JP2006119103A - Method for classifying pathologic conditions of renal disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method using human urine as a specimen and classifying pathologic conditions into five types which are tubulointerstitial injury, glomerular injury, without the accompanying cellular infiltration, glomerular injury accompanied by cellular infiltration, glomerular injury and tubulointerstitial injury accompanied by cellular infiltration, and non-glomerular hematuria, using three kinds analysis results of erythrocyte forms, immunocyte characters and protein fractional pathological conditions. <P>SOLUTION: Three kinds of analytical results obtained by urine analyses are classified into five kinds of pathologic conditions, in accordance with classification method shown in Figure 1, and they are utilized for predicting the diseased portions in kidney and treating renal diseases. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

Technical field related to methods of classifying kidney disease pathology based on urinalysis results

Conventionally, urine erythrocyte morphology has been examined with a phase contrast microscope. Detection sensitivity was low, and it was difficult to determine whether it was due to renal glomerular disease or urinary tract disease. Recently, the automatic urine erythrocyte morphology test apparatus using laser scattered light as disclosed in Patent Documents 1, 2, and 3 has become capable of finely classifying morphologies and is being used on a daily basis. Diagnosis of kidney disease is not only made from the morphological measurement results of these blood, but has also been tried together with diagnosis by another test method. It is a pathological classification method for classifying glomerular renal disorder or tubular renal disorder by urine protein fraction as shown in Patent Document 4. As a method for examining surface characteristics of lymphocytes, whether or not infiltration of inflammatory cells is active has been established by a flow cytometry method (FACS method) using a fluorescently labeled antibody (Non-patent Document 1). . Furthermore, regarding the diagnosis of kidney disease, in addition to these examination methods, kidney tissue was collected (referred to as renal biopsy) and finally used in combination with pathological examination methods. However, renal biopsy is an invasive examination, and complications including death cases have been reported, and due to recent medical circumstances, severely applicable cases have been selected for conducting this examination.
JP-A-8-240520 Japanese Patent Laid-Open No. 11-295207 Japanese Patent Laid-Open No. 11-118793 Japanese Patent Application No. 2003-367424 J Am Soc Nephrol 12: 2636-2644,2001

In the diagnosis of kidney disease, doctors have independently diagnosed each test result as described in the prior art until now, but in the present invention, these test results are combined to make non-invasive. The purpose was to make use of it for the diagnosis of renal diseases.

Until now, doctors have comprehensively diagnosed based on the results of these examination methods. In the present invention, these examination methods are useful for diagnosis by classifying the pathological conditions of renal diseases by computer image analysis and statistical methods. A method that can provide information is presented. In order to solve this problem, a method has been invented in which a number of clinical cases are used, the respective examination methods are compared and examined, and the pathological condition of renal disease can be grasped based on clinical diagnosis by renal biopsy. As a result, an approximate diagnosis can be made without performing a renal biopsy. Details are as follows.

In general, red blood cells and proteins do not appear due to kidney function, and the appearance of urinary red blood cells, urinary immune cells, and urine protein itself is a predictive disorder of renal function, that is, renal disease. Among the kidney diseases, IgA nephropathy, membranous nephropathy, crescent nephritis, focal glomerulosclerosis and the like are known as renal glomerular disorders. Examples of renal tubular disorders include Fancony syndrome and interstitial nephritis.

Renal biopsy is widely used for definite diagnosis of kidney disease. Renal biopsy is a method in which a needle is punctured toward a patient's kidney, a renal tissue is taken out, a histopathological specimen is prepared, and a pathological diagnosis is performed by a specialist after staining. There are also reports of complications, and considerable cost, time, and skill were required for collecting tissues, preparing pathological tissue specimens, and reading and judging the specimens.

In addition, some of the patient's organs are removed, causing damage to surrounding tissues, resulting in complications related to bleeding such as gross hematuria and hematoma formation. If bleeding is severe, blood pressure drops, shock, and death May lead to.

The present invention is based on the results of many examination methods such as the aforementioned Patent Documents 1, 2, 3 and 4 and Non-Patent Document 1, and the results of detailed examination of clinical examples of renal biopsy and highly sensitive urinary protein electrophoresis images. By combining various types of non-invasive urine testing methods, the disease was classified into 5 groups, and the pathological analysis of the algorithm specific to that group was performed by a computer, and it was concluded that it was very useful.

The three types of inspection methods are as follows.

The first examination method is that urine red blood cells are deformed by determining the red blood cell morphology by an automatic urine red blood cell morphology test using laser scattered light as shown in Patent Documents 1 and 2 and 3, or Categorize whether or not The deformation of erythrocytes is considered to occur when the erythrocytes are partially crushed because the erythrocytes pass through pathological changes such as rupture of the glomerular capillaries. Further, when passing through the tubule after that, the shape is changed to a confetti shape due to a change in osmotic pressure of urine. On the other hand, normal erythrocytes without deformation means bleeding from the lower urinary tract without such changes.

The next test method is a test of immune cells in urine, which is called FACS method and analyzes the appearance of cell surface and classifies it as effector cells or non-effector cells.

Effector cells are surface antigens of immune cells such as monocytes and lymphocytes that are negative for CD62L, and lymphocytes are simultaneously CD45RO positive and CD45RA negative. These effector cells are activated cells that cause tissue damage, and nephritic infiltrating cells also show the same phenotype. Urinary effector cells are thought to have appeared in urine as a result of renal inflammatory cells damaging renal tissue, such as by destroying renal glomerular capillaries (Non-patent Document 1). On the other hand, non-effector cells are positive for CD62L, and lymphocytes are simultaneously negative for CD45RO and positive for CD45RA. Harmless non-activated cells, the majority of immune cells in the peripheral blood of healthy individuals exhibit this phenotype.

The third method is to perform electrophoresis of patient urine protein using cellulose acetate membrane as a support. After detecting the ultra-trace amount of protein with silver stain, it is based on the method shown in Patent Document 4. Classify into glomerular damage pattern, tubule damage pattern, and mixed pattern.

The glomerular injury pattern refers to a group characterized in that the fraction values of the albumin fraction and the β fraction appear significantly higher than the fraction values of the other disease groups, respectively.

The tubule injury pattern is that the fraction value of the albumin fraction is significantly lower than the fraction value of the albumin fraction of the group mainly composed of renal glomerular disorders and is sandwiched between them instead of the β fraction. A group characterized by the fact that another protein fraction appears significantly darker.

The mixed pattern refers to a pattern in which both the glomerular damage pattern and the tubular damage pattern are seen.

The pathological analysis algorithm for urinary disease combining three types of testing methods was as follows.

Non-effector that is an immune cell in urine, with R1 when there are red blood cells in the urine and the shape of the red blood cells is normal, R2 when the shape is deformed, and R3 when no red blood cells are seen in the urine The case where cells were seen was designated as I1, the case where effector cells were seen as I2, and the case where immune cells were not found in urine as I3. In the urine protein pathological classification, the glomerular damage pattern is P1, the tubular damage pattern is P2, and the mixed type is P3.

According to the above combination, the following A to E type damaged sites can be examined.

Type A tubulointerstitial disorder: a combination of R3, I2, and P2, and related diseases include tubulointerstitial nephritis.

Since there is no glomerular injury such as glomerular capillary destruction / rupture, there is no deformed red blood cell of R3, and infiltration of effector cells into the tubulointerstitium and consequent destruction of the tubules is considered, and I2 effector cells appear. As a result, urinary protein tubule injury pattern of P2 is shown, and tubular proteinuria appears.

B-type glomerular disorder without cell infiltration: a combination of R3, I3, and P1, and related diseases include micro-variable, membranous nephropathy, and glomerular hypertrophy.

Since there is no glomerular damage such as destruction or rupture of glomerular capillaries by inflammatory cells, there is no deformed erythrocyte of R3, and there is a problem of protein leakage from the glomeruli, indicating a urinary protein glomerular damage pattern of P1. In addition, since there is no effector cell of I3, there is no inflammatory cell infiltration in renal tissue.

Glomerular disorder with type C cell infiltration: Diseases related to the combination of R2, I2, and P1 include IgA nephropathy and crescent-forming nephritis such as ANCA-related nephritis.

Since there are glomerular disorders such as destruction and rupture of glomerular capillaries by inflammatory cells, there are deformed erythrocytes of R2, and there is also a disorder of protein leakage from the glomeruli. There are also I2 effector cells, suggesting inflammatory cell infiltration in renal tissue.

D-tubulointerstitial disorder with cell infiltration: Diseases associated with R2, I2, P2 or P3 combinations include ANCA-associated nephritis with tubulointerstitial nephritis (TIN).

R2 deformed red blood cells due to glomerular damage such as destruction and rupture of glomerular capillaries by inflammatory cells, P2 or P3 urine protein tubule injury pattern only, urine protein tubule injury pattern and urine protein In order to show a glomerular damage pattern, there is a reabsorption disorder of small molecule protein, and then protein leaks from the glomerulus as the damage progresses. In addition, this type suggests inflammatory cell infiltration in kidney tissue, and I2 effector cells are also found.

Type E non-glomerular hematuria ・ ・ ・ R1 and I1 related diseases include urinary tract bleeding and idiopathic renal bleeding.

Since there is no glomerular damage such as glomerular capillary destruction / rupture, there is no deformed red blood cell of R3, red blood cells are peripheral blood types, and urine immune cells are also peripheral blood types, so there are non-effector cells of I1.

Until now, renal biopsy has been considered essential for diagnosis. This is invasive because the patient's kidney must be pierced with a biopsy needle and the kidney tissue must be collected, which naturally hurts the patient. It also damages normal cells. In addition, the collected kidney tissue requires several days for the preparation of a pathological specimen by a specialist engineer, and further requires microscopic diagnosis of the specimen by a specialist.

The role played by the present invention mainly has two aspects. One of them is that early detection of a renal disease is possible, and patients who need treatment can be quickly extracted. Up to now, due to constraints such as time constraints, cost constraints, and too many patients, patients who have been left without a renal biopsy, which was the only diagnostic test, will be saved.

Secondly, it is possible to perform a rough pathological diagnosis of kidney disease instead of a renal biopsy. Renal biopsy is dangerous and costly and time consuming, but by using the present invention to start treatment before the disease progresses without performing renal biopsy, it is possible to accelerate disease healing or start dialysis Can be delayed.

Subdivided diagnostic technology is based on the accumulation of various specialized technologies such as IgA nephropathy, dialysis, diabetes, glomerulonephropathy, and autoimmune diseases. Although this situation should not be overlooked, the present invention supports these diagnoses from the aspect.

Early treatment by early diagnosis dramatically improves the cure rate. Moreover, even when the disease progresses without reaching the therapeutic effect, the start time of the artificial dialysis can be delayed and the deterioration of the disease state can be prevented, so that the medical cost can be reduced as a result.

After collecting urine, erythrocyte morphology is determined by the urine erythrocyte analyzer described in Patent Documents 1, 2, and 3, and then urine sediment and supernatant are centrifuged. Surface traits of spheres and monocytes were analyzed by FACS method. The supernatant was subjected to urine protein fractionation by cellulose acetate membrane electrophoresis, classified by the method of Patent Document 4, and further classified by the above-described algorithm of the present invention.

Non-effector that is an immune cell in urine, with R1 when there are red blood cells in the urine and the shape of the red blood cells is normal, R2 when the shape is deformed, and R3 when no red blood cells are seen in the urine The case where cells were seen was designated as I1, the case where effector cells were seen as I2, and the case where immune cells were not found in urine as I3. In the classification of urinary protein pathology, the glomerular damage pattern was P1, the tubular damage pattern was P2, and the mixed type was P3.
According to the above combinations, the A to E type lesions could be classified as shown in FIG. It should be noted that there is no problem even if the names of the A to E type faulty sites are changed to unique names.

Urinalysis pathology classification table

Claims (6)

According to human urine analysis results, tubulointerstitial disorders, glomerular disorders without cell infiltration, glomerular disorders with cell infiltration, glomerular and tubulointerstitial disorders with cell infiltration, non- How to classify the five pathologies of glomerular hematuria 2. The method for grasping the pathophysiology of tubulointerstitial disorder is characterized in that urine erythrocytes are not significantly present, effector type immune cells are significantly present, and urinary protein electrophoretic images show tubular images. Classification of disease states As a method of grasping the pathology of glomerular injury without cell infiltration, it is characterized by the absence of urine red blood cells, the absence of effector-type immune cells, and the urinary protein electrophoresis image showing glomerular image The pathological classification method of claim 1 As a method of grasping glomerular injury accompanied by cell infiltration, urine erythrocytes are significantly deformed, there are significant number of effector type immune cells, and urine protein electrophoresis image shows glomerular image The pathological classification method of claim 1 As a method of grasping the pathophysiology of glomerular injury and tubulointerstitial injury associated with cell infiltration, urine erythrocytes are significantly present and deformed, there are a significant number of effector-type immune cells, and urine protein electrophoresis images 2. The pathological classification method according to claim 1, characterized in that it shows a mixed or tubular image Non-glomerular hematuria as a method of understanding the pathology of urine erythrocytes is significantly present and untransformed, non-effector immune cells are significant, and urine protein electrophoresis images show the same image as serum The pathological classification method of claim 1 characterized by

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