{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T21:55:53Z","timestamp":1791410153815,"version":"4.3.3"},"reference-count":48,"publisher":"Walter de Gruyter GmbH","issue":"4","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["BM"],"published-print":{"date-parts":[[2015,8]]},"abstract":"<jats:p>Type 1 diabetes (T1D) is a chronic autoimmune disease characterised by excessive immune reactions against auto-antigens of pancreatic \u03b2-cells. Restoring auto-antigen tolerance remains the superior therapeutic strategy. Oral auto-antigen administration uses the tolerogenic nature of the gut-associated immune system to induce antigen-specific tolerance. However, due to gastric degradation, proper mucosal product delivery often imposes a challenge. Recombinant <jats:italic>Lactococcus lactis<\/jats:italic> have proven to be effective and safe carriers for gastrointestinal delivery of therapeutic products: <jats:italic>L. lactis<\/jats:italic> secreting diabetes-associated auto-antigens in combination with interleukin (IL)-10 have demonstrated therapeutic efficacy in a well-defined mouse model for T1D. Here, we describe the construction of recombinant <jats:italic>L. lactis<\/jats:italic> secreting the 65 kDa isoform of glutamic acid decarboxylase (GAD65) and tyrosine phosphatase-like protein ICA512 (IA-2), two major T1D-related auto-antigens. Attempts to secrete full size human GAD65 and IA-2 protein by <jats:italic>L. lactis<\/jats:italic> were unsuccessful. Trimming of GAD65 and IA-2 was investigated to optimise antigen secretion while maintaining sufficient bacterial growth. GAD65<jats:sub>370-575<\/jats:sub> and IA-2<jats:sub>635-979<\/jats:sub> showed to be efficiently secreted by recombinant <jats:italic>L. lactis<\/jats:italic>. Antigen secretion was verified by immunoblotting. Plasmid-derived GAD65 and IA-2 expression was combined in single strains with human IL-10 expression, a desired combination to allow tolerance induction. This study reports the generation of recombinant <jats:italic>L. lactis<\/jats:italic> secreting two major diabetes-related auto-antigens: human GAD65 and IA-2, by themselves or combined with the anti-inflammatory cytokine human IL-10. Prohibitive sequence obstacles hampering antigen secretion were resolved by trimming the full size proteins.<\/jats:p>","DOI":"10.3920\/bm2014.0083","type":"journal-article","created":{"date-parts":[[2015,1,10]],"date-time":"2015-01-10T02:04:26Z","timestamp":1420855466000},"page":"591-602","source":"Crossref","is-referenced-by-count":12,"title":["Trimming of two major type 1 diabetes driving antigens, GAD65 and IA-2, allows for successful expression in Lactococcus lactis"],"prefix":"10.1163","volume":"6","author":[{"given":"S.","family":"Robert","sequence":"first","affiliation":[{"name":"Clinical and Experimental Endocrinology (CEE), KU Leuven, Herestraat 49 bus 902, 3000 Leuven, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Van Huynegem","sequence":"additional","affiliation":[{"name":"ActoGeniX NV, Technologiepark 4, 9052 Zwijnaarde, 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