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Universitätsfrauenklinik, 74 Tübingen, and *II. Medizinische Universitätsklinik, Düsseldorf, Germany

(Received 9 May 1975)

We have reported previously changes of plasma dehydroepiandrosterone (DHA) and plasma dehydroepiandrosterone sulphate (DHAS) before and during pregnancy (Nieschlag, Walk & Schindler, 1974). There are strong indications that labour represents a stressful stimulus to the endocrine system. This is reflected in a rise of maternal plasma corticotrophin (ACTH), corticosteroids, free fatty acids, insulin, androgens and oestrogens (Migneon, Kenny & Taylor, 1968; Rivarola, Forest & Migneon, 1968; Lefebvre, Chapedelaine & Bolté, 1970; Nieschlag, Wombacher, Kremer & Martin, 1970; Kuwabara, Kihara, Arai & Sakamoto, 1971; Kauppila, Tuimala & Haapalahti, 1974). Steroid changes in cord blood during labour have also been noticed (Arai, Kuwabara, Kihara, Okinaga & Sakamoto, 1972). Determinations of DHA and DHAS concentrations in maternal blood during the course of labour are rare and do show a variety of results (Lefebvre et al. 1970; Gandy, 1971). Therefore, we

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L Monetini, F Barone, L Stefanini, A Petrone, T Walk, G Jung, R Thorpe, P Pozzilli, and MG Cavallo

Enhanced cellular immune response to bovine beta-casein has been reported in patients with type 1 diabetes. In this study we aimed to establish beta-casein-specific T cell lines from newly diagnosed type 1 diabetic patients and to characterise these cell lines in terms of phenotype and epitope specificity. Furthermore, since sequence homologies exist between beta-casein and putative beta-cell autoantigens, reactivity to the latter was also investigated. T cell lines were generated from the peripheral blood of nine recent onset type 1 diabetic patients with different HLA-DQ and -DR genotypes, after stimulation with antigen pulsed autologous irradiated antigen presenting cells (APCs) and recombinant human interleukin-2 (rhIL-2). T cell line reactivity was evaluated in response to bovine beta-casein, to 18 overlapping peptides encompassing the whole sequence of beta-casein and to beta-cell antigens, including the human insulinoma cell line, CM, and a peptide from the beta-cell glucose transporter, GLUT-2. T cell lines specific to beta-casein could not be isolated from HLA-matched and -unmatched control subjects. beta-Casein T cell lines reacted to different sequences of the protein, however a higher frequency of T cell reactivity was observed towards the C-terminal portion (peptides B05-14, and B05-17 in 5/9 and 4/9 T cell lines respectively). Furthermore, we found that 1 out of 9 beta-casein-specific T cell lines reacted also to the homologous peptide from GLUT-2, and that 3 out of 4 of tested cell lines reacted also to extracts of the human insulinoma cell line, CM. We conclude that T cell lines specific to bovine beta-casein can be isolated from the peripheral blood of patients with type 1 diabetes; these cell lines react with multiple and different sequences of the protein particularly towards the C-terminal portion. In addition, reactivity of beta-casein T cell lines to human insulinoma extracts and GLUT-2 peptide was detected, suggesting that the potential cross-reactivity with beta-cell antigens deserves further investigation.