{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T23:07:40Z","timestamp":1773702460174,"version":"3.50.1"},"reference-count":19,"publisher":"Oceanside Publications Inc.","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["allergy asthma proc"],"published-print":{"date-parts":[[2025,3,1]]},"abstract":"Background:<\/jats:bold> Allergen immunotherapy (AIT) is the most effective treatment for atopic allergic diseases, aiming to induce regulatory T cells (Treg) that modify the immune response to specific allergens, which leads to long-term tolerance and reduced symptoms. Enhancing Treg\n activity is crucial for improving immunotherapy outcomes. In a previous murine model study, we examined the effects of a synthetic methylated DNA oligodeoxynucleotide (ODN) from the Bl-T2 m5C motif of Bifidobacterium longum<\/jats:italic> subsp. infantis<\/jats:italic>. The ODN that contains the methylated\n BI-T2 m5C motif (methylated ODNA) sequence conjugated with ovalbumin induced Treg production, whereas ODN that contains the unmethylated BI-T2 m5C motif (unmethylated ODNB) induced proinflammatory responses, which demonstrated the potential of methylated ODNs for AIT. Objective:<\/jats:bold>\n In building on these results, this study explored the effects of methylated and nonmethylated DNA motifs from B. longum<\/jats:italic> subsp. infantis<\/jats:italic> on inflammation and Treg induction, while investigating the dose-response relationships of methylated Cytosine-phosphate-Guanine (CpG) ODNs\n for optimal Treg stimulation in clinical applications. Methods:<\/jats:bold> Serum levels of IL-17A, IL-4, IL-10, and transforming growth factor beta (TGF-\u03b2) were measured by enzyme linked immunosorbent assay (ELISA), and flow cytometry assessed splenic Treg populations in BALB\/c\n mice receiving graded doses of methylated or unmethylated ODNs. Mice were immunized intraperitoneally with a single 100-\u03bcg dose (plan A) or multiple 25 \u03bcg (plan B) or 100 \u03bcg (plan C) doses. Calf thymic DNA served as a positive control, with phosphate-buffered saline solution and alum\n as negative controls. Results:<\/jats:bold> Methylated ODNs significantly increased CD25+<\/jats:sup>FOXP3+<\/jats:sup> Tregs compared with unmethylated ODNs and controls. Plan A (100 \u03bcg) elevated serum IL-10, which indicated effective Treg induction, whereas plan B (four 25 \u03bcg\n doses) did not activate Tregs. Plan C (multiple 100 \u03bcg doses) reduced Treg responses, which highlighted a critical dosing threshold for optimal Treg induction. Conclusion:<\/jats:bold> This study demonstrated the potential of methylated DNA motifs as therapeutic agents in AIT. The\n dose-response relationships of methylated CpG ODNs from B. longum<\/jats:italic> pave the way for clinical applications that target Treg activity in allergic diseases.<\/jats:p>","DOI":"10.2500\/aap.2025.46.250001","type":"journal-article","created":{"date-parts":[[2025,2,27]],"date-time":"2025-02-27T04:31:02Z","timestamp":1740630662000},"page":"98-104","source":"Crossref","is-referenced-by-count":7,"title":["Dose-response studies of methylated and nonmethylated CpG ODNs from Bifidobacterium<\/i> longum subsp. infantis<\/i> for optimizing Treg cell stimulation"],"prefix":"10.2500","volume":"46","author":[{"given":"Dongmei","family":"Li","sequence":"first","affiliation":[{"name":"From the Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, D.C."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Idalia","family":"Cruz","sequence":"additional","affiliation":[{"name":"Department of Animal Shared Resource, Georgetown University Medical Center, Washington, D.C., and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sharareh","family":"Sorkhabi","sequence":"additional","affiliation":[{"name":"From the Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, D.C."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia L.","family":"Foley","sequence":"additional","affiliation":[{"name":"Department of Animal Shared Resource, Georgetown University Medical Center, Washington, D.C., and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie","family":"Wagner","sequence":"additional","affiliation":[{"name":"From the Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, D.C."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph A.","family":"Bellanti","sequence":"additional","affiliation":[{"name":"From the Department of Microbiology and Immunology, Georgetown University Medical 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