PMC9868302_01
(A) Bilateral ground-glass opacities with crazy-paving aspects and more consolidated components, greater at right lower lobe. (B) Extension of pulmonary opacities at the left basal lobe compared to the previous imaging.
fmed-09-1062450-g001.jpg
PMC9868302_02_fmed-09-1062450-g001.jpg
F1
PMC9868302_02
(A) Bilateral ground-glass opacities with crazy-paving aspects and more consolidated components, greater at right lower lobe. (B) Extension of pulmonary opacities at the left basal lobe compared to the previous imaging.
fmed-09-1062450-g001.jpg
PMC9868302_02_fmed-09-1062450-g001.jpg
F1
(A) Interstitial pneumonia with peripheral small nodules. Postero-basal and para-scissural opacities. (B) Ground-glass opacities at the bases of the upper lobes dorsally, with partial periscissural consolidative area. Moderate bilateral pleural effusion with associated disventilatory area.
fmed-09-1062450-g002.jpg
PMC9868302_02_fmed-09-1062450-g002.jpg
F2
Due to the recurrent fever up to 38 C starting at the beginning of February 2022, he underwent a chest CT scan with findings of interstitial pneumonia with peripheral small nodules and ground glass opacities (Figure 2A).
Immunofluorescence analyses of two patients with RT-PCR SARS-CoV-2 positive viremia and bronchoalveolar lavage fluid. (A,B) Representative examples of CD4+ and CD8+ T subset percentages on peripheral CD3+ T cells derived from before and after therapies (A) and frequency of circulating CD4+ Treg (CD4 + FoxP3 + CD25 +) on circulating CD4+ T population derived from before and after therapies (B). (C-G) Multidimensional data reduction analysis. t-SNE analysis of maturation, senescence, exhaustion, effector and activation markers of circulating CD4+ and CD8+ T cells on merged samples derived before (red clusters) and after (bleu clusters) therapies. In particular: the analysis of maturation and exhaustion marker expression in CD4+ (C) and CD8+ (D) T subsets; the analysis of maturation, senescence and granzyme effector molecule in CD8+ T cells (E); the evaluation of cycling cells by ki-67 marker in CD8+ (F) and CD4+ (G) T subsets.
fmed-09-1062450-g003.jpg
PMC9868302_02_fmed-09-1062450-g003.jpg
F3
Firstly, we observed a decrease in the percentage of circulating CD4+ T lymphocytes and a consequent elevation of the CD8+ counterpart in both patients (Figure 3A) after therapies. As a consequence of this drastic decrease in circulating TCD4+, we also recorded an important reduction in the frequency of circulating CD4 + FoxP3 + CD25 high Treg lymphocytes after therapies (Figure 3B). T cells (ki-67+, CD45RA-CCR7-) in samples before therapies (Figure 3F); (b) CD4+ T cell comparison also showed a cluster of ki-67+ cells in samples before therapies which exhibits expression of CCR7, CXCR5 markers, showing a presence of T helper follicular (TFH) cells only in samples before therapies (Figure 3G). Therefore, the analysis of the CD4+ and CD8+ T cell maps analyzed before and after combination therapy in RNAemic and non-RNAemic samples, respectively, would seem to highlight immunological patterns indicative of a resolution of the infection in the samples derived after therapies, such as the decrease of EOMES expression, the activation status and the reduction of granzyme level.