{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T05:31:04Z","timestamp":1792215064486,"version":"4.4.0"},"reference-count":44,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T00:00:00Z","timestamp":1666742400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation (FAPESP)","doi-asserted-by":"publisher","award":["2018\/18538-0"],"award-info":[{"award-number":["2018\/18538-0"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation (FAPESP)","doi-asserted-by":"publisher","award":["2019\/26074-7"],"award-info":[{"award-number":["2019\/26074-7"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation (FAPESP)","doi-asserted-by":"publisher","award":["2020\/09113-6"],"award-info":[{"award-number":["2020\/09113-6"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation (FAPESP)","doi-asserted-by":"publisher","award":["2020\/11307-3"],"award-info":[{"award-number":["2020\/11307-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation (FAPESP)","doi-asserted-by":"publisher","award":["IRG#123660"],"award-info":[{"award-number":["IRG#123660"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Institutional Research Grant from MD Anderson Cancer Center, Houston, TX","award":["2018\/18538-0"],"award-info":[{"award-number":["2018\/18538-0"]}]},{"name":"Institutional Research Grant from MD Anderson Cancer Center, Houston, TX","award":["2019\/26074-7"],"award-info":[{"award-number":["2019\/26074-7"]}]},{"name":"Institutional Research Grant from MD Anderson Cancer Center, Houston, TX","award":["2020\/09113-6"],"award-info":[{"award-number":["2020\/09113-6"]}]},{"name":"Institutional Research Grant from MD Anderson Cancer Center, Houston, TX","award":["2020\/11307-3"],"award-info":[{"award-number":["2020\/11307-3"]}]},{"name":"Institutional Research Grant from MD Anderson Cancer Center, Houston, TX","award":["IRG#123660"],"award-info":[{"award-number":["IRG#123660"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Chimeric antigen receptors (CARs) redirect T cells to recognize a specific target. CAR components play a pivotal role in antigen specificity, structure stability, expression on cell surface, and induction of cellular activation, which together determine the success of CAR T-cell therapy. CAR products targeting B-cell lymphoma encouraged the development of new CAR applications beyond cancer. For example, our group developed a CAR to specifically target glucuronoxylomannan (GXM) in the capsule of Cryptococcus species, called GXMR-CAR or GXMR-IgG4-28\u03b6. Cryptococcus are fungi that cause the life-threatening disease cryptococcosis, and GXMR-IgG4-28\u03b6 redirected T cells to target yeast and titan cell forms of Cryptococcus spp. Here, we replaced the IgG4-hinge and CD28-transmembrane domains from GXMR-CAR with a CD8\u03b1 molecule as the hinge\/transmembrane and used CD28 or 4-1BB molecules as co-stimulatory domains, creating GXMR-8-28\u03b6 and GXMR-8-BB\u03b6, respectively. Jurkat cells expressing GXMR-CAR containing CD8\u03b1 as the hinge\/transmembrane improved the CAR expression and induced a tonic signaling. GXMR-8-28\u03b6 and GXMR-8-BB\u03b6 induced high levels of IL-2 and up-regulation of CD69 expression in the presence of reference strains of C. neoformans and C. gattii. Moreover, GXMR-8-28\u03b6 and GXMR-8-BB\u03b6 showed increased strength in response to incubation with clinical isolates of Cryptococcuss spp., and 4-1BB co-stimulatory domain triggered a more pronounced cellular activation. Dasatinib, a tyrosine kinase inhibitor, attenuated the GXMR-CAR signaling cascade\u2019s engagement in the presence or absence of its ligand. This study optimized novel second-generation GXMR-CARs containing the CD8-hinge\/transmembrane domain that improved CAR expression, antigen recognition, and signal strength in T-cell activation.<\/jats:p>","DOI":"10.3390\/cells11213386","type":"journal-article","created":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T09:59:37Z","timestamp":1666778377000},"page":"3386","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Modification of Hinge\/Transmembrane and Signal Transduction Domains Improves the Expression and Signaling Threshold of GXMR-CAR Specific to Cryptococcus spp."],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6121-3825","authenticated-orcid":false,"given":"Matheus Henrique","family":"dos Santos","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 140490-900, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele Proc\u00f3pio","family":"Machado","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 140490-900, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pappanaicken R.","family":"Kumaresan","sequence":"additional","affiliation":[{"name":"Department of Hematopoietic Biology and Malignancy, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5017-6539","authenticated-orcid":false,"given":"Thiago Aparecido","family":"da Silva","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo, Ribeir\u00e3o Preto 140490-900, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1158\/2159-8290.CD-12-0548","article-title":"The basic principles of chimeric antigen receptor design","volume":"3","author":"Sadelain","year":"2013","journal-title":"Cancer Discov."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1007\/s00262-012-1254-0","article-title":"Targeted immunotherapy of cancer with CAR T cells: Achievements and challenges","volume":"61","author":"Gilham","year":"2012","journal-title":"Cancer Immunol. 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