Application of IL-40 in manufacturing of detection kit for evaluating or predicting prognosis of severe pneumonia patient

Abstract

The invention discloses application of IL-40 in manufacturing of a detection kit for evaluating or predicting prognosis of a severe pneumonia patient, and the application comprises application of the IL-40 serving as a severe pneumonia prognosis biomarker in manufacturing of the detection kit for evaluating or predicting prognosis of the severe pneumonia patient. The detection kit is used for detecting IL-40 in serum of a patient, and a detection result is used for evaluating the prognosis risk of the severe pneumonia patient. When the expression quantity of the IL-40 in the serum is greater than or equal to 1.244 ng / mL, the prognosis of the severe pneumonia patient is poor, and when the expression quantity of the IL-40 in the serum is 1t; 1.244 ng / mL, which shows that the prognosis of the severe pneumonia patient is better.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of severe pneumonia prognosis, and particularly relates to application of IL-40 in manufacturing a detection kit for evaluating or predicting the prognosis of a severe pneumonia patient. BACKGROUND

[0002] Severe pneumonia (SP) is a serious respiratory disease that threatens human health worldwide in recent years. Its pathogenesis is complex, and it is difficult to treat. It has a high morbidity and mortality, and the elderly and infants are high-risk groups. Among them, the incidence of SP is about 145 cases per 100,000 people, and the 30-day mortality rate is about 27%. Even the mortality rate within 1 year can be as high as 47%. At the same time, SP also brings a huge economic burden to patients and society, and the United States needs to spend about $17 billion per year. Clinical studies have shown that the initiating factor of severe pneumonia death is pathogen infection. If not effectively controlled, the disease will mostly develop rapidly, leading to abnormal enhancement of the body's immune system and oxidative stress response, causing damage to alveolar and vascular endothelial cells, which can cause severe lung tissue damage, often leading to acute respiratory distress syndrome (ARDS), and some patients can also be complicated with sepsis, septic shock, multiple organ dysfunction syndrome, and other complications. Abnormal activation of inflammatory cells and the coagulation system can also aggravate the disease development process and even death. In view of the important position of infection in severe pneumonia death, the new strategy for treating severe pneumonia is to accurately assess infection and disease outcome as soon as possible, and develop targeted anti-infection and organ support treatment measures to effectively reduce mortality.

[0003] The occurrence and development of SP are accompanied by changes in a variety of prognostic biomarkers. The existing SP-related evaluation indicators in the clinic include two parts: clinical scoring systems and biomarkers. The scoring systems mainly include CURB-65 score, Pneumonia Severity Index (PSI), acute physiology and chronic health evaluation (APACHE II) score, and sequential organ failure assessment (SOFA) score. However, the above scoring systems involve a large number of indicators (liver, kidney function, nervous system GCS score, use of vasoactive drugs, and a series of indicators), and are cumbersome to operate. The biomarkers mainly include procalcitonin (PCT), C-reaction protein (CRP), and Interleukin-6 (IL-6), which have been widely used in clinical practice due to their simplicity, low cost, and other advantages. However, previous studies have found that the sensitivity and specificity of the above indicators for predicting prognosis are not high. In summary, there is no biomarker with high specificity and sensitivity that can quickly predict the prognosis of severe pneumonia, which greatly limits the early treatment of severe pneumonia. Therefore, finding new markers for rapid prognosis prediction and developing a refined treatment plan are of great significance for improving the prognosis of severe pneumonia patients. SUMMARY

[0004] In view of the deficiencies of the prior art, the purpose of the present application is to provide an application of IL-40 in the manufacture of a detection kit for evaluating or predicting the prognosis of severe pneumonia patients, and specifically to provide the use of IL-40 as a prognostic biomarker in severe pneumonia, to provide a new biomarker for predicting the prognosis and efficacy of severe pneumonia, and to provide a new evaluation method for reducing the mortality rate of severe pneumonia treatment.

[0005] In one embodiment, the present application provides a detection kit for evaluating or predicting the prognosis of severe pneumonia patients, which is used to detect IL-40 as a prognostic biomarker for severe pneumonia, and the detection results are used to evaluate the prognosis of the severe pneumonia patients. Preferably, the detection kit described above detects IL-40 in serum.

[0006] The detection kit of the application detects IL-40 in serum, and when the expression amount of IL-40 in serum is greater than or equal to 1.244 ng / mL, the prognosis of the severe pneumonia patient is poor, and the patient has a high risk of death; otherwise, when the expression amount of IL-40 in serum is less than 1.244 ng / mL, the prognosis of the severe pneumonia patient is better.

[0007] In another embodiment, the application provides an application of IL-40 as a prognosis biomarker of severe pneumonia in manufacturing a detection kit for evaluating or predicting the prognosis of a severe pneumonia patient, which is used for detecting IL-40 in serum of the severe pneumonia patient, and when the expression amount of IL-40 in serum is greater than or equal to 1.244 ng / mL, the prognosis of the severe pneumonia patient is poor, and the patient has a high risk of death; otherwise, when the expression amount of IL-40 in serum is less than 1.244 ng / mL, the prognosis of the severe pneumonia patient is better.

[0008] In still another embodiment, the application provides a method for detecting IL-40 in serum of a severe pneumonia patient, which is used as a biomarker of severe pneumonia for the prognosis of the severe pneumonia patient, and the method comprises the following steps: 1) using the detection kit of claim 1 to detect the content of IL-40 in serum of the severe pneumonia patient; 2) according to the detection result, determining the expression amount of IL-40, and when the expression level of IL-40 in serum is greater than or equal to 1.244 ng / mL, it indicates that the prognosis of the severe pneumonia patient is poor, and the patient has a high risk of death; otherwise, when the expression level of IL-40 in serum is less than 1.244 ng / mL, it indicates that the prognosis of the severe pneumonia patient is better.

[0009] The application has the following beneficial effects: Firstly, the application researches and finds that the content of IL-40 in serum of a clinical severe pneumonia patient is significantly higher than that of a non-severe pneumonia patient and a healthy person, and the expression of IL-40 in serum of a severe pneumonia shock patient and a death patient is also significantly higher than that of a non-shock patient and a survival patient.

[0010] Secondly, the expression of IL-40 is positively correlated with common prognosis biomarkers (CRP, PCT, SOFA score and IL-6) of severe pneumonia, which indicates that the content of IL-40 in serum of a clinical severe pneumonia patient is positively correlated with the severity of the disease.

[0011] Thirdly, further through the ROC curve to evaluate the prediction efficiency of IL-40 on the death of patients with severe pneumonia, it is found that the area under the curve of IL-40 is the largest, which is obviously larger than other commonly used indexes WBC, CRP, PCT, SOFA score, APACHE II score, IL-6 and Lym, which shows that IL-40 has good prediction value for the death risk of patients with severe pneumonia, and the cutoff value is 1.244 ng / mL.

[0012] Finally, through the Kaplan-Meier survival curve, it is found that the death risk of patients with severe pneumonia with the expression amount of IL-40 in serum being greater than or equal to the cutoff value (1.244 ng / mL) is significantly higher than that of patients with the expression amount of IL-40 being less than the cutoff value (1.244 ng / mL).

[0013] Therefore, the present application takes IL-40 as a new biomarker of severe pneumonia, which is used for the prognosis risk evaluation of severe pneumonia and reflects the severity of the disease, and provides a new evaluation method for the treatment of severe pneumonia and related diseases. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 It is a detection result graph of the expression amount of IL-40 in blood of clinical severe pneumonia patients and healthy controls in Example 1.

[0015] Figure 2 It is a detection result graph of the expression amount of IL-40 in blood of clinical severe pneumonia shock patients and non-shock patients in Example 1.

[0016] Figure 3 It is a detection result graph of the expression amount of IL-40 in blood of clinical severe pneumonia survivors and death patients in Example 1.

[0017] Figure 4 It is a correlation analysis result graph of the expression amount of IL-40 in blood of clinical severe pneumonia patients and SOFA score in Example 1.

[0018] Figure 5 It is a correlation analysis result graph of the expression amount of IL-40 in blood of clinical severe pneumonia patients and IL-6 score in Example 1.

[0019] Figure 6 It is a correlation analysis result graph of the expression amount of IL-40 in blood of clinical severe pneumonia patients and CRP score in Example 1.

[0020] Figure 7 It is a correlation analysis result graph of the expression amount of IL-40 in blood of clinical severe pneumonia patients and PCT score in Example 1.

[0021] Figure 8Figure 2 shows the ROC curve of the serum marker IL-40 in predicting the death of patients with severe pneumonia.

[0022] Figure 9 Figure 3 shows the Kaplan-Meier survival curve of the serum marker IL-40 in predicting the death of patients with severe pneumonia. DETAILED DESCRIPTION

[0023] The following examples are provided to further illustrate the present application. However, the following examples are provided only for the purpose of further illustrating the present application and are not intended to limit the present application. It will be understood by those skilled in the art that equivalent substitutions or corresponding modifications of the present content are still within the scope of the present application.

[0024] The present application focuses on the application value of interleukin-40 in predicting the prognosis of severe pneumonia.

[0025] IL-40, as one of the new members of the interleukin family, is encoded by the C17orf99 gene of stromal cells and only expressed in mammals, and its main function is to participate in the immune response mechanism and maintain B cell homeostasis. In recent years, many studies have shown that IL-40 plays an important role in autoimmune diseases. For example, the serum IL-40 level of patients with rheumatoid arthritis is up-regulated, and decreases after B cell depletion therapy. The serum and salivary gland IL-40 of patients with primary Sjogren's syndrome is elevated, and the expression of IL-40 in tissues is related to the expression of cytokines IL-4 and TGF-β. IL-40 shows up-regulated levels in the serum of patients with type 2 diabetes. However, the occurrence and development of severe pneumonia involve complex inflammatory disorders and immune disorders, and the correlation between the elevated expression level of IL-40 and the occurrence and development of severe pneumonia has not been reported.

[0026] The application adopts ELISA method to detect the serum of 104 cases of severe pneumonia patients, 52 cases of non-severe pneumonia patients and 86 cases of healthy persons. First, it is found that the IL-40 content in the serum of severe pneumonia patients is significantly higher than that of non-severe pneumonia patients and healthy people, and the IL-40 content in severe pneumonia shock and death patients is also significantly higher than that of non-shock and survival, and the above results have statistical differences. Second, the application also makes correlation analysis on the serum IL-40 level of severe pneumonia patients and common prognostic indicators CRP, PCT, SOFA score, APACHE II score and IL-6, and finds that the IL-40 content is positively correlated with CRP, PCT, SOFA score and IL-6, which has statistical difference. Third, the ROC curve is used to evaluate the prediction performance of IL-40 on the prognosis of severe pneumonia, and compared with WBC, CRP, PCT, SOFA score, APACHE II score, IL-6 and Lym, it is found that IL-40 has good diagnostic performance for severe pneumonia, and the area under the curve of IL-40 for death patient group is the largest, which is significantly higher than other indicators, and the cutoff value is 1.244 ng / mL. Finally, through the Kaplan-Meier survival curve, it is found that the death risk of severe pneumonia patients with serum IL-40 expression amount ≥cutoff value (1.244 ng / mL) is significantly higher than that of patients with IL-40 expression amount <cutoff value (1.244 ng / mL). The above results show that IL-40 can be used as an index for auxiliary prediction of death risk of severe pneumonia patients, i.e. a biomarker, and its prediction and evaluation performance for the risk degree of severe pneumonia patients is better than that of common indicators such as WBC, CRP, PCT, SOFA score, APACHE II score, IL-6 and Lym, and the threshold value of IL-40 for predicting death risk is 1.244 ng / mL.

[0027] Example 1 Correlation between IL-40 expression amount in serum of severe pneumonia patients and disease severity Serum specimens of severe pneumonia patients, non-severe pneumonia patients and healthy persons (clinical characteristics are shown in Table 1) were collected from the First Affiliated Hospital of Chongqing Medical University on the day of admission, and the expression amount of IL-40 was detected by using ELISA kit (purchased from Mybiosource company, item number MBS2905409-96T), and the specific operation steps strictly refer to the kit instruction manual. The patients all meet the diagnostic criteria of the international severe pneumonia diagnosis and treatment guidelines for the disease, and are excluded from having malignant tumor, HIV infection, autoimmune disease and the like. SPSS 20.0 software is used for statistical analysis, and GraphPad Prism 8.0 software is used for drawing, and the results are shown in Table 1 and Figures 1-7 .

[0028] Table 1. Clinical characteristics of patients and healthy subjects

[0029] Figure 1 The results of detection of IL-40 expression in blood of patients with severe pneumonia, patients with non-severe pneumonia and healthy people in the control group in the present embodiment are shown, Figure 2 The results of detection of IL-40 expression in blood of patients with severe pneumonia shock and non-shock patients in the present embodiment are shown, Figure 3 The results of detection of IL-40 expression in blood of patients with severe pneumonia survival and death in the present embodiment are shown, Figure 4 The results of correlation analysis of IL-40 expression in blood of patients with severe pneumonia and SOFA score in the present embodiment are shown, Figure 5 The results of correlation analysis of IL-40 expression in blood of patients with severe pneumonia and IL-6 score in the present embodiment are shown, Figure 6 The results of correlation analysis of IL-40 expression in blood of patients with severe pneumonia and CRP score in the present embodiment are shown, Figure 7 The results of correlation analysis of IL-40 expression in blood of patients with severe pneumonia and PCT score in the present embodiment are shown.

[0030] As shown in Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 and Figure 7 The results show that the IL-40 level is significantly increased in the serum of patients with severe pneumonia compared with patients with non-severe pneumonia and healthy people, and the IL-40 content in patients with severe pneumonia shock and death is also significantly higher than that in non-shock and survival patients, in addition, the IL-40 content is positively correlated with the commonly used prognostic indicators CRP, PCT, SOFA score and IL-6. The above results all suggest that IL-40 is closely related to the severity of severe pneumonia and can be used as a prognostic indicator or biomarker of severe pneumonia, which has important prognostic value in severe pneumonia.

[0031] Example 2 ROC curve analysis and comparison of the prediction efficiency of serum IL-40 for the prognosis of severe pneumonia The laboratory indicators and clinical score indicators of patients with severe pneumonia and healthy volunteers in the control group were collected through the HIS system, including WBC, CRP, PCT, SOFA score, APACHE II score, IL-6 and Lym. The predictive efficiency of IL-40 for the prognosis of severe pneumonia was evaluated by ROC curve analysis, and compared with WBC, CRP, PCT, SOFA score, APACHE II score, IL-6 and Lym. The ROC curve was drawn by GraphPad Prism 8.0 software, and the statistical difference of the area under the ROC curve was compared by SPSS 20.0 software. The results are shown in Table 2 and Figure 1

[0032] Figure 2 The results shown in Table 2 and

[0033] The results shown in Table 2 and Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figures 8-9 Figure 8 Figure 9 The results shown in Table 2 and

[0034] ​In summary, the present application finds that the expression amount of IL-40 in the body of a clinical severe pneumonia patient is significantly up-regulated compared with that of a non-severe pneumonia patient and a healthy person, and is also highly expressed in shock patients and death patients compared with non-shock patients and survivors of severe pneumonia. At the same time, the expression of IL-40 is positively correlated with the commonly used prognostic biomarkers (CRP, PCT, SOFA score and IL-6) of severe pneumonia, which indicates that the content of IL-40 in the serum of a clinical severe pneumonia patient is positively correlated with the severity of the disease. Further analysis finds that the prediction performance of IL-40 for death patients with severe pneumonia is obviously better than that of other commonly used indicators by evaluating the ROC curve, which indicates that IL-40 has good prediction value for the death risk of severe pneumonia patients, and the cutoff value is 1.244 ng / mL. Finally, it is found by Kaplan-Meier survival curve that the death risk of severe pneumonia patients with the expression amount of IL-40 in the serum being greater than or equal to the cutoff value (1.244 ng / mL) is significantly higher than that of patients with the expression amount of IL-40 being less than the cutoff value (1.244 ng / mL). Therefore, the present application finds that IL-40 can be used as a new biomarker for severe pneumonia to reflect the severity of severe pneumonia and the prognosis risk assessment of severe pneumonia, and provides a new evaluation method for the treatment of severe pneumonia and related diseases.

[0035] The above examples are only illustrative of the principles and effects of the present application, and are not intended to limit the present application. Any person skilled in the art can modify or change the above examples without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes completed by those skilled in the art without departing from the spirit and technical idea of the present application should be covered by the claims of the present application.

Claims

1. The use of interleukin-40 (IL-40) as a prognostic biomarker for severe pneumonia in the manufacture of a diagnostic kit for assessing or predicting the prognosis of patients with severe pneumonia, the diagnostic kit being used to detect IL-40 in the serum of said patients with severe pneumonia.

2. The application as described in claim 1, characterized in that, When the serum IL-40 expression level is ≥1.244 ng / mL, the prognosis of patients with severe pneumonia is poor and the risk of death is high. Conversely, when the serum IL-40 expression level is <1.244 ng / mL, the prognosis of patients with severe pneumonia is better.

3. A diagnostic kit for assessing or predicting the prognosis of severe pneumonia, characterized in that, The kit is used to detect IL-40, which serves as a prognostic biomarker for severe pneumonia. The detection results are used to assess or predict the prognosis of patients with severe pneumonia. When the serum IL-40 expression level is ≥1.244 ng / mL, the prognosis of patients with severe pneumonia is poor, and the risk of death is high. When the serum IL-40 expression level is <1.244 ng / mL, the prognosis of patients with severe pneumonia is better.

4. The detection kit as described in claim 3, characterized in that, The detection of IL-40 refers to the detection of IL-40 in serum.

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