Application of neutrophil CD64 as early prediction biomarker of adult Stetier disease macrophage activation syndrome

Abstract

The invention discloses a neutrophil CD64 as a novel early prediction biomarker of an adult Stetill disease macrophage activation syndrome. The invention provides application of neutrophile granulocyte CD64, neutrophile granulocyte CD64 combined ferritin and / or sIL-2R or a detection reagent of the neutrophile granulocyte CD64 combined ferritin and / or sIL-2R in preparation of a product for predicting or diagnosing adult Stetill disease macrophage activation syndrome. Logistic regression analysis shows that nCD64 index increase is a risk factor (OR = 1.073, p = 0.003) of the MAS. ROC curve analysis shows that the nCD64 index can reliably distinguish AOSD patients (AUC = 0.877; the critical value is equal to 32.09; plt; 0.001), and the combined use of the nCD64 index, the ferritin and the soluble IL-2R shows a relatively strong prediction value.

Description

Technical Field

[0001] The present invention relates to application of neutrophil CD64 as an early prediction biomarker for Still's disease macrophage activation syndrome in adults, and belongs to the technical field of biomedical detection. Background Art

[0002] Adult-onset Still's disease (AOSD) is widely recognized as an autoinflammatory disorder. This autoinflammatory disease typically presents with transient rash, polyarthritis, sore throat, elevated C-reactive protein (CRP) levels, and increased neutrophil counts. Although many patients with AOSD experience mild symptoms, some may develop severe complications, including macrophage activation syndrome (MAS), acute respiratory distress syndrome, diffuse alveolar hemorrhage, fulminant hepatitis, and disseminated intravascular coagulation.

[0003] MAS is a key complication of severe AOSD, characterized by fever, pancytopenia, hepatitis, hyperferritinemia, and multi-organ dysfunction. The incidence of MAS in patients with AOSD is estimated to be between 10% and 19%, and the mortality rate of MAS associated with AOSD is approximately 10% to 20%. Patients with AOSD and MAS typically require aggressive treatment early in the disease, including high-dose intravenous pulse glucocorticoids, to prevent life-threatening visceral complications.

[0004] Currently, no specific biomarkers have been identified that can distinguish between patients with adult-onset Still's disease (AOSD) with macrophage activation syndrome (MAS) and those without MAS. This situation highlights the urgent need for biomarkers that can stratify patients who may be at risk for MAS. CD64, also known as Fc gamma receptor I (FcγRI), is a high-affinity receptor for the Fc region of immunoglobulin G (IgG) that is primarily expressed on monocytes, macrophages, and activated neutrophils (Li S, Huang X, Chen Z, Zhong H, Peng Q, Deng Y, et al. Neutrophil CD64 expression as a biomarker in the early diagnosis of bacterial infection: a meta-analysis. Int J Infect Dis. 2013; 17(1): e12-23.). During infection, inflammatory signals activate CD64, leading to a significant upregulation of its expression on the surface of activated neutrophils within 4 to 6 hours (

[10] de Jong E, de Lange DW, Beishuizen A, vande Ven PM, Girbes AR, Huisman A. Neutrophil CD64 expression as a longitudinal biomarker for severe disease and acute infection in critically ill patients. Int J Lab Hematol. 2016; 38(5): 576–84.). Therefore, the expression level of CD64 on neutrophils (nCD64) can be used as an indicator during the inflammatory response. However, whether it can be used as an early predictive biomarker for patients with AOSD and MAS has not been reported. Summary of the Invention

[0005] The purpose of the present invention is to provide an application of neutrophil CD64 as an early predictive biomarker for Still's disease macrophage activation syndrome in adults.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides the use of a biomarker or a detection reagent thereof in the preparation of a predictive, diagnostic or prognostic product for Still's disease macrophage activation syndrome in adults, wherein the biomarker is selected from neutrophil CD64 or a combination of neutrophil CD64 and a second biomarker, and the second biomarker is selected from ferritin and / or sIL-2R.

[0008] Preferably, the detection reagent at least includes a reagent for detecting the expression level of CD64 (nCD64) on neutrophils.

[0009] Preferably, the detecting the expression level of CD64 on neutrophils is detecting the expression level of CD64 on neutrophils by flow cytometry.

[0010] Preferably, the detection of the expression level of CD64 on neutrophils is the detection of the relative expression level of CD64 on neutrophils, and the detection of the relative expression level of CD64 on neutrophils uses the expression level of CD64 on monocytes as a positive control and the expression of CD64 on lymphocytes as a negative control.

[0011] Preferably, the detection of the expression level of CD64 on neutrophils is to detect the relative expression level of CD64 on neutrophils, and the detection of the relative expression level of CD64 on neutrophils is to detect the nCD64 index, and the calculation formula of the nCD64 index is: (neutrophil CD64 expression / lymphocyte CD64 MFI) / (monocyte CD64 MFI / neutrophil CD64 MFI).

[0012] Preferably, the detection reagent includes at least antibodies to human CD14, CD45 and CD64.

[0013] Preferably, the detection reagent further comprises a reagent for detecting a second biomarker.

[0014] Preferably, the reagent used to detect the second marker includes anti-ferritin antibody (for detecting ferritin) and / or anti-sIL-2R antibody (for detecting sIL-2R).

[0015] In a second aspect, the present invention provides the use of biomarkers in constructing a predictive or diagnostic model for Still's disease macrophage activation syndrome in adults, wherein the biomarkers are selected from neutrophil CD64, ferritin and sIL-2R, and the predictive or diagnostic model is a nomogram model constructed based on nCD64 index, ferritin and sIL-2R.

[0016] In a third aspect, the present invention provides a method for obtaining a predicted probability of Still's disease macrophage activation syndrome in adults, comprising the following steps:

[0017] The steps of respectively detecting the subject's nCD64 index, ferritin level and sIL-2R level; inputting the detected nCD64 index, ferritin level and sIL-2R level into a pre-constructed prediction model to obtain a predicted probability; the prediction model is a nomogram model.

[0018] In a fourth aspect, the present invention provides a prediction system or device for Still's disease macrophage activation syndrome in adults, comprising a detection module and a prediction module, wherein the detection module is used to detect the subject's nCD64 index, ferritin level, and sIL-2R level; and the prediction module is used to input the nCD64 index, ferritin level, and sIL-2R level quickly obtained by the detection module into a pre-constructed nomogram model to obtain a predicted probability.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention uses the nCD64 index or the combination of the nCD64 index and other biomarkers (ferritin, sIL-2R) as an effective method to distinguish patients with adult-onset Still's disease (AOSD) with macrophage activation syndrome (MAS) and those without MAS. Receiver operating characteristic (ROC) curve analysis shows that the nCD64 index or the combination has good predictive / diagnostic value. The biomarker or combination of biomarkers described in the present invention is particularly beneficial in clinical settings because it promotes timely diagnostic evaluation, thereby allowing earlier intervention and better management of MAS. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 . Schematic diagram of flow cytometry (gating strategy) detection of nCD64 index;

[0022] Figure 2 .nCorrelation between CD64 index and AOSD disease activity, including:

[0023] A. Comparison of CD64 mRNA expression in neutrophils between AOSD patients and healthy controls (HC); Comparison of nCD64 index according to different disease manifestations in AOSD patients; B. Reduction of nCD64 index in 16 AOSD patients after treatment; C. nCD64 index was positively correlated with Pouchot score, AST, LDH, and CD8% in AOSD patients; correlations were assessed using the Spearman test; * = p < 0.05, ** = p < 0.01, *** = p < 0.001; HC, healthy control; AOSD, adult-onset Still's disease; nCD64 index, neutrophil CD64 index; LDH, lactate dehydrogenase; CD8%, CD8 + T cell percentage.

[0024] Figure 3 Comparison of nCD64 index, laboratory and immunological characteristics between the group without MAS and the group with MAS in the training group.

[0025] Figure 4 The nCD64 index was elevated in patients with AOSD and MAS, including:

[0026] A. Observed incidence of MAS in low, intermediate, and high nCD64 index groups; B. Changes in nCD64 index in eight AOSD patients with MAS during multiple follow-up visits; C. Correlation between nCD64 index and NK%, fibrinogen (FIB), soluble interleukin-2 receptor (sIL-2R), triglycerides (TG), platelets (PLT), and hemoglobin (HB) in AOSD patients; D. Logistic regression analysis of risk factors for MAS; correlations were assessed using the Spearman test. The dotted line in B indicates the threshold level at which 90% of MAS episodes occurred in these eight patients. The solid line indicates the cutoff value predicting the occurrence of MAS.

[0027] Figure 5 Comparison of the CD64 index with other biomarkers in predicting AOSD with MAS:

[0028] A. Receiver operating characteristic (ROC) curves of the nCD64 index, sIL-2R, ferritin, and CRP for differentiating AOSD patients with MAS from those without MAS; B. Receiver operating characteristic (ROC) curves for the combination of sIL-2R, ferritin, and nCD64 index for identifying MAS; C. Nomogram for predicting AOSD with MAS.

[0029] Figure 6 .nCD64 index is significantly correlated with prognosis, including:

[0030] AB. Correlation between the nCD64 index and length of hospital stay and maximum glucocorticoid dose; C. Comparison of the nCD64 index with other biomarkers in predicting AOSD with MAS; Comparison of the cumulative event rate of MAS in AOSD patients with an nCD64 index greater than 32.09 and less than 32.09. DETAILED DESCRIPTION

[0031] To make the present invention more clearly understood, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0032] The experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or the conditions recommended by the manufacturers. The materials and reagents used in the following examples, unless otherwise specified, were obtained from commercial sources.

[0033] Macrophage activation syndrome (MAS) is a serious and potentially life-threatening complication of adult-onset Still's disease (AOSD), so there is a need to identify sensitive and specific biomarkers for early diagnosis. The experimental results of the present invention showed that compared with the healthy control group, the expression of CD64 mRNA in neutrophils of AOSD patients was significantly increased (p = 0.029). The neutrophil CD64 index (nCD64 index) was positively correlated with multiple key clinical manifestations, including splenomegaly, sore throat, pulmonary infiltration and pericarditis. Effective treatment resulted in a rapid and significant decrease in the nCD64 index (p < 0.001). Logistic regression analysis showed that an elevated nCD64 index was a risk factor for MAS (OR = 1.073, p = 0.003). Receiver operating characteristic (ROC) curve analysis demonstrated that the nCD64 index reliably distinguished patients with AOSD who presented with MAS (AUC = 0.877; cutoff value = 32.09; p < 0.001), and the combined use of the nCD64 index, ferritin, and soluble IL-2R demonstrated strong predictive value. Correlations were also observed with hospitalization duration (r = 0.382, p < 0.001) and maximum glucocorticoid dose (r = 0.326, p = 0.003). Kaplan-Meier analysis revealed that patients with an nCD64 index greater than 32.09 had a significantly higher cumulative incidence of MAS (p < 0.001). These results suggest that the nCD64 index may serve as a biomarker for early identification of MAS risk in patients with AOSD, facilitating timely diagnosis and management.

[0034] Example

[0035] 1. Materials and Methods

[0036] 1. Patient

[0037] This example included 118 patients with adult-onset Still's disease (AOSD), including 83 in the training group and 35 in the validation group, as well as 19 healthy controls (HC). All participants were diagnosed and admitted to the Department of Rheumatology and Immunology at Renji Hospital in Shanghai, China, between April 2019 and October 2024. Demographic data, clinical information, and laboratory results from the study were systematically organized into a single database. Table 1 provides a detailed overview of the characteristics of the training group. Patients included in this example met Yamaguchi's criteria (N. Mahroum, H. Mahagna, H. Amital Diagnosis and Classification of Adult Still's Disease, J. Autoimmun., 48–49 (2014), pp. 34–37, 10.1016 / j.jaut.2014.01.011). Patients with infections, tumors, or other autoimmune diseases were excluded. Macrophage activation syndrome (MAS) was diagnosed according to the HLH-2004 criteria (Henter JI, Horne A, Arico M et al. HLH-2004: diagnostic and therapeutic guidelines for hemophagocytic lymphohistiocytosis. Pediatr Blood Cancer 2007;48:124–31). All healthy controls had no history of rheumatic or other diseases and were age- and sex-matched with the patients. Informed consent was obtained from all participants in accordance with the recommendations of the Declaration of Helsinki. The experimental protocol for this example was approved by the Institutional Review Committee of Renji Hospital (approval number: 2016-083).

[0038] 2. Data Collection

[0039] Clinical characteristics, including demographic data, symptoms, signs, complications, systemic scores, laboratory data, immunological test results, and treatment regimens were recorded. Hepatic dysfunction was assessed based on the presence of hepatomegaly and elevated liver enzyme levels, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transpeptidase (GGT). Treatment was continuously tailored to each patient, and all treatment measures during hospitalization were carefully documented. Methylprednisolone doses were converted to the corresponding prednisone doses, and the doses of other glucocorticoids were standardized for comparison.

[0040] 3. Quantitative Real-time PCR

[0041] Neutrophils were isolated from AOSD patients and healthy controls by Percoll density gradient centrifugation. Total RNA was extracted using Trizol reagent. mRNA expression levels were normalized to human β-actin as an internal control. The following primers were used for qRT-PCR: human CD64: forward primer 5′-ATACAGGTGCCAGAGAGGTCTC-3′, reverse primer 5′-CCAGCTTATCCTTCCACGCATG-3′.

[0042] 4. Measurement of nCD64 Index

[0043] Whole blood samples were collected using EDTA as an anticoagulant and the nCD64 index was measured by flow cytometry (gating strategy). For a flow chart of the gating strategy, see Figure 1 . The reagents used to assess the nCD64 index include antibodies to human CD14, CD45, and CD64. The assay procedure involves adding 50 μL of EDTA-treated whole blood to an experimental tube and adding 5 μL of each antibody. After thorough mixing, the sample is incubated in the dark for 15 minutes. Subsequently, red blood cells are lysed for analysis by flow cytometry. The mean fluorescence intensity (MFI) of CD64 expression on neutrophils, lymphocytes, and monocytes in the experimental sample is recorded. The nCD64 index is calculated as follows: (neutrophil CD64 MFI / lymphocyte CD64 MFI) / (monocyte CD64 MFI / neutrophil CD64 MFI). The calculation of the nCD64 index takes into account inherent inter-individual variability to ensure accuracy and stability of the measurement. Given that CD64 expression is consistently high on monocytes, this serves as a positive control for the nCD64 index, while lower expression on lymphocytes serves as a negative control.

[0044] 5. Statistical Analysis

[0045] All statistical analyses were performed using GraphPad Prism version 9.0.0 and SPSS software. Normally distributed data were expressed as mean ± standard deviation (SD), while non-normally distributed data were expressed as median and interquartile range. Unpaired t-test or one-way analysis of variance (ANOVA) was used for normally distributed data; Mann-Whitney U test or Kruskal-Wallis test was used for non-parametric data. Count data were compared using chi-square (χ 2 ) test. Correlations between variables were assessed using the Spearman rank correlation coefficient. Receiver operating characteristic (ROC) curve analysis was used to determine the optimal cutoff value for distinguishing AOSD patients with and without MAS. The significance level for all statistical tests was set at p < 0.05.

[0046] 2. Results

[0047] 1. Baseline characteristics of AOSD patients

[0048] A total of 83 patients with adult-onset Still's disease (AOSD) were included in this example, of whom 48 (57.8%) were AOSD patients with macrophage activation syndrome (MAS) (AOSD-MAS). Table 1 shows a comparison between AOSD patients without MAS and those with MAS. There were no significant differences between the two groups in terms of fever, rash, arthralgia or arthritis, pleurisy, etc. Compared with patients with MAS, patients without MAS had significantly lower frequencies of muscle pain (p = 0.040), sore throat (p = 0.008), splenomegaly (p = 0.008), lymphadenopathy (p < 0.001), pulmonary infiltration (p = 0.001), pericarditis (p = 0.028), and hepatomegaly or abnormal liver function (p = 0.032). Laboratory tests showed significantly higher levels of ALT (p=0.021), AST (p=0.003), lactate dehydrogenase (LDH) (p<0.001), triglycerides (p=0.002), and ferritin (p<0.001) in AOSD patients with MAS, while significantly lower levels of hemoglobin (p=0.014) and platelets (p=0.037) compared with those without MAS. AOSD patients with MAS also had significantly higher Pouchot scores than those without MAS (p<0.001).

[0049] Table 1. Demographic, clinical, and laboratory characteristics of patients in the training group

[0050]

[0051]

[0052] MAS, macrophage activation syndrome; WBC, white blood cell; ALT, alanine aminotransferase; AST, aspartate aminotransferase; LDH, lactate dehydrogenase; CRP, C-reactive protein; ESR, erythrocyte sedimentation rate.

[0053] 2. Manifestation of increased nCD64 index in active AOSD

[0054] First, this example demonstrates that the expression level of nCD64 mRNA in AOSD patients is higher than that in healthy controls (p = 0.029). This example also observed that the nCD64 index was significantly elevated in AOSD patients with splenomegaly (p = 0.008), sore throat (p = 0.031), pulmonary infiltration (p < 0.001), and pericarditis (p = 0.002) (see Figure 2A). In addition, during follow-up, the nCD64 index of AOSD patients was significantly reduced after effective treatment (p<0.001) (see Figure 2 B). Subsequently, this example evaluated the relationship between the nCD64 index and laboratory inflammatory markers and Pouchot score of AOSD. The results showed that the nCD64 index was positively correlated with laboratory parameters of disease severity, such as Pouchot score (r = 0.482, p < 0.001), AST (r = 0.354, p < 0.001), LDH (r = 0.437, p < 0.001) and CD8% (r = 0.411, p < 0.001) (see Figure 2 C) These results suggest that there is a close correlation between the nCD64 index and disease activity in AOSD patients.

[0055] 3. nCD64 Index and Immunological Findings in AOSD Patients with MAS

[0056] This example further analyzed the nCD64 index and immunological data of 83 patients in the training group. Compared with patients without MAS, the nCD64 index of AOSD patients with MAS was significantly increased (p < 0.001). + T cell % (p=0.010), CD8 + T cell % (p=0.002), CD3 + HLA - DR + / CD3 + % (p=0.006), CD8 + HLA - DR + / CD8 + % (p=0.027), CD8 + CD38 + / CD8 + %(p<0.001), CD4 + HLA - DR + / CD4 + % (p = 0.002), sIL-2R (p < 0.001) and IL-10 (p = 0.018) levels were significantly higher, while NK cell % (p < 0.001) and CD4 + / CD8 + The ratio (p = 0.021) was significantly lower in AOSD patients with MAS than in patients without MAS. Figure 3 .

[0057] 4. nCD64 index as a potential biomarker for AOSD with MAS

[0058] Patients with adult-onset Still's disease (AOSD) and macrophage activation syndrome (MAS) require intensive treatment and may have a poor prognosis. Figure 3 As shown, nCD64 index was significantly elevated in AOSD patients with MAS. Therefore, the present invention investigated the potential of nCD64 index as an early diagnostic marker for active AOSD patients experiencing MAS.

[0059] According to the decreasing order of nCD64 index at admission, AOSD patients were divided into three groups using the tertile method: high nCD64 index group (n=27), medium nCD64 index group (n=27), and low nCD64 index group (n=27). We observed a significant difference in the incidence of MAS among the three groups (p<0.001) (see Figure 4 In addition, we also found that the nCD64 index increased significantly during MAS episodes compared with non-MAS episodes (see Figure 4 B). When investigating the correlation between the nCD64 index and laboratory parameters of MAS, we found that the nCD64 index was positively correlated with sIL-2R (r = 0.319, p = 0.015) and triglycerides (TG) (r = 0.397, p < 0.001). The nCD64 index was also strongly correlated with lower levels of platelets (r = -0.388, p < 0.001), hemoglobin (r = -0.202, p = 0.089), fibrinogen (FIB) (r = -0.280, p = 0.019), and NK cell percentage (r = -0.373, p = 0.001) (see Figure 4 C). Meanwhile, logistic regression analysis showed that elevated nCD64 index (OR=1.073, p=0.003) was identified as a risk factor for the occurrence of MAS (see Figure 4 D). These data demonstrate the validity of the nCD64 index as a biomarker in patients with AOSD and MAS.

[0060] 5. Comparison of nCD64 Index with Other MAS Markers

[0061] There is a lack of specific biomarkers for the differential diagnosis of MAS. Next, the present invention evaluated the effectiveness of the nCD64 index as a biomarker for AOSD patients with MAS and compared it with other potential markers such as CRP, ferritin and sIL-2R. In the training group, ROC curve analysis showed that nCD64 index (AUC = 0.877, p < 0.001), ferritin (AUC = 0.833, p < 0.001) and sIL-2R (AUC = 0.866, p < 0.001) were able to effectively distinguish AOSD patients with MAS and those without MAS. In contrast, CRP performed as a less effective diagnostic biomarker in identifying AOSD patients with MAS (AUC = 0.553, p = 0.427) (see Figure 5 A). In the combined analysis of biomarkers, we first performed binary logistic regression to integrate the nCD64 index and other indicators, and as a result, a new composite index was developed. The corresponding ROC curve analysis was then performed to evaluate its performance. The results showed that combining other biomarkers with the nCD64 index significantly improved the diagnostic accuracy of detecting MAS. In the training group, the combination of ferritin and nCD64 index significantly improved the predictive performance compared with ferritin alone (AUC = 0.942, p < 0.001). Similarly, the combination of sIL-2R and nCD64 index also significantly improved the predictive accuracy compared with sIL-2R alone (AUC = 0.944, p < 0.001) (see Figure 5 B). Subsequently, the present invention developed a nomogram combining nCD64 index, ferritin and sIL-2R for assessing the risk of AOSD patients progressing to MAS (see Figure 5 C). Both ferritin and sIL-2R refer to serum levels. Serum ferritin can be detected using an enzyme-linked immunosorbent assay (ELISA), and serum soluble interleukin-2 receptor (sIL-2R) can be detected using a double-antibody sandwich ELISA. Detection reagents include the corresponding antibodies.

[0062] In the training set, receiver operating characteristic (ROC) curve analysis showed that the cutoff value for the nCD64 index was 32.09, with a sensitivity of 0.792 and a specificity of 0.971. The cutoff value for ferritin was 1894 ng / ml, with a sensitivity of 0.714 and a specificity of 0.857. The cutoff value for sIL-2R was 1363 U / ml, with a sensitivity of 0.688 and a specificity of 0.943. In the validation set, the nCD64 index also demonstrated good predictive performance (sensitivity = 0.786, specificity = 0.905, positive predictive value (PPV) = 0.846, negative predictive value (NPV) = 0.864). The nCD64 index had better predictive performance than sIL-2R (sensitivity = 0.750, specificity = 0.829, PPV = 0.857, NPV = 0.707). In contrast, ferritin demonstrated good sensitivity but poor specificity (sensitivity = 0.857, specificity = 0.476, PPV = 0.522, NPV = 0.833) (see Table 2). Overall, these results suggest that the nCD64 index and its combination with ferritin and sIL-2R provide strong predictive value.

[0063] Table 2. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the predicted probability at the optimal cutoff value

[0064]

[0065]

[0066] 6. Association of nCD64 Index with Prognosis of AOSD

[0067] In this example, the treatment of AOSD patients during hospitalization was recorded. AOSD patients with macrophage activation syndrome (MAS) received intravenous dexamethasone (p < 0.001), intravenous immunoglobulin (p < 0.001), and VP-16 (p < 0.001) more frequently than those without MAS. The maximum glucocorticoid dose in the group without MAS was significantly lower than that in the group with MAS (p < 0.001). There was no significant difference in the frequency of CsA, tacrolimus, and anti-IL-6R medications between the two groups.

[0068] When exploring the correlation between nCD64 index and prognosis, we found that nCD64 index was significantly associated with hospitalization time (r = 0.382, p < 0.001) (see Figure 6 A) and maximum glucocorticoid dose (r = 0.326, p = 0.003) (see Figure 6B) was positively correlated. To further evaluate the prognostic value of nCD64 index in AOSD patients, we grouped 83 patients according to the pre-determined optimal cutoff value of nCD64 index (32.09). Among these patients, 48 ​​(57.8%) experienced MAS during the clinical course. Kaplan-Meier analysis showed that the cumulative incidence of MAS within 28 days of hospitalization was significantly higher in patients with nCD64 index greater than 32.09 than in patients with nCD64 index less than 32.09 (p < 0.001) (see Figure 6 C). The mean time from hospital admission to MAS onset in AOSD patients with a higher nCD64 index was 3.59 ± 4.93 days.

[0069] The above description is only a preferred embodiment of the present invention and does not constitute any formal or substantial limitation to the present invention. It should be noted that ordinary technicians in this technical field can make several improvements and supplements without departing from the present invention, and these improvements and supplements should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a biomarker or a detection reagent thereof in the preparation of a product for prediction, diagnosis or prognosis assessment of adult-onset Still's disease macrophage activation syndrome, characterized in that: The biomarker is selected from neutrophil CD64 or a combination of neutrophil CD64 and a second biomarker selected from ferritin and / or sIL-2R.

2. The use according to claim 1, characterized in that The detection reagent at least includes a reagent for detecting the expression level of CD64 on neutrophils.

3. The use according to claim 2, characterized in that The detecting of the expression level of CD64 on neutrophils is detecting the expression level of CD64 on neutrophils by flow cytometry.

4. The use according to claim 2, characterized in that The detection of the expression level of CD64 on neutrophils is to detect the relative expression level of CD64 on neutrophils. The detection of the relative expression level of CD64 on neutrophils uses the expression level of CD64 on monocytes as a positive control and the expression of CD64 on lymphocytes as a negative control.

5. The use according to claim 4, characterized in that The detection of the expression level of CD64 on neutrophils is to detect the relative expression level of CD64 on neutrophils, and the detection of the relative expression level of CD64 on neutrophils is to detect the nCD64 index. The calculation formula of the nCD64 index is: (neutrophil CD64 expression / lymphocyte CD64 MFI) / (monocyte CD64 MFI / neutrophil CD64 MFI).

6. The use according to any one of claims 1 to 5, characterized in that The detection reagent at least includes antibodies to human CD14, CD45 and CD64.

7. The use according to claim 6, characterized in that The detection reagents also include reagents used to detect a second biomarker.

8. Use of a biomarker in constructing a predictive or diagnostic model for Still's disease macrophage activation syndrome in adults, characterized in that: The biomarkers are selected from neutrophil CD64, ferritin and sIL-2R, and the prediction or diagnostic model is a nomogram model constructed based on nCD64 index, ferritin and sIL-2R.

9. A method for obtaining a predicted probability of Still's disease macrophage activation syndrome in adults, characterized in that: The following steps are involved: The steps of respectively detecting the subject's nCD64 index, ferritin level and sIL-2R level; inputting the detected nCD64 index, ferritin level and sIL-2R level into a pre-constructed prediction model to obtain a predicted probability; the prediction model is a nomogram model.

10. A prediction system or device for Still's disease macrophage activation syndrome in adults, characterized in that: The system comprises a detection module and a prediction module. The detection module is used to detect the subject's nCD64 index, ferritin level and sIL-2R level. The prediction module is used to input the nCD64 index, ferritin level and sIL-2R level quickly obtained by the detection module into a pre-built nomogram model to obtain a predicted probability.

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