{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T06:04:45Z","timestamp":1790057085291,"version":"4.0.1"},"reference-count":61,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004735","name":"Natural Science Foundation of Hunan Province","doi-asserted-by":"publisher","award":["2025JJ50650"],"award-info":[{"award-number":["2025JJ50650"]}],"id":[{"id":"10.13039\/501100004735","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100017695","name":"Health Commission of Hunan Province","doi-asserted-by":"publisher","award":["D202310008129"],"award-info":[{"award-number":["D202310008129"]}],"id":[{"id":"10.13039\/100017695","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Mycoplasma pneumoniae<\/jats:italic>\n                      (Mp) remains a major cause of community-acquired pneumonia in children and adolescents, and no vaccine against this pathogen  is currently available. This study designed and evaluated P1Tx, a novel multi-epitope vaccine candidate derived from the immunodominant regions of the P1 adhesin and the community-acquired respiratory distress syndrome (CARDS) toxin.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Immunoinformatics approaches were used to select immunodominant epitopes from the P1 adhesin and the CARDS toxin of\n                      <jats:italic>Mp<\/jats:italic>\n                      , which were assembled into a 263-amino acid chimeric construct designated P1Tx. The construct was expressed in\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      , and the recombinant protein was characterized for physicochemical properties and predicted structural stability. Its immunoreactivity was assessed using sera from\n                      <jats:italic>Mp<\/jats:italic>\n                      -infected patients. BALB\/c mice received three subcutaneous immunizations with P1Tx, and humoral and cellular immune responses were evaluated by antibody endpoint titers and flow cytometric analysis of germinal center B cells and T cell subsets. The functional activity of immune sera was tested by measuring its capacity to block\n                      <jats:italic>Mp<\/jats:italic>\n                      adhesion to human bronchial epithelial cells and to neutralize CARDS toxin-induced cytotoxicity. Protective efficacy was assessed in mice challenged intranasally with the\n                      <jats:italic>Mp<\/jats:italic>\n                      M129 strain, with pulmonary bacterial burden and histopathological changes determined after challenge.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      P1Tx displayed favorable physicochemical characteristics and structural stability and reacted strongly with sera from\n                      <jats:italic>Mp<\/jats:italic>\n                      -infected patients. Immunized mice developed high-titer, IgG1-dominant antibody responses (endpoint titers up to 1:50,000) accompanied by marked germinal center B cell expansion. Compared with the P1C subunit vaccine, P1Tx elicited substantially stronger cellular immunity, with significant expansion of CD4+ and CD8+ effector and memory T cell populations. Sera from P1Tx-immunized mice inhibited\n                      <jats:italic>Mp<\/jats:italic>\n                      adhesion to bronchial epithelial cells and neutralized CARDS toxin-mediated cytotoxicity in vitro. Following intranasal challenge, P1Tx-immunized mice showed a 98.5% reduction in pulmonary bacterial burden and a 52.4% reduction in lung histopathological scores relative to the PBS group.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      P1Tx confers protection against\n                      <jats:italic>Mp<\/jats:italic>\n                      infection through complementary mechanisms of adhesion blockade and toxin neutralization, supporting its further development as a vaccine candidate for the prevention of\n                      <jats:italic>Mp<\/jats:italic>\n                      infection.\n                    <\/jats:p>\n                  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