key: cord-1036736-99j84ovg authors: Wang, J.; Dong, X.; Zhang, B.; Yang, X.; Li, Z.; Wang, X.; Zuo, S. title: Characteristics of lymphocyte subsets and their predicting values for the severity of COVID-19 patients date: 2020-05-05 journal: nan DOI: 10.1101/2020.05.01.20086421 sha: c3e671130963eb7f8e0bf2c91caefffb2522a8c5 doc_id: 1036736 cord_uid: 99j84ovg Severe COVID-19 patients showed worse clinical outcomes compared to mild and moderate patients. However, effective indicators are still lacking to predict the severity of the disease. In the present study, we retrospectively analyzed the clinical and laboratory data of 16 COVID-19 patients and found that the absolute counts of three T-cells (CD3+, CD4+, and CD8+) were significantly lower in the moderate and severe patients than those in mild patients and were significantly lower in severe patients than in moderate patients on admission. With the recovery of the COVID-19, serum levels of inflammatory biomarkers (CRP, PCT, and IL6) of moderate and severe patients gradually decreased. In contrast, the counts of lymphocytes and their subsets including CD3+, CD4+, and CD8+ T cells gradually increased in severe patients, and eventually showed comparable levels with moderate patients. ROC analysis showed that the counts of CD3+, CD4+, and CD8+ T-cells with AUC > 0.9 have potential values for predicting the severity of COVID-19 patients. In conclusion, the reduction of CD3+, CD4+, and CD8+ T-cells is related to the severity of COVID-19 and dynamic detection of these T-lymphocyte subsets may help predict the outcome of the patients. were mild, 4 were moderate, and 1 was severe). Except for one patient who is a native Wuhan and have no history of contact with the seafood market. Four patients were 1 4 family clustered, of which three patients were mild and one patient was moderate. The 1 5 median time from the onset of symptoms to admission was 6 days (2 days for mild 1 6 patients, 7 days for moderate patients, and 7.5 days for severe patients). Three patients 1 7 had hypertension (one in the mild group, two in the severe group), and three patients had a fever, of which the mild, moderate and severe groups included 3, 5, and 4 2 0 patients, respectively. Of the 16 patients, 7 had a cough, 6 had sputum production, 2 2 1 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint had chest tightness, 2 had shortness of breath, 6 had fatigue, 1 had a headache, 1 had a 1 sore throat, and 2 had body aches. CT imaging of 6 patients was normal, and the other 2 10 patients were abnormal. Patients with abnormal CT imaging showed abnormalities 3 in the subpleural, pulmonary base, and bronchoalveolar segments, which is consistent 4 with atypical upper respiratory symptoms. Although the CT scan showed abnormal 5 lesions close to the pleura, no pleural fluid was found in all patients, indicating that 6 the pleura was not affected, which is consistent with the absence of chest pain in all 7 patients. Three patients were treated with antibiotics (piracillin and sulbactam, 8 moxifloxacin, and meropenem). All patients received antiviral therapy with inhaled 9 IFN-α for 7 days. Fourteen patients received KALETRA [(lopinavir/ritonavir), BID, 1 0 500 mg/day], of which 12 patients completed a 14-day course, and one severe patient 1 1 was discontinued because of vomiting after 2 doses of medication, and another severe 1 2 patient was stopped due to hepatic dysfunction. Five patients received 1 3 immunoglobulin (400 mg/kg/day), of which one was a moderate patient and four were was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. in both moderate and severe groups. PCT levels continued to be lower from 6 to 21 2 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint days in the moderate group, whereas it was initially higher and then gradually 1 decreased in the severe group. In the severe group, IL6 levels peaked at days 8-11 but 2 then continued to decline. In the moderate group, IL6 showed a downward trend in 3 fluctuations. Except for days 12-14, there was no significant difference in IL6 levels 4 between the two groups. At days 6-8 after disease onset, the LC counts in the 5 moderate group were higher than that of the severe group, but there was no statistical 6 difference. In the following days, the LC counts of the moderate group gradually 7 reached their highest levels and kept close to normal levels (1.5 × 10 9 cells/L). The 8 LC counts in the severe group gradually decreased, reached its lowest value on days 9 12-14, and then gradually increased. On days 18-20, LC counts in the severe group 1 0 reached a comparable level to the moderate group. However, there were statistically 1 1 significant differences in LC counts between the two groups over three consecutive Neutrophil counts, and NLRs between the two groups throughout the course of the 1 4 disease. Next, we analyzed the dynamic changes of CD3 + , CD4 + and CD8 + T-cells in the 1 6 moderate and the severe COVID-19 patients. As shown in Figure 2 , the CD3 + , CD4 + 1 7 and CD8 + T-cell counts in the moderate group were significantly higher than that of subsets including CD3 + , CD4 + , and CD8 + T-cells in the severe group gradually 2 0 increased. With the disease recovery, all these cells reached a comparable level to the 2 1 moderate group. There were no significant differences in the percentage of CD3 + , 2 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint CD4 + and CD8 + T-cells, CD4/CD8 ratio between the two groups throughout the 1 course of the disease. To investigate the association between these indicators, we performed To investigate the potential predictive value of these parameters, a receiver operating was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. to mild and moderated patients on admission. We also found that the lymphocytes Other studies have attributed the activation of the hypothalamus-pituitary-adrenal 2 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. research is needed to explore the corresponding mechanism in detail. Effective predictors can help doctors provide appropriate supportive care for patients 1 4 with severe COVID-19. In this study, we found that four variables including study, patients aged 50 years or older and NLR equal to or greater than 3.13 may 2 0 serve as a high-risk factor for COVID-19 patients. In our study, we found that NLRs 2 1 for all severe patients all exceeded 5, and NLRs for moderate patients ranged from 3 2 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint to 4. Moreover, the AUC of NLR for predicting severe patients is less than 0.9, 1 indicating poor predictive performance. Another study showed that N8R (neutrophils 2 to CD8 + T-cell ratio) has better performance with a higher AUC value than NLR in 3 the ROC curve analysis, and may serve as a more powerful factor than NLR for 4 predicting the severity in COVID-19 patients [19] . However, neutrophils are 5 susceptible to many factors, and its ratio to CD8 + T cells will mask the actual 6 reduction of T cells. Therefore, we believe that using direct counting of T cells may 7 more accurately reflect the patient's immune status and thus have better predictive 8 performance. T cells increase reactively [21] . In this study, we found that the CD4 + and CD8 + T was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. usually be observed in patients with severe COVID-19[9]。Of course, the mechanism 2 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. In conclusion, the counts of CD3 + , CD4 + , and CD8 + T-cells are significantly reduced The lines represent mean, error bar represents SD. Significance between different 1 0 groups is tested by the Wilcoxon signed-rank test. Asterisk (*) indicates significant 1 1 (P<0.05) differences between the two groups in various time points. The correlation coefficient (R) was calculated using Spearman's correlation method. R All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint The best cutoff values of the parameters with higher areas under the curve (AUC) were 1 calculated from the ROC curves. 2 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint Lymphopenia and 2 neutrophilia in SARS are related to the prevailing serum cortisol Persistent lymphopenia after diagnosis of sepsis predicts mortality The authors disclose no conflicts of interest. The data that support the findings of this study are available from the corresponding 1 9 author upon reasonable request. All rights reserved. No reuse allowed without permission.was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. e/naming-the-coronavirus-disease-(covid-2019)-and-the-virus-that-causes-it. All rights reserved. No reuse allowed without permission.was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which this version posted May 5, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which this version posted May 5, 2020. . https://doi.org/10.1101/2020.05.01.20086421 doi: medRxiv preprint