key: cord-0905461-o2unit6k authors: Yu, Yang; Xu, Na; Cheng, Qi; Deng, Fei; Liu, Meiqin; Zhu, Airu; Min, Yuan-Qin; Zhu, Dan; Huang, Wenbo; Feng, Xu; Jing, Xizhong; Chen, Ying; Yue, Daoyuan; Fan, Yawei; Shu, Chang; Guan, Qing; Yang, Zifeng; Zhao, Jincun; Song, Wenjun; Guo, Deyin; Liu, Huanliang; Zhao, Jindong; Lan, Ping; Shi, Zhengli; Liu, Yingfang; Chen, Xiaoping; Liang, Huanhuan title: IFP35 as a promising biomarker and therapeutic target for the syndromes induced by SARS-CoV-2 or influenza virus date: 2021-12-03 journal: Cell Rep DOI: 10.1016/j.celrep.2021.110126 sha: 66b0226f800a50f1d7947793999d95172cfca932 doc_id: 905461 cord_uid: o2unit6k Previous studies showed that the high mortality caused by viruses such as SARS-CoV-2 and influenza virus, primarily results from the complications of a cytokine storm. Therefore, it is critical to identify the key factors participating in the cytokine storm. Here, we demonstrate that the interferon-induced protein 35 (IFP35), plays an important role in the cytokine storm induced by SARS-CoV-2 and influenza virus infection. We find that the levels of serum IFP35 in SARS-CoV-2 patients correlates with the severity of the syndrome. Using mouse model and cell assays, we show that IFP35 is released by lung epithelial cells and macrophage after SARS-CoV-2 or influenza virus infection. In addition, we show that administration of neutralizing antibody against IFP35 considerably reduce lung injuries, and thus the mortality rate of mice exposed to viral infection. In conclusion, our findings suggest that IFP35 serves both as a biomarker and a therapeutic target in virus-induced syndromes. . 142 Blood IFP35 level reflects the severity of COVID-19 patients 143 To study whether IFP35 is involved in the pathogenic process of COVID-19, we 144 tested the concentrations of blood IFP35 in the patients of Group 1 using an 145 enzyme-linked immunosorbent assay (ELISA). As shown in Figure 1D , the blood 146 IFP35 in 40 uninfected donors were below 150 pg/ml, the kit detection limit (average 147 at 46 ± 16 pg/ml). Similar results were also observed in the 29 not-severely-ill 148 patients, with only six patients' blood IFP35 were higher than 150 pg/ml (average at the average reach a high level of 950 ± 135 pg/ml. For comparison, we also measured 152 the serum IL-6 levels, which was previously recognized as biomarker indicating the 153 severity of COVID-19 (Liu et al., 2020) . Our results showed that the serum IL-6 154 levels in not-severely-ill and severely-ill patients were 14 ± 5 pg/ml and 66 ± 9 pg/ml, 155 respectively ( Figure 1E ), which was consistent with previous report. 156 To evaluate the accuracy of IFP35 to predict the severity of COVID-19, we 157 obtained the ROC curves of IFP35 and other clinically available biomarkers, 158 including CRP, PCT, TNF and IL-6. As shown in Figure 1F , the area under the curve 159 of IFP35 is 0.81. The optimal cutoff value of the serum IFP35 concentration is 131.7 Blood IFP35 represents a biomarker of COVID-19 165 To study the characteristics of blood IFP35 as biomarker of COVID-19, we performed 166 statistical analysis for the clinical parameters of the 112 COVID-19 patients in group1. 167 Firstly, our binary logistic regression analysis clearly showed that IFP35 was the main 168 factor (P=0.001) associated with the severity of disease (Figure 2A ). In addition, we 169 performed principal-component analysis (PCA) to study the correlation between the eigenvalues. These two components are responsible for 50.97% of the overall 173 variance. PC1 comprised four main variables (CRP, LDH, IL-6 and Neutrophil) and 174 was responsible for 39.06% of the total variability. PC2 comprised the main variable 175 IFP35 and was responsible for 11.91% of total variability (Table S7) To examine whether serum concentrations of IFP35 change during the course of the 183 disease, we conducted a follow-up study by randomly selected eight COVID-19 184 patients in Group 1 and measured the concentrations of IFP35 and IL-6 in their sera 185 both before and after treatment (Table S3) Next, we analyzed the correlation between IFP35 serum levels and treatment 196 outcome using 38 patients in Group 2 (Table S2 ). In the sera of 11 deceased patients 197 in this group, the concentrations of IFP35 reached an average of 4683 ± 1233 pg/ml. Notably, 10 of the 11 deceased patients exhibited serum IFP35 levels ranging from 199 1046 pg/ml to 12435 pg/ml, which was above 950 pg/ml, the average level for 200 severely-ill patients in Group 1 ( Figure 3D ). In contrast, the average levels of serum 201 IFP35 in the improved patients were measured as 137 ± 41 pg/ml, which were close to 202 those levels detected in uninfected donors. In contrast to IFP35, IL-6 levels showed 203 no clear correlation with the treatment outcome ( Figure 3E ). These results 204 demonstrated that IFP35 is related with the development of disease, which may be 205 used as a marker to predict the prognosis of patients. The pro-inflammatory action of IFP35 in influenza infected mouse model 244 We speculated that the released IFP35 enhances the inflammatory response during Figure 5B ). According to the clinical score, Ifp35 knockout delayed the peak 260 incidence of mice from day 7 to day 9. At the most severe stage, the clinical score of 261 the wild type mice was 4.22 (day 7), whereas the clinical score of the Ifp35 -/mice 262 was 3.05 (day 9). Consistently, the survival rate of the Ifp35 -/mice was 50% (6/12), 263 which was higher than that of wild type mice (25%, 3/12) ( Figure 5C ). To inspect the lung injury caused by influenza virus, we performed hematoxylin-265 eosin (H&E) staining of the lung tissues collected at the day 3. As shown in Figure 266 5D, the lung tissues of wild type mice were severely damaged following infection Figure 5E ). Next, we measured the 278 concentrations of TNF and IL-6 in the sera of influenza virus infected mice. Our 279 results showed that the mean values of serum TNF and IL-6 in wild type mice was 280 578 ± 63 pg/ml and 255 ± 105 pg/ml, which was higher than that in the Ifp35 -/mice 281 with 377 ± 39 pg/ml (P<0.01) and 67 ± 11 pg/ml (P<0.01) ( Figure 5F and 5G). To study whether released IFP35 exacerbated the severity of the inflammatory 298 J o u r n a l P r e -p r o o f response, we developed a neutralizing antibody against IFP35 (Anti-IFP35). The activity of the antibody was tested in both SARS-CoV-2 and influenza viruses 300 induced sepsis models, using a mouse IgG (mIgG) as negative control. 301 We developed a severe sepsis model by infecting mice with 2LD50 of PR8 virus. The process of influenza virus infected mouse model is shown in Figure 6A . Briefly, ). Therefore, we speculate that the down-regulated TLR-4 will be beneficial for 401 COVID-19 patients. Although our results showed that IFP35 possesses a number of strengths in clinical, 403 the present study still has many questions that should be further addressed. For 404 example, we found exceptions that some severely-ill patients (15/83) have low serum 405 IFP35. 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