Marty Wulferink

Heinrich-Heine-Universität Düsseldorf, Düsseldorf, North Rhine-Westphalia, Germany

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Publications (14)46.08 Total impact

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    ABSTRACT: A 34-year-old female patient with a three year history of generalized granuloma annulare was treated systemically with dapsone (DADPS). Six weeks after the onset of treatment, the patient developed an extensive tonsillitis of the base of the tongue with fever and malaise. Routine laboratory work showed a leukocytopenia with agranulocytosis. Further investigation revealed a marked decrease of the enzyme activity of N-acetyltransferase 2, which plays an important role in dapsone metabolism. Treatment included the cessation of dapsone, antibiotic coverage, and G-CSF leading to the rapid improvement of symptoms and normalization of leukocyte counts. Dapsone-induced angina agranulocytotica is a rare event and is interpreted as an idiosyncratic reaction. Depending on genetic polymorphisms of various enzymes, dapsone can be metabolized to immunologically or toxicologically relevant intermediates. Because of the risk of severe hematologic reactions, dapsone should only be employed for solid indications and with appropriate monitoring.
    Der Hautarzt 08/2005; 56(7):673-7. · 0.50 Impact Factor
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    ABSTRACT: Wir berichten ber eine 34-jhrige Patientin mit einem seit 3Jahren bestehenden disseminierten Granuloma anulare, das systemisch mit Dapson (DADPS) behandelt wurde. Sechs Wochen nach Therapiebeginn entwickelte die Patientin eine Zungengrundtonsillitis mit Fieber und schwerem Krankheitsgefhl. Im Routinelabor zeigte sich eine Leukozytopenie (700/l) mit Agranulozytose. In der weitergehenden Labordiagnostik zeigte sich bei der Patientin eine extreme Aktivittsminderung der N-Azetyltransferase2, dem wichtigsten Dapson metabolisierenden Enzym. Nach Absetzen von Dapson, antibiotischer Abdeckung und Therapie der Leukozytopenie mit Granulozyten-Kolonie-stimulierendem Faktor (G-CSF) kam es zu einer raschen Besserung der Symptomatik und Normalisierung der Leukozytenzahl. Die Dapson-induzierte Agranulozytose ist ein seltenes Ereignis und wird als Idiosynkrasiereaktion aufgefasst. Dapson kann ber verschiedene Enzyme zu immunologisch bzw. toxikologisch relevanten Intermediaten metabolisiert werden, sodass mglicherweise genetische Polymorphismen dieser Enzyme zu einer individuell unterschiedlichen Disposition gegenber Arzneimittelreaktionen fhren. Da unter einer Dapson-Behandlung potenziell schwere hmatologische Arzneimittelnebenwirkungen auftreten knnen, sollten eine strenge Indikationsstellung vor sowie engmaschige Laborkontrollen whrend der Therapie erfolgen.A 34-year-old female patient with a three year history of generalized granuloma annulare was treated systemically with dapsone (DADPS). Six weeks after the onset of treatment, the patient developed an extensive tonsillitis of the base of the tongue with fever and malaise. Routine laboratory work showed a leukocytopenia with agranulocytosis. Further investigation revealed a marked decrease of the enzyme activity of N-acetyltransferase 2, which plays an important role in dapsone metabolism. Treatment included the cessation of dapsone, antibiotic coverage, and G-CSF leading to the rapid improvement of symptoms and normalization of leukocyte counts. Dapsone-induced angina agranulocytotica is a rare event and is interpreted as an idiosyncratic reaction. Depending on genetic polymorphisms of various enzymes, dapsone can be metabolized to immunologically or toxicologically relevant intermediates. Because of the risk of severe hematologic reactions, dapsone should only be employed for solid indications and with appropriate monitoring.
    Der Hautarzt 06/2005; 56(7):673-678. · 0.50 Impact Factor
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    ABSTRACT: Although the ability of CD4+CD25+ T suppressor (Ts) cells to prevent experimental autoimmune diseases has been described, nothing is known concerning their role and mechanism of action in xenobiotic-induced autoimmunity. Procainamide, mercuric chloride, and gold(I) are three xenobiotics that can induce autoimmune reactions in humans and rodents. After the induction of IgG1 antinuclear autoantibodies (ANA) in mice treated with either of the above xenobiotics, adoptive transfer of their CD4+CD25+ T cells completely prevented ANA formation in recipients treated with the same xenobiotic; transfer of CD8+ T cells was ineffective. Furthermore, xenobiotic-primed CD4+CD25+ T cells could also partially prevent ANA formation in recipients treated with a different xenobiotic. CD4+CD25- T cells from xenobiotic-treated donors failed to suppress, but induced de novo IgG1 ANA formation in untreated recipients. Our findings suggest that during xenobiotic treatment T cell reactivity may spread from xenobiotic-induced, nucleoprotein-related neoantigens to peptides of the unaltered nucleoproteins.
    European Journal of Immunology 02/2004; 34(1):36-46. · 4.97 Impact Factor
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    ABSTRACT: The onset or exacerbation of psoriasis, a T-cell-dependent skin disease with autoimmune features, can be triggered by drugs such as antimalarials and beta-blockers. Xenobiotics may also play a role in idiopathic psoriasis. It has been hypothesized that different metabolic efficiencies caused by variant alleles of xenobiotic metabolizing enzymes could lead to the accumulation of xenobiotics or their reactive metabolites in target organs. Subsequently, neoantigens or cryptic peptides could be presented and initiate an aggressive T cell response. In this context, we analyzed a broad array of xenobiotic metabolizing enzymes in up to 327 Caucasian psoriasis patients and compared them to 235 control persons. Alleles tested include four phase I and three phase II enzymes. Significantly more carriers of the variant alleles of CYP1A1 (alleles *2A and *2C) were found in healthy controls than in patients, suggesting a protective role for these alleles. No significant difference between patients and controls could be found, however, for the other phase I alleles 1B1*1 and 1B1 *3, 2C19*1A and 2C19*2A, and 2E1*1A and 2E1*5B. Of the variant alleles coding for phase II enzymes only GSTM1, but not GSTT1 or NQOR, correlated with a risk to contract psoriasis. Some combinations of phase I and phase II enzymes suggested protective or risk-associated effects. Interestingly, heterozygosity for CYP2C19 alleles *1A and *2A was associated with increased risk for "late onset" psoriasis, whereas this genotype was protective for psoriatic arthritis. This is the first large-scale study on these enzymes and the results obtained support the concept that different activities of metabolizing enzymes can contribute to disease etiology and progression.
    Journal of Investigative Dermatology 06/2003; 120(5):765-70. · 6.19 Impact Factor
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    Marty Wulferink, Sabine Dierkes, Ernst Gleichmann
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    ABSTRACT: To analyze T cell cross-reactivity to para-compounds, we established CD4(+) T cell hybridomas from mice immunized with adducts of self-globin and one of three different para-compounds: p-aminophenol, p-phenylenediamine, or Bandrowski's base. Some of the hybridomas obtained reacted not only to the immunizing antigen, but also to metabolically related para-compounds, bound to the same protein, thus suggesting formation of common metabolites. Other hybridomas cross-reacted to globin adducts of metabolically unrelated para-compounds, which denotes them as truly cross-reactive cells whose TCR failed to distinguish among the different haptens. One of these hybridomas also reacted against a non-haptenated, cryptic peptide of hemoglobin but not to the full-length native protein. As this hybridoma reacted even more strongly to the respective peptide after it was haptenated, recognition of the native, cryptic peptide was apparently due to true cross-reactivity. To conclude, true T cell cross-reactivity to haptens does occur, as well as the formation of a common reactive metabolite, and T cell recognition of cryptic self-peptides may underlie cross-sensitization to chemicals.
    European Journal of Immunology 06/2002; 32(5):1338-48. · 4.97 Impact Factor
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    ABSTRACT: The most basic arylamine, aniline, belongs to a class of compounds notorious for inducing allergic and autoimmune reactions. In 1981 in Spain, many people succumbed to toxic oil syndrome (TOS), a disease caused by ingestion of cooking oil contaminated with aniline. Indirect evidence points toward an immune pathogenesis of TOS driven by T lymphocytes, but it is unclear to which antigens these cells could react. Here, using the popliteal lymph node (PLN) assay in mice, we analyzed the sensitizing potential of aniline, its metabolites, and some of the aniline-coupled lipids detected in the contaminated cooking oil. Whereas aniline itself and its non-protein-reactive metabolites nitrobenzene, p-aminophenol and N-acetyl-p-aminophenol, failed to elicit PLN responses, its reactive metabolites nitrosobenzene and N-hydroxylaniline did. The aniline-coupled lipids, namely, linoleic anilide and linolenic anilide, and a mixture of fatty acid esters of 3-(N-phenylamino)-1,2-propanediol, all implicated in TOS, induced significant PLN responses, whereas the respective aniline-free lipids, linoleic acid, linolenic acid, and triolein, did not. Hence, the aniline moiety plays a crucial role in the immunogenicity of the aniline-coupled lipids of TOS. PLN responses to the reactive aniline metabolites and the one aniline-coupled lipid that was tested, linolenic anilide, were T-cell-dependent. Secondary PLN responses to nitrosobenzene were detectable not only after priming with nitrosobenzene but, in some experiments, also after priming with linolenic anilide. This suggests that the aniline moiety was cleaved from the aniline-coupled lipid and metabolized to the intermediate nitrosobenzene that generated the prospective neoantigens. Consistent with this, in lymphocyte proliferation tests in vitro, T cells primed to nitrosobenzene reacted in anamnestic fashion to white bone marrow cells (WBMCs) pulsed with aniline. Hence, we propose that aniline is a prohapten that can be metabolized by WBMCs, which form neoantigens that are recognized by T cells. The possible significance of these findings for the pathogenesis of TOS is discussed.
    Chemical Research in Toxicology 05/2001; 14(4):389-97. · 3.67 Impact Factor
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    ABSTRACT: A 32-year-old patient developed an anaphylactic reaction minutes after oral intake of acetaminophen-containing tablets (Doregrippin)). Scratch testing of the whole preparation was positive in contrast with the negative results obtained with pure acetaminophen. Therefore, scratch tests with the remaining drug components were performed and showed polyvinylpyrrolidone (PVP) to be the aetiological agent. Furthermore, specific IgE antibodies against PVP were demonstrated using a dot blot technique, thus ruling out a pseudo-allergic reaction. This case underlines the necessity to consider not only the active ingredient, but also additives as the causative agent.
    British Journal of Dermatology 12/2000; 143(5):1055-8. · 3.76 Impact Factor
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    ABSTRACT: Procainamide (PA) may cause drug-induced lupus, and its reactive metabolites, hydroxylamine-PA (HAPA) and nitroso-PA, are held responsible for this. Here, we show that N-oxidation of PA to these metabolites can take place in macrophages and lead to formation of neoantigens that sensitize T cells. Murine peritoneal macrophages (PMvarphi), exposed to PA in vitro, generated neoantigens related to HAPA as indicated by (1) their capacity to elicit a specific recall response of HAPA-primed T cells in the adoptive transfer popliteal lymph node (PLN) assay and (2) the appearance of metabolite-bound protein in PA-pulsed PMvarphi, as determined by Western blot. Analysis of five phase I enzymes that might be responsible for HAPA formation by PMvarphi pointed to prostaglandin H synthase-2 (PGHS-2) as a likely candidate. Experimental evidence that PA can be oxidized to HAPA by PGHS was obtained by exposing PA to PGHS in vitro. The resulting metabolites were identified by mass spectral analysis and covalent protein binding in ELISA. In vitro, PA exposure of PMvarphi of slow acetylator A/J and fast acetylator C57BL/6 mice failed to show significant strain differences in enzyme mRNA expression, enzyme activities, or formation of HAPA-related neoantigens. By contrast, after long-term PA treatment in vivo only in slow acetylators the PMvarphi harbored HAPA-related neoantigens and T cells were sensitized to them. PMvarphi of fast acetylator C57BL/6 mice only contained HAPA-related neoantigens, and their T cells were only sensitized to them if, in addition to long-term PA treatment, their donors had received injections of phorbol myristate acetate (PMA), a known enhancer of oxidative enzymes in phagocytes. In conclusion, PA treatment leads to N-oxidation of PA by enzymes, in particular PGHS-2, present in antigen-presenting cells (APC) and, hence, to generation of neoantigens which sensitize T cells. The enhanced neoantigen formation and T cell sensitization seen in slow acetylators might be explained by their higher concentration of PA substrate that is available for extrahepatic N-oxidation in APC.
    Chemical Research in Toxicology 07/1999; 12(6):488-500. · 3.67 Impact Factor
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    ABSTRACT: Using the popliteal lymph node (PLN) assay in mice, we studied the sensitizing potential of benzene and its metabolites. Whereas benzene and phenol failed to induce a PLN reaction, catechol and hydroquinone induced a moderate, and p-benzoquinone a vigorous response. Following a single injection of the reactive metabolite p-benzoquinone (100 nmol/mouse), cellularity in the draining PLN was increased > 15-fold, and reverted back to normal only after approximately 100 days. Although the PLN response was T cell-dependent, flow cytometric analysis revealed that the increased cellularity in the PLN after a single injection of p-benzoquinone was mainly due to an increase in B cells. Mice primed to p-benzoquinone and challenged with a small dose of p-benzoquinone (0.1 nmol/mouse) mounted a secondary PLN reaction, indicating hapten specificity of the reaction; this was confirmed by results obtained in the adoptive transfer PLN assay. An unexpected finding was the secondary PLN response to benzene (1 nmol/mouse) observed in mice primed to p-benzoquinone. This finding suggests that some of the benzene (at least 10%) was locally converted into p-benzoquinone, which then elicited the secondary response observed. In conclusion, the reactive intermediate metabolites hydroquinone and p-benzoquinone can act as haptens and sensitize; their precursors, benzene and phenol, may be considered as prohaptens.
    Archive für Toxikologie 05/1999; 73(3):159-67. · 5.22 Impact Factor
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    ABSTRACT: Induction of allergic and autoimmune reactions by drugs and other chemicals constitutes a major public health problem. Elucidation of the underlying mechanisms might help improve diagnostic tools and therapeutic approaches. Here, Peter Griem and colleagues focus on several aspects of neoantigen formation by xenobiotics: metabolism of xenobiotics into reactive, haptenic metabolites; polymorphisms of metabolizing enzymes; induction of costimulatory signals; and sensitization of T cells.
    Immunology Today 03/1998; 19(3):133-41. · 9.49 Impact Factor
  • Journal of Dermatological Science - J DERMATOLOGICAL SCI. 01/1998; 16.
  • Toxicology Letters 10/1996; 88:55-55. · 3.15 Impact Factor
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    Marty Bernardus Franciscus Wulferink
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    ABSTRACT: This thesis aims towards a better understanding of the mechanisms that lead to chemical-induced adverse immune effects. We focussed thereby on the initial induction stage of the immune reaction that consists of three major steps: (i) formation of neoantigen, (ii) processing and presentation of the neoantigen, and (iii) recognition of peptide-MHC-complex by the T cell. Chemical induced allergy or autoimmunity is often observed after administration of certain drugs or after contact with industrial or environmental chemicals. T cells play a central role in the development of drug-induced adverse immune reactions. A major difficulty in studying T-cell reactions to sensitizing chemicals is the fact that in most cases the ultimate neoantigen recognized by "drug-specific" T cells is unknown. As most T cells can only recognize peptides on MHC molecules, chemicals have to bind to a protein carrier in order to be recognized by T cells. Parts of this neoantigen are presented on MHC molecules on the surface of the APC and can be engaged by T cells. In order to bind to proteins, most chemicals have to be metabolized into protein-reactive intermediates, called haptens. Although the liver is the main organ for metabolic transformation, it does not play a major role in the induction of chemical induced adverse immune effects. In contrast, cells from the immune system itself, like neutrophils and monocytes have been shown to be involved in drug-metabolism prior to drug-induced adverse immune effects. In this thesis we have tried to elucidate some of the mechanisms involved in chemical induced adverse immune effects. Some of the steps possibly involved in the formation of neoantigens and subsequent T cell recognition are discussed. The hypothesis that extrahepatic metabolism plays an important role in the formation of neoantigen with the prohaptens procainamide and aniline could be substantiated, and the processing of protein adducts, presentation of neoantigens and subsequent recognition of either haptenated peptides or cryptic peptides by T cells was shown. Furthermore, this thesis elucidates some of the mechanisms involved in cross-sensitization and discusses the possible implications of T-cell cross-reactivity in chemical induced allergy and autoimmunity.
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    Marty Wulferink, Sabine Dierkes, Ernst Gleichmann
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    ABSTRACT: We have adapted the sandwich ELISA to rapidly screen a large number of T-cell hybridomas. The method, called CellELISA is similar to the sandwich ELISA, but instead of transferring the supernatants in cytokine-coated 96-well plates, the cells are cultured directly in sterile, cytokine-coated wells. After 24 hours, the plates are washed, incubated with the detection antibody and developed using ortho-phenyldiamine substrate. Compared with the cytokine bioassay, where supernatants are tested on cytokine-dependent ConA blasts, the CellELISA is as sensitive, but does not require radioactivity and takes one day less. Compared with the sandwich ELISA, the CellELISA is more sensitive, especially in the lower range of cytokine secretion. We conclude, that the CellELISA is a suitable method for rapid screening and measuring specific reactions of T-cell hybridomas.