{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T13:48:46Z","timestamp":1791985726588,"version":"4.4.0"},"reference-count":36,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R24AI192252, R01AI172154, 75N93019C00064"],"award-info":[{"award-number":["R24AI192252, R01AI172154, 75N93019C00064"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI125045"],"award-info":[{"award-number":["R01AI125045"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Infections due to\n                      <jats:italic toggle=\"yes\">Cryptococcus<\/jats:italic>\n                      species are estimated to cause over 100,000 deaths a year. No cryptococcal vaccines are approved for human use. We have shown that vaccine formulations consisting of\n                      <jats:italic toggle=\"yes\">C. neoformans<\/jats:italic>\n                      chitin deacetylase 2 (Cda2) and Cda1 adjuvanted with Cationic Adjuvant Formulation 01 (CAF01) protect mice against experimental cryptococcosis by a mechanism dependent upon CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells. Here, using overlapping peptide libraries, we mapped epitopes in Cda2 and Cda1 that stimulated interferon-gamma (IFN\u03b3) release from splenocytes of C57BL\/6 and DR4 mice immunized with these proteins. DR4 mice lack mouse major histocompatibility class II (MHC-II) proteins and express chimeric MHC-II proteins with the specificity of the human HLA-DR allele, HLA-DRB1*04:01. Experimental results were then compared with MHC-II binding predictions using the Immune Epitope Database NetMHCIIpan 4.1 BA MHC-II epitope prediction tool. Unique epitopes in Cda2 and Cda1 were discovered for each mouse strain, some of which were not predicted to bind well to MHC-II. CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells were responsible for cytokine release as IFN\u03b3 production was lost if CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells were depleted from the splenocyte population. Finally, we tested a hybrid recombinant protein that consisted of portions of Cda2 and Cda1 containing the MHC-II\/CD4\n                      <jats:sup>+<\/jats:sup>\n                      T-cell epitopes that induce CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells in DR4 mice. When administered as a CAF01-adjuvanted vaccine, the hybrid protein protected DR4 mice from an otherwise lethal\n                      <jats:italic toggle=\"yes\">C. neoformans<\/jats:italic>\n                      pulmonary challenge. These results provide a proof of principle regarding the utility of MHC-II\/CD4\n                      <jats:sup>+<\/jats:sup>\n                      T-cell epitope mapping in cryptococcal vaccine development.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The development of vaccines to prevent cryptococcosis, a fungal disease which causes over 100,000 deaths annually, is a public health priority. We have demonstrated that adjuvanted vaccines containing two\n                      <jats:italic toggle=\"yes\">Cryptococcus neoformans<\/jats:italic>\n                      protein antigens, chitin deacetylase (Cda)2 and Cda1, protect mice against experimental cryptococcosis by a mechanism dependent upon CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells. In this study, we determined which 15-amino-acid peptides of Cda2 and Cda1 stimulate CD4\n                      <jats:sup>+<\/jats:sup>\n                      T cells in C57BL\/6 and DR4 mice; the latter mouse strain has human major histocompatibility class II (MHC-II) specificity. We then synthesized a small hybrid protein containing those portions of Cda2 and Cda1 that strongly stimulated CD4\n                      <jats:sup>+<\/jats:sup>\n                      T-cell responses in DR4 mice. As an adjuvanted vaccine, the hybrid Cda2\/1 protein protected DR4 mice from a\n                      <jats:italic toggle=\"yes\">C. neoformans<\/jats:italic>\n                      pulmonary challenge. Thus, CD4\n                      <jats:sup>+<\/jats:sup>\n                      T-cell\/MHC-II epitope mapping experimentally validated the design of a protective multi-epitope protein for use in cryptococcal vaccines.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.02176-26","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T13:12:21Z","timestamp":1791983541000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["A protective hybrid protein vaccine informed by mapping major histocompatibility class II epitopes in\n                    <i>Cryptococcus neoformans<\/i>\n                    chitin deacetylases that stimulate CD4\n                    <sup>+<\/sup>\n                    T-cell responses"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5749-3112","authenticated-orcid":true,"given":"Charles A.","family":"Specht","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1762-0076","authenticated-orcid":false,"given":"Lorena V. N.","family":"Oliveira","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-5268-0646","authenticated-orcid":false,"given":"Diana","family":"Carlson","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2680-4854","authenticated-orcid":false,"given":"Maureen M.","family":"Hester","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5762-439X","authenticated-orcid":false,"given":"Satya","family":"Sahu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6872-3975","authenticated-orcid":false,"given":"Gabriel Kristian","family":"Pedersen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0417ye583","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Vaccine Research, Statens Serum Institut","place":["Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3799-3064","authenticated-orcid":true,"given":"Stuart M.","family":"Levitz","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/0464eyp60","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Medicine, University of Massachusetts Chan Medical School","place":["Worcester, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"e_1_3_6_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(22)00499-6"},{"key":"e_1_3_6_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(23)00692-8"},{"key":"e_1_3_6_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-021-00294-8"},{"key":"e_1_3_6_5_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1011115"},{"key":"e_1_3_6_6_2","doi-asserted-by":"publisher","DOI":"10.3390\/jof10120840"},{"key":"e_1_3_6_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jinf.2025.106521"},{"key":"e_1_3_6_8_2","doi-asserted-by":"publisher","DOI":"10.3390\/jof11120886"},{"key":"e_1_3_6_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1012220"},{"key":"e_1_3_6_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-023-00606-0"},{"key":"e_1_3_6_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2019.10.051"},{"key":"e_1_3_6_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01872-17"},{"key":"e_1_3_6_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.01746-24"},{"key":"e_1_3_6_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-022-00707-2"},{"key":"e_1_3_6_15_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20250444"},{"key":"e_1_3_6_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201545930"},{"key":"e_1_3_6_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2023.03.009"},{"key":"e_1_3_6_18_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1155679"},{"key":"e_1_3_6_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-4141(200209)32:9<2510::AID-IMMU2510>3.0.CO;2-Q"},{"key":"e_1_3_6_20_2","doi-asserted-by":"publisher","DOI":"10.3389\/fgene.2023.866407"},{"key":"e_1_3_6_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz1029"},{"key":"e_1_3_6_22_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-082119-124838"},{"key":"e_1_3_6_23_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae1092"},{"key":"e_1_3_6_24_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1014162"},{"key":"e_1_3_6_25_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.183.6.2635"},{"key":"e_1_3_6_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.03367-21"},{"key":"e_1_3_6_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.smim.2018.10.003"},{"key":"e_1_3_6_28_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1004701"},{"key":"e_1_3_6_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-64034-5"},{"key":"e_1_3_6_30_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.03151"},{"key":"e_1_3_6_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2015.12.018"},{"key":"e_1_3_6_32_2","doi-asserted-by":"publisher","DOI":"10.2217\/fmb-2021-0247"},{"key":"e_1_3_6_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.humimm.2026.111788"},{"key":"e_1_3_6_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2012.09.039"},{"key":"e_1_3_6_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00566-12"},{"key":"e_1_3_6_36_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1012876"},{"key":"e_1_3_6_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.71.9.4831-4841.2003"}],"container-title":["mBio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.02176-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.02176-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T13:12:27Z","timestamp":1791983547000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/mbio.02176-26"}},"subtitle":[],"editor":[{"given":"Marcio","family":"Rodrigues","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,10,14]]},"references-count":36,"alternative-id":["10.1128\/mbio.02176-26"],"URL":"https:\/\/doi.org\/10.1128\/mbio.02176-26","relation":{},"ISSN":["2150-7511"],"issn-type":[{"value":"2150-7511","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,14]]},"assertion":[{"value":"2026-09-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-10-14","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e02176-26"}}