Application of monocyte subsets in monitoring disease activity in nephrotic syndrome

Abstract

The present invention proposes a method for monitoring the disease activity of idiopathic nephrotic syndrome (INS) based on changes in the proportion of monocyte subpopulations. The CD14++CD16+ intermediate monocyte subpopulation in the patient's peripheral blood is detected by flow cytometry, and its proportion in monocytes at different stages of the disease is monitored to evaluate the disease state of INS in real time. This method overcomes the limitations of traditional clinical indicators in reflecting the activity state of the disease, provides more sensitive and specific biomarkers, helps to guide individualized treatment, optimize prognosis evaluation, and provide a new perspective for the study of the pathogenesis of INS. The present invention provides innovative technical means for the clinical diagnosis and treatment of INS and disease monitoring, and has important clinical application value.

Description

Technical Field

[0001] The present invention relates to diseases, and in particular to the application of monocyte subsets in monitoring the disease activity of nephrotic syndrome Background Art

[0002] Idiopathic Nephrotic Syndrome (INS) is a common glomerular disease in children, and its clinical manifestations mainly include massive proteinuria, hypoalbuminemia, edema and hyperlipidemia. The pathogenesis of INS is complex and is generally believed to be closely related to immune disorders. In recent years, with the deepening of immunological research, the role of monocytes in autoimmune diseases has gradually attracted attention. As innate immune cells, monocytes can respond to infection and tissue damage, produce inflammatory responses, and play an important role in antigen presentation and cytokine production.

[0003] Among the subpopulation classifications of monocytes, intermediate monocytes (CD14++CD16+) in human peripheral blood have received special attention due to their significant pro-inflammatory characteristics. The CD14++CD16+ monocyte subpopulation exhibits active pro-inflammatory effects in a variety of immune diseases, participating in and exacerbating inflammatory responses by secreting a variety of inflammatory factors and chemokines. However, in INS, the specific roles of different subpopulations of monocytes and their relationship with the active state of the disease remain unclear.

[0004] At present, the diagnosis of INS mainly relies on clinical manifestations and laboratory tests, such as urine protein quantification, serum albumin level, blood lipid detection, etc. However, these indicators have certain limitations in reflecting the activity of the disease. Although urine protein quantification can reflect the degree of damage to the glomerular filtration membrane, it cannot directly reflect the state of the immune system. Although serum albumin level is related to the severity of the disease, its change is relatively lagging and is affected by multiple factors. Therefore, it is of great clinical significance to find more sensitive and specific biomarkers to monitor the disease activity of INS.

[0005] In order to solve the above problems, the applicant proposed the application of monocyte subsets in monitoring the disease activity of nephrotic syndrome. Summary of the invention

[0006] The object of the present invention is to provide an application of monocyte subpopulations in monitoring the disease activity of nephrotic syndrome, so as to solve the problems in the prior art.

[0007] To achieve the above object, the present invention provides the following technical solution: a method for monitoring the disease activity state of idiopathic nephrotic syndrome, the method comprising:

[0008] Detect the proportion of intermediate monocyte subsets in the peripheral blood of INS patients;

[0009] The disease activity status of INS is reflected according to the changes in the proportion of the monocyte subsets. Among them, the proportion of the CD14++CD16+ monocyte subsets increases significantly during the active stage of the disease and is significantly positively correlated with the urine protein level. The proportion decreases after the disease is relieved.

[0010] Optionally, it also includes detecting the ratio of classical monocyte subsets and non-classical monocyte subsets, and comprehensively analyzing the changes in the ratios of these monocyte subsets to more comprehensively monitor the disease activity status of INS.

[0011] Optionally, the detection is performed by flow cytometry, using specific antibodies to mark CD14 and CD16, and distinguishing different monocyte subsets accordingly.

[0012] Optionally, the method further comprises collecting clinical indicators of INS patients, including blood albumin level, blood creatinine level and urine protein level, and performing correlation analysis between changes in monocyte subset ratios and these clinical indicators to further confirm the disease activity status.

[0013] Optionally, the correlation analysis shows that the ratio of CD14++CD16+ monocyte subsets is significantly negatively correlated with blood albumin levels, and significantly positively correlated with urine protein levels.

[0014] A reagent composition for monitoring the disease activity state of idiopathic nephrotic syndrome, comprising a specific antibody for detecting CD14++CD16+ monocyte subpopulations in peripheral blood and a reagent for flow cytometer analysis.

[0015] Optionally, specific antibodies for detecting classical monocyte subsets and non-classical monocyte subsets are also included.

[0016] Optionally, the reagent composition further comprises reagents and / or equipment for collecting and analyzing clinical indicators of INS patients.

[0017] A method for preparing the reagent composition according to claim 6 or 7, the method comprising:

[0018] Provides specific antibodies for labeling CD14, CD16, and possibly other monocyte subset markers in peripheral blood;

[0019] The antibody is combined with reagents required for flow cytometric analysis to form a reagent composition capable of detecting monocyte subpopulations.

[0020] Beneficial effects: 1. The present invention provides a new, non-invasive means of monitoring disease activity in the clinic by monitoring the changes in the proportion of monocyte subsets in the peripheral blood of INS patients, especially the proportion of CD14++CD16+ monocyte subsets. This method can reflect the disease activity status of INS patients in real time, which helps doctors to adjust the treatment plan in time and improve the treatment effect.

[0021] 2. Improved the accuracy of disease activity judgment:

[0022] Compared with traditional clinical indicators (such as urine protein quantification, serum albumin level, etc.), the proportion of monocyte subsets as a biomarker has higher sensitivity and specificity. By combining traditional indicators with monitoring of monocyte subset proportions, clinicians can significantly improve their ability to judge disease activity and guide the adjustment of treatment plans and prognosis judgment.

[0023] 3. Guide individualized treatment:

[0024] Different INS patients have different responses to treatment, and individualized treatment is the key to improving the efficacy. The present invention can evaluate the patient's immune status by monitoring the proportion of monocyte subsets, providing a basis for the formulation of individualized treatment plans. For example, for patients with a significantly increased proportion of CD14++CD16+ monocyte subsets, more active immunomodulatory treatment may be required.

[0025] 4. Optimize prognostic assessment:

[0026] The method of the present invention can also be used for the prognosis assessment of INS patients. By continuously monitoring the changes in the proportion of monocyte subsets, the progression trend of the disease can be predicted, providing guidance for the long-term management of patients. This is of great significance for improving the quality of life of patients and reducing the incidence of complications.

[0027] 5. Promote scientific research progress:

[0028] The implementation of the present invention not only provides a new tool for clinical diagnosis and treatment, but also provides a new perspective for the study of the pathogenesis of INS. By deeply studying the role of monocyte subsets in INS, it is expected to reveal new mechanisms of the disease, provide targets for new drug development, and promote scientific research progress in the field of INS treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Changes in the proportion of CD14+ monocyte subsets in children with INS;

[0030] Figure 2 Relationship between different subsets of CD14+ monocytes and urine protein. DETAILED DESCRIPTION

[0031] The following describes the preferred embodiments of the present invention with reference to the drawings in the specification, so that the technical content is clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0032] In the drawings, components with the same structure are indicated by the same numerical reference numerals, and components with similar structures or functions are indicated by similar numerical reference numerals. The size and thickness of each component shown in the drawings are arbitrarily shown, and the present invention does not limit the size and thickness of each component. In order to make the illustration clearer, the thickness of the components is appropriately exaggerated in some places in the drawings.

[0033] Example 1

[0034] The present invention provides a technical solution for monitoring the disease activity state of idiopathic nephrotic syndrome (INS) based on the change of monocyte subset ratio. The specific technical solution is as follows:

[0035] 1. Sample Collection and Processing

[0036] Step 1.1 Sample collection

[0037] Peripheral blood samples were collected from patients with INS or suspected INS.

[0038] At the same time, peripheral blood samples from healthy individuals were collected as a control group.

[0039] Step 1.2 Sample processing

[0040] Peripheral blood samples were collected using ethylenediaminetetraacetic acid (EDTA) anticoagulant tubes to prevent blood clotting.

[0041] Anticoagulated whole blood samples were lysed to obtain single-cell suspensions.

[0042] The cell pellet was washed with phosphate-buffered saline (PBS) and resuspended into a single-cell suspension at an appropriate concentration.

[0043] 2.Monocyte Subpopulation Detection

[0044] Step 2.1 Antibody labeling

[0045] Use specific antibodies (such as CD14 and CD16 antibodies) to label monocyte subsets in single cell suspensions.

[0046] After antibody labeling, incubate at room temperature for a certain period of time (eg, 20 minutes) to allow the antibody to fully bind to the cell surface antigen.

[0047] Step 2.2 Flow cytometry detection

[0048] The labeled cell suspension was detected by flow cytometry.

[0049] Flow cytometry can distinguish cell subsets with different fluorescent labels, thereby accurately measuring the proportions of CD14++CD16+ (intermediate monocytes), CD14++CD16- (classical monocytes) and CD14+CD16++ (non-classical monocytes).

[0050] 3. Data analysis and disease monitoring

[0051] Step 3.1 Data Collection

[0052] The proportion data of different monocyte subsets in each sample were recorded and collated.

[0053] At the same time, the patients' clinical indicator data were collected, including urine protein level, serum albumin level, blood creatinine level, etc.

[0054] Step 3.2 Correlation analysis

[0055] Statistical software (such as GraphPad Prism) was used to perform correlation analysis between the proportion of monocyte subsets and clinical indicators.

[0056] Special attention was paid to the correlation between the proportion of CD14++CD16+ monocyte subsets and urine protein levels and serum albumin levels.

[0057] Step 3.3 Assessment of disease activity

[0058] The disease activity of INS was assessed based on changes in the proportion of CD14++CD16+ monocyte subsets.

[0059] A significantly higher ratio usually indicates active disease, while a lower ratio may indicate remission.

[0060] 4. Kit and computer program application

[0061] Step 4.1 Kit preparation

[0062] A kit containing CD14 and CD16 specific antibodies, buffers, and controls was developed for the detection of monocyte subsets by flow cytometry.

[0063] The kit may further contain antibodies for detecting other monocyte subsets to provide a more comprehensive monitoring.

[0064] Step 4.2 Computer program development

[0065] Development of computer programs for receiving, analyzing, and processing monocyte subset proportion data.

[0066] The program can automatically assess the disease activity status of INS and output monitoring results and reports.

[0067] 5. Clinical application and verification

[0068] Step 5.1 Clinical validation

[0069] Verify the accuracy and reliability of this technical solution in different medical institutions and patient groups.

[0070] Collect feedback and continuously optimize and improve technical solutions.

[0071] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the attached claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0072] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A method for monitoring the disease activity state of idiopathic nephrotic syndrome, characterized in that, The method includes: Detect the proportion of intermediate monocyte subsets in the peripheral blood of INS patients; The disease activity status of INS is reflected according to the changes in the proportion of the monocyte subsets. Among them, the proportion of the CD14++CD16+ monocyte subsets increases significantly during the active stage of the disease and is significantly positively correlated with the urine protein level. The proportion decreases after the disease is relieved.

2. The method according to claim 1, characterized in that It also includes detecting the ratio of classical monocyte subsets and non-classical monocyte subsets, and comprehensively analyzing the changes in the ratios of these monocyte subsets to more comprehensively monitor the disease activity status of INS.

3. The method according to claim 1 or 2, characterized in that: The detection is performed by flow cytometry, using specific antibodies to label CD14 and CD16, and different monocyte subsets are distinguished accordingly.

4. The method according to claim 1 or 2, characterized in that: The method also includes collecting clinical indicators of INS patients, blood albumin level, blood creatinine level and urine protein level, and performing correlation analysis between changes in monocyte subset ratios and these clinical indicators to further confirm the disease activity status.

5. The method according to claim 4, characterized in that The correlation analysis showed that the proportion of CD14++CD16+ monocyte subsets was significantly negatively correlated with the blood albumin level, and significantly positively correlated with the urine protein level.

6. A reagent composition for monitoring the disease activity state of idiopathic nephrotic syndrome, characterized in that: The reagent composition comprises a specific antibody for detecting CD14++CD16+ monocyte subpopulations in peripheral blood and a reagent for flow cytometer analysis.

7. The reagent composition according to claim 6, characterized in that Also included are specific antibodies for the detection of classical and nonclassical monocyte subsets.

8. The reagent composition according to claim 6 or 7, characterized in that The reagent composition also includes reagents and / or equipment for collecting and analyzing clinical indicators of INS patients.

9. A method for preparing the reagent composition according to claim 6 or 7, characterized in that: The method includes: Provides specific antibodies for labeling CD14, CD16, and possibly other monocyte subset markers in peripheral blood; The antibody is combined with reagents required for flow cytometric analysis to form a reagent composition capable of detecting monocyte subpopulations.

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