search by epitope characteristics such as the infectious agent the epitope is derived from, or the kind of immune response involved in its recognition. We expect that this comprehensive capture of epitope specific immune receptor information will provide new insights into receptor-epitope interactions, and facilitate the development of novel tools that help in the analysis of receptor repertoire data.
- Published
- 2018
- Full Text
- View/download PDF
25. Development of a novel clustering tool for linear peptide sequences.
- Author
-
Dhanda SK, Vaughan K, Schulten V, Grifoni A, Weiskopf D, Sidney J, Peters B, and Sette A
- Subjects
- Animals, Mice, Rats, Algorithms, Databases, Protein, Epitopes chemistry, Epitopes genetics, Peptides chemistry, Peptides genetics, Sequence Analysis, Protein methods
- Abstract
Epitopes identified in large-scale screens of overlapping peptides often share significant levels of sequence identity, complicating the analysis of epitope-related data. Clustering algorithms are often used to facilitate these analyses, but available methods are generally insufficient in their capacity to define biologically meaningful epitope clusters in the context of the immune response. To fulfil this need we developed an algorithm that generates epitope clusters based on representative or consensus sequences. This tool allows the user to cluster peptide sequences on the basis of a specified level of identity by selecting among three different method options. These include the 'clique method', in which all members of the cluster must share the same minimal level of identity with each other, and the 'connected graph method', in which all members of a cluster must share a defined level of identity with at least one other member of the cluster. In cases where it is not possible to define a clear consensus sequence with the connected graph method, a third option provides a novel 'cluster-breaking algorithm' for consensus sequence driven sub-clustering. Herein we demonstrate the tool's clustering performance and applicability using (i) a selection of dengue virus epitopes for the 'clique method', (ii) sets of allergen-derived peptides from related species for the 'connected graph method' and (iii) large data sets of eluted ligand, major histocompatibility complex binding and T-cell recognition data captured within the Immune Epitope Database (IEDB) with the newly developed 'cluster-breaking algorithm'. This novel clustering tool is accessible at http://tools.iedb.org/cluster2/., (© 2018 The Authors. Immunology Published by John Wiley & Sons Ltd.)
- Published
- 2018
- Full Text
- View/download PDF
26. Peanut-specific T cell responses in patients with different clinical reactivity.
- Author
-
Birrueta G, Tripple V, Pham J, Manohar M, James EA, Kwok WW, Nadeau KC, Sette A, Peters B, and Schulten V
- Subjects
- Adult, Cells, Cultured, Cohort Studies, Epitope Mapping, Female, Humans, Immunoglobulin E blood, Interferon-gamma blood, Interleukin-5 blood, Male, Peanut Hypersensitivity blood, Receptors, Immunologic blood, Receptors, Prostaglandin blood, Antigens, Plant immunology, Arachis immunology, Peanut Hypersensitivity immunology, T-Lymphocytes immunology
- Abstract
Whole extract or allergen-specific IgE testing has become increasingly popular in the diagnosis of peanut allergy. However, much less is known about T cell responses in peanut allergy and how it relates to different clinical phenotypes. CD4+ T cells play a major role in the pathophysiology of peanut allergy as well as tolerance induction during oral desensitization regimens. We set out to characterize and phenotype the T cell responses and their targets in peanut sensitized patients. Using PBMC from peanut-allergic and non-allergic patients, we mapped T cell epitopes for three major peanut allergens, Ara h 1, 2 and 3 (27 from Ara h 1, 4 from Ara h 2 and 43 from Ara h 3) associated with release of IFNγ (representative Th1 cytokine) and IL5 (representative Th2 cytokine). A pool containing 19 immunodominant peptides, selected to account for 60% of the total Ara h 1-3-specific T cell response in allergics, but only 20% in non-allergics, was shown to discriminate T cell responses in peanut-sensitized, symptomatic vs non-symptomatic individuals more effectively than peanut extract. This pool elicited positive T cell responses above a defined threshold in 12/15 sensitized, symptomatic patients, whereas in the sensitized but non-symptomatic cohort only, 4/14 reacted. The reactivity against this peptide pool in symptomatic patients was dominated by IL-10, IL-17 and to a lesser extend IL-5. For four distinct epitopes, HLA class II restrictions were determined, enabling production of tetrameric reagents. Tetramer staining in four donors (2 symptomatic, 2 non-symptomatic) revealed a trend for increased numbers of peanut epitope-specific T cells in symptomatic patients compared to non-symptomatic patients, which was associated with elevated CRTh2 expression whereas cells from non-symptomatic patients exhibited higher levels of Integrin β7 expression. Our results demonstrate differences in T cell response magnitude, epitope specificity and phenotype between symptomatic and non-symptomatic peanut-sensitized patients. In addition to IgE reactivity, analysis of peanut-specific T cells may be useful to improve our understanding of different clinical manifestations in peanut allergy., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2018
- Full Text
- View/download PDF
27. Urinary Peptides As a Novel Source of T Cell Allergen Epitopes.
- Author
-
da Silva Antunes R, Pham J, McMurtrey C, Hildebrand WH, Phillips E, Mallal S, Sidney J, Busse P, Peters B, Schulten V, and Sette A
- Subjects
- Adult, Allergens urine, Animals, Asthma blood, Epitopes, T-Lymphocyte immunology, Female, Humans, Immunoglobulin E blood, Immunoglobulin E immunology, Male, Middle Aged, Peptides urine, Proteome analysis, Proteome immunology, Proteomics methods, Rhinitis, Allergic blood, T-Lymphocytes immunology, Allergens immunology, Asthma immunology, Mice urine, Peptides immunology, Rhinitis, Allergic immunology
- Abstract
Mouse allergy in both laboratory workers and in inner-city children is associated with allergic rhinitis and asthma, posing a serious public health concern. Urine is a major source of mouse allergens, as mice spray urine onto their surroundings, where the proteins dry up and become airborne on dust particles. Here, we tested whether oligopeptides that are abundant in mouse urine may contribute to mouse allergic T cell response. Over 1,300 distinct oligopeptides were detected by mass spectrometry analysis of the low molecular weight filtrate fraction of mouse urine (LoMo). Posttranslationally modified peptides were common, accounting for almost half of total peptides. A pool consisting of 225 unique oligopeptides of 13 residues or more in size identified within was tested for its capacity to elicit T cell reactivity in mouse allergic donors. Following 14-day in vitro stimulation of PBMCs, we detected responses in about 95% of donors tested, directed against 116 distinct peptides, predominantly associated with Th2 cytokines (IL-5). Peptides from non-urine related proteins such as epidermal growth factor, collagen, and Beta-globin accounted for the highest response (15.9, 9.1, and 8.1% of the total response, respectively). Peptides derived from major urinary proteins (MUPs), kidney androgen-regulated protein (KAP), and uromodulin were the main T cell targets from kidney or urine related sources. Further ex vivo analysis of enrichment of 4-1BB expressing cells demonstrated that LoMo pool-specific T cell reactivity can be detected directly ex vivo in mouse allergic but not in non-allergic donors. Further cytometric analysis of responding cells revealed a bone fide memory T cell phenotype and confirmed their Th2 polarization. Overall, these data suggest that mouse urine-derived oligopeptides are a novel target for mouse allergy-associated T cell responses, which may contribute to immunopathological mechanisms in mouse allergy.
- Published
- 2018
- Full Text
- View/download PDF
28. Allergen and Epitope Targets of Mouse-Specific T Cell Responses in Allergy and Asthma.
- Author
-
Schulten V, Westernberg L, Birrueta G, Sidney J, Paul S, Busse P, Peters B, and Sette A
- Subjects
- Adult, Allergens immunology, Animal Technicians, Animals, Animals, Laboratory immunology, Asthma blood, Epitope Mapping, Epitopes, T-Lymphocyte immunology, Female, Humans, Immunodominant Epitopes immunology, Interferon-gamma immunology, Interferon-gamma metabolism, Interleukin-5 immunology, Interleukin-5 metabolism, Male, Middle Aged, Occupational Diseases blood, Proteins immunology, Rhinitis, Allergic blood, T-Lymphocytes metabolism, Asthma immunology, Mice immunology, Occupational Diseases immunology, Rhinitis, Allergic immunology, T-Lymphocytes immunology
- Abstract
Mouse allergy has become increasingly common, mainly affecting laboratory workers and inner-city households. To date, only one major allergen, namely Mus m 1, has been described. We sought to identify T cell targets in mouse allergic patients. PBMC from allergic donors were expanded with either murine urine or epithelial extract and subsequently screened for cytokine production (IL-5 and IFNγ) in response to overlapping peptides spanning the entire Mus m 1 sequence, peptides from various Mus m 1 isoforms [major urinary proteins (MUPs)], peptides from mouse orthologs of known allergens from other mammalian species and peptides from proteins identified by immunoproteomic analysis of IgE/IgG immunoblots of mouse urine and epithelial extracts. This approach let to the identification of 106 non-redundant T cell epitopes derived from 35 antigens. Three major T cell-activating regions were defined in Mus m 1 alone. Moreover, our data show that immunodominant epitopes were largely shared between Mus m 1 and other MUPs even from different species, suggesting that sequence conservation in different allergens is a determinant for immunodominance. We further identified several novel mouse T cell antigens based on their homology to known mammalian allergens. Analysis of cohort-specific T cell responses revealed that rhinitis and asthmatic patients recognized different epitope repertoires. Epitopes defined herein can be formulated into an epitope "megapool" used to diagnose mouse allergy and study mouse-specific T cell responses directly ex vivo . This analysis of T cell epitopes provides a good basis for future studies to increase our understanding of the immunopathology associated with MO-allergy and asthma.
- Published
- 2018
- Full Text
- View/download PDF
29. Allergen-specific immunotherapy modulates the balance of circulating Tfh and Tfr cells.
- Author
-
Schulten V, Tripple V, Seumois G, Qian Y, Scheuermann RH, Fu Z, Locci M, Rosales S, Vijayanand P, Sette A, Alam R, Crotty S, and Peters B
- Subjects
- Adult, Aged, Female, Humans, Hypersensitivity pathology, Male, Middle Aged, T-Lymphocytes, Helper-Inducer pathology, Allergens administration & dosage, Hypersensitivity immunology, Hypersensitivity therapy, Immunotherapy, Phleum, T-Lymphocytes, Helper-Inducer immunology
- Published
- 2018
- Full Text
- View/download PDF
30. The Identification of Allergen-Derived T Cell Epitopes.
- Author
-
Schulten V and Sette A
- Subjects
- Allergens chemistry, Amino Acid Sequence, Cells, Cultured, Cytokines metabolism, Data Analysis, Epitopes, T-Lymphocyte chemistry, Humans, Lymphocyte Activation, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets metabolism, Th2 Cells immunology, Th2 Cells metabolism, Allergens immunology, Epitope Mapping methods, Epitopes, T-Lymphocyte immunology
- Abstract
Type I allergy is a disease primarily mediated by immunoglobulin E (IgE) and T helper type 2 (Th2) cells. The role of Th2 and other T cell subsets in the pathology of allergic disease as well as induction of tolerance has become an area of intense research over the last decades. Studying allergen-specific T cells to gain a better understanding of their contribution to allergic pathology and how they are modulated by allergen-specific immunotherapy requires knowledge of the allergens targeted by these cells. Identification of T cell epitopes in allergy can be achieved by a variety of methods. In this chapter, we will focus on a technique named FluoroSpot, which relies on the detection of cytokines secreted by T cells in response to stimulation with an antigen (allergen), such as timothy grass (TG) extract or an allergen-derived peptide, for which the cell is specific. We will describe how to overcome the challenge of detecting rare, TG-specific, T cells that occur at low frequency in the blood by using an in vitro expansion culture and subsequent mapping of the precise T cell epitope using FluoroSpot.
- Published
- 2018
- Full Text
- View/download PDF
31. Unique phenotypes and clonal expansions of human CD4 effector memory T cells re-expressing CD45RA.
- Author
-
Tian Y, Babor M, Lane J, Schulten V, Patil VS, Seumois G, Rosales SL, Fu Z, Picarda G, Burel J, Zapardiel-Gonzalo J, Tennekoon RN, De Silva AD, Premawansa S, Premawansa G, Wijewickrama A, Greenbaum JA, Vijayanand P, Weiskopf D, Sette A, and Peters B
- Subjects
- Adolescent, Adult, Aged, CD4-Positive T-Lymphocytes classification, CD8-Positive T-Lymphocytes classification, CD8-Positive T-Lymphocytes immunology, Cells, Cultured, Core Binding Factor Alpha 3 Subunit biosynthesis, Gene Expression Profiling, Granzymes biosynthesis, Heterogeneous-Nuclear Ribonucleoproteins biosynthesis, Humans, Immunologic Memory immunology, Male, Middle Aged, Perforin biosynthesis, Receptors, CCR7 metabolism, Signaling Lymphocytic Activation Molecule Family biosynthesis, T-Box Domain Proteins biosynthesis, Young Adult, CD4-Positive T-Lymphocytes immunology, Cytomegalovirus immunology, Dengue Virus immunology, Herpesvirus 4, Human immunology, Leukocyte Common Antigens metabolism, Receptors, G-Protein-Coupled metabolism
- Abstract
The expression of CD45RA is generally associated with naive T cells. However, a subset of effector memory T cells re-expresses CD45RA (termed TEMRA) after antigenic stimulation with unknown molecular characteristics and functions. CD4 TEMRA cells have been implicated in protective immunity against pathogens such as dengue virus (DENV). Here we show that not only the frequency but also the phenotype of CD4 TEMRA cells are heterogeneous between individuals. These cells can be subdivided into two major subsets based on the expression of the adhesion G protein-coupled receptor GPR56, and GPR56
+ TEMRA cells display a transcriptional and proteomic program with cytotoxic features that is distinct from effector memory T cells. Moreover, GPR56+ TEMRA cells have higher levels of clonal expansion and contain the majority of virus-specific TEMRA cells. Overall, this study reveals the heterogeneity of CD4 TEMRA cells and provides insights into T-cell responses against DENV and other viral pathogens.- Published
- 2017
- Full Text
- View/download PDF
32. Experimental validation of the RATE tool for inferring HLA restrictions of T cell epitopes.
- Author
-
Paul S, Arlehamn CSL, Schulten V, Westernberg L, Sidney J, Peters B, and Sette A
- Subjects
- Female, Humans, Male, South Africa, Alleles, Epitopes, T-Lymphocyte genetics, Epitopes, T-Lymphocyte immunology, Genotyping Techniques instrumentation, Genotyping Techniques methods, HLA Antigens genetics, HLA Antigens immunology, Histocompatibility Testing instrumentation, Histocompatibility Testing methods
- Abstract
Background: The RATE tool was recently developed to computationally infer the HLA restriction of given epitopes from immune response data of HLA typed subjects without additional cumbersome experimentation., Results: Here, RATE was validated using experimentally defined restriction data from a set of 191 tuberculosis-derived epitopes and 63 healthy individuals with MTB infection from the Western Cape Region of South Africa. Using this experimental dataset, the parameters utilized by the RATE tool to infer restriction were optimized, which included relative frequency (RF) of the subjects responding to a given epitope and expressing a given allele as compared to the general test population and the associated p-value in a Fisher's exact test. We also examined the potential for further optimization based on the predicted binding affinity of epitopes to potential restricting HLA alleles, and the absolute number of individuals expressing a given allele and responding to the specific epitope. Different statistical measures, including Matthew's correlation coefficient, accuracy, sensitivity and specificity were used to evaluate performance of RATE as a function of these criteria. Based on our results we recommend selection of HLA restrictions with cutoffs of p-value < 0.01 and RF ≥ 1.3. The usefulness of the tool was demonstrated by inferring new HLA restrictions for epitope sets where restrictions could not be experimentally determined due to lack of necessary cell lines and for an additional data set related to recognition of pollen derived epitopes from allergic patients., Conclusions: Experimental data sets were used to validate RATE tool and the parameters used by the RATE tool to infer restriction were optimized. New HLA restrictions were identified using the optimized RATE tool.
- Published
- 2017
- Full Text
- View/download PDF
33. It's a lot of work to be nonallergic.
- Author
-
Sette A and Schulten V
- Subjects
- Animals, Epithelial Cells, Hypersensitivity, Inflammation, Dermatophagoides pteronyssinus, Pyroglyphidae
- Published
- 2017
- Full Text
- View/download PDF
34. Immunodominance in allergic T-cell reactivity to Japanese cedar in different geographic cohorts.
- Author
-
Oseroff C, Pham J, Frazier A, Hinz D, Sidney J, Paul S, Greenbaum JA, Vita R, Peters B, Schulten V, and Sette A
- Subjects
- Adolescent, Adult, Alleles, Amino Acid Sequence, Cohort Studies, Female, HLA Antigens genetics, HLA Antigens immunology, Humans, Immunoglobulin E immunology, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear metabolism, Lymphocyte Activation immunology, Male, Middle Aged, Peptides immunology, Pollen immunology, Rhinitis, Allergic, Seasonal genetics, Rhinitis, Allergic, Seasonal metabolism, T-Lymphocytes metabolism, Young Adult, Allergens immunology, Antigens, Plant immunology, Cryptomeria adverse effects, Epitopes, T-Lymphocyte immunology, Rhinitis, Allergic, Seasonal immunology, T-Lymphocytes immunology
- Abstract
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., (Published by Elsevier Inc.)
- Published
- 2016
- Full Text
- View/download PDF
35. 17q21 asthma-risk variants switch CTCF binding and regulate IL-2 production by T cells.
- Author
-
Schmiedel BJ, Seumois G, Samaniego-Castruita D, Cayford J, Schulten V, Chavez L, Ay F, Sette A, Peters B, and Vijayanand P
- Subjects
- B-Lymphocytes metabolism, Base Sequence, Binding Sites genetics, Cells, Cultured, Enhancer Elements, Genetic genetics, Humans, Introns genetics, Promoter Regions, Genetic genetics, Protein Binding, Risk Factors, Asthma genetics, Asthma immunology, CCCTC-Binding Factor metabolism, Chromosomes, Human, Pair 17 genetics, Genetic Predisposition to Disease, Interleukin-2 biosynthesis, Polymorphism, Single Nucleotide genetics, T-Lymphocytes metabolism
- Abstract
Asthma and autoimmune disease susceptibility has been strongly linked to genetic variants in the 17q21 haploblock that alter the expression of ORMDL3; however, the molecular mechanisms by which these variants perturb gene expression and the cell types in which this effect is most prominent are unclear. We found several 17q21 variants overlapped enhancers present mainly in primary immune cell types. CD4
+ T cells showed the greatest increase (threefold) in ORMDL3 expression in individuals carrying the asthma-risk alleles, where ORMDL3 negatively regulated interleukin-2 production. The asthma-risk variants rs4065275 and rs12936231 switched CTCF-binding sites in the 17q21 locus, and 4C-Seq assays showed that several distal cis-regulatory elements upstream of the disrupted ZPBP2 CTCF-binding site interacted with the ORMDL3 promoter region in CD4+ T cells exclusively from subjects carrying asthma-risk alleles. Overall, our results suggested that T cells are one of the most prominent cell types affected by 17q21 variants.- Published
- 2016
- Full Text
- View/download PDF
36. Reply.
- Author
-
Westernberg L, Schulten V, Sette A, and Peters B
- Published
- 2016
- Full Text
- View/download PDF
37. T-cell epitope conservation across allergen species is a major determinant of immunogenicity.
- Author
-
Westernberg L, Schulten V, Greenbaum JA, Natali S, Tripple V, McKinney DM, Frazier A, Hofer H, Wallner M, Sallusto F, Sette A, and Peters B
- Subjects
- Adult, Allergens genetics, Antigens, Plant genetics, Antigens, Plant immunology, Conserved Sequence, Epitopes, T-Lymphocyte genetics, Evolution, Molecular, Female, Gene Expression Profiling, Humans, Immunoglobulin E blood, Immunoglobulin E immunology, Male, Middle Aged, Poaceae genetics, Poaceae immunology, Pollen genetics, Pollen immunology, Rhinitis, Allergic, Seasonal immunology, Sequence Analysis, DNA, T-Lymphocytes immunology, T-Lymphocytes metabolism, Transcriptome, Young Adult, Allergens immunology, Cross Reactions immunology, Epitopes, T-Lymphocyte immunology
- Abstract
Background: Patients with pollen allergies are frequently polysensitized. Pollens contain epitopes that are conserved across multiple species., Objective: We sought to demonstrate that cross-reactive T cells that recognize conserved epitopes show higher levels of expansion than T cells recognizing monospecific epitopes because of more frequent stimulation., Method: RNA was sequenced from 9 pollens, and the reads were assembled de novo into more than 50,000 transcripts. T-cell epitopes from timothy grass (Phleum pratense) were examined for conservation in these transcripts, and this was correlated to their ability to induce T-cell responses. T cells were expanded in vitro with P pratense-derived peptides and tested for cross-reactivity to pollen extracts in ELISpot assays., Results: We found that antigenic proteins are more conserved than nonimmunogenic proteins in P pratense pollen. Additionally, P pratense epitopes that were highly conserved across pollens elicited more T-cell responses in donors with grass allergy than less conserved epitopes. Moreover, conservation of a P pratense peptide at the transcriptomic level correlated with the ability of that peptide to trigger T cells that were cross-reactive with other non-P pratense pollen extracts., Conclusion: We found a correlation between conservation of peptides in plant pollens and their T-cell immunogenicity within P pratense, as well as their ability to induce cross-reactive T-cell responses. T cells recognizing conserved epitopes might be more prominent because they can be stimulated by a broader range of pollens and thereby drive polysensitization in allergic donors. We propose that conserved peptides could potentially be used in diagnostic or immunomodulatory approaches that address the issue of polysensitization and target multiple pollen allergies., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
38. Transcriptional Profiling of Th2 Cells Identifies Pathogenic Features Associated with Asthma.
- Author
-
Seumois G, Zapardiel-Gonzalo J, White B, Singh D, Schulten V, Dillon M, Hinz D, Broide DH, Sette A, Peters B, and Vijayanand P
- Subjects
- Asthma genetics, Cell Separation, Enzyme-Linked Immunospot Assay, Gene Expression Profiling, Gene Regulatory Networks, Humans, Lymphocyte Activation genetics, Lymphocyte Activation immunology, Polymerase Chain Reaction, Rhinitis, Allergic genetics, Asthma immunology, Rhinitis, Allergic immunology, Th2 Cells immunology, Transcriptome immunology
- Abstract
Allergic asthma and rhinitis are two common chronic allergic diseases that affect the lungs and nose, respectively. Both diseases share clinical and pathological features characteristic of excessive allergen-induced type 2 inflammation, orchestrated by memory CD4(+) T cells that produce type 2 cytokines (Th2 cells). However, a large majority of subjects with allergic rhinitis do not develop asthma, suggesting divergence in disease mechanisms. Because Th2 cells play a pathogenic role in both these diseases and are also present in healthy nonallergic subjects, we performed global transcriptional profiling to determine whether there are qualitative differences in Th2 cells from subjects with allergic asthma, rhinitis, and healthy controls. Th2 cells from asthmatic subjects expressed higher levels of several genes that promote their survival as well as alter their metabolic pathways to favor persistence at sites of allergic inflammation. In addition, genes that enhanced Th2 polarization and Th2 cytokine production were also upregulated in asthma. Several genes that oppose T cell activation were downregulated in asthma, suggesting enhanced activation potential of Th2 cells from asthmatic subjects. Many novel genes with poorly defined functions were also differentially expressed in asthma. Thus, our transcriptomic analysis of circulating Th2 cells has identified several molecules that are likely to confer pathogenic features to Th2 cells that are either unique or common to both asthma and rhinitis., (Copyright © 2016 by The American Association of Immunologists, Inc.)
- Published
- 2016
- Full Text
- View/download PDF
39. The identification of potentially pathogenic and therapeutic epitopes from common human allergens.
- Author
-
Schulten V, Oseroff C, Alam R, Broide D, Vijayanand P, Peters B, Sette A, Schulten, Véronique, Oseroff, Carla, Alam, Rafeul, Broide, David, Vijayanand, Pandurangan, Peters, Bjoern, and Sette, Alessandro
- Published
- 2013
- Full Text
- View/download PDF
40. The MegaPool Approach to Characterize Adaptive CD4+ and CD8+ T Cell Responses.
- Author
-
da Silva Antunes R, Weiskopf D, Sidney J, Rubiro P, Peters B, Lindestam Arlehamn CS, Grifoni A, and Sette A
- Subjects
- Humans, Enzyme-Linked Immunospot Assay, Peptides, CD4-Positive T-Lymphocytes, CD8-Positive T-Lymphocytes, Epitopes, T-Lymphocyte
- Abstract
Epitopes recognized by T cells are a collection of short peptide fragments derived from specific antigens or proteins. Immunological research to study T cell responses is hindered by the extreme degree of heterogeneity of epitope targets, which are usually derived from multiple antigens; within a given antigen, hundreds of different T cell epitopes can be recognized, differing from one individual to the next because T cell epitope recognition is restricted by the epitopes' ability to bind to MHC molecules, which are extremely polymorphic in different individuals. Testing large pools encompassing hundreds of peptides is technically challenging because of logistical considerations regarding solvent-induced toxicity. To address this issue, we developed the MegaPool (MP) approach based on sequential lyophilization of large numbers of peptides that can be used in a variety of assays to measure T cell responses, including ELISPOT, intracellular cytokine staining, and activation-induced marker assays, and that has been validated in the study of infectious diseases, allergies, and autoimmunity. Here, we describe the procedures for generating and testing MPs, starting with peptide synthesis and lyophilization, as well as a step-by-step guide and recommendations for their handling and experimental usage. Overall, the MP approach is a powerful strategy for studying T cell responses and understanding the immune system's role in health and disease. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Generation of peptide pools ("MegaPools") Basic Protocol 2: MegaPool testing and quantitation of antigen-specific T cell responses., (© 2023 Wiley Periodicals LLC.)
- Published
- 2023
- Full Text
- View/download PDF
41. Association between specific timothy grass antigens and changes in TH1- and TH2-cell responses following specific immunotherapy.
- Author
-
Schulten V, Tripple V, Sidney J, Greenbaum J, Frazier A, Alam R, Broide D, Peters B, and Sette A
- Subjects
- Antigens, Plant chemistry, Cytokines immunology, Female, Humans, Immunoglobulin E immunology, Male, Peptides chemistry, Plant Proteins chemistry, Th1 Cells pathology, Th2 Cells pathology, Antigens, Plant administration & dosage, Desensitization, Immunologic, Hypersensitivity drug therapy, Hypersensitivity immunology, Hypersensitivity pathology, Peptides administration & dosage, Phleum chemistry, Plant Proteins administration & dosage, Th1 Cells immunology, Th2 Cells immunology
- Abstract
Background: Different populations of T cells are involved in the pathogenesis of allergic diseases., Objective: We investigated changes in TH-cell populations in patients with allergies after specific immunotherapy (SIT)., Methods: PBMCs were isolated from patients with allergies who received SIT and those who did not (controls). We tested the ability of peptides from 93 timothy grass (TG) proteins to induce T-cell responses (cytokine production). We used ELISPOT and staining assays for intracellular cytokines to measure the production of IL-4, IL-5, IL-13, IFN-γ, and IL-10., Results: Compared with PBMCs from controls, PBMCs from patients who received SIT produced lower levels of TH2 cytokines on incubation with several different TG peptides. These data were used to select 20 peptides to be tested in an independent cohort of 20 patients with allergies who received SIT and 20 controls. We again observed a significant decrease in the production of TH2 cytokines, and an increase in the production of the TH1 cytokine IFN-γ, in PBMCs from the validation groups. These changes correlated with improved symptoms after SIT. Immunization with this selected pool of peptides (or their associated antigens) could protect a substantial proportion of the population from TG allergy., Conclusions: We observed a significant decrease in the production of TH2 cytokines by PBMCs from patients who received SIT for TG allergy compared to those who did not. These changes might be used to monitor response to therapy. The decrease occurred in response to antigens that elicit little (if any) IgE responses; these antigens might be developed for use in immunotherapy., (Copyright © 2014 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
42. Immune Cell Molecular Pharmacodynamics of Lanreotide in Relation to Treatment Response in Patients with Gastroenteropancreatic Neuroendocrine Tumors.
- Author
-
Alaklabi, Sabah, Maguire, Orla, Pattnaik, Harsha, Zhang, Yali, Chow, Jacky, Wang, Jianmin, Minderman, Hans, and Iyer, Renuka
- Subjects
THERAPEUTIC use of antineoplastic agents ,GASTROINTESTINAL tumors ,IN vitro studies ,NF-kappa B ,CHEMOKINES ,PROTEINS ,HORMONES ,T cells ,RESEARCH funding ,CELL physiology ,CELL proliferation ,APOPTOSIS ,IMMUNE system ,CANCER patients ,TREATMENT effectiveness ,IN vivo studies ,TRANSCRIPTION factors ,CELLULAR signal transduction ,DESCRIPTIVE statistics ,PANCREATIC tumors ,ENERGY metabolism ,NEUROENDOCRINE tumors ,SOMATOSTATIN ,GENE expression profiling ,PROTEOLYTIC enzymes ,CYTOKINES ,DATA analysis software - Abstract
Simple Summary: Somatostatin analogs like lanreotide are considered first-line agents for the treatment of gastroenteropancreatic neuroendocrine tumors (NET). Although its effects on tumor proliferation and hormonal regulation are somewhat understood, its influence on the immune system has not been well elucidated. We used a double-pronged approach to understand the role of lanreotide in immune system regulation in healthy donor T cells in vitro as well as in vivo in cells obtained from 17 NET patients. We looked at cytokine signaling and differential gene expression to elucidate lanreotide's role in the treatment of NET through additional novel immune pathways. The CLARINET trial led to the approval of lanreotide for the treatment of patients with gastroenteropancreatic neuroendocrine tumors (NETs). It is hypothesized that lanreotide regulates proliferation, hormone synthesis, and other cellular functions via binding to somatostatin receptors (SSTR1–5) present in NETs. However, our knowledge of how lanreotide affects the immune system is limited. In vitro studies have investigated functional immune response parameters with lanreotide treatment in healthy donor T cell subsets, encompassing the breadth of SSTR expression, apoptosis induction, cytokine production, and activity of transcription factor signaling pathways. In our study, we characterized in vitro immune mechanisms in healthy donor T cells in response to lanreotide. We also studied the in vivo effects by looking at differential gene expression pre- and post-lanreotide therapy in patients with NET. Immune-focused gene and protein expression profiling was performed on peripheral blood samples from 17 NET patients and correlated with clinical response. In vivo, lanreotide therapy showed reduced effects on wnt, T cell receptor (TCR), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signaling in CD8+ T cells in responders compared to non-responders. Compared to non-responders, responders showed reduced effects on cytokine and chemokine signaling but greater effects on ubiquitination and proteasome degradation genes. Our results suggest significant lanreotide pharmacodynamic effects on immune function in vivo, which correlate with responses in NET patients. This is not evident from experimental in vitro settings. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
43. Strategies and delivery systems for cell-based therapy in autoimmunity.
- Author
-
Puccetti, Matteo, Costantini, Claudio, Schoubben, Aurélie, Giovagnoli, Stefano, and Ricci, Maurizio
- Published
- 2024
- Full Text
- View/download PDF
44. Heterogeneous IL-9 Production by Circulating Skin-Tropic and Extracutaneous Memory T Cells in Atopic Dermatitis Patients.
- Author
-
García-Jiménez, Irene, Sans-de San Nicolás, Lídia, Curto-Barredo, Laia, Bertolín-Colilla, Marta, Sensada-López, Eloi, Figueras-Nart, Ignasi, Bonfill-Ortí, Montserrat, Guilabert-Vidal, Antonio, Ryzhkova, Anna, Ferran, Marta, Damiani, Giovanni, Czarnowicki, Tali, Pujol, Ramon M., and Santamaria-Babí, Luis F.
- Subjects
HOUSE dust mites ,IMMUNOLOGIC memory ,ATOPIC dermatitis ,ECONOMIC stimulus ,INTERLEUKIN-9 ,T cells - Abstract
Interleukin (IL)-9 is present in atopic dermatitis (AD) lesions and is considered to be mainly produced by skin-homing T cells expressing the cutaneous lymphocyte-associated antigen (CLA). However, its induction by AD-associated triggers remains unexplored. Circulating skin-tropic CLA
+ and extracutaneous/systemic CLA− memory T cells cocultured with autologous lesional epidermal cells from AD patients were activated with house dust mite (HDM) and staphylococcal enterotoxin B (SEB). Levels of AD-related mediators in response to both stimuli were measured in supernatants, and the cytokine response was associated with different clinical characteristics. Both HDM and SEB triggered heterogeneous IL-9 production by CLA+ and CLA− T cells in a clinically homogenous group of AD patients, which enabled patient stratification into IL-9 producers and non-producers, with the former group exhibiting heightened HDM-specific and total IgE levels. Upon allergen exposure, IL-9 production depended on the contribution of epidermal cells and class II-mediated presentation; it was the greatest cytokine produced and correlated with HDM-specific IgE levels, whereas SEB mildly induced its release. This study demonstrates that both skin-tropic and extracutaneous memory T cells produce IL-9 and suggests that the degree of allergen sensitization reflects the varied IL-9 responses in vitro, which may allow for patient stratification in a clinically homogenous population. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
45. In Silico Identification of Peanut Peptides Suitable for Allergy Immunotherapy in HLA-DRB1*03:01-Restricted Patients.
- Author
-
Doytchinova, Irini, Atanasova, Mariyana, Sotirov, Stanislav, and Dimitrov, Ivan
- Subjects
PEANUT allergy ,HLA histocompatibility antigens ,ALLERGIES ,PEPTIDES ,IMMUNOLOGICAL tolerance ,PEANUTS - Abstract
Peanut allergy, a prevalent and potentially severe condition affecting millions worldwide, has been linked to specific human leukocyte antigens (HLAs), suggesting increased susceptibility. Employing an immunoinformatic strategy, we developed a "logo model" based on amino acid frequencies in the peptide binding core and used it to predict peptides originating from 28 known peanut allergens binding to HLA-DRB1*03:01, one of the susceptibility alleles. These peptides hold promise for immunotherapy in HLA-DRB1*03:01 carriers, offering reduced allergenicity compared to whole proteins. By targeting essential epitopes, immunotherapy can modulate immune responses with minimal risk of severe reactions. This precise approach could induce immune tolerance with fewer adverse effects, presenting a safer and more effective treatment for peanut allergy and other allergic conditions. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
46. Allergy-associated T cell epitope repertoires are surprisingly diverse and include non-IgE reactive antigens.
- Author
-
Frazier A, Schulten V, Hinz D, Oseroff C, Sidney J, Peters B, and Sette A
- Abstract
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.
- Published
- 2014
- Full Text
- View/download PDF
47. New strategies for allergen T cell epitope identification: going beyond IgE.
- Author
-
Schulten V, Peters B, and Sette A
- Subjects
- Antigens, Plant immunology, Desensitization, Immunologic, Humans, Hypersensitivity, Immediate immunology, Hypersensitivity, Immediate therapy, Immunoglobulin E immunology, Phleum immunology, Allergens immunology, Epitope Mapping methods, Epitopes, T-Lymphocyte immunology
- Abstract
Background: Type I allergy and allergic asthma are common diseases in the developed world associated with IgE antibodies and Th2 cell reactivity. To date, the only causative treatment for allergic disease is specific immunotherapy (SIT)., Method: Here, we review recent works from our laboratory focused on identifying human T cell epitopes associated with allergic disease and their potential use as biomarkers or therapeutic targets for SIT. In previous studies, we have mapped T cell epitopes associated with the major 10 timothy grass (Tg) allergens, defined on the basis of human IgE reactivity by ELISPOT., Results: Interestingly, in about 33% of allergic donors, no T cell epitopes from overlapping peptides spanning the entire sequences of these allergens were identified despite vigorous T cell responses to the Tg extract. Using a bioinformatic-proteomic approach, we identified a set of 93 novel Tg proteins, many of which were found to elicit IL-5 production in T cells from allergic donors despite lacking IgE reactivity. Next, we assessed T cell responses to the novel Tg proteins in donors who had been treated with subcutaneous SIT. A subset of these proteins showed a strong reduction of IL-5 responses in donors who had received subcutaneous SIT compared to allergic donors, which correlated with patients' self-reported improvement of allergic symptoms., Conclusion: A bioinformatic-proteomic approach has successfully identified additional Tg-derived T cell targets independent of IgE reactivity. This method can be applied to other allergies potentially leading to the discovery of promising therapeutic targets for allergen-specific immunotherapy., (© 2014 S. Karger AG, Basel.)
- Published
- 2014
- Full Text
- View/download PDF
48. An update on recent developments and highlights in food allergy.
- Author
-
Locke A, Hung L, Upton JEM, O'Mahony L, Hoang J, and Eiwegger T
- Subjects
- Humans, Food, Allergens, Immunotherapy, Desensitization, Immunologic, Food Hypersensitivity therapy, Peanut Hypersensitivity
- Abstract
While both the incidence and general awareness of food allergies is increasing, the variety and clinical availability of therapeutics remain limited. Therefore, investigations into the potential factors contributing to the development of food allergy (FA) and the mechanisms of natural tolerance or induced desensitization are required. In addition, a detailed understanding of the pathophysiology of food allergies is needed to generate compelling, enduring, and safe treatment options. New findings regarding the contribution of barrier function, the effect of emollient interventions, mechanisms of allergen recognition, and the contributions of specific immune cell subsets through rodent models and human clinical studies provide novel insights. With the first approved treatment for peanut allergy, the clinical management of FA is evolving toward less intensive, alternative approaches involving fixed doses, lower maintenance dose targets, coadministration of biologicals, adjuvants, and tolerance-inducing formulations. The ultimate goal is to improve immunotherapy and develop precision-based medicine via risk phenotyping allowing optimal treatment for each food-allergic patient., (© 2023 The Authors. Allergy published by European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.)
- Published
- 2023
- Full Text
- View/download PDF
49. A strategy to determine HLA class II restriction broadly covering the DR, DP, and DQ allelic variants most commonly expressed in the general population.
- Author
-
McKinney DM, Southwood S, Hinz D, Oseroff C, Arlehamn CS, Schulten V, Taplitz R, Broide D, Hanekom WA, Scriba TJ, Wood R, Alam R, Peters B, Sidney J, and Sette A
- Subjects
- Alleles, Antigen Presentation, B-Lymphocytes immunology, Cells, Cultured, Epitopes immunology, HLA-DP Antigens immunology, HLA-DQ Antigens immunology, HLA-DR Antigens immunology, Histocompatibility Antigens Class II immunology, Humans, Peptide Fragments genetics, Peptide Fragments immunology, T-Lymphocytes immunology, Genetic Variation genetics, Genetics, Population, HLA-DP Antigens genetics, HLA-DQ Antigens genetics, HLA-DR Antigens genetics, Histocompatibility Antigens Class II genetics
- Abstract
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.
- Published
- 2013
- Full Text
- View/download PDF
50. Previously undescribed grass pollen antigens are the major inducers of T helper 2 cytokine-producing T cells in allergic individuals.
- Author
-
Schulten V, Greenbaum JA, Hauser M, McKinney DM, Sidney J, Kolla R, Lindestam Arlehamn CS, Oseroff C, Alam R, Broide DH, Ferreira F, Grey HM, Sette A, and Peters B
- Subjects
- Allergens immunology, Antibodies immunology, Electrophoresis, Gel, Two-Dimensional, Epitopes immunology, Gene Expression Profiling, Humans, Hypersensitivity genetics, Immunoblotting, Immunoglobulin E immunology, Immunologic Memory immunology, Molecular Sequence Data, Plant Extracts immunology, Plant Proteins immunology, Proteomics, Tissue Donors, Antigens, Plant immunology, Hypersensitivity immunology, Interleukin-5 biosynthesis, Phleum immunology, Pollen immunology, Th2 Cells immunology
- Abstract
T cells play an important role in the pathogenesis of allergic diseases. However, the proteins considered as potential immunogens of allergenic T-cell responses have traditionally been limited to those that induce IgE responses. Timothy grass (TG) pollen is a well-studied inhaled allergen for which major IgE-reactive allergens have also been shown to trigger T helper 2 (Th2) responses. Here we examined whether other TG pollen proteins are recognized by Th2 responses independently of IgE reactivity. A TG pollen extract was analyzed by 2D gel electrophoresis and IgE/IgG immunoblots using pooled sera from allergic donors. Mass spectrometry of selected protein spots in combination with de novo sequencing of the whole TG pollen transcriptome identified 93 previously undescribed proteins for further study, 64 of which were not targeted by IgE. Predicted MHC binding peptides from the previoulsy undescribed TG proteins were screened for T-cell reactivity in peripheral blood mononuclear cells from allergic donors. Strong IL-5 production was detected in response to peptides from several of the previously undescribed proteins, most of which were not targeted by IgE. Responses against the dominant undescribed epitopes were associated with the memory T-cell subset and could even be detected directly ex vivo after Th2 cell enrichment. These findings demonstrate that a combined unbiased transcriptomic, proteomic, and immunomic approach identifies a greatly broadened repertoire of protein antigens targeted by T cells involved in allergy pathogenesis. The discovery of proteins that induce Th2 cells but are not IgE reactive may allow the development of safer immunotherapeutic strategies.
- Published
- 2013
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.