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. 2023 Apr 15;14:2164. doi: 10.1038/s41467-023-37835-9

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Overview of study cohort (n = 69), and b the timeline of longitudinal sampling. Patients are grouped based on peak disease severity, including mild (green), moderate (orange) and severe (red). Individual patients are color-coded based on daily disease severity. PaO2 is expressed in kPa. c Gating strategy to identify cTfh cells in PBMCs by flow cytometry. From single, live CD3⁺ CD4⁺ T cells, memory CD4⁺ T cells were identified as CD45RA⁻. From memory CD4⁺ T cells, cTfh cells were identified as CXCR5⁺ cells. cTfh subsets were identified as cTfh1 (CXCR3⁺ CCR6⁻), cTfh2 (CXCR3⁻ CCR6⁻) and cTfh17 (CXCR3^- CCR6⁺). Activated cTfh and its subsets were identified with ICOS⁺ CD38⁺ expressing. Patients are grouped based on peak disease severity, including mild (green), moderate (orange) and severe (red). d Bar charts show the frequency of bulk and activated cTfh, cTfh1, cTfh2, and cTfh17 cells from COVID-19 patients with acute disease (Acute, full circle), 3 months convalescence (3 months, half circle) and 8 months convalescence (8 months, open circle) with median. Dots are individual samples color-coded according to peak disease severity. Dotted lines show the median frequency with 95% Cl (gray area) of prepandemic healthy controls. X axis shows the number of patients in each bar. During acute disease, 9, 22, and 33 individual samples from 9 mild, 14 moderate and 18 severe patients, respectively, were analyzed using two-sided Generalized Estimating Equations (GEE) to account for the intra-person correlations inherent to repeated measures and assess statistically significant differences without adjusting for multiple comparisons. During 3 and 8 months convalescence, only one sample from each patient was analyzed and two-sided Kruskal–Wallis with Dunn’s multiple comparisons test was used to assess statistically significant differences. P < 0.05 was considered to be a significant difference. *P values <0.05 are listed above each comparison. Source data are provided as a Source Data file.