{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T23:16:05Z","timestamp":1785798965433,"version":"3.56.0"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,4,30]],"date-time":"2025-04-30T00:00:00Z","timestamp":1745971200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Mycobacterium tuberculosis (Mtb)<\/jats:italic> presents a significant global health threat, and the existing treatments have notable limitations. V\u03b39V\u03b42 T cells activated by HMBPP can inhibit the growth of intracellular Mtb. Additionally, vitamin C (VC) promotes the differentiation and proliferation of related T cells. However, it remains uncertain whether VC can enhance the expansion of V\u03b39V\u03b42 T cells within PBMCs activated by HMBPP and rIL-2, and the underlying mechanism of the inhibitory effect of the expanded T cells on intracellular <jats:italic>Mtb<\/jats:italic> has not been elucidated.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Venous blood was collected from healthy individuals, and PBMCs were subsequently isolated. In vitro, V\u03b39V\u03b42 T cells were selectively expanded with HMBPP, rIL-2, and VC. Flow cytometry was utilized to analyze the purities and phenotypes of V\u03b39V\u03b42 T cells, while cell counts were performed to determine the total number of viable cells. Magnetic bead sorting was employed to purify V\u03b39V\u03b42 T cells. <jats:italic>Mtb<\/jats:italic> strains were cultured, and macrophage infection models derived from THP1 cells were established. Co-culture experiments were conducted with <jats:italic>Mtb<\/jats:italic>-infected macrophages and V\u03b39V\u03b42 T cells, and the number of intracellular bacteria was quantified through CFU counting. The levels of cytokines were measured using the CBA method and flow cytometry. Statistical analysis was carried out using GraphPad Prism and SPSS software.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>VC (70 \u03bcM) significantly enhances the expansion of V\u03b39V\u03b42 T cells within PBMCs during primary HMBPP activation in the presence of rIL-2, with higher induction rates and total cell proliferation. By day 14 of induction, V\u03b39V\u03b42 T cells expanded with HMBPP, VC, and rIL-2 exhibited the central memory (10-20%) and the effector memory phenotypes (75-90%). Furthermore, these expanded T cells effectively inhibited the growth of intracellular virulent <jats:italic>Mtb<\/jats:italic> strain (H37Rv) in a cell-contact-dependent manner. The inhibitory effect was associated with an up-regulated production of TNF-\u03b1 and IFN-\u03b3, and a down-regulated expression of IL-10 and IL-17A during <jats:italic>Mtb<\/jats:italic> infection. <\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study demonstrates that VC enhances the proliferative expansion of V\u03b39V\u03b42 T cells in PBMCs primarily stimulated with HMBPP and rIL-2. The expanded V\u03b39V\u03b42 T cells are capable of effectively inhibiting the growth of virulent H37Rv strain, likely through the secretion of TNF-\u03b1 and IFN-\u03b3. These findings provide a novel direction for tuberculosis treatment research.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2025.1533277","type":"journal-article","created":{"date-parts":[[2025,4,30]],"date-time":"2025-04-30T05:42:42Z","timestamp":1745991762000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["V\u03b39V\u03b42 T cells expanded with vitamin C combined with HMBPP in vitro inhibit intracellular Mycobacterium tuberculosis growth"],"prefix":"10.3389","volume":"15","author":[{"given":"Shuyan","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guohui","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanxin","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teng","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaoxiang","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuju","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenwen","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoliang","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,4,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"580","DOI":"10.1128\/iai.01262-15","article-title":"Mycobacterium-specific \u03b39\u03b42 T cells mediate both pathogen-inhibitory and CD40 ligand-dependent antigen presentation effects important for tuberculosis immunity","volume":"84","author":"Abate","year":"2016","journal-title":"Infect. 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