{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T06:28:57Z","timestamp":1766125737280,"version":"3.48.0"},"reference-count":57,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T00:00:00Z","timestamp":1766102400000},"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. Immunol."],"abstract":"<jats:p>\n                    Virus-specific CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells typically undergo T helper (Th) 1 differentiation and contribute to a type 1 immune response in infection with lymphocytic choriomeningitis virus (LCMV). Using this model pathogen, we performed an in-depth analysis of the quantitative expression stability of the Th1 key transcription factor T-bet. Previously, it was shown that virus-specific Th1 cells arising in acute infections expressed T-bet at distinct intensities and maintained their T-bet expression differences after viral clearance as memory cells for weeks in the steady state. However, it was unclear whether differential T-bet expression was associated with heterogeneity inside the Th1 population and if the quantitative T-bet memory, particularly of those cells expressing T-bet at low levels, could withhold the strong and continuous stimulation present during chronic infection. Using T-bet-ZsGreen reporter mice, virus-specific Th1 cells were characterized phenotypically at protein, RNA, and DNA\/chromatin accessibility levels. The Th1 cells arising during acute LCMV Armstrong infection showed a continuous spectrum of T-bet expression, ranging from cells with very high T-bet to cells with low T-bet. Even though the cells with low T-bet expression clearly possessed Th1 characteristics, they additionally showed certain T follicular helper (Tfh)-like features at protein and RNA level. When virus-specific Th1 cells were sorted according to T-bet-ZsGreen reporter expression intensity, adoptively transferred, and rechallenged by infecting the host animals with the chronic LCMV Clone 13 strain, they maintained quantitative differences in T-bet reporter and IFN-\u03b3 expression levels. The progeny of the former T-bet\n                    <jats:sup>low<\/jats:sup>\n                    cells still included a subpopulation with a mild Tfh-associated phenotype. Independent of their past and present T-bet expression level, all virus-reactive CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells acquired phenotypic signs of exhaustion as characterized by upregulation of PD-1, LAG3, and TOX and vast absence of effector cytokine co-expression in the chronic infection environment. Collectively, our findings highlight the heterogeneity of T-bet\n                    <jats:sup>+<\/jats:sup>\n                    antiviral CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells and the stability of quantitative differences in individual virus-specific CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells during chronic viral infection.\n                  <\/jats:p>","DOI":"10.3389\/fimmu.2025.1716422","type":"journal-article","created":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T06:25:10Z","timestamp":1766125510000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Stable characteristics of intrapopulation heterogeneity in virus-specific Th1 cells during chronic viral challenge 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