JP5275211B2 - Determination method using interstitial pneumonia marker - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a marker for interstitial pneumonia capable of assisting in more accurately performing the detection diagnosis of interstitial pneumonia, the evaluation and elapse observation of disease activity, especially an aftercare prospect without performing histopathological retrieval because the execution of surgical pulmonary biopsy is often difficult in relation to interstitial pneumonia and capable of simply performing measurement. <P>SOLUTION: In the method for determining interstitial pneumonia, the amount of IL-8 in blood is used as the index for analyzing or evaluating the determination or diagnosis of intestitial pneumonia, the specification of the kind of intestitial pneumonia, a degree of the advance possibility of the sympton of intestitial pneumonia, a risk degree, the pathophysiological state of intestitial pneumonia and/or the tendency of treatment responsiveness. <P>COPYRIGHT: (C)2011,JPO&amp;INPIT

Description

  The present invention relates to a marker for determining the pathology of interstitial lung disease, particularly interstitial pneumonia, and a method for determining the pathology of interstitial pneumonia using the marker.

Interstitial Pneumonia (IP) is a general term for inflammatory diseases of the interstitium of the lung, that is, the alveolar septum (alveolar septum). From the obvious etiology to the unknown etiology Various types are included, some of which develop alveolar fibrosis, which progresses to present respiratory failure and follow a terrible course of exacerbation and inability to treat. Here, the lung interstitium includes alveolar septum in the narrow sense, lobular stroma in the broad sense, and the vicinity of the pleura. Interstitial pneumonia differs in its treatment method and outlook of the course of illness depending on its type, so it is considered that discrimination of the type is very important.
Interstitial pneumonia is caused by chronic diseases such as rheumatoid arthritis and polydermatomyositis and other collagen diseases (autoimmune diseases), occupational and daily dust (dust), mold, pet hair and feathers. It is known that there are various inhalations, medicines prescribed in hospitals, herbal medicines, health foods such as supplements, special infections, etc., but those that cannot identify a clear cause are idiopathic interstitial pneumonia (Ideopathic Interstitial Pneumonia, IIP) and pathologically classified as follows.

Idiopathic pulmonary fibrosis (IPF) is a pathological finding of normal interstitial pneumonia (UIP), clinical course is chronic, reaction to steroids is poor, acute exacerbation due to colds, etc. (Stable medical conditions worsen rapidly), the course and prognosis of the disease is extremely poor, it does not recover completely, and care should be taken not to worsen the medical condition. .
Non-specific interstitial pneumonia (NSIP) is further subdivided into groups I, II, and III, group I has no fibrosis, group II has fibrosis, and honeycombs None, Group III is a type with both fibrosis and honeycombing, its clinical course is subacute to chronic, its response to steroids is relatively good, and its course and prognosis are generally good, but rare Is that there is a defect. In the case of this disease, there is improvement in the condition and complete recovery, but the prognosis is not good compared with idiopathic organized pneumonia (COP).
The clinical course of acute interstitial pneumonia (AIP) is acute, the reaction to steroids is poor, and the course and prognosis of the disease are very poor, but in rare cases it may be fully recovered. In the case of this disease, there is a high fever in the beginning, and a symptom of a cold usually appears and respiratory difficulty progresses frequently. Moreover, when recovering, it is natural recovery.

Cryptogenic Organizing Pneumonia (COP), formerly called Bronchiolitis Obliterans Organizing Pneumonia (BOOP), has a sub-acute clinical course and no response to steroids. It is good and the course and prognosis of the disease are good. In this disease, there is an improvement in the condition and complete recovery.
Respiratory Bronchiolitis-associated Interstitial Lung Disease (RB-ILD) and exfoliative interstitial Pneumonia (DIP) have subclinical or chronic clinical courses The disease has a good course and prognosis. In this disease, there is improvement in the condition and complete recovery.
Lymphphocytic Interstitial Pneumonia (LIP) is often associated with collagen disease or blood disease, and is known to be extremely rare clinically.
Here, chronic means that the disease progresses yearly, subacute means that the disease progresses monthly, and acute means that the disease progresses weekly. It is something to do.

Of these idiopathic interstitial pneumonia, IPF, NSIP, and OP (Organizing Pneumonia) account for the majority, followed by RB-ILD and DIP, which are highly associated with smoking, followed by LIP. Like unspecified AIP, it is considered extremely rare clinically.
Currently, disease classification is performed by comparing and contrasting pathology (mainly examination of lung tissue collected by biopsy) and clinical features (clinical findings, responsiveness to treatment, course of disease). Yes. However, because it is difficult to perform lung biopsy on all humans from the patient's perspective, it is possible to more accurately discriminate between cases with good prospects for disease progress and those with other conditions, such as blood tests. It is requested to do.
Examples of prior art patent documents relating to the diagnosis of lung diseases related to interstitial pneumonia include Patent Documents 1 to 4 and the like. Moreover, nonpatent literatures 1-3 etc. can be mentioned about the serum marker etc. of interstitial pneumonia.

JP-A-5-312087 JP 2005-30852 Special Table 2002-540801 Special table 2006-519588

Shuko Kono, “Markers for Diagnosis and Treatment of Interstitial Pneumonia”, Clinician, Vol. 24, No. 12, pp.2412-2415 (1998) Shuko Kono and Junichi Kondo, Latest Medicine, Vol. 56, No. 11, pp.2521-2528 (2001) Noriyoshi Ogawa et al., Rheumatism, 43 (1): pp.19-28 (2003)

  As noted above, surgical lung biopsy is often difficult, at least with a good outlook on the course of the disease, such as by blood tests, without histopathological search, and otherwise There is a need to more accurately distinguish cases. KL-6 / MUC1, SP-A, SP-D, etc. are known to be useful as serum markers for interstitial pneumonia, but assessment of disease activity, follow-up, especially prognosis There is a need to find markers that can help to do more accurately.

The present inventors have extensively searched for a technique that can easily, more accurately and reliably perform the cause, progress prediction and discrimination of interstitial pneumonia using blood markers that can be easily measured. As a result, it was found that there is a close relationship between the amount of interleukin-8 (IL-8) in the blood and interstitial pneumonia. In particular, the present inventors have found that the amount of IL-8 in the blood of a patient suffering from interstitial pneumonia has a large correlation in judging the course of the disease. For example, the amount of IL-8 in the blood, especially plasma IL-8, correlates with the degree of inflammation of the lung interstitium and the severity of alveolar fibrosis, arterial blood associated with the severity of the disease It was found that there was a correlation with the measured value of oxygen partial pressure (PaO ₂ ). Thus, the present invention has been reached.
The present invention provides a method for determining or diagnosing interstitial pneumonia, characterized by using the amount of IL-8 in blood, particularly the amount of IL-8 in plasma as an index. The reagent containing the antibody against IL-8 can be used as a determination or diagnosis agent for interstitial pneumonia and a determination or diagnosis kit for interstitial pneumonia.

From one aspect, the present invention provides the following aspects.
[1] A method for detecting or diagnosing interstitial pneumonia, comprising quantitatively measuring interleukin 8 (IL-8) in a test sample obtained from a subject, and determining the amount of IL-8 in the test sample , Detection or diagnosis of interstitial pneumonia, identification of the type of interstitial pneumonia, degree of possibility of progression of interstitial pneumonia, high or low risk, pathophysiological status and / or therapeutic response of interstitial pneumonia A method characterized by analyzing or evaluating sex trends.
[2] A method for detecting or diagnosing interstitial pneumonia, which is an inflammatory disease in which cell infiltration is observed in the alveolar septum (alveolar space) that is the interstitium of the alveolar region, and the change in the alveolar space is Quantitatively measure interleukin 8 (IL-8) in a test sample obtained from a subject suspected of having a disease that is the main pathological condition, and determine the amount of IL-8 in the test sample. The above [1], which is used as an index for detecting or diagnosing interstitial pneumonia in comparison with the amount of IL-8 in a healthy subject who does not show symptoms of interstitial pneumonia. ] The method of description.
[3] The method is a method for detecting or diagnosing the severe transition of interstitial pneumonia, and the amount of IL-8 in the test sample is determined in a healthy subject who does not show symptoms of interstitial pneumonia. The method according to [1] or [2] above, wherein the method is used as an index for detecting or diagnosing the severe transition of interstitial pneumonia in the subject in comparison with the amount of IL-8.
[4] Suspected of having an inflammatory disease in which cell infiltration is observed in the alveolar septum (alveolar septum), which is the stroma of the alveolar region, and whose alteration is the main pathological condition When the amount of IL-8 in a test sample obtained from a subject is greater than a threshold value, it is used as an index indicating the severity of interstitial pneumonia in the subject. The method according to any one of [1] to [3].
[5] The method according to any one of [1] to [4] above, wherein the test sample is plasma.
[6] Detection or diagnosis of interstitial pneumonia by measuring plasma IL-8 in patients with collagen disease, or differential diagnosis between interstitial pneumonia and drug-induced pneumonia in patients with collagen disease The method according to any one of [1] to [5] above.
[7] Differential diagnosis between severe interstitial pneumonia such as idiopathic interstitial pneumonia and chronic hypersensitivity pneumonitis, and / or severe interstitial pneumonia such as idiopathic interstitial pneumonia and pneumoconiosis The method according to any one of [1] to [5], wherein differential diagnosis is performed.
[8] The method according to any one of [1] to [5] above, wherein a differential diagnosis between severe interstitial pneumonia such as idiopathic interstitial pneumonia and bacterial pneumonia is performed.
[9] KL-6 or SP-D in blood is quantitatively measured, and the obtained KL-6 or SP-D measurement value is used together with the plasma IL-8 measurement value. ] The method as described in any one of [8].
[10] An agent for detecting or diagnosing interstitial pneumonia, comprising an antibody against IL-8 as an active ingredient.

By using the amount of IL-8 in the blood, especially plasma IL-8 as an indicator, differential diagnosis with higher accuracy of interstitial pneumonia is possible. Can evaluate and analyze disease activity, follow-up, etc., and differential diagnosis between interstitial pneumonia and drug-induced pneumonia in patients with rheumatoid arthritis, and general interstitial pneumonia and chronic hypersensitivity It is useful for differential diagnosis from pneumonia, pneumoconiosis, and bacterial pneumonia.
Other objects, features, excellence and aspects of the present invention will be apparent to those skilled in the art from the following description. However, it is understood that the description of the present specification, including the following description and the description of specific examples and the like, show preferred embodiments of the present invention and are presented only for explanation. I want. Various changes and / or modifications (or modifications) within the spirit and scope of the present invention disclosed herein will occur to those skilled in the art based on the following description and knowledge from other parts of the present specification. Will be readily apparent. All patent documents and references cited herein are cited for illustrative purposes and are not to be construed as a part of this specification. is there.

Plasma IL-8 values for each disease. Serum KL-6 levels for each disease. Serum SP-D value for each disease. ROC analysis using a case (RA-NonIP) that did not occur in patients with rheumatoid arthritis (RA), which is a type of collagen disease, compared to a case that caused interstitial pneumonia (RA-IP). ROC analysis of patients with rheumatoid arthritis (RA), which is a type of collagen disease, with interstitial pneumonia (RA-IP) and cases without RA (RA-NonIP) and healthy subjects . Correlation between IL-8 and KL-6 values. Correlation between IL-8 and SP-D values. Changes in plasma IL-8 levels due to treatment of patients with rheumatoid arthritis, a type of collagen disease. A) Cases in which symptoms were not changed by treatment, B) Cases in which symptoms were improved by treatment.

The present invention provides a technique capable of easily, more accurately and reliably performing the cause, progress prediction and discrimination of interstitial pneumonia using blood marker IL-8 that can be easily measured. The present invention also relates to a technique for more accurately performing interstitial pneumonia disease activity assessment, follow-up, and in particular, prognostic outlook using the blood marker IL-8.
In the present invention, the amount of IL-8 in the blood of a patient suffering from interstitial pneumonia accompanied by alveolar fibrosis is a healthy person or an interstitial pneumonia patient who has not caused alveolar fibrosis, It is also found to be significantly higher than In addition, the amount of IL-8 in the blood is useful for finding insidious onset in IIP, finding asymptomatic patients, determining progression, determining exacerbation, and determining severity, and correlate with them. Is also found.

In one preferred embodiment of the present invention, the amount of IL-8 in the plasma of a patient suffering from interstitial pneumonia associated with alveolar fibrosis does not cause a healthy person or alveolar fibrosis. Based on the knowledge that it is significantly higher than patients with pneumonia. In addition, the amount of IL-8 in plasma can be determined very accurately, particularly in the discovery of insidious onset in IIP, detection of asymptomatic patients, determination of progression, determination of exacerbation, and determination of severity. It is useful in enabling and is based on the knowledge that it correlates with them.
According to one preferred embodiment of the present invention, the amount of IL-8 in the blood, particularly plasma IL-8, is used as a correlate with the degree of inflammation of the lung stroma and the severity of alveolar fibrosis. The present invention also provides a technique used as a use and a use indicating the severity of the disease.

  Interleukin-8 (IL-8) is a type of cytokine or chemokine, and is a basic polypeptide (protein consisting of 72 amino acid residues) with a molecular weight of 8 kDa that selectively acts on neutrophils and T lymphocytes. It is a chemotactic factor. Producing cells are known to be peripheral blood monocytes, tissue macrophages, NK cells, fibroblasts, vascular endothelial cells, etc., and stimulated with mitogens such as LPS and krestin, IL-1 and TNF. -8 is produced. The biological activity of IL-8 is chemotactic for neutrophils and T lymphocytes, and has increased adhesion of leukocytes to vascular endothelial cells and activated neutrophil function.

In the present invention, the amount of IL-8 in blood can be measured using a commercially available IL-8 measuring reagent or IL-8 assay kit. In an exemplary embodiment, antibodies to IL-8 can be used to measure the amount of IL-8 in the blood.
Examples of the antibody against IL-8 used in the present invention include an antibody against IL-8 or a fragment thereof, and an antibody fragment derived from the antibody. The antibody against IL-8 is not particularly limited as long as it specifically recognizes IL-8.
The antibody against IL-8 may be a polyclonal antibody obtained from serum obtained from animals such as mice, rats, rabbits, goats, horses, etc., or obtained from a hybridoma culture such as a mouse hybridoma. It may be a monoclonal antibody. Moreover, the antibody produced using gene recombination techniques etc. may be sufficient.

The antibody against IL-8 can be labeled or immobilized as necessary. The method and means for labeling the antibody and the method and means for detecting it are not limited in any way, and known methods and means can be applied. Among these, labels include enzymes such as horseradish peroxidase (HRP) and alkaline phosphatase, fluorescein (for example, fluorescein isothiocyanate, FITC), rhodamine, DyLight ^™ (Thermo Fisher Scientific), fluorescent substances such as phycobiliprotein, ³² P Binds the antibody (or antibody fragment) to a radioactive substance such as ¹²⁵ I, a chemiluminescent substance such as an acridinium ester, an electrochemiluminescent substance such as a ruthenium complex, or a labeling substance selected from a biotin-avidin system, etc. Is done. The solid phase is performed by binding an anti-IL-8 antibody (or antibody fragment) to an appropriate solid phase.

  As the solid phase, any solid phase commonly used in immunochemical measurement methods can be used. For example, polystyrene, polyethylene, polypropylene, polyester, polyacrylonitrile, polyacrylate, Teflon (registered trademark), etc. Polymers such as fluorine resin, polyacetal, cross-linked dextran, polysaccharide, and other paper, glass, metal, agarose, and combinations thereof can be used. For example, tray shape, spherical shape, fiber shape, rod shape, disc shape, container Various shapes such as shapes, cells, test tubes, and the like can be mentioned. In a typical embodiment, for example, a plate such as a 96-well microtiter plate made of polystyrene, an amino group-bonded microtiter plate, particles such as magnetic particles, and various kinds of beads including fine particles can be used. The method and means for immobilizing the antibody are not limited in any way, and known methods and means can be applied. In order to immobilize the anti-IL-8 antibody, for example, a buffer containing the antibody may be added onto the carrier and incubated.

  Various techniques known in the art can be used to measure the amount of IL-8 in the blood, but in a typical embodiment an immunoassay (immunoassay) can be used, for example, a radioisotope Immunoassay (RIA method), enzyme immunoassay (EIA method: Engvall, E, Methods in Enzymol, 70, 419-439 (1980)), fluorescent antibody method, plaque method, spot method, agglutination method, octalonony (Ouchterlony) ) Etc. ("Hybridoma method and monoclonal antibody", published by R & D Planning, pages 30-53, March 5, 1982) used in general immunochemical assay methods Can be used.

  As these measurement methods, an appropriate assay method can be appropriately selected from various viewpoints, but the ELISA method can be preferably used from the viewpoint of sensitivity, simplicity, etc., for example, it recognizes a specific epitope of IL-8. Anti-IL-8 antibody (first antibody) and an anti-IL-8 antibody (second antibody) that recognizes an epitope different from the epitope recognized by this antibody are preferably used it can. In the ELISA method, the first reagent containing the immobilized first anti-IL-8 antibody and the second reagent containing the labeled second anti-IL-8 antibody can be preferably used. .

  The present invention provides an agent for detecting, determining or diagnosing interstitial pneumonia containing an antibody against IL-8 as an active ingredient. The antibody against IL-8 can be appropriately used in a form suitable for a method for measuring IL-8. And this can be used to measure the amount of IL-8 in samples including blood samples, for example, samples such as plasma and plasma, and by using this amount as an index, detection of interstitial pneumonia, Determination and / or diagnosis can be made. In a typical embodiment, a first reagent containing a solid-phased first anti-IL-8 antibody and a second reagent containing a labeled second anti-IL-8 antibody are contained. An agent for detecting, determining or diagnosing interstitial pneumonia is provided.

Specifically, the diagnosis of interstitial pneumonia may include differentiation between interstitial pneumonia accompanied by interstitial inflammation and alveolar fibrosis and other lung diseases. Here, “other lung diseases” may be construed to basically refer to lung diseases not accompanied by alveolar fibrosis, such as granulomatous lung diseases such as alveolar pneumonia and sarcoidosis. It is done. More specifically, when differentiating interstitial pneumonia with interstitial inflammation or alveolar fibrosis from other lung diseases, if the IL-8 value in the sample shows a cut-off value or higher, the interstitial inflammation Or interstitial pneumonia with alveolar fibrosis may be diagnosed.
The IL-8 value in the sample is useful for monitoring the pathophysiology of interstitial pneumonia and evaluating treatment responsiveness. By using the IL-8 value as an index, it is possible to detect, determine and / or diagnose interstitial pneumonia with good sensitivity and accuracy. In particular, monitoring of the pathological condition of interstitial pneumonia and treatment response Evaluation can be accurately performed with high sensitivity.

  According to the present invention, a method for detecting or diagnosing interstitial pneumonia is provided, and in the method for detecting or diagnosing interstitial pneumonia, cell infiltration occurs in the alveolar septum (alveoli) that is the stroma of the alveolar region. Quantitative measurement of IL-8 in a test sample obtained from a subject who is suspected of having a disease that is a major pathological change in the alveolar septum As an index for detecting or diagnosing interstitial pneumonia in the subject by comparing the amount of IL-8 in the sample with the amount of IL-8 in healthy subjects who do not show symptoms of interstitial pneumonia It is characterized by being used.

  Further, the method is a method for detecting or diagnosing the severe transition of interstitial pneumonia, wherein the amount of IL-8 in the test sample is determined in a healthy person who does not show symptoms of interstitial pneumonia. In contrast to the amount of IL-8, it can also be used as an index for detecting or diagnosing the severe transition of interstitial pneumonia in the subject. Furthermore, the method is suspected to have an inflammatory disease in which cell infiltration is observed in the alveolar septum (alveolar septum), which is the stroma of the alveolar region, and in which the alteration of the alveolar septum is the main pathological condition. A case where the amount of IL-8 in a test sample obtained from a subject having the above is greater than a threshold is used as an index indicating the severity of interstitial pneumonia in the subject. It is also possible to do.

Serum KL-6 can be measured, for example, by Shunobu Kawano, et al .: “Diagnostic ability for interstitial pneumonia KL-6 measurement kit ED066 by electrochemiluminescence immunoassay”, Clinical and Research, 75 (5): 217 -222, 1998 and Satoshi Kobayashi, et al .: “Study on setting of reference range and cutoff value of serum marker KL-6 antigen for interstitial pneumonia”, Clinical Pathology, 44: 653-658, 1996, etc. Can be implemented.
Serum SP-D is measured, for example, by Naoto Nagae, et al., Medicine and Pharmacy, 36 (4): pp.803-808 (1996) and Yasuto Honda, et al., Medicine and Pharmacy, 36 (4): pp It can be implemented with reference to .809-815 (1996).

The disease marker means a finding that is useful for diagnosis or follow-up of the disease. The usefulness of the marker includes sensitivity, specificity, correct diagnosis rate, positive predictive value, and negative predictive value. It is thought that it must be evaluated by one usefulness index.
The IL-8 level in the sample is useful for differential diagnosis of interstitial pneumonia, evaluation of disease activity, follow-up, etc. Quantitative analysis of IL-8 in the blood, especially plasma IL-8, and using the obtained IL-8 measurements, identify the type of interstitial pneumonia, symptoms of interstitial pneumonia Methods can be constructed to analyze or evaluate the degree of progression, the level of risk, and the pathophysiological state and / or treatment response trends of interstitial pneumonia. In particular, using an individual-derived specimen, it is possible to analyze an individual's medical condition, symptom, and tendency toward the above problem. Quantitative analysis of blood IL-8, especially plasma IL-8, and using the obtained IL-8 measurements, accurate diagnosis of interstitial pneumonia, evaluation of malignancy, progression of symptoms A method can be provided for assessing and / or analyzing potential risks.

  In a preferred embodiment of the invention, the measurement is performed using blood as a sample sample. Then, IL-8 level in blood, particularly plasma IL-8 level, is used as a disease marker for interstitial pneumonia. Using plasma IL-8 measurement as an index, it is possible to achieve a highly accurate test in the determination and diagnosis of interstitial pneumonia. In addition, with regard to the diagnosis and / or differentiation of interstitial pneumonia, and further follow-up, it is possible to achieve a test with higher accuracy by utilizing the correlation between plasma IL-8 level and serum KL-6 level. Furthermore, similarly, regarding the diagnosis and / or differentiation of interstitial pneumonia and the follow-up, if the correlation between plasma IL-8 level and serum KL-6 level and / or serum SP-D level is used, Highly accurate inspection can be achieved.

  According to the present invention, by using the plasma IL-8 value as an index of patients suspected of having interstitial pneumonia, (1) detecting or diagnosing whether or not a collagen disease patient has interstitial pneumonia. Furthermore, it can be used and useful for evaluation and analysis such as severe transition of interstitial pneumonia, evaluation of disease activity, and follow-up. (2) It can also be used for differential diagnosis of interstitial pneumonia and drug-induced pneumonia in patients with collagen disease such as rheumatoid arthritis. For example, IL-8 is high and KL-6 or SP- If D is also high, it is interstitial pneumonia, and IL-8 is normal, whereas if KL-6 or SP-D is high, it can be used for differential diagnosis of drug-induced pneumonia .

  Furthermore, by using the plasma IL-8 level as an index for patients suspected of having interstitial pneumonia, (3) severe interstitial pneumonia such as idiopathic interstitial pneumonia and chronic hypersensitivity pneumonitis For example, if IL-8 is high and KL-6 or SP-D is high, severe interstitial pneumonia such as idiopathic interstitial pneumonia or its suspicion On the other hand, if IL-8 is normal but KL-6 or SP-D is high, chronic hypersensitivity pneumonitis or its suspicion is very strong Can be used for differential diagnosis. In addition, (4) can be used and useful for differential diagnosis between severe interstitial pneumonia such as idiopathic interstitial pneumonia and pneumoconiosis, for example, IL-8 is high and KL-6 or SP- If D is also high, general interstitial pneumonia or its suspicion is very strong, while IL-8 is normal but KL-6 or SP-D is high, pneumoconiosis It is useful because it can be used for differential diagnosis. Further, (5) it can be used and useful for differential diagnosis between severe interstitial pneumonia such as idiopathic interstitial pneumonia and bacterial pneumonia, for example, IL-8 is high and KL-6 or If SP-D is also high, interstitial pneumonia or its suspicion is very strong, while IL-8 is high, but KL-6 and SP-D are low, bacterial Differential diagnosis such as pneumonia is useful.

  Previously, there has been a report on serum IL-8 measurement in patients with collagen disease such as rheumatoid arthritis, but the serum does not have the correct value. Cannot diagnose and / or differentiate interstitial pneumonia. Therefore, using the IL-8 value in this plasma as an index is of great clinical significance and is a surprising result that cannot be predicted in the past. In addition, when a patient with collagen disease including rheumatism develops any interstitial lung inflammation symptoms during treatment, it is also useful in assisting diagnosis of interstitial pneumonia by measuring plasma IL-8. Moreover, since KL6 and SP-D levels also rise, it is of great clinical significance to use both as indicators.

  Furthermore, it is also useful to measure both KL-6, SP-D, and IL-8, and use them for the diagnosis assistance of the above (2) to (5). In particular, IL-8 is a bacterial inflammation and is produced by peripheral blood monocytes, tissue macrophages, NK cells, etc., so it can be distinguished from drug and bacterial properties, and the treatment method changes depending on the cause Therefore, utilizing the technology of the present invention has great clinical significance. In addition, chronic hypersensitivity pneumonitis is difficult to distinguish from idiopathic interstitial pneumonia on an image, and it is difficult to distinguish unless it is a pathological diagnosis such as surgery. And a significant part of chronic hypersensitivity pneumonitis may be misdiagnosed as idiopathic interstitial pneumonia. However, use of the technology of the present invention contributes to the reduction of such problems.

The present invention will be described in detail with reference to the following examples, which are provided merely for the purpose of illustrating the present invention and for reference to specific embodiments thereof. These exemplifications are for explaining specific specific embodiments of the present invention, but are not intended to limit or limit the scope of the invention disclosed in the present application. In the present invention, it should be understood that various embodiments based on the idea of the present specification are possible.
All examples were performed or can be performed using standard techniques, except as otherwise described in detail, and are well known and routine to those skilled in the art. .

Targeting respiratory disease groups such as patients with emphysema, bronchiectasis, bronchial asthma, bacterial pneumonia, and interstitial pneumonia being treated, collagen disease groups such as patients with rheumatoid arthritis (RA), and healthy individuals (Normal) Plasma IL-8, serum KL-6 and serum SP-D were measured. Interstitial pneumonia was diagnosed as having a ground glass shadow and granular mesh, ring-shaped shadow or histopathological evidence on chest radiographs and high-resolution lung CT (HRCT). In addition to images or histological findings, the activity was determined by subjective symptoms such as dry cough, dyspnea during exertion, chest auscultation findings, arterial blood gas analysis, respiratory function test, and gallium scintigraphy.
IL-8 was measured by the EIA method (see J. Immunology, Vol. 147, pp. 2187-2195 (1991)), and the results are shown in Table 1 and FIG. If the cut-off value (Cutoff) is tentatively set to 7.6 pg / mL, all healthy subjects (Normal) show less than the cut-off value. Emphysema, bronchiectasis, bronchial asthma, chronic hypersensitivity pneumonitis, eosinophilic Most of the respiratory diseases such as pneumonia and pneumoconiosis showed below cut-off values. All cases of rheumatoid arthritis (RA), which is a type of collagen disease, did not have interstitial pneumonia (RA-Non IP), and methotrexate pneumonia (MTX pneumonia), a drug-induced pneumonia of collagen disease, was also cut. Less than off value. IL-8 showed high values in patients with acute intersensitivity pneumonitis, bacterial pneumonia, idiopathic interstitial pneumonia (IIP), collagen disease including rheumatoid arthritis and interstitial pneumonia (RA-IP) Etc.

Serum KL-6 was measured by electrochemiluminescence immunoassay (ECLIA) (Shuno Kawano et al .: Clinical and Research 75,1167-1172,1998), and serum SP-D was measured by EIA kit (Makoto Tanaka) Other: Medicine and pharmacy 59, 439-446, 2008). The results are shown in Table 1, FIG. 2 and FIG. Serum KL-6 has a cutoff value of 500 U / mL, and serum SP-D has a cutoff value of 110 ng / mL. Serum KL-6 levels were less than the cut-off value in healthy subjects (Normal) except for one case, and most of them were cut-off values in respiratory diseases such as emphysema, bronchiectasis, bronchial asthma, and bacterial pneumonia Less than. Rheumatoid arthritis (RA) did not have interstitial pneumonia (RA-Non IP), except for one case, showed a cutoff value. KL-6 showed a cut-off value or higher due to chronic hypersensitivity pneumonitis, acute hypersensitivity pneumonitis, idiopathic interstitial pneumonia (IIP), pneumoconiosis, rheumatoid arthritis (RA) and interstitial pneumonia Case (RA-IP) and methotrexate pneumonia (MTX pneumonia), a drug-induced pneumonia of collagen disease.
Serum SP-D values are less than the cut-off value except for 1 case in healthy individuals (Normal). In respiratory diseases such as emphysema, bronchiectasis, bronchial asthma, eosinophilic pneumonia, bacterial pneumonia, Most of them showed less than the cutoff value. Rheumatoid arthritis (RA) did not have interstitial pneumonia (RA-Non IP), and most of them showed a cut-off value. SP-D showed a cut-off value or higher for chronic hypersensitivity pneumonitis (all cases), acute hypersensitivity pneumonitis, pneumoconiosis (2 of 3 cases), idiopathic interstitial pneumonia (IIP), rheumatoid arthritis (RA) was part of a case of interstitial pneumonia (RA-IP). In one case, methotrexate pneumonia (MTX pneumonia), a drug-induced pneumonia of collagen disease, showed a cut-off value or more in one case.

  In chronic hypersensitivity pneumonitis and pneumoconiosis, plasma IL-8 was less than the cut-off value in all cases, but in KL-6, the cut-off value was exceeded in all cases, and in SP-D, excluding one case of pneumoconiosis It was over the off value. In particular, KL-6 showed a high value of 3,000 U / mL or more in chronic hypersensitivity pneumonitis. In bacterial pneumonia, IL-8 was more than the cut-off value in 4 out of 5 cases, but all cases were less than the cut-off value in KL-6 and SP-D (only 1 case was measured). Did not rise. In methotrexate pneumonia (MTX pneumonia), which is drug-induced pneumonia, plasma IL-8 was below the cut-off value, but in one case of KL-6 and SP-D, the cut-off value was exceeded.

  In patients with rheumatoid arthritis (RA), which is a type of collagen disease, cases with interstitial pneumonia (RA-IP: 10 cases) versus non-complicated cases (RA-NonIP: 43 cases) Receiver Operating Characteristic (ROC) analysis was performed. The results are shown in FIG. In Table 2, SE represents standard deviation, and CI represents confidence interval. FIG. 4 is an ROC curve. True positive rate is true positive rate (TPR) and shows sensitivity (Sensitivity), False positive rate is false positive rate (FPR) and 1-specificity (1-Specificity). ). From Table 2 and Fig. 4, plasma IL-8 measurement is used as an index, which is much more accurate in determining and diagnosing interstitial pneumonia in patients with collagen disease than KL-6 and SP-D. It was shown to be a high test.

  In addition, in patients with rheumatoid arthritis (RA), which is a type of collagen disease, cases with interstitial pneumonia (RA-IP: 10 cases) compared with cases without RA (43 cases) with RA Receiver operating characteristic ROC analysis using human specimens (20 cases) was performed in the same manner as described above. The results are shown in FIG. In Table 3 and Fig. 5 as well, measured IL-8 in plasma is used as an index, which is much more accurate in determining and diagnosing interstitial pneumonia in patients with collagen disease than KL-6 or SP-D. It was shown to be a high test.

  FIG. 6 shows Scatter Plot with Passing & Bablok Fit, and is a scatter diagram obtained by analyzing by applying the Passing-Bablok method. FIG. 6 shows the results of analyzing the correlation between plasma IL-8 levels and serum KL-6 levels using healthy subjects and rheumatoid arthritis cases that are a type of collagen disease. FIG. 7 shows plasma IL levels. The result of having analyzed the correlation of -8 value and serum SP-D value is shown. The plasma IL-8 value showed a high correlation with the serum KL-6 value of 0.810, but showed a low correlation with the serum SP-D value of 0.383.

  In addition, we investigated the therapeutic effects and changes in plasma IL-8 levels in patients with rheumatoid arthritis, a type of collagen disease, with interstitial pneumonia (FIGS. 8-A and -B). In cases where the symptoms were improved by treatment, plasma IL-8 levels were all significantly reduced, and in cases where there were no changes in symptoms, plasma IL-8 levels remained almost unchanged.

From the above results, by using plasma IL-8 level as an index for patients with suspected interstitial pneumonia,
(1) Diagnosis of interstitial pneumonia and monitoring of patient treatment by measuring plasma IL-8 in patients with collagen disease
(2) Interstitial pneumonia (high IL-8, high KL-6 and SP-D) and drug-induced pneumonia (normal IL-8, high KL-6 and SP-D) in patients with collagen disease Differential diagnosis
(3) Chronic hypersensitivity pneumonitis (IL-8 is normal, high KL-6 and SP-D) and interstitial pneumonia such as idiopathic interstitial pneumonia (IL-8 is high, KL-6 and SP -D high value) differential diagnosis
(4) Pneumoconiosis (IL-8 is normal, KL-6 and SP-D are high) and interstitial pneumonia such as idiopathic interstitial pneumonia (IL-8 is high, KL-6 and SP-D are high) Differential diagnosis
(5) Differentiation between bacterial pneumonia (IL-8 is high, KL-6 normal) and interstitial pneumonia such as idiopathic interstitial pneumonia (IL-8 is high, KL-6 and SP-D are high) Useful for diagnosis.

  Previously, serum IL-8 measurement in patients with collagen disease has been reported, but the serum does not have the correct value, that is, if the serum IL-8 value is used as an indicator, it is correct for interstitial pneumonia in patients with collagen disease. Cannot diagnose and / or differentiate. Therefore, it is of great clinical significance to use IL-8 level in this plasma as an index. In addition, when patients with collagen disease including rheumatoid arthritis have any symptoms of interstitial lung inflammation during treatment, measurement of plasma IL-8 is also useful for diagnosis assistance such as the cause and treatment policy of interstitial pneumonia It is. In interstitial pneumonia, KL-6 levels and SP-D levels also increase, so it is also clinically significant to use plasma IL-8 levels and their markers simultaneously as indicators.

  Furthermore, it is also useful to measure both KL-6, SP-D, and IL-8 at the same time, and use them for the diagnosis assistance of the above (2) to (5). In particular, IL-8 is bacterial inflammation and is produced by peripheral blood monocytes, tissue macrophages, NK cells, etc., so it can be distinguished from drug and bacterial properties, and there are also different treatment methods depending on the cause. It is expected to change and has great clinical significance. In addition, chronic hypersensitivity pneumonitis is difficult to distinguish from idiopathic interstitial pneumonia on the image, and it is difficult to distinguish unless it is a pathological diagnosis such as surgery. A significant part of chronic hypersensitivity pneumonitis may be misdiagnosed as idiopathic interstitial pneumonia, and this problem can be solved with a simple method by using plasma IL-8 as an indicator. is there.

In accordance with the present invention, by using the amount of IL-8 in blood, especially the amount of IL-8 in plasma as an indicator, it is possible to use a blood marker that can be easily measured to predict and discriminate the progress of interstitial pneumonia. Making it easier, more accurate and reliable. According to the present invention, differential diagnosis, which has been difficult in the past, such as interstitial pneumonia and drug-induced pneumonia in patients with rheumatoid arthritis, without performing surgical lung biopsy and without histopathological search, etc. Differential diagnosis, and differential diagnosis between interstitial pneumonia such as idiopathic interstitial pneumonia, chronic hypersensitivity pneumonitis, pneumoconiosis, and even bacterial pneumonia is a simple and more accurate testing method Is possible.
It will be apparent that the invention may be practiced otherwise than as particularly described in the foregoing description and examples. Many modifications and variations of the present invention are possible in light of the above teachings, and thus are within the scope of the claims appended hereto.

Claims (8)

Including quantitative measurement of interleukin 8 (IL-8) in plasma obtained from a subject and quantitative measurement of KL-6 and / or SP-D in blood obtained from the subject , morbidity interstitial pneumonia or drug pneumonia, specific types, the degree of likelihood of symptoms progression, level of risk, in a way that provides an indication of the tendency of pathophysiological conditions and / or therapeutic response There,
When the measured value of IL-8 and the measured value of KL-6 and / or SP-D are higher than the respective threshold values, it becomes an index of the possibility of suffering from interstitial pneumonia,
When the measured value of IL-8 is the same as or lower than the threshold value, and the measured value of KL-6 and / or SP-D is higher than the threshold value, it is an indicator of the possibility of developing drug-induced pneumonia. Method. Including quantitative measurement of interleukin 8 (IL-8) in plasma obtained from a subject and quantitative measurement of KL-6 and / or SP-D in blood obtained from the subject , Interstitial pneumonia, chronic hypersensitivity pneumonitis or pneumoconiosis morbidity, identification of type, degree of symptom progression, high or low risk, pathophysiological status and / or trend of treatment responsiveness A method of providing ,
When the measured value of IL-8 and the measured value of KL-6 and / or SP-D are higher than the respective threshold values, it becomes an index of the possibility of suffering from interstitial pneumonia,
If the measured value of IL-8 is equal to or lower than the threshold value and the measured value of KL-6 and / or SP-D is higher than the threshold value, the possibility of chronic hypersensitivity pneumonitis or pneumoconiosis An indicator, a method. Including quantitative measurement of interleukin 8 (IL-8) in plasma obtained from a subject and quantitative measurement of KL-6 and / or SP-D in blood obtained from the subject , morbidity interstitial pneumonia or bacterial pneumonia, the type of a particular degree of likelihood of symptoms progression, level of risk, in a way that provides an indication of the tendency of pathophysiological conditions and / or therapeutic response There,
When the measured value of IL-8 and the measured value of KL-6 and / or SP-D are higher than the respective threshold values, it becomes an index of the possibility of suffering from interstitial pneumonia,
When the measured value of IL-8 is higher than the threshold value, and the measured value of KL-6 and / or SP-D is the same value or lower than the threshold value, it is an indicator of the possibility of bacterial pneumonia. Method. The subject is suspected of having an inflammatory disease in which cell infiltration is observed in the alveolar septum (alveolar septum), which is the interstitium of the alveolar region, and the alteration of the alveolar septum is the main pathological condition The measured values of IL-8 in plasma obtained from the subjects are compared with the measured values of IL-8 in healthy subjects who do not show symptoms of interstitial pneumonia. The method according to any one of claims 1 to 3 , wherein the method is used as an index of morbidity of qualitative pneumonia. The measured value of IL-8 in the plasma is compared with the measured value of IL-8 in healthy subjects who do not show symptoms of interstitial pneumonia. the method according to any one of claims 1-4, characterized by an indicator. The subject is suspected of having an inflammatory disease in which cell infiltration is observed in the alveolar septum (alveolar septum), which is the interstitium of the alveolar region, and the alteration of the alveolar septum is the main pathological condition and which, claim 1-5, characterized in that the indicator of the severity of interstitial pneumonia when the measured value of IL-8 in plasma obtained from a subject is higher than the threshold value The method according to 1. The method according to any one of claims 1 to 6 , wherein the subject is a collagen disease patient. Interstitial pneumonia, drug-induced pneumonia, chronic hypersensitivity pneumonitis, pneumoconiosis or bacterial pneumonia characterized by containing an antibody against KL-6 and / or SP-D and an antibody against IL-8 as active ingredients Detection or diagnostic agent.

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