Inflamm. res. 54 (2005) 415–419 1023-3830/05/100415-5 DOI 10.1007/s00011-005-1362-9 Inflammation Research © Birkhäuser Verlag, Basel, 2005 Original Research Papers Th1 and Th2 cell responses of type 1 diabetes patients and healthy controls to human heat-shock protein 60 peptides AA437-460 and AA394-408 A. Szebeni 1 , N. Schloot 2 , V. Kecskeméti 1 , N. Hosszúfalusi 3 , P. Pánczél 3 , Z. Prohászka 3 , G. Füst 3 , K Uray 4 , F Hudecz 4, 5 , G. Meierhoff 2 1 Department of Pharmacology and Pharmacotherapy, Semmelweis University, PO Box 370, 1445 Budapest, Hungary, Fax: ++ 00 361 210 4412, e-mail: szeband@pharma.sote.hu 2 German Diabetes Clinic, German Diabetes Center, Leibniz Institute at the Heinrich-Heine-University Düsseldorf, Auf’m Hennekamp 65, 40225 Düsseldorf 3 IIIrd Department of Internal Medicine, Semmelweis University, Budapest and Research Group of Metabolism and Atherosclerosis, Hungarian Academy of Sciences, Kutvolgyi ut 4., 1125, Budapest, Hungary 4 Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Budapest 112, PO Box 32, 1518, Hungary 5 Department of Organic Chemistry, Loránd Eötvös University, Budapest 112, PO Box 32, 1518, Hungary Received 24 November 2004; returned for revision 22 December 2004; accepted by I. Ahnfelt-Rønne 20 May 2005 Abstract. Rationale: Type 1 diabetes mellitus (T1) is considered to be an immune mediated disease. Based on previous findings it might be suggested that heat shock protein 60 (Hsp60) could be involved in the mediation of the development of the disease. Furthermore a bias toward Th1 immune response was observed in T1D patients where the level of Th1 cytokines was elevated, while the level of Th2 was decreased. Aim of the study: To determine Th1 (IFN-g) and Th2 (IL-13) cytokine levels in T1 diabetic and control subjects as well as to determine whether there is a shift towards Th1 or Th2 immune response. Materials and methods: ELISPOT (Enzyme-linked Immu- noSPOT) analysis was employed to differentiate antigen spe- cific T-cell responses of a Th1 (IFN-g) or Th2 (IL-13) type. 11 T1 diabetic patients and 9 healthy controls were investi- gated. For T-cell stimulation, we used a polyclonal mitogen or Tetanus toxoid (TT) as positive controls and two peptide antigens Hsp60 AA394–408 and Hsp60 AA437–460. Results: In case of Hsp60 AA437–460 we found significantly decreased Th2 response in patients, although there was no significant difference in Th1 response. In case of Hsp60 AA394–408 and positive controls there was no significant difference. Conclusion: Comparing the control and diabetic subjects a significant shift towards Th1 response in T1 diabetes mellitus for Hsp60 AA437–460 was observed. Key words: Interferon-g – Interleukin-13 – Diabetes – Heat shock protein 60 – p277 Introduction Numerous studies suggested that T1 diabetes mellitus can be considered an immune system mediated disease [1] leading to destruction of beta cells in the pancreas and resulting in the lack of insulin production. In the past decades several studies proposed new cellular mechanisms of the beta cell destruction process [2]. Since T1 diabetes is the result of an autoimmune process, the disease could potentially be influenced by the modifica- tion of the inflammatory autoimmune process. Therefore it is important to identify the target antigens to develop an effec- tive immunotherapy. As candidate targets in the autoimmune process, the fam- ily of 60 kD heat shock proteins (Hsp60, Hsp65) in beta cells could be considered. Antibodies and T-cells reactive to Hsp60 were detected in mice with autoimmune diabetes induced by the beta cell toxin, streptozotocin (STZ) [3]. T-cells reactive to a peptide antigen, corresponding to a conserved Hsp60 domain of amino acids 437–460 have been observed in STZ susceptible male mice of the C57BL/KsJ strain [3]. More- over, both Hsp65 reactive T-cells and antibodies have been detected in NOD/Lt mice in the course of the autoimmune insulitis [4]. In extension of these results, vaccination with different Hsps (Hsp60, Hsp65) and Hsp peptide fragments, like p277 (our Hsp60 AA437–460 peptide), prevented the de- velopment of low-dose STZ induced diabetes in C57BL/KSJ Correspondence to: A. Szebeni