CN119716052A - Application of plasmacytoid dendritic cells as a marker in prognostic products for pediatric sepsis - Google Patents
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Abstract
本发明涉及医学免疫学技术领域，特别涉及浆细胞样树突状细胞作为标志物在儿童脓毒症预后产品中的应用。本发明基于流式细胞术检测患者外周血中pDC水平，并基于其表达水平评估患者的预后。pDC表达水平与儿童脓毒症评分（Phoenix和pSOFA评分）呈显著正相关，与脓毒症患儿的死亡率呈正相关。当pDC的表达高于0.96%时，提示患者具有较高的死亡风险。本发明方法能够在儿童脓毒症的早期筛查中提供准确的预后评估，并具有高特异性和敏感性，帮助医生及时调整治疗方案，优化个性化治疗，具有良好的实际应用价值。The present invention relates to the technical field of medical immunology, and in particular to the use of plasmacytoid dendritic cells as markers in children's sepsis prognosis products. The present invention detects the level of pDC in the peripheral blood of patients based on flow cytometry, and evaluates the prognosis of patients based on its expression level. The pDC expression level is significantly positively correlated with the children's sepsis score (Phoenix and pSOFA scores), and is positively correlated with the mortality rate of children with sepsis. When the expression of pDC is higher than 0.96%, it indicates that the patient has a higher risk of death. The method of the present invention can provide accurate prognostic evaluation in the early screening of children's sepsis, and has high specificity and sensitivity, helping doctors to adjust treatment plans in time, optimize personalized treatment, and has good practical application value.
Description
Technical Field
The invention relates to the technical field of medical immunology, in particular to a prognosis evaluation and application method of children sepsis of plasma cell-like dendritic cells (pDC).
Background
Sepsis is a systemic inflammatory response syndrome caused by infection, leading mainly to multiple organ dysfunction, especially in pediatric patients, with higher morbidity and mortality （Singer M, et al. The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3). Jama 2016;315;Karakike E, et al. The early change of SOFA score as a prognostic marker of 28-day sepsis mortality: analysis through a derivation and a validation cohort. Critical Care 2019;23;Rudd KE, et al. Global, regional, and national sepsis incidence and mortality, 1990-2017: analysis for the Global Burden of Disease Study. Lancet 2020;395:200-211）. despite the significant progress of modern medicine in the treatment of sepsis, how to accurately predict the prognosis of an infant and provide personalized treatment remains a major challenge in clinical treatment due to rapid progression and complex clinical manifestations of sepsis.
Current prognostic assessment of sepsis relies primarily on clinical scoring systems such as pSOFA (sepsis-related organ failure assessment) and Phoenix sepsis scores. These systems are based primarily on patient organ dysfunction and help clinicians assess the severity of the condition. However, these methods often lack timely reflection of the immune system status, particularly in patients with sepsis in the early stages of immune imbalance. This makes it difficult for a physician to identify patients with severely impaired immune function at an early stage of the disease, thereby missing a timely opportunity for intervention.
Plasmacytoid dendritic cells (pdcs) are an important cell type in the human immune system responsible for initiating antiviral immune responses and mediating immune regulatory functions in viral infections and systemic inflammatory responses. pDC plays an important immunomodulatory role in sepsis, and dynamic changes in pDC levels can reflect the immune status of sepsis patients and their ability to clear pathogens, with dynamic changes in pDC being closely related to the progression of the patient's course. When pDC levels rise abnormally, patients are often prompted to face severe immune imbalances and may lead to exacerbation of the immunosuppressive response, which is closely related to mortality in sepsis patients.
Existing prognostic sepsis assessment approaches rely primarily on the patient's organ function scoring system and conventional inflammatory markers, but these methods often fail to effectively reflect the state of the patient's immune system, especially early immunosuppressive phenomena. Immune imbalance in sepsis patients is one of the leading causes of death, but conventional clinical markers such as C-reactive protein and procalcitonin are less sensitive in assessing immunosuppressive states. Furthermore, while the role of type I interferon in regulating immune responses has been widely studied, its dynamic relationship with pDC and its role in the prognosis of sepsis has not been fully explored.
Therefore, development of a detection method based on immunoregulatory cells is urgently needed, and the immune status and the disease progress of a patient suffering from sepsis in children can be accurately estimated, so that doctors can be helped to make more accurate prognosis judgment in the early stage of the disease.
Disclosure of Invention
In view of this, the present invention provides a method for prognosis evaluation of pediatric sepsis based on pDC (plasmacytoid dendritic cells) using flow cytometry to detect pDC levels in peripheral blood of a patient and determining its optimal threshold value of 0.96% based on ROC curve. When the patient's level of pDC is above this value, it is suggested that it is at higher risk of death. The method is innovative in that the method has high specificity and sensitivity, and can provide scientific basis for early risk screening and personalized treatment of sepsis patients.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an application of plasmacytoid dendritic cells serving as markers in preparation of a pediatric sepsis prognosis product.
In some embodiments of the invention, the above criteria for applying the prognosis include indicating that the patient is at a higher risk of death when the proportion of plasmacytoid dendritic cells to total lymphocytes is greater than 0.96.
In some embodiments of the invention, the plasmacytoid dendritic cells used above are peripheral blood-derived plasmacytoid dendritic cells.
The invention also provides the use of antibodies in the preparation of a pediatric sepsis prognosis product, including anti-CD 3 antibodies, anti-CD 19 antibodies, anti-CD 20 antibodies, anti-CD 14 antibodies, anti-CD 16 antibodies, anti-CD 56 antibodies, anti-CD 123 antibodies, and anti-HLA-DR antibodies.
In some embodiments of the invention, the above application obtains the ratio of plasmacytoid dendritic cells in the sample to be tested to total lymphocytes by flow cytometry based on the antibodies, resulting in a prognostic result;
the prognosis result judging rule comprises that when the ratio is higher than 0.96, the patient is prompted to have higher death risk.
In some embodiments of the present invention, the sample to be tested is a peripheral blood sample.
The invention also provides a kit for the prognosis of childhood sepsis, which takes the plasmacytoid dendritic cells as markers.
In some embodiments of the invention, the above-described kit captures cells in a peripheral blood sample with a specific antibody, learns the ratio of plasmacytoid dendritic cells in total lymphocytes, and then predicts based on the ratio;
The prognosis determination rule comprises that when the ratio is higher than 0.96, the patient is prompted to have higher death risk;
The specific antibodies include an anti-CD 3 antibody, an anti-CD 19 antibody, an anti-CD 20 antibody, an anti-CD 14 antibody, an anti-CD 16 antibody, an anti-CD 56 antibody, an anti-CD 123 antibody, and an anti-HLA-DR antibody.
In some embodiments of the invention, the diagnostic reagent described above comprises a specific antibody combination for detecting plasmacytoid dendritic cells (pDC) in peripheral blood of a patient, is detected in conjunction with flow cytometry, and is used to assess the prognosis of the patient.
In some embodiments of the invention, the above diagnostic kit further comprises reagents for sample processing, buffers, and fluorescent dyes for antibody labeling, in order to accurately detect the expression level of pDC in flow cytometry.
In some embodiments of the invention, the above diagnostic kit comprises an antibody to CD3, an antibody to CD19, an antibody to CD14, an antibody to CD16, an antibody to CD123, and an antibody to HLA-DR, in order to identify and determine pDC subpopulations of patients.
In some embodiments of the invention, the above diagnostic kit is suitable for early risk assessment of patients with sepsis, in particular for early screening and prognosis of pediatric sepsis.
The invention also provides a device for prognosis of sepsis in children, which takes the plasmacytoid dendritic cells as markers and is coated with specific antibodies for capturing the plasmacytoid dendritic cells.
In some embodiments of the invention, the specific antibodies of the above device include an anti-CD 3 antibody, an anti-CD 19 antibody, an anti-CD 20 antibody, an anti-CD 14 antibody, an anti-CD 16 antibody, an anti-CD 56 antibody, an anti-CD 123 antibody, and an anti-HLA-DR antibody.
The invention also provides an immune monitoring system for prognosis risk assessment of sepsis in children, based on prognosis of plasmacytoid dendritic cell expression level;
The prognosis criteria include that the plasma cell-like dendritic cell expression level is greater than 0.96, suggesting that the patient has a higher risk of death;
The expression level of the plasmacytoid dendritic cells is the proportion of the plasmacytoid dendritic cells to the total lymphocytes.
In some embodiments of the invention, the above-described immune monitoring system determines the prognosis of a patient by detecting the expression level of pDC in the patient's peripheral blood, in combination with a specific threshold, and helps the physician to adjust the treatment regimen by dynamically monitoring immune changes in disease progression.
In some embodiments of the invention, the above-described immunomonitoring system comprises:
the sample collection device is used for collecting a peripheral blood sample;
The flow cell detection module is used for detecting the percentage of pDC in total lymphocytes;
And a data analysis module for analyzing the prognosis risk of the patient according to the percentage.
In some embodiments of the invention, the data analysis module of the above-described immune monitoring system further analyzes the patient's prognostic risk in conjunction with ROC curve and clinical scoring system.
In some embodiments of the invention, the above-described immune monitoring system can be used to dynamically monitor patient pDC levels for real-time prognostic evaluation at different stages of pediatric sepsis treatment.
The invention also provides a detection method for pediatric sepsis prognosis risk assessment, based on the detection of plasmacytoid dendritic cells (pDC) in the peripheral blood of a patient, by analyzing the expression level of pDC cells, assessing the prognosis of pediatric sepsis patients, wherein when the expression level of pDC is higher than 0.96%, the patient is prompted to have a higher risk of death.
In some embodiments of the invention, the method of detecting pDC by the above detection method is flow cytometry, comprising:
s1, collecting a peripheral blood sample of a patient;
S2, obtaining the percentage of pDC in total lymphocytes through flow cytometry;
s3, determining the immune state of the patient according to the detection result, and judging the prognosis of the patient.
In some embodiments of the invention, the step S3 further comprises determining prognosis of the patient in combination with ROC curve analysis.
In some embodiments of the invention, the optimal threshold value for pDC is 0.96% for the above detection method, indicating that the patient is at a higher risk of mortality when pDC expression in the patient's peripheral blood is above this value.
In some embodiments of the invention, the above-described test methods further comprise dynamic monitoring of the patient at various time points, specifically by taking samples during 24 hours, 48 hours and 72 hours, respectively, of the course of the disease.
In some embodiments of the invention, the above-described detection methods may be further combined with other clinical scoring systems (e.g., pSOFA and Phoenix scores) to improve the accuracy of sepsis prognosis evaluation.
In some embodiments of the invention, the above-described detection methods are applicable to pediatric sepsis patients and may be used for prognosis evaluation of sepsis patients of other ages.
By monitoring the level of pDC, the invention can find out high-risk patients in early sepsis and help doctors to make timely treatment decisions. pDC is an important cell for immune regulation in sepsis patients, and its level is closely related to the severity of sepsis. Compared with traditional inflammation indexes (such as C-reactive protein and procalcitonin), the detection of the pDC has higher specificity and sensitivity.
The invention provides the dynamic monitoring of the pDC at multiple time points, can reflect the change of the immune state of a patient in real time, and is beneficial to predicting the disease course development and adjusting the treatment scheme.
The invention is suitable for early screening and prognosis evaluation of patients with sepsis, and is particularly suitable for patients with children sepsis. Through the detection of pDC, high-risk patients can be identified in time, and the sepsis related mortality rate is reduced. In addition, the method can be combined with other prognosis evaluation tools to provide reference basis for personalized treatment. With the development of detection technology, the method is expected to be widely applied in clinical practice and becomes a standard tool for sepsis diagnosis and management.
In summary, according to the sepsis prognosis evaluation method based on the pDC level, patients with high risk sepsis can be identified early by detecting the proportion of pDC in peripheral blood, and the immunological status change of the patients can be tracked in real time by dynamic monitoring. The method is simple and convenient to operate, has high specificity and sensitivity, can provide a reliable prognosis evaluation means for clinic, and provides scientific basis for the establishment of personalized treatment schemes.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 illustrates child in-group nano-row criteria and group groupings;
FIG. 2 shows the percentage of pDC in infants suffering from sepsis;
FIG. 3 shows analysis of correlation of levels of pDC in sepsis patients with Phoenix score and pSOFA score;
FIG. 4 shows the predictive efficacy of ROC curve analysis of pDC versus nosocomial death in sepsis patients;
FIG. 5 shows pDC versus the death outcome of an infant with sepsis;
FIG. 6 shows that pDC can be dynamically monitored over 72 hours.
Detailed Description
The invention discloses a pDC-based children sepsis prognosis evaluation method, which can be realized by appropriately improving process parameters by a person skilled in the art based on the content of the present disclosure. It is expressly noted that all such similar substitutions and modifications will be apparent to those skilled in the art, and are deemed to be included in the present invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the relevant art that variations and modifications can be made in the methods and applications described herein, and in the practice and application of the techniques of this invention, without departing from the spirit or scope of the invention.
The invention provides a pDC-based children sepsis prognosis evaluation method. The method judges and evaluates the prognosis risk of the patient by detecting the pDC level in the peripheral blood of the patient.
By dynamically monitoring the change trend of the pDC, a doctor can grasp the immune state of a patient in real time and adjust the treatment strategy in time. The invention can also be combined with the existing clinical scoring systems (such as Phoenix and pSOFA scoring) to further improve the accuracy of prognosis evaluation.
The specific technical scheme is as follows:
1. collecting a peripheral blood sample of the child sepsis infant for immune cell analysis;
2. Detecting the percentage of pDC in peripheral blood by flow cytometry, the level of which is closely related to the immune status of sepsis;
3. Prognosis risk assessment, when pDC levels are higher than 0.96%, suggesting that the patient has a higher risk of mortality;
4. dynamically monitoring the level of pDC at different time points (24 hours, 48 hours and 72 hours), and carrying out continuous prognosis evaluation by combining the clinical manifestation and the scoring system of the patient;
5. The method of the invention can be combined with other clinical scoring systems (such as pSOFA, phoenix scoring) to further improve the accuracy of prognosis prediction. By combining immune cell detection with organ function scoring, a more comprehensive risk assessment is provided.
In some embodiments of the invention, patients diagnosed with pediatric sepsis are enrolled as subjects, and their peripheral blood samples are taken within 6 hours of patient admission.
In some embodiments of the invention, the anti-CD 3 antibody, anti-CD 19 antibody, anti-CD 20 antibody, anti-CD 14 antibody, anti-CD 16 antibody, anti-CD 56 antibody, anti-CD 123 antibody, and anti-HLA-DR antibody are subjected to flow cytometry analysis ,CD3-CD19-CD20-CD14-CD16-CD56-CD123+HLA-DR+ to represent pDC cells, and the proportion of pDC in peripheral blood is determined. The expression of pDC was detected by antibody staining in combination with a flow cytometer.
In some embodiments of the invention, the ROC curve is used to evaluate the relationship of pDC levels to sepsis prognosis. From the analysis results, when pDC levels were higher than 0.96%, the patient was judged to be at higher risk of death.
In some embodiments of the invention, blood samples are taken from the patient at different time points (24 hours, 48 hours, 72 hours) for dynamic monitoring of pDC. By monitoring its changes over the course of the disease, the patient's immune response is assessed and clinical intervention strategies are guided.
The present invention focuses on methods of assessing prognosis of pediatric sepsis patients by exploring plasma cell-like dendritic cell (pDC) levels. By detecting the proportion of pDC in peripheral blood, patients at high risk for sepsis can be identified early and the change in immune status of the patients can be tracked in real time by dynamic monitoring.
The present inventors examined 99 sepsis infants using flow cytometry, divided them into Non-Remote and Remote groups, and two subgroups of survivinal and Non-survivinal groups, and followed for 28 days to evaluate mortality. Flow cytometry analyzed the percentage of pDC cells, which was found to be significantly higher in the Remote group than in the Non-Remote group and significantly higher in the dead group than in the surviving group. The pDC expression level was significantly positively correlated with sepsis-related scores. Infants with sepsis may be at risk of death when the percentage of pDC cells in human peripheral blood lymphocytes is higher than 0.96%. pDC was not different between the death group and the inventory group for 72 hours. The present application provides a method for assessing the prognosis of a sepsis patient by detecting pDC levels.
The study crowd receives 102 cases of sepsis infants meeting inclusion exclusion criteria during the period of 7 months of 2022 to 4 months of 2024 and receives treatment by PICUs of affiliated children hospitals of Chongqing medical university. The inclusion criteria were (1) diagnosis meeting the definition of "severe sepsis" or "septic shock" in International PEDIATRIC SEPSIS consensus conference in 2005 (2) age above 28 days and below 18 years, and (3) patient guardian signed informed consent. The exclusion criteria were (1) autoimmune disease, hematological malignancy, immunodeficiency disease patients, and (2) infants who had undergone immunotherapy (hormone impact, IVIG, immunosuppressant, mab, etc.) or blood purification therapy (hemodiafiltration, perfusion, plasmapheresis) prior to group entry. Whereas the Society of advanced medical science in the united states (Society of CRITICAL CARE MEDICINE, SCCM) of 2024 has issued new standards of Phoenix sepsis for pediatric sepsis and septic shock, the present application analyzed only 99 of 102 infants who met the new standards of Phoenix sepsis in 2024. The 2024 standard infant patients were classified into Remote group and Non-Remote group according to the presence or absence of organ dysfunction far from the primary infection site. The infant patients were classified into Survival groups and Non-survivinal groups according to their hospital fates. Partial immune indexes of the early (3 days before the course of disease) and late (3 days before the course of disease) different hospital outcome infants (survivinal (e) group/nonsurvival (e) group) and the different hospital outcome infants (survivinal (l) group/nonsurvival (l) group) are analyzed according to blood sampling time. Basic information (sex, age), vital signs, test results (blood routine, biochemical, etiological, inflammatory index), phoenix sepsis score, treatment regimen, hospital outcome and 28-day survival were recorded for all infants. Peripheral blood of the infants was obtained within 6 hours of the group. Some patients additionally obtained blood samples 24 hours, 48 hours and 72 hours after their inclusion in the group.
Sample collection blood samples of the group of infants were collected using ethylenediamine tetraacetic acid (EDTA) anticoagulation tube, and Peripheral Blood Mononuclear Cells (PBMCs) were immediately isolated by density gradient centrifugation using human lymphocyte separation solution for flow cytometry.
Flow cytometry detection by adding 1640 medium containing 2% fetal bovine serum to freshly extracted Peripheral Blood Mononuclear Cells (PBMCs) and mixing well, adding 100. Mu.L of the cell mixture to flow tubes, with 10 6 cells per tube. Centrifuging, removing supernatant, adding anti-CD 3 antibody, anti-CD 19 antibody, anti-CD 20 antibody, anti-CD 14 antibody, anti-CD 16 antibody, anti-CD 56 antibody, anti-CD 123 antibody and anti-HLA-DR antibody, staining flow cell, and incubating at room temperature for 30 min. Cell detection was performed using a flow cytometer and data was analyzed using flow cytometry software (e.g., flowJo). The ratio of pDC to total lymphocytes (obtained using FSC-A and SSA-A gates of the flow cytometer) was determined and the percentage of pDC was recorded.
PDC level evaluation the test results showed that pDC was a percentage of lymphocytes, suggesting that patients had a higher risk of death when pDC expression levels were higher than 0.96%. The threshold is based on the analysis of the working characteristic curve (ROC) of the subject, and has good sensitivity and specificity.
Dynamic monitoring in order to improve the monitoring effect on patient prognosis, the present invention suggests blood sample collection and detection at various time points during sepsis, usually detecting pDC levels again 24, 48 and 72 hours after admission. By dynamically monitoring the change trend of pDC, the recovery or deterioration of the immune function of the patient is evaluated, and the disease progress is further judged. pDC levels were not different for 72 hours of pDC expression between the death group and the survival group.
In combination with clinical scoring systems the methods of the invention may be used in combination with existing sepsis clinical scoring systems such as pSOFA and Phoenix scores. By comparing the detection result of pDC with clinical scores, the accuracy of prognosis evaluation can be improved. If the patient has a pDC level above 0.96% and the clinical score indicates that the condition is severe, the patient is more confident that the patient is at high risk of mortality and is thus taking more aggressive treatment.
It should be understood that one or more of the expressions ". The expressions" individually include each of the objects recited after the expressions and various combinations of two or more of the recited objects unless otherwise understood from the context and usage. The expression "and/or" in combination with three or more recited objects should be understood as having the same meaning unless otherwise understood from the context.
The use of the terms "comprising," "having," or "containing," including grammatical equivalents thereof, should generally be construed as open-ended and non-limiting, e.g., not to exclude other unrecited elements or steps, unless specifically stated otherwise or otherwise understood from the context.
It should be understood that the order of steps or order of performing certain actions is not important so long as the application remains operable. Furthermore, two or more steps or actions may be performed simultaneously.
The use of any and all examples, or exemplary language such as "e.g." comprising "or" including "in this document is intended merely to better illuminate the application and does not pose a limitation on the scope of the application. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the application.
Furthermore, the numerical ranges and parameters setting forth the present application are approximations that may vary as precisely as possible in the exemplary embodiments. However, any numerical value inherently contains certain standard deviations found in their respective testing measurements. Accordingly, unless explicitly stated otherwise, it is to be understood that all ranges, amounts, values and percentages used in this disclosure are modified by "about". As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a particular value or range.
Abbreviations involved in the present application ：WBC,white blood cell;ANC,absolute neutrophil count;ALC,absolute lymphocyte count;CRP,C-reactive protein;PCT,procalcitonin;ALT,alanine transaminase;APTT,activated partial thromboplastin time;INR,international normalized ratio;OFI,organ failure index;PSS,phoenix sepsis score;pSOFA,pediatric sequential organ failure assessment;D,day;CNS,central nervous system;UTI,urinary tract infection;GCC,glucocorticoid;CRRT,continuous renal replacement therapy;IVIG,intravenous immunoglobulin.
Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all commercially available and commercially available.
The invention is further illustrated by the following examples.
EXAMPLE 1 pDC level detection in sepsis infant
The samples to be tested are 99 cases of infants suffering from sepsis (figure 1) according to the nano-row standard, and the clinical characteristics are shown in table 1. Of these, non-Remote group (28, 28.3%), remote group (71, 71.7%), surviving group (87, 87.9%), dead group (12, 12.1%) the primary infection site of sepsis infant was mainly respiratory tract (67, 67.7%), followed by digestive tract (20, 20.2%) and central nervous system (9, 9.1%). The infant was mainly diagnosed with severe pneumonia (63 cases, 63.6%). All cases of death in the present invention were hospital deaths, with half of the children dying within 3 days after admission (6, 50.0%).
TABLE 1 clinical characterization of surviving and dying groups
Data are expressed in terms of n (number, percent), mean ± standard deviation, or median (first quartile, third quartile).
Materials are fluorescent-labeled anti-CD 3 (APC-Cy 7) antibodies, anti-CD 19 (APC) antibodies, anti-CD 14 (PE-Cy 7) antibodies, anti-CD 20 (APC) antibodies, anti-CD 16 (BB 700) antibodies, CD56 (APC-Cy 7) antibodies, anti-CD 123 (PE) antibodies and anti-HLA-DR (FITC) antibodies purchased from BD Pharmingen, and human lymphocyte separation solution (Cedarlane) purchased from Beijing daceae Biotechnology Co.
The experimental method comprises the following steps:
Taking 1 mL anticoagulated peripheral blood sample for peripheral blood mononuclear cell separation, and detecting the separated PBMCs by flow cytometry.
Mu.L of 10 6 PBMCs were taken, 1. Mu.L of each of the APC-Cy7 anti-CD 3 antibody, APC anti-CD 19 antibody, APC anti-CD 20 antibody, PE-Cy7 anti-CD 14 antibody, BB700 anti-CD 16 antibody, APC-Cy7 anti-CD 56 antibody, PE anti-CD 123 antibody and FITC anti-HLA-DR was added, incubated at room temperature for 30 min, 1 mL of PBS was added, 500g of PBS was centrifuged for 5 min, and after washing once, 200. Mu. LPBS was added for suspension, and the results were run on a flow machine and analyzed.
The result analysis shows that the percentage of pDC of the sepsis infant is shown in the figures 2A-2C. Fig. 2A shows the gate strategy for pDC in this example. The results in fig. 2B show that pDC is expressed significantly higher in the Remote group than in the nonremote group. The results in fig. 2C show that pDC cells are elevated in the dead group. Taken together, the sepsis infant who progressed and died had highly expressed pDC cells.
Example 2 levels of pDC in sepsis patients were inversely correlated with Phoenix and pSOFA scores
The new standard of Phoenix for diagnosis of pediatric sepsis and septic shock by the Society of medicine of CRITICAL CARE MEDICINE, SCCM in the united states of america, 2024, score Phoenix sepsis of 2 minutes or more, indicating the potential life threatening organ dysfunction of the respiratory, cardiovascular, clotting and/or nervous systems of suspected or diagnosed infected infants.
In sepsis diagnostic standard sepsis 3.0.0, pSOFA score is an important index for diagnosing sepsis, and a large number of study data show that when pSOFA score is more than or equal to 2, patients with ICU infection or suspected infection are diagnosed with sepsis, and pSOFA reflects the degree of multiple organ dysfunction of patients with multiple organ functional status syndrome (MODS) and is closely related to the hospitalization rate of the patients.
Fig. 3A-3C show correlation analysis of the levels of pDC in the sepsis infant in this example with Phoenix score and pSOFA score. As shown in fig. 3A, the correlation coefficient r=0.39, p < 0.0001 of the percent pDC change and Phoenix Sepsis Scores (PSS) -4 score of the sepsis infant. The results in FIG. 3B show that the correlation coefficient r=0.37 and P=0.0002 between the percentage of pDC and the score of PSS-8 in the sepsis infants. Fig. 3C correlation analysis found that the correlation coefficient r=0.38, p=0.0001 of pDC expression level and pSOFA score of pediatric sepsis patients. Taken together, the levels of pDC in sepsis infants were significantly positively correlated with Phoenix scores and pSOFA scores.
EXAMPLE 3 analysis of ROC Curve found that pDC had good predictive efficacy against nosocomial death in infants with sepsis
Laboratory indicators and clinical data of the sepsis infant are collected. The diagnostic efficacy of pDC against nosocomial mortality in infants with sepsis was analyzed using ROC curves.
Fig. 4 shows the predicted efficacy of pDC on nosocomial death for the ROC curve analysis of sepsis patients in this example. As shown in fig. 4 and table 2, ROC curve analysis found that pDC had good predictive efficacy against nosocomial death of sepsis infants, with an area under the curve of 0.7632 (95% CI: 0.5951-0.9313, P < 0.005), an optimal threshold of 0.96%, at this time sensitivity of 81.8%, specificity of 77.9%. From the above statistical results, the area under the curve of the pDC for the diagnosis of sepsis in children is 0.7632, the diagnosis efficiency is good, and the statistical significance is achieved, so that the pDC has good prediction efficiency on the hospital death of the sepsis infant.
TABLE 2
EXAMPLE 4 pDC to sepsis infant mortality outcome relationship
Further observing the death outcome of pDC and the sepsis infant, and making a Kaplan-Meier survival curve of the sepsis infant. The results in FIG. 5 show that the mortality increases in the lower expression group of the high expression group, with the pDC expression level being 0.96%, and the difference is statistically significant.
EXAMPLE 5 pDC did not have stable predicted efficacy over 72 hours
To explore whether pDC as a potential predictor has stable predictive value in the short term, changes in pDC in sepsis children with different prognosis in different stages were analyzed and the level of pDC in 16 sepsis children within 72 hours was randomly monitored. The results in fig. 6A show that the percentage of pDC is increased in the early-death group of sepsis compared to the survival group (P < 0.05). Fig. 6B shows that pDC expression was not significantly different between the two groups in the late-death group of sepsis infants. The results in fig. 6C show no significant difference in pDC expression for 72 hours between the death group and the survival group.
In summary, the application provides a sepsis prognosis evaluation method based on plasmacytoid dendritic cells (pDC), which can effectively evaluate the prognosis of a patient suffering from sepsis by detecting the level of pDC in peripheral blood through flow cytometry and combining a dynamic monitoring and clinical scoring system. The method has high specificity and sensitivity, is particularly suitable for early risk assessment of patients with children sepsis, and provides scientific basis for the establishment of clinical personalized treatment schemes.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
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| OLIVIER GUISSET 等: "Decrease in circulating dendritic cells predicts fatal outcome in septic shock", 《INTENSIVE CARE MED》, vol. 33, 8 November 2006 (2006-11-08), pages 148 - 152, XP019471500 * |
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