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  • 1.
    Akdis, M
    et al.
    SIAF, Switzerland.
    Verhagen, J
    SIAF, Switzerland.
    Taylor, A
    SIAF, Switzerland.
    Karamloo, F
    SIAF, Switzerland.
    Karagiannidis, C
    SIAF, Switzerland.
    Crameri, R
    SIAF, Switzerland.
    Thunberg, Sarah
    Karolinska Institutet.
    Deniz, G
    SIAF, Switzerland.
    Valenta, R
    SIAF, Switzerland.
    Fiebig, H
    SIAF, Switzerland.
    Kegel, C
    SIAF, Switzerland.
    Disch, R
    SIAF, Switzerland.
    Schmidt-Weber, C B
    SIAF, Switzerland.
    Blaser, K
    SIAF, Switzerland.
    Akdis, C A
    SIAF, Switzerland.
    Immune responses in healthy and allergic individuals are characterized by a fine balance between allergen-specific T regulatory 1 and T helper 2 cells2004In: Journal of Experimental Medicine, ISSN 0022-1007, E-ISSN 1540-9538, Vol. 199, no 11, p. 1567-1575Article in journal (Refereed)
    Abstract [en]

    The mechanisms by which immune responses to nonpathogenic environmental antigens lead to either allergy or nonharmful immunity are unknown. Single allergen-specific T cells constitute a very small fraction of the whole CD4(+) T cell repertoire and can be isolated from the peripheral blood of humans according to their cytokine profile. Freshly purified interferon-gamma-, interleukin (IL)-4-, and IL-10-producing allergen-specific CD4(+) T cells display characteristics of T helper cell (Th)1-, Th2, and T regulatory (Tr)1-like cells, respectively. Tr1 cells consistently represent the dominant subset specific for common environmental allergens in healthy individuals; in contrast, there is a high frequency of allergen-specific IL-4-secreting T cells in allergic individuals. Tr1 cells use multiple suppressive mechanisms, IL-10 and TGF-beta as secreted cytokines, and cytotoxic T lymphocyte antigen 4 and programmed death 1 as surface molecules. Healthy and allergic individuals exhibit all three allergen-specific subsets in different proportions, indicating that a change in the dominant subset may lead to allergy development or recovery. Accordingly, blocking the suppressor activity of Tr1 cells or increasing Th2 cell frequency enhances allergen-specific Th2 cell activation ex vivo. These results indicate that the balance between allergen-specific Tr1 cells and Th2 cells may be decisive in the development of allergy.

  • 2. Emelie, Radestad
    et al.
    Emma, Watz
    Sarah, Thunberg
    KTH, School of Engineering Sciences (SCI), Applied Physics. Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Stockholm, Sweden.
    Mikael, Sundin
    Jonas, Mattsson
    Uhlin, Michael
    KTH, School of Engineering Sciences (SCI), Applied Physics. Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Stockholm, SwedenKarolinska Univ Hosp, Dept Clin Immunol & Transfus Med, Huddinge, Sweden.
    Individualization of hematopoietic stem cell transplantation using alpha/ beta depletion2017In: Scandinavian Journal of Immunology, ISSN 0300-9475, E-ISSN 1365-3083, Vol. 86, no 4, p. 338-338Article in journal (Other academic)
  • 3.
    Gafvelin, G
    et al.
    Karolinska institutet.
    Thunberg, Sarah
    Karolinska Institutet.
    Kronqvist, M
    Karolinska institutet.
    Gronlund, H
    Karolinska institutet.
    Gronneberg, R
    Karolinska institutet.
    Troye-Blomberg, M
    Karolinska institutet.
    Akdis, M
    SIAF, switzerland.
    Fiebig, H
    Allergopharma, Germany.
    Purohit, A
    Hôpitaux Universitaires de Strasbourg, France.
    Horak, F
    Medical University of Vienna, Austria.
    Reisinger, J
    Medical University of Vienna, Austria.
    Niederberger, V
    Medical University of Vienna, Austria.
    Akdis, C A
    SIAF Switzerland.
    Cromwell, O
    Allergopharma, Germany.
    Pauli, G
    Hôpitaux Universitaires de Strasbourg, France.
    Valenta, R
    Medical University of Vienna, Austria.
    van Hage, M
    Karolinska institutet.
    Cytokine and antibody responses in birch-pollen-allergic patients treated with genetically modified derivatives of the major birch pollen allergen Bet v 12005In: International Archives of Allergy and Immunology, ISSN 1018-2438, E-ISSN 1423-0097, Vol. 138, no 1, p. 59-66Article in journal (Refereed)
    Abstract [en]

    Background: Recently, recombinant hypoallergenic derivatives of the major birch pollen allergen, Bet v 1, were used to treat birch-pollen-allergic patients in a double-blind, placebo-controlled, multi-centre immunotherapy study. The aim of this study was to evaluate the effects of vaccination with aluminium-hydroxide-adsorbed recombinant Bet v 1 derivatives versus placebo on T-cell, cytokine and antibody responses in a subgroup of patients. Methods: Blood was drawn from patients of the Swedish centre (n=27; rBet v 1 fragments: n=10; rBet v 1 trimer: n=8, and placebo-aluminium hydroxide: n=9) before the start and after completion of the treatment. PBMC were stimulated with rBet v 1 and analysed for cytokine (IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-gamma)-secreting cells by ELISpot. Bet v 1-specific antibody levels in serum (IgG(1-4), IgE and IgA) were measured by ELISA. Skin prick tests with defined Bet v 1 concentrations were performed before and 10-11 months after the beginning of the study. Results: Bet v 1-specific IgG levels, consisting of IgG 1, IgG 2 and IgG 4, were significantly increased after treatment with recombinant allergen derivatives. Treatment with rBet v 1 trimer led to a significant (p<0.05) reduction of Bet v 1-reactive IL-5- and IL-13-producing cells, reflecting a reduced Th2 response. In addition, a decreased number of Bet v 1-reactive IL-4 producing (p=0.07) and an increase of IL-12-producing (p=0.06) cells was noted in the trimer-treated patients. In contrast to placebo, active treatment resulted in significantly reduced immediate-type skin reactions to Bet v 1 even 10-11 months after treatment. Conclusion: Vaccination with recombinant hypoallergenic Bet v 1 derivatives induces a Bet v 1-specific IgG response and leads to reduced skin reactivity in allergic patients. A reduction of Bet v 1-specific Th2 responses was observed in trimer-treated patients, which may reflect the intrinsic property of this allergen derivative. Copyright (C) 2005 S. Karger AG, Basel.

  • 4.
    Grundstroem, J.
    et al.
    Karolinska Institutet, Sweden.
    Linton, L.
    Karolinska Institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Forsslund, H.
    Karolinska Institutet, Sweden.
    Janczewska, I.
    Karolinska Institutet, Sweden.
    Befrits, R.
    Karolinska university Hospital, Sweden.
    van Hage, M.
    Karolinska Institutet, Sweden.
    Gafvelin, G.
    Karolinska Institutet, Sweden.
    Eberhardson, M.
    Karolinska Institutet, Sweden.
    Altered immunoregulatory profile during anti-tumour necrosis factor treatment of patients with inflammatory bowel disease2012In: Clinical and Experimental Immunology, ISSN 0009-9104, E-ISSN 1365-2249, Vol. 169, no 2, p. 137-147Article in journal (Refereed)
    Abstract [en]

    Inflammatory bowel disease (IBD) can be treated effectively by anti-tumour necrosis factor (TNF) therapy. We set out to investigate the unclear immunoregulatory mechanisms of the treatment. Thirty-four patients with IBD treated with anti-TNF were included. Lymphocytes from peripheral blood and intestinal biopsies were analysed by flow cytometry. Regulation of antigen-stimulated proliferation was analysed by blocking of interleukin (IL)-10, transforming growth factor (TGF)-beta or depletion of CD25+ cells in peripheral blood mononuclear cell cultures. No changes in CD4+CD25+, CD25+TNF-RII+ or CD4+CD25+forkhead box protein 3 (FoxP3+) T cells could be observed in peripheral blood after, in comparison to before, 6 weeks of treatment. The suppressive ability of CD4+CD25+ cells did not change. There was an initial decrease of CD4+CD25+ cells in intestinal mucosa after 2 weeks of treatment, followed by an increase of these cells from weeks 2 to 6 of treatment (P < 0.05). This was accompanied by an increased percentage of CD69+ cells among these cells after 6 weeks of treatment compared to before treatment (P < 0.01). There was also an increase of mucosal T helper type1 cells from weeks 2 to 6 (P < 0.05). In addition, CD25+TNF-RII+ cells in the mucosa were decreased after 6 weeks of treatment compared to before treatment (P < 0.05). Before treatment, peripheral blood mononuclear cell baseline proliferation was increased when IL-10 was blocked (P < 0.01), but not after. In CD25+ cell-depleted cultures proliferation increased after treatment (P < 0.05). Our data indicate that anti-TNF treatment leads to an induction of effector T cells. Anti-TNF therapy has no significant impact on regulatory T cells in IBD, although the composition of regulatory T cell subsets may change during treatment.

  • 5.
    Kaiser, Liselotte
    et al.
    Karolinska Institutet, Sweden.
    Velickovic, Tanja Cirkovic
    Karolinska Institutet, Sweden.
    Badia-Martinez, Daniel
    Karolinska Institutet, Sweden.
    Adedoyin, Justus
    Karolinska Institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Hallen, Dan
    Electrum, Sweden.
    Berndt, Kurt
    Karolinska Institutet, Sweden.
    Gronlund, Hans
    Karolinska Institutet, Sweden.
    Gafvelin, Guro
    Karolinska Institutet, Sweden.
    van Hage, Marianne
    Karolinska Institutet, Sweden.
    Achour, Adnane
    Karolinska Institutet, Sweden.
    Structural characterization of the tetrameric form of the major cat allergen Fel d 12007In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 370, no 4, p. 714-727Article in journal (Refereed)
    Abstract [en]

    Felis domesticus allergen 1(Fel d 1) is a 35 kDa tetrameric glycoprotein formed by two heterodimers which elicits IgE responses in 95% of patients with allergy to cat. We have previously established in vitro conditions for the appropriate folding of recombinant Fel d 1 using a direct linkage of chain I to chain 2 (construct Fel d 1 (1 + 2)) and chain 2 to chain 1 (construct Fel d 1 (2 + 1)). Although the crystal structure of Fel d 1 (2 + 1) revealed a striking structural similarity to that of uteroglobin, a steroid-inducible cytokine-like molecule with anti-inflammatory and immunomodulatory properties, no functional tetrameric form of Fel d I could be identified. Here we present the crystal structure of the Fel d I (1 +2) tetramer at 1.6 angstrom resolution. Interestingly, the crystal structure of tetrameric Fel d I reveals two different calcium-binding sites. Symmetrically positioned on each side of the Fel d 1 tetramer, the external Ca2+ -binding sites correspond to a putative Ca2+- binding site previously suggested for uteroglobin. The second Ca2+-binding site lies within the dimerization interface, stabilizing the formation of the Fel d 1 tetramer, and inducing important local conformational changes that directly govern the shape of two water-filled cavities. The crystal structure suggests a potential portal for an unknown ligand. Alternatively, the two cavities could be used by the allergen as a conditional inner space allowing for the spatial rearrangement of centrally localized side-chains, such as Asp130, without altering the overall fold of the molecule. The striking structural similarity of the major cat allergen to uteroglobin, coupled to the identification in the present study of a common Ca2+ -binding site, let us speculate that Fel d I could provoke an allergic response through the modulation of phospholipase A2, by sequestering Ca ions in a similar manner as previously suggested for uteroglobin. (c) 2007 Elsevier Ltd. All rights reserved.

  • 6.
    Lundstrom, Susanna L.
    et al.
    Karolinska Institutet, Sweden.
    Yang, Jun
    University of California, USA.
    Kallberg, Henrik J.
    Karolinska Institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Gafvelin, Guro
    Karolinska Institutet, Sweden.
    Haeggstrom, Jesper Z.
    Karolinska Institutet, Sweden.
    Gronneberg, Reidar
    Karolinska Institutet, Sweden.
    Grunewald, Johan
    Karolinska Institutet, Sweden.
    van Hage, Marianne
    Karolinska Institutet, Sweden.
    Hammock, Bruce D.
    University of California, USA.
    Eklund, Anders
    Karolinska Institutet, Sweden.
    Wheelock, Asa M.
    Karolinska Institutet, Sweden.
    Wheelock, Craig E.
    Karolinska Institutet, Sweden.
    Allergic Asthmatics Show Divergent Lipid Mediator Profiles from Healthy Controls Both at Baseline and following Birch Pollen Provocation2012In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 7, no 3Article in journal (Refereed)
    Abstract [en]

    Background: Asthma is a respiratory tract disorder characterized by airway hyper-reactivity and chronic inflammation. Allergic asthma is associated with the production of allergen-specific IgE and expansion of allergen-specific T-cell populations. Progression of allergic inflammation is driven by T-helper type 2 (Th2) mediators and is associated with alterations in the levels of lipid mediators. Objectives: Responses of the respiratory system to birch allergen provocation in allergic asthmatics were investigated. Eicosanoids and other oxylipins were quantified in the bronchoalveolar lumen to provide a measure of shifts in lipid mediators associated with allergen challenge in allergic asthmatics. Methods: Eighty-seven lipid mediators representing the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 (CYP) metabolic pathways were screened via LC-MS/MS following off-line extraction of bronchoalveolar lavage fluid (BALF). Multivariate statistics using OPLS were employed to interrogate acquired oxylipin data in combination with immunological markers. Results: Thirty-two oxylipins were quantified, with baseline asthmatics possessing a different oxylipin profile relative to healthy individuals that became more distinct following allergen provocation. The most prominent differences included 15-LOX-derived omega-3 and omega-6 oxylipins. Shared-and-Unique-Structures (SUS)-plot modeling showed a correlation (R-2 = 0.7) between OPLS models for baseline asthmatics ((RY)-Y-2[cum] = 0.87, Q(2)[cum] = 0.51) and allergen-provoked asthmatics ((RY)-Y-2[cum] = 0.95, Q(2)[cum] = 0.73), with the majority of quantified lipid mediators and cytokines contributing equally to both groups. Unique structures for allergen provocation included leukotrienes (LTB4 and 6-trans-LTB4), CYP-derivatives of linoleic acid (epoxides/diols), and IL-10. Conclusions: Differences in asthmatic relative to healthy profiles suggest a role for 15-LOX products of both omega-6 and omega-3 origin in allergic inflammation. Prominent differences at baseline levels indicate that non-symptomatic asthmatics are subject to an underlying inflammatory condition not observed with other traditional mediators. Results suggest that oxylipin profiling may provide a sensitive means of characterizing low-level inflammation and that even individuals with mild disease display distinct phenotypic profiles, which may have clinical ramifications for disease.

  • 7.
    Marits, Per
    et al.
    Karolinska Institutet, Sweden.
    Wikstrom, Ann-Charlotte
    Karolinska Institutet, Sweden.
    Popadic, Dusan
    University of Belgrade, Serbia.
    Winqvist, Ola
    Karolinska institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Evaluation of T and B lymphocyte function in clinical practice using a flow cytometry based proliferation assay2014In: Clinical Immunology, ISSN 1521-6616, E-ISSN 1521-7035, Vol. 153, no 2, p. 332-342Article in journal (Refereed)
    Abstract [en]

    The golden standard for functional evaluation of immunodeficiencies is the incorporation of [H-3]-thymidine in a proliferation assay stimulated with mitogens. Recently developed whole blood proliferation assays have the advantage of parallel lymphocyte lineage analysis and in addition provide a non-radioactive alternative. Here we evaluate the Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) in a comparison with [H-3]-thymidine incorporation in four patients with severe combined immunodeficiency. The threshold for the mininium number of lymphocytes required for reliable responses in FASCIA is determined together with reference values from 100 healthy donors when stimulated with mitogens as well as antigen specific stimuli. Finally, responses against PWM and SEA + SEB stimuli are conducted with clinically relevant immunomodulatory compounds. We conclude that FASCIA is a rapid, stable and sensitive functional whole blood assay that requires small amounts of whole blood that can be used for reliable assessment of lymphocyte reactivity in patients. (C) 2014 Elsevier Inc. All rights reserved.

  • 8.
    Mustjoki, S.
    et al.
    University of Helsinki, Finland.
    Richter, J.
    Skåne university hospital, Sweden.
    Barbany, G.
    Karolinska institutet, Sweden.
    Ehrencrona, H.
    Skåne university hospital, Sweden.
    Fioretos, T.
    Skåne university hospital, Sweden.
    Gedde-Dahl, T.
    Oslo university hospital, Norway.
    Gjertsen, B. T.
    Haukeland university hospital, Norway.
    Hovland, R.
    Haukeland university hospital, Norway.
    Hernesniemi, S.
    University of Helsinki, Finland.
    Josefsen, D.
    The noewegian radium hospital, Norway.
    Koskenvesa, P.
    University of Helsinki, Finland.
    Dybedal, I.
    Oslo university hospital, Norway.
    Markevarn, B.
    Umeå university hospital, Sweden.
    Olofsson, T.
    Skåne university hospital, Sweden.
    Olsson-Stromberg, U.
    Uppsala university hospital, Sweden.
    Rapakko, K.
    Oulo university hospital, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Stockholm, Sweden.
    Stenke, L.
    Karolinska Institutet, Sweden.
    Simonsson, B.
    Uppsala university hospital.
    Porkka, K.
    Norwegian University of Science and Technology, Norway.
    Hjorth-Hansen, H.
    Norwegian University of Science and Technology, Norway.
    Grp, Nordic CML Study
    Impact of malignant stem cell burden on therapy outcome in newly diagnosed chronic myeloid leukemia patients2013In: Leukemia, ISSN 0887-6924, E-ISSN 1476-5551, Vol. 27, no 7, p. 1520-1526Article in journal (Refereed)
    Abstract [en]

    Chronic myeloid leukemia (CML) stem cells appear resistant to tyrosine kinase inhibitors (TKIs) in vitro, but their impact and drug sensitivity in vivo has not been systematically assessed. We prospectively analyzed the proportion of Philadelphia chromosome-positive leukemic stem cells (LSCs, Ph+CD34+CD38=) and progenitor cells (LPCs, Ph+CD34+CD38+) from 46 newly diagnosed CML patients both at the diagnosis and during imatinib or dasatinib therapy (ClinicalTrials.gov NCT00852566). At diagnosis, the proportion of LSCs varied markedly (1-100%) between individual patients with a significantly lower median value as compared with LPCs (79% vs 96%, respectively, P = 0.0001). The LSC burden correlated with leukocyte count, spleen size, hemoglobin and blast percentage. A low initial LSC percentage was associated with less therapy-related hematological toxicity and superior cytogenetic and molecular responses. After initiation of TKI therapy, the LPCs and LSCs rapidly decreased in both therapy groups, but at 3 months time point the median LPC level was significantly lower in dasatinib group compared with imatinib patients (0.05% vs 0.68%, P = 0.032). These data detail for the first time the prognostic significance of the LSC burden at diagnosis and show that in contrast to in vitro data, TKI therapy rapidly eradicates the majority of LSCs in patients.

  • 9. Neimert-Andersson, T.
    et al.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Swedin, L.
    Karolinska Institutet, Sweden.
    Wiedermann, U.
    Medical University Vienna, Austria.
    Jacobsson-Ekman, G.
    Karolinska Institutet, Sweden.
    Dahlen, S. -E
    Karolinska Institutet, Sweden.
    Scheynius, A.
    Karolinska Institutet, Sweden.
    Gronlund, H.
    Karolinska Institutet, Sweden.
    van Hage, M.
    Karolinska Institutet, Sweden.
    Gafvelin, G.
    Karolinska Institutet, Sweden.
    Carbohydrate-based particles reduce allergic inflammation in a mouse model for cat allergy2008In: Allergy. European Journal of Allergy and Clinical Immunology, ISSN 0105-4538, E-ISSN 1398-9995, Vol. 63, no 5, p. 518-526Article in journal (Refereed)
    Abstract [en]

    Background: Allergen-specific immunotherapy (ASIT) is the only treatment of allergic disease that gives long-lasting relief of symptoms. However, concerns for safety and efficiency have highlighted the need for improvement of the therapy. We have previously suggested carbohydrate-based particles (CBPs) as a novel adjuvant and allergen carrier for ASIT. Our aim of this study was to evaluate the therapeutic potential of CBPs in ASIT, employing a mouse model for cat allergy. Methods: BALB/c mice were subcutaneously immunized with the recombinant (r) cat allergen Fel d 1 followed by intranasal challenge with cat dander extract (CDE). The sensitized mice were therapeutically treated with rFel d 1 covalently coupled to CBPs (CBP-rFel d 1). Airway hyper-reactivity (AHR), infiltration of leucocytes in bronchoalveolar lavage (BAL) fluid, allergen-specific serum immunoglobulin levels and in vitro splenocyte responses were evaluated. Results: Mice treated with CBP-rFel d 1 showed reduced features of allergic inflammation. They responded with (i) significantly decreased AHR and infiltration of eosinophils in BAL fluid after CDE challenge, (ii) the serum level of rFel d 1-specific IgE was reduced and the level of IgG(2)a was more pronounced after CBP-rFel d 1 treatment, and (iii) there was also a tendency of decreased allergen-specific cellular response. Conclusions: Carbohydrate-based particles are effective tools as adjuvant and allergen carriers for use in ASIT and constitutes a promising strategy to improve allergy treatment.

  • 10. Nilsson, J.
    et al.
    Granrot, I.
    Mattsson, J.
    Omazic, B.
    Uhlin, Michael
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
    Thunberg, Sarah
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
    Functionality testing of stem cell grafts to predict infectious complications after allogeneic hematopoietic stem cell transplantation2017In: Vox Sanguinis, ISSN 0042-9007, E-ISSN 1423-0410, Vol. 112, no 5, p. 459-468Article in journal (Refereed)
    Abstract [en]

    Background and Objectives Allogeneic hematopoietic stem cell transplantation (HSCT) is a routine clinical procedure performed to treat patients with haematological malignancies, primary immune deficiencies or metabolic disorders. Infections during lymphopenia after allogeneic HSCT are associated with high mortality and morbidity. Typical infectious agents are Epstein-Barr virus, cytomegalovirus, herpes simplex virus, varicella-zoster virus and fungi. The study aim was to evaluate whether measurement of the responses of antigen-specific T-cells, recognizing infectious pathogens would correlate to protective functions in the stem cell recipient post-transplant. Materials and MethodsTwenty-one grafts were analysed by flow cytometry and cells were stimulated in vitro with relevant infectious antigens, followed by evaluation of T-cell proliferation and cytokine production. Results were compared to the recipients' clinical records 1-year post-transplantation. ResultsWe show that an extensive repertoire of transferred antigen-specific T-cells from allogeneic donor grafts against infectious agents, involved in post-transplant infections, are linked to an absence of infectious complications for the recipient up-to 1-year post-transplant. The protective effect was associated with antigen-specific T-cell proliferation and IL-1 secretion. Conclusion Our results suggest that assaying T-cell function before HSCT could determine individual risks for infectious complications and thus aid in clinical decision-making regarding prophylactic and pre-emptive anti-infective therapy.

  • 11.
    Radestad, E.
    et al.
    Karolinska institutet, Sweden.
    Wikell, H.
    Karolinska institutet, Sweden.
    Engstrom, M.
    Karolinska university hospital, Sweden.
    Watz, E.
    Karolinska university hospital, Sweden.
    Sundberg, B.
    Karolinska institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Uzunel, M.
    Karolinska Institutet, Sweden.
    Mattsson, J.
    Karolinska Institutet, Sweden.
    Uhlin, M.
    Karolinska Institutet, Sweden.
    Alpha/Beta T-Cell Depleted Grafts as an Immunological Booster to Treat Graft Failure after Hematopoietic Stem Cell Transplantation with HLA-Matched Related and Unrelated Donors2014In: Journal of Immunology Research, ISSN 2314-8861, E-ISSN 2314-7156Article in journal (Refereed)
    Abstract [en]

    Allogeneic hematopoietic stem cell transplantation is associated with several complications and risk factors, for example, graft versus host disease (GVHD), viral infections, relapse, and graft rejection. While high levels of CD3+ cells in grafts can contribute to GVHD, they also promote the graft versus leukemia (GVL) effect. Infusions of extra lymphocytes from the original stem cell donor can be used as a treatment after transplantation for relapse or poor immune reconstitution but also they increase the risk for GVHD. In peripheral blood, 95% of T-cells express the alpha beta T-cell receptor and the remaining T-cells express the gamma delta. T-cell receptor. As alpha beta T-cells are the primary mediators of GVHD, depleting them from the graft should reduce this risk. In this pilot study, five patients transplanted with HLA-matched related and unrelated donors were treated with alpha beta T-cell depleted stem cell boosts. The majority of gamma delta T-cells in the grafts expressed V delta 2 and/or V gamma 9. Most patients receiving alpha beta-depleted stem cell boosts increased their levels of white blood cells, platelets, and/or granulocytes 30 days after infusion. No signs of GVHD or other side effects were detected. A larger pool of patients with longer follow-up time is needed to confirm the data in this study.

  • 12.
    Radestad, Emelie
    et al.
    Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Div Transplantat Surg, Stockholm, Sweden..
    Sundin, Mikael
    Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Div Pediat, Stockholm, Sweden.;Karolinska Univ Hosp, Astrid Lindgren Childrens Hosp, Hematol Immunol HSCT Sect, Stockholm, Sweden..
    Torlen, Johan
    Karolinska Univ Hosp, Cell Therapy & Allogene Stem Cell Transplantat, Stockholm, Sweden.;Karolinska Inst, Dept Oncol Pathol, Stockholm, Sweden..
    Thunberg, Sara
    KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences (SCI), Applied Physics.
    Önfelt, Björn
    KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Ljungman, Per
    Karolinska Univ Hosp, Cell Therapy & Allogene Stem Cell Transplantat, Stockholm, Sweden.;Karolinska Inst, Div Hematol, Dept Med, Stockholm, Sweden..
    Watz, Emma
    Karolinska Inst, Dept Clin Sci Intervent & Technol CLINTEC, Div Transplantat Surg, Stockholm, Sweden.;Karolinska Univ Hosp, Dept Clin Immunol & Transfus Med, Stockholm, Sweden..
    Mattsson, Jonas
    Karolinska Inst, Dept Oncol Pathol, Stockholm, Sweden.;Princess Margaret Canc Ctr, Div Med Oncol & Hematol, Toronto, ON, Canada.;Univ Toronto, Toronto, ON, Canada..
    Uhlin, Michael
    KTH, School of Engineering Sciences (SCI), Applied Physics, Biophysics.
    Individualization of Hematopoietic Stem Cell Transplantation Using Alpha/Beta T-Cell Depletion2019In: Frontiers in Immunology, ISSN 1664-3224, E-ISSN 1664-3224, Vol. 10, article id 189Article in journal (Refereed)
    Abstract [en]

    Allogeneic hematopoietic stem cell transplantation (HSCT) is associated with several potentially lethal complications. Higher levels of CD3+ T-cells in the graft have been associated with increased risk of graft-versus-host disease (GVHD), but also beneficial graft-versus-leukemia effect and reduced infections. To tackle post-transplant complications, donor lymphocyte infusions have been used but with an increased risk of GVHD. To reduce this risk, we performed depletion of alpha beta T-cells and treated 12 patients post-HSCT suffering from infections and/or poor immune reconstitution. The alpha beta T-cell depleted cell products were characterized by flow cytometry. The median log depletion of alpha beta T-cells was -4.3 and the median yield of gamma delta T-cells was 73.5%. The median CD34+ cell dose was 4.4 x 10(6)/kg. All 12 patients were alive 3 months after infusion and after 1 year, two patients had died. No infusion-related side effects were reported and no severe acute GVHD (grade III-IV) developed in any patient post-infusion. Overall, 3 months after infusion 11 out of 12 patients had increased levels of platelets and/or granulocytes. In conclusion, we describe the use of alpha beta T-cell depleted products as stem cell boosters with encouraging results.

  • 13.
    Sundin, Mikael
    et al.
    Karolinska university hospital, Sweden.
    Tesi, Bianca
    Karolinska Institutet, Sweden.
    Bohme, Maria Sund
    Karolinska university hospital, Sweden.
    Bryceson, Yenan T.
    Karolinska Institutet, Sweden.
    Putsep, Katrin
    Karolinska Institutet, Sweden.
    Chiang, Samuel C.
    Karolinska Institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Winiarski, Jacek
    Karolinska university hospital, Sweden.
    Wikstorm, Ann-Charlotte
    Karolinska Institutet, Sweden.
    Novel STAT3 Mutation Causing Hyper-IgE Syndrome: Studies of the Clinical Course and Immunopathology2014In: Journal of Clinical Immunology, ISSN 0271-9142, E-ISSN 1573-2592, Vol. 34, no 4, p. 469-477Article in journal (Refereed)
    Abstract [en]

    Purpose Reporting a clinical case with a novel mutation in the signal transducer and activator of transcription 3 (STAT3) gene resulting in autosomal dominant hyper-immunoglobulin E syndrome (AD-HIES). Here we also had the opportunity to perform in-depth immunologic investigations to further understand the immunopathology of this primary immunodeficiency. Methods The patient, a baby boy, was clinically assessed according to the scoring system developed by Grimbacher et al. and STAT3 was investigated by DNA sequencing. Immunologic work-up consisted of lymphocyte phenotyping and proliferation assays, measurement of soluble mediators and routine investigations. Results According to the Grimbacher score the patient was likely to have AD-HIES and a novel heterozygous STAT3 mutation (c.1110-3C>A), causing a splice error, was identified. Lymphocyte phenotyping revealed decreased numbers of interleukin (IL)-17 producing T-helper lymphocytes and aberrant B-lymphocyte subsets. Proliferative in vitro lymphocyte responses against C. albicans, staphylococcal enterotoxins and pokeweed mitogen were supernormal at presentation, whereas only the elevated response to pokeweed mitogen persisted. The soluble mediators IL-5, -10, -12, -13, -15 and granulocyte colony stimulatory factor were elevated in serum. Conclusion A novel heterozygous STAT3 mutation causing defective splicing of exon 12 was identified. Lymphocyte phenotyping revealed deranged subpopulations. Despite the clinical picture with severe C. albicans and staphylococcal infections, the patient’s lymphocytes mounted responses to these pathogens. The hypereosinophilia and high immunoglobulin E levels might partly be explained by elevated IL-5 and -13 as a result of lack of negative feedback from defective STAT3 signaling.

  • 14.
    Thielen, Noortje
    et al.
    VU University Medical Center, The Netherlands.
    Richter, Johan
    Skåne university hospital, Sweden.
    Baldauf, Matthias
    Medical University of Innsbruck, Austria.
    Barbany, Gisela
    Karolinska institutet, Sweden.
    Fioretos, Thoas
    Lund university, Sweden.
    Giles, Francis
    RobertH. Lurie Comprehensive Cancer Center of Northwestern University, USA.
    Gjertsen, Bjorn-Tore
    University of Bergen, Norway.
    Hochhaus, Andreas
    Universit€atsklinikum Jena, Germany.
    Schuurhuis, Gerrit Jan
    VU University Medical Center, The Netherlands.
    Sopper, Sieghart
    Innsbruck Medical University and Tyrolean Cancer Research Institute, Austria.
    Stenke, Leif
    Karolinska university hospital, Sweden.
    Thunberg, Sarah
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics. Karolinska university hospital, Sweden.
    Wolf, Dominik
    University Clinic Bonn, Germany.
    Ossenkoppele, Gert
    VU University Medical Center, The Netherlands.
    Porkka, Kimmo
    Helsinki University Hospital, Finland.
    Janssen, Jeroen
    VU University Medical Center, The Netherlands.
    Mustjoki, Satu
    University of Helsinki, Finland.
    Leukemic Stem Cell Quantification in Newly Diagnosed Patients With Chronic Myeloid Leukemia Predicts Response to Nilotinib Therapy2016In: Clinical Cancer Research, ISSN 1078-0432, E-ISSN 1557-3265, Vol. 22, no 16, p. 4030-4038Article in journal (Refereed)
    Abstract [en]

    Purpose: Leukemic stem cells (LSCs) may harbor important resistance to tyrosine kinase inhibitors in chronic myelogenous leukemia (CML). We identified Philadelphia chromosome (Ph)-positive CD34(+)CD38(-) bone marrow cells (here denoted LSCs) and addressed their response-predictive value in patients with CML (n = 48) subjected to nilotinib in the ENEST1st trial (NCT01061177). Experimental design: Two flow cytometry-based cell sorting methods were used with multiparameter-directed CD45-(MPFC) and BCR-ABL1 probe-linked (FISH) identification of Ph-positive cells, respectively. Results: We observed a positive correlation between the proportion of LSCs at diagnosis and established prognostic markers (blast count, spleen size, Sokal score, and hemoglobin). Conversely, a high LSC burden predicted for an inferior molecular response at 3 (MPFC and FISH), 6 (MPFC), 9 (FISH), and 15 months (FISH). During nilotinib therapy, the proportion of LSCs decreased rapidly. At 3 months, a median of only 0.3% LSCs remained among CD34(+)CD38(-) cells, and in 33% of the patients the LSC clone was not detectable anymore (FISH). The response kinetics was similar in LSC fractions as it was in the progenitor and unseparated bone marrow cell fractions. Conclusions: The proportion of LSCs at diagnosis, as analyzed by two independent methodologies, reflects the biology of the disease and appeared as a prognostic and response-predictive marker in patients with CML subjected to first-line nilotinib therapy. (C) 2016 AACR.

  • 15.
    Thunberg, Sarah
    et al.
    Karolinska Institutet.
    Akdis, M.
    SIAF, Switzerland.
    Akdis, C. A.
    SIAF, Switzerland.
    Gronneberg, R.
    Karolinska institutet, Sweden.
    Malmstrom, V.
    Karolinska institutet, Sweden.
    Trollmo, C.
    Karolinska institutet, Sweden.
    van Hage, M.
    Karolinska institutet, Sweden.
    Gafvelin, G.
    Karolinska institutet, Sweden.
    Immune regulation by CD4(+)CD25(+) T cells and interleukin-10 in birch pollen-allergic patients and non-allergic controls2007In: Clinical and Experimental Allergy, ISSN 0954-7894, E-ISSN 1365-2222, Vol. 37, no 8, p. 1127-1136Article in journal (Refereed)
    Abstract [en]

    CD4(+)CD25(+) regulatory T (Treg) cells and the cytokines IL-10 or TGF-beta play key roles in the maintenance of T cell homeostasis and tolerance to infectious and non-infectious antigens such as allergens. To investigate the regulation of immune responses to birch pollen allergen compared with influenza antigen by Treg cells obtained from birch pollen-allergic patients and non-allergic controls. Peripheral blood was collected from 10 birch pollen-allergic patients and 10 non-allergic healthy controls. CD4(+)CD25(+) and CD4(+)CD25(-) cells isolated by magnetic-activated cell sorting were co-cultured and stimulated with birch pollen extract or influenza vaccine in the absence or presence of anti-IL-10 or soluble TGF-beta RII. CD4(+)CD25(+) cells from non-allergic controls were able to suppress influenza antigen and birch pollen stimulated effector cell proliferation, whereas CD4(+)CD25(+) cells from allergic patients suppressed influenza antigen-, but not birch pollen-stimulated proliferation. The production of Th1 cytokines, but not Th2 cytokines, was suppressed by CD4(+)CD25(+) cells from both allergic patients and controls, upon stimulation with birch pollen extract. Neutralization of IL-10 led to significantly increased production of IFN-gamma in cultures with CD4(+)CD25(-) T effector cells. In addition, six-fold higher concentrations of TNF-alpha were detected after neutralization of IL-10 in both CD4(+)CD25(-) and CD4(+)CD25(+) cell cultures from allergic patients and controls. We demonstrate that the allergen-specific suppressive function of CD4(+)CD25(+) cells from allergic patients is impaired compared with non-allergic controls. Moreover, neutralization of IL-10 enhances the production of TNF-alpha, suggesting counter-acting properties of IL-10 and TNF-alpha, where IL-10 promotes tolerance and suppression by Treg cells and TNF-alpha promotes inflammatory responses.

  • 16.
    Thunberg, Sarah
    et al.
    Karolinska Institutet, Sweden.
    Gafvelin, G.
    Karolinska Institutet, Sweden.
    Nord, M.
    Karolinska Institutet, Sweden.
    Gronneberg, R.
    Karolinska Institutet, Sweden.
    Grunewald, J.
    Karolinska Institutet, Sweden.
    Eklund, A.
    Karolinska Institutet, Sweden.
    van Hage, M.
    Karolinska Institutet, Sweden.
    Allergen provocation increases TH2-cytokines and FOXP3 expression in the asthmatic lung2010In: Allergy. European Journal of Allergy and Clinical Immunology, ISSN 0105-4538, E-ISSN 1398-9995, Vol. 65, no 3, p. 311-318Article in journal (Refereed)
    Abstract [en]

    P>Background: Allergic asthma is caused by allergen-specific IgE and T-helper cell (Th) type 2 responses towards airborne allergens. The objective of this study was to investigate local and systemic regulatory mechanisms in the early asthmatic response to bronchial allergen provocation. Methods: Birch pollen-allergic patients with mild asthma (n = 13) and healthy nonallergic controls (n = 14) were subjected to bronchoalveolar lavage (BAL) and blood sampling. On patients BAL was performed twice: without preceding provocation (’before samples’) and 24 h after bronchial provocation with birch pollen allergen. Lymphocytes in BAL and peripheral blood mononuclear cells (PBMCs) were phenotyped by multi-colour flow cytometry and cytokines measured by cytometric bead array. Proliferation and secreted cytokines were analysed in allergen-stimulated PBMCs, CD25+ depleted PBMCs and PBMCs with IL-10 neutralizing antibodies. Results: The numbers of CD69+ and FOXP3+ lymphocytes were higher in BAL after compared with before allergen provocation in asthmatic patients. Moreover, allergen provocation increased expression of FOXP3 in CD4+CD25bright cells. The cytokine profile in BAL fluid from asthmatics revealed higher levels of IL-5, compared with the controls, and an increase in IL-5, IL-6, IL-9 and IL-10 after allergen provocation. Pollen allergen stimulated PBMC cultures from asthmatic patients produced elevated levels of IL-5 and IL-13 compared with the controls, which were not affected by depletion of CD25+ cells or IL-10 neutralization. Conclusion: Despite an increase in CD4+CD25bright cells expressing high levels of FOXP3 in response to bronchial allergen provocation, asthmatic patients exhibit enhanced levels of Th2 cytokines in the lung, which may indicate an inability among infiltrating cells to suppress Th2 responses.

  • 17.
    Thunberg, Sarah
    et al.
    Karolinska Institutet, Sweden.
    Neimert-Andersson, T.
    Karolinska Institutet, Sweden.
    Cheng, Q.
    Karolinska Institutet, Sweden.
    Wermeling, F.
    Karolinska Institutet, Sweden.
    Bergstrom, U.
    Uppsala university, Sweden.
    Swedin, L.
    Karolinska Institutet, Sweden.
    Dahlen, S. -E
    Karolinska Institutet, Sweden.
    Arner, E.
    Karolinska Institutet, Sweden.
    Scheynius, A.
    Karolinska Institutet, Sweden.
    Karlsson, M. C. I.
    Karolinska Institutet, Sweden.
    Gafvelin, G.
    Karolinska Institutet, Sweden.
    van Hage, M.
    Karolinska Institutet, Sweden.
    Gronlund, H.
    Karolinska Institutet, Sweden.
    Prolonged antigen-exposure with carbohydrate particle based vaccination prevents allergic immune responses in sensitized mice2009In: Allergy. European Journal of Allergy and Clinical Immunology, ISSN 0105-4538, E-ISSN 1398-9995, Vol. 64, no 6, p. 919-926Article in journal (Refereed)
    Abstract [en]

    Defined particles carrying tightly bound allergens at high density have been suggested as alternatives in allergy vaccination. Carbohydrate based particles (CBP), sized 2 mu m, provide a platform for covalent coupling of allergens. To investigate the mechanisms of antigen presentation by CBP, as well as cellular and humoral responses after vaccination with the major cat allergen Fel d 1, covalently coupled to CBP. Mice (n = 10/group) were subcutaneously vaccinated with CBP-rFel d 1, CBP or phosphate buffer saline (PBS) before sensitization with rFel d 1 and challenged with cat dander extract. Fluorescent and (75)Se-radiolabeled tracking of allergens and particles were performed with flow cytometry and whole-body autoradiography. Humoral, cellular and regulatory immune responses were analyzed by ELISA and flow cytometry. Cytokines were measured in bronchoalveolar lavage fluid and splenocyte cultures. CBP-rFel d 1 prevented induction of airway inflammation and induced allergen-specific T-cell anergy. CBP-rFel d 1 also induced rapid IgM and IgG1-responses compared with soluble rFel d 1. Particles were phagocytosed by antigen-presenting cells and transported to draining lymph nodes and spleen. Moreover, antigen coupled to CBP remained longer at the injection site compared with alum. Covalent coupling of rFel d 1 to CBP induces rapid antibody production, prevents induction of allergic immune responses and systemic allergen spreading. Thus, CBP comprise several attractive adjuvant features for use in allergy vaccination. Prolonged allergen exposure through covalent coupling to particles suitable for phagocytosis, provides an adjuvant for safer and efficient allergy vaccination.

  • 18.
    Thunberg, Sarah
    et al.
    KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab. Karolinska Inst, Sweden.
    Uhlin, M.
    Nfelt, B.
    KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Differentiation and expansion of gamma delta T-cells by Zolodronic acid efficiently activates cytotoxicity in vitro2016In: European Journal of Immunology, ISSN 0014-2980, E-ISSN 1521-4141, Vol. 46, p. 154-154Article in journal (Other academic)
  • 19.
    Uhlin, Michael
    et al.
    Karolinska Institutet, Sweden.
    Sairafi, Darius
    Karolinska Institutet, Sweden.
    Berglund, Sofia
    Karolinska Institutet, Sweden.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Gertow, Jens
    Karolinska Institutet, Sweden.
    Ringden, Olle
    Karolinska Institutet, Sweden.
    Uzunel, Mehmet
    Karolinska Institutet, Sweden.
    Remberger, Mats
    Karolinska Institutet, Sweden.
    Mattsson, Jonas
    Karolinska Institutet, Sweden.
    Mesenchymal Stem Cells Inhibit Thymic Reconstitution After Allogeneic Cord Blood Transplantation2012In: Stem Cells and Development, ISSN 1547-3287, E-ISSN 1557-8534, Vol. 21, no 9, p. 1409-1417Article in journal (Refereed)
    Abstract [en]

    Cord blood (CB) as a source of stem cells has been a successful addition to the field of allogeneic stem cell transplantation (ASCT). The increased human leukocyte antigen (HLA) permissiveness of CB grafts has made it possible for more patients to undergo treatment. The drawback is that patients suffer from a longer period of compromised immunity. We analyzed T-cell receptor excision circles (TRECs), immunoglobulin G (IgG), and immunoglobulin M (IgM) levels after cord blood transplantation (CBT) in 50 patients transplanted at our center. These immunological parameters were compared retrospectively with clinical factors and complications. We found that TREC levels after CBT were lower in adults, patients with myeloablative conditioning, and in patients with a lower nucleated cell dose in the graft. In addition mesenchymal stem cells (MSC) as co-infusion at the time of CBT had a negative effect on TREC reconstitution. This was found to be associated with decreased overall survival for this patient category. Reduced IgM and IgG levels post-CBT were associated with a major ABO mismatch, and infusion of MSCs. Our results highlight the importance of close monitoring of the immune reconstitution after CBT. In addition it shows a potentially new suppressive effect of MSCs on the immune system.

  • 20.
    Velickovic, Tanja Cirkovic
    et al.
    Faculty of Chemistry, Serbia.
    Thunberg, Sarah
    Karolinska Institutet, Sweden.
    Polovic, Natalija
    Faculty of Chemistry, Serbia.
    Neimert-Andersson, Theresa
    Karolinska Institutet, Sweden.
    Gronlund, Hans
    Karolinska Institutet, Sweden.
    van Hage, Marianne
    Karolinska Institutet, Sweden.
    Gafvelin, Guro
    Karolinska Institutet, Sweden.
    Low levels of endotoxin enhance allergen-stimulated proliferation and reduce the threshold for activation in human peripheral blood cells2008In: International Archives of Allergy and Immunology, ISSN 1018-2438, E-ISSN 1423-0097, Vol. 146, no 1, p. 1-10Article in journal (Refereed)
    Abstract [en]

    Background: Endotoxins, comprised of bacterial cell wall lipopolysaccharides (LPS), have been reported to have both protective and exacerbating effects on the development and maintenance of allergic disease in humans and on markers of allergic inflammation in animal models of allergy. In this study, we investigated the effect of low concentrations of LPS on human peripheral blood mononuclear cells (PBMC) stimulated with the major cat allergen Fel d 1. Methods: Extensive purification of recombinant (r) Fel d 1 yielded essentially endotoxin-free rFel d 1 (0.2 ng LPS/mg protein). PBMCs prepared from 15 subjects having IgE to cat (>0.7 kU(A)/l) and 8 subjects IgE negative to cat were stimulated with 2, 10 or 25 mu g/ml of rFel d 1 in the presence or absence of 50 pg/ml LPS. Proliferation was measured after 7 days of culture and supernatants were analyzed for IFN gamma, IL-5 and IL-10. Results: LPS (50 pg/ml) increased rFel d 1-stimulated proliferation of PBMCs both from subjects IgE-positive and subjects negative to cat allergens. PBMCs from 13 of the subjects did not proliferate in response to stimulation with 2 and 10 mu g/ml rFel d 1 alone but did so in the presence of LPS. Moreover, LPS increased the levels of rFel d 1-stimulated IFN gamma in cultures from cat-negative subjects, IL-5 from cat-positive subjects and IL-10 from both groups. Conclusion: Very low doses of LPS enhance proliferation and decrease the apparent threshold level for cell activation, prompting careful evaluation of allergen stimulated T cell activation in vitro. Copyright (C) 2007 S. Karger AG, Basel.

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