Team Anna Nopp
Investigating the immune modulatory role and function of leukocytes in inflammation.
Inflammation is a protective response that involves immune cells, blood vessels, and molecular mediators. The purpose of inflammation is to eliminate the initial cause of cell injury, clear out necrotic cells and tissues damaged from the original insult and the inflammatory process, and to initiate tissue repair. Inflammation can be classified as either acute or chronic. Acute inflammation is the initial response of the body to harmful stimuli and is achieved by the increased movement of plasma and leukocytes, especially granulocytes, from the blood into the injured tissues. Prolonged inflammation, known as chronic inflammation, leads to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells, and is characterized by simultaneous destruction and healing of the tissue from the inflammatory process. Leukocytes are a fundamental part of the inflammatory process. Leukocytes exert however their main physiological and pathophysiological effects extravascular, when they have adhered to the endothelium and transmigrated to the intima sub-endothelia or to the interstitial compartment. The importance of analysing leukocyte recruitment and function at the site where the cells exercise its essential function is of essential importance to position laboratory and clinical findings in a patient benefit perspective.
Anna Nopp, PhD, Associate Professor, Group Leader
The research is focused on the role of basophils in inflammation and as a diagnostic tool for allergy, inflammation and autoimmunity.
Joachim Lundahl, MD, Assistant Director of Karolinska University Laboratory, PhD, Professor
The research is focused on inflammation in general with emphasis on the innate cellular response.
S.G.O. Johansson, MD, PhD, Professor
The research is focused on IgE-mediated allergy.
Zekiye Cansu, BMA
The work includes several immunological
Zenib Aljadi, MD, PhD-student
The research is focused on developing a new method (microfluidic chip) to study basophil functions in inflammation.
Ladan Mansouri, MD, PhD-student
The research is focused on inflammation and functionality of lymphocytes in patients with chronic kidney disease.
Frida Kalm, MSc, PhD-student
The research is focused on improving allergy diagnostics using microfluidic chip technology.
Senka Sendic, MD, PhD-student
The research is focused on inflammation in patients with chronic kidney disease, mainly IgA nephropathy and SLE nephritis.
Leukocytes in inflammation
Leukocyte functionality in chronic kidney disease
Patients with renal failure or chronic kidney disease (CKD) have a greater risk of morbidity and mortality due to infections and cardiovascular diseases, compared to the general population. These emerge from an immunodeficiency, as well as a high inflammatory state in CKD patients. Previous studies of human leukocyte functions in CKD patients have been mainly carried out ex vivo on cells from the peripheral circulation. Leukocytes exert however their main physiological and pathophysiological effects extravascular, when they have adhered to the endothelium and transmigrated to the intima sub-endothelia or to the interstitial compartment. Therefore our projects aim to study recruitment and activation of leukocytes, focusing on how the inflammatory process affects the biological activity of immune cells and how different degrees of kidney failure affect these processes. Analysing leukocyte recruitment and function at the inflammatory site, where the cells execute their essential functions, is of great importance to place the laboratory and clinical findings in a patient benefit perspective.
An important conceptual finding is the recently demonstrated gene regulation of transmigrated neutrophilic granulocytes, which indicates that these cells are capable of expressing a different profile of genes and protein synthesis, essential for survival and apoptosis. Our research group has previously studied how gene expression in neutrophils and monocytes changed in connection with transmigration into an inflammatory site, in the sub-endothelial environment, in patients with kidney disease compared to healthy individuals.
T lymphocytes are considered to play an important pathophysiological role in CKD patients and aberration in the functionality of these cells contributes to increased susceptibility to infections, and decreased response to vaccination. We have shown that T-cells in patients have a different capacity of cytokine production compared to healthy individuals. Moreover, a different profile of T-cell subpopulation in CKD patients may drive diverse features in chronic kidney disease.
A subpopulation of granulocytes, whose role in pato-mechanism of inflammatory diseases is not fully understood, is the basophilic granulocytes. Basophils are able to infiltrate the inflamed tissue in several inflammatory diseases. Recent development of new analytical tools for basophil function in vivo, including basophil deficient mice, has identified pivotal roles for basophils in autoimmune diseases, like systemic lupus erythematosus (SLE). In lupus-like nephritis mice models, basophils have been shown to be activated by deposition of IgE-anti-dsDNA immune complexes in the kidney tissue resulting in up-regulation of adhesion molecules. This regulates cell transmigration into lymphoid organs. Upregulation of MHC class II confers the basophils antigen-presenting properties and production of IL-4 results in a Th2 phenotype. However, since much of this new information has been studied in basophil mice modes it needs to be investigated in human basophils to permit clinical conclusions.
Basophil involvement in IgE-mediated allergy
Allergic diseases are common health problems and have increased over the last fifty years. Allergic inflammation is often mediated through IgE-antibodies (IgE-ab) and diagnosis of is usually based on medical history and analysis of the presence of allergen-specific IgE-ab in skin (SPT) or serum. In many cases, however, the clinical significance of the presence of IgE-ab in skin (positive skin test) or in serum is uncertain and needs to be confirmed by an in vivo challenge, e.g. bronchial or oral, with the suspected allergen. A challenge, however, is very resource and time consuming. Nor is it completely harmless and may be associated with side effects, i.e. general allergic reactions of varying severity. There are in the light of the above, a need to develop alternative diagnostics methods for diagnosis and monitoring of treatment efficacy.
One way to study basophil function is by stimulating basophils, in vitro, with several doses of an allergen and measure the minimum amount of allergen that stimulates the basophil, CD-sens. The stimulated basophils are then stained with monoclonal antibodies, CD203c to identify basophils, and CD63 to measure activity, and then analysed by flow cytometry. CD63 is located in the same granules as histamine and is released during the activation process (Figure 1).
|Figure 1. When an allergen, such as cat, crosslink membrane-bound IgE antibodies the basophil becomes activated and upregulates the expression of activity marker CD63 and release inflammatory mediators such as histamine, IL-4 and IL-13. CD203c is constitutively expressed on the basophil cell surface and is used to identify the cell.|
We have shown that CD-sens is a useful complement for complex allergen challenges in vivo with good correlations between CD-sens and SPT-titration, bronchial-, nasal-, and oral- challenge. CD-sens can also be used to monitor treatment with anti-IgE (omalizumab, XolairÒ) and specific immunotherapy. CD-sens with flow cytometric detection is since 2010 implemented both into clinical practice and research for diagnosis and monitoring of treatment efficacy.
Development of future diagnostic tools
A major drawback with the current ways to diagnose allergic, inflammatory and autoimmune diseases driven by basophil is the lack of instantaneous, simple, quantitative and patient-near diagnostics. One way to address these needs is a diagnostic test based on a cell-on-chip system.
Since a few years back, new technologies have been developed to effectively isolate a highly purified population of cells without affecting cell sensitivity and reactivity. The technology, cell-on-chip, is based on the principle that blood is passed over a microchip that is coated with antibodies directed against a specific cell population which will be stuck on the chip. Based on this technology, we have previously developed a number of micro fabricated chips for efficient separation and isolation of cells for a variety of applications. For isolation of cell populations, methods that combine the molecular specificity of monoclonal antibodies with well-defined flow in micro channels have been developed. The channels are coated with monoclonal antibodies specific for the cell of interest.
15 selected publications
The clinical utility of basophil activation testing in diagnosis and monitoring of allergic disease.
Allergy 2015 Nov;70(11):1393-405
Basophil allergen threshold sensitivity and component-resolved diagnostics improve hazelnut allergy diagnosis.
Clin. Exp. Allergy 2015 Sep;45(9):1412-8
Activation of basophils is a new and sensitive marker of biocompatibility in hemodialysis.
Artif Organs 2014 Nov;38(11):945-53
The discovery of immunoglobulin e and its role in allergy.
Chem Immunol Allergy 2014 ;100():150-4
Leukocyte proliferation and immune modulator production in patients with chronic kidney disease.
PLoS ONE 2013 ;8(8):e73141
False-positive penicillin immunoassay: an unnoticed common problem.
J. Allergy Clin. Immunol. 2013 Jul;132(1):235-7
Activation of Wnt/β-catenin pathway in monocytes derived from chronic kidney disease patients.
PLoS ONE 2013 ;8(7):e68937
In-vivo extravasation induces the expression of interleukin 1 receptor type 1 in human neutrophils.
Clin. Exp. Immunol. 2012 Apr;168(1):105-12
Basophil allergen threshold sensitivity, CD-sens, IgE-sensitization and DBPCFC in peanut-sensitized children.
Allergy 2012 Feb;67(2):242-7
History of the world allergy organization: time to change!
World Allergy Organ J 2011 Nov;4(11):184-7
Basophil allergen threshold sensitivity, CD-sens, is a measure of allergen sensitivity in asthma.
Clin. Exp. Allergy 2011 Aug;41(8):1091-7
Basophil allergen threshold sensitivity: a useful approach to anti-IgE treatment efficacy evaluation.
Allergy 2006 Mar;61(3):298-302
Passive IgE-sensitization by blood transfusion.
Allergy 2005 Sep;60(9):1192-9
Immunoglobulin E. A new class of human immunoglobulin.
Immunochemistry 1968 Jul;5(4):327-8