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EuropePMC; 2021.
Preprint in English | EuropePMC | ID: ppcovidwho-313420


Aim: Exosomes are frequently used for characterization of diseases, e.g., viral infections, but have not been reported for COVID-19. The aims of our study were to determine exosomes’ novel applications for SARS-CoV-2 RNA infection and prognosis of COVID-19 disease.Methods: From 15 COVID-19 patients and five healthy controls, 32 serum samples were collected by clinicians in Wuhan, China. Exosomes were purified, their RNAs (exoRNAs) were isolated and analyzed using RT-PCR for the presence of the N and/or ORF1ab viral genes, and the human RNaseP gene as reference.Results: Exosomes showed their typical lipid bilayer structure between 30 to 150 nm in size. No viral RNAs were detected from the 5 healthy controls. From 4 patients (26.67% of the patients), only the viral N but not the ORF1ab genes was detected. However, intact viral particles were not observed in exosome samples from the four positive patients. Importantly, expression of the N gene was detected during the active or early-stages of the disease, and the expression levels gradually decreased from the critical to the mild patients (in reverse to their CT values): Patient 1 /critical/ progress/ (CT-value: 34.74), Patient 9/severe/progress (CT-value: 37.65), Patient 11/severe /progress (CT-value: 37.16), and Patient 15/mild/early stage (CT-value: 38.44). Among them, only patient 1 had received stem cell transfusion. Patients 9 and 11 demonstrated recurrence of SARS-CoV-2. Patient 15 was the earliest phase patient with the N gene.Conclusion: SARS-CoV-2 RNAs were detected in exosomes which came only from the 15 COVID-19 patients but not from the 5 healthy controls. In addition, the presence of the viral N gene showed association with severity in the disease. Our novel discovery indicates that SARS-CoV-2 RNAs in serum exosomes can be used to complement existing assays for characterization of the SARS-CoV-2 infection and for prognosis of the COVID-19 disease.Funding Statement: This study was funded by National Natural Science Foundation of China (Grant No. 81702273), the Key Project for Anti-2019 Novel Coronavirus Pneumonia from the National Key Research and Development Program of China (Grant No. 2020YFC0845500), the National Key Research and Development Program of China (Grant No. 2018YFE0204500), Science and technology key project of Guangdong Province: Study on the source and epidemiology of COVID-19 (No.2020B111107001), the Scientific research project of COVID-19 epidemic prevention and control in Guangdong universities (No.2020KZDZX1087), and the Zhongnan Hospital of Wuhan University Science, Technology and Innovation Seed Fund, Project znpy2018117 and znpy2019064.Declaration of Interests: We declare no competing interests.Ethics Approval Statement: The study was reviewed and approved by Medical Ethics Committee Zhongnan Hospital of Wuhan University in the Scientific Research project No 2020066.

Cytokine ; 143: 155523, 2021 07.
Article in English | MEDLINE | ID: covidwho-1163610


Cytokines play pleiotropic, antagonistic, and collaborative in viral disease. The high morbidity and mortality of coronavirus disease 2019 (COVID-19) make it a significant threat to global public health. Elucidating its pathogenesis is essential to finding effective therapy. A retrospective study was conducted on 71 patients hospitalized with COVID-19. Data on cytokines, T lymphocytes, and other clinical and laboratory characteristics were collected from patients with variable disease severity. The effects of cytokines on the overall survival (OS) and event-free survival (EFS) of patients were analyzed. The critically severe and severe patients had higher infection indexes and significant multiple organ function abnormalities than the mild patients (P < 0.05). IL-6 and IL-10 were significantly higher in the critically severe patients than in the severe and mild patients (P < 0.05). IL-6 and IL-10 were closely associated with white blood cells, neutrophils, T lymphocyte subsets, D-D dimer, blood urea nitrogen, complement C1q, procalcitonin C-reactive protein. Moreover, the IL-6 and IL-10 levels were closely correlated to dyspnea and dizziness (P < 0.05). The patients with higher IL-10 levels had shorter OS than the group with lower levels (P < 0.05). The older patients with higher levels of single IL-6 or IL-10 tended to have shorter EFS (P < 0.05), while the patients who had more elevated IL-6 and IL-10 had shorter OS (P < 0.05). The Cox proportional hazard model revealed that IL-6 was the independent factor affecting EFS. IL-6 and IL-10 play crucial roles in COVID-19 prognosis.

COVID-19/blood , COVID-19/pathology , Interleukin-10/blood , Interleukin-6/blood , T-Lymphocyte Subsets/immunology , Adult , Age Factors , Aged , Aging , Blood Coagulation Factors/analysis , COVID-19/mortality , COVID-19/therapy , Cytokine Release Syndrome/pathology , Female , Humans , Lymphocyte Count , Male , Middle Aged , Retrospective Studies , SARS-CoV-2/immunology , Severity of Illness Index , Survival Analysis , T-Lymphocyte Subsets/cytology , Thromboembolism/pathology , Treatment Outcome