RU1777083C - Method for predicting development stages of sclerodermia systematics - Google Patents

Abstract

The invention relates to immunodiagnostics of an active pathological process in systemic scleroderma (SJS). The purpose of the invention is to increase accuracy, ensure early diagnosis, accelerate the method and reduce the complexity. For testing, 2-3 ml of blood is taken from the ulnar vein. A density gradient of ficoll-verographin (1.077 g / ml) is used to isolate lymphocytes. Quantification of antigen-binding lymphocytes (ASLs) reacting with a skin antigen is carried out using the indirect rosette method. Tu suppressors and T) m-helpers are quantified in the EA rosette test. Determination results. Tu - lymphocytes and ASL are expressed as a relative indicator as a percentage of the number of lymphocytes circulating in the peripheral blood. The ratio of ASL to the product of T and lymphocytes is calculated, expressing it in conditional AR L ASL x 100% (unit) Analytes / units. The lease showed that in patients with SJS in stage 1: 11.8 units. at 24.2 conventional units in patients with stage II SSD. 34..0-79.5 conventional units, 1 tab. sl c

Description

The invention relates to immunodiagnostics of an active pathological process in systemic scleroderma (SJS).

A known method for diagnosing the stages of development of SJS by assessing the clinical picture of the disease, taking into account individual laboratory and instrumental data (N.G. Guseva, Systemic scleroderma. - M., Medicine, - 1975, 271 s). The stages of development of SSD are distinguished in accordance with the working classification of SSD adopted in our country (N.G. Guseva, 1975). Stage of the initial manifestations (1) is characterized by the presence of articular syndrome, Raynaud's syndrome or skin lesions; differs from the developed stage (generalization - It and terminal - III) in the absence of visceral pathology, polysyndromism and polysystemic lesion. The relative diagnostic value are laboratory

data that are mainly important for determining the degree of disease activity; determination of ESR, SRV, seromucoid, O.1 -, γ-globulins, RF in average titers, urinalysis, hydroxyproline in blood plasma and urine, determination of immunoglobulins of classes G, AM, morphological examination skin biopsies. The accumulation of ANA, an increase in CEC with a diffuse, rapidly progressive SJS were established. To diagnose visceral pathology, a number of instrumental studies are also necessary (X-ray examination of the organs of the chest, gastrointestinal tract, joints, if necessary, endoscopic, ultrasound, ECG, study of the function of external respiration, kidneys) With far-reaching changes with the development of severe fibrotic -sclerotic or vascular-necrotic lesions leading to

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Xj

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to failure of certain organs, they diagnose the terminal stage.

The purpose of the invention is to increase accuracy, ensure early diagnosis, speed up the method of reducing the complexity.

The proposed method is carried out as follows.

For research, blood is taken from the cubital vein (2-3 ml of blood in tubes containing heparin - 25 IU per 1 ml of liquid). The selection of lymphocytes is carried out according to the method of A. Boym (Baym A. - Separation blood lencocytes. Granulocytes and lymphocytes. Tissue Antigens. - 1974. - No. 4. - p. 269).

Regulatory T-subpopulations (Tj, suppressors and T-helpers) are defined by L. Moretta e.a. (J. Jmmunol. 1976. - No. 117, - p. 2171-74) in the modification of F.Yu. Gariba, M.V. Lie Hall (Normative materials on the content of individual subpopulations of lymphocytes in human peripheral blood in an age-related aspect. Guidelines. Tashkent, 1983. - p. 4-7). The principle of the method consists in the formation of rosettes with T-lymphocytes with red blood cells loaded with class G or M- anti-RBC antibodies and is based on the detection of T-lymphocytes carrying receptors for Fc - a fragment of immunoglobulin G (Tf-suppressor cells) and M (T / V helper cells).

The quantification of antigen-binding lymphocytes (ASL) was carried out on the basis of the indirect rosette method (F.Yu. Garib et al. Clinical value of the determination of antigen-binding cells in patients with typhoid fever and other diseases. Methodical recommendations. Tashkent. 1983. - p. 3-8). The principle of the method is the formation of rosettes consisting of a centrally located sensitized lymphocyte and attached red blood cells pre-loaded with skin antigen.

Tj determination results. -, T / α-lymphocytes, ASL were expressed as a relative indicator as a percentage of the number of lymphocytes circulating in the peripheral blood.

The coefficient of degree of autoimmunization (KSAI) calculated by

formula: KSAI-FSh- x 100% (conventional units)

y x Analysis showed that in healthy individuals KSAI

averages (M ± t) - 1.4-0.16 conv.ed., at M ± 2aot 0.74 to 2.5 srvc. The stage of the initial manifestations of SJS (1) is characterized by an CLI equal to an average of 16.9 ± 0.9 conventional units, at M ± 2 st from 8.7 to 25.1 standard units, the Generalized stage (II) is characterized by

KSAI on average equal to 61.0 ± 3.1 conventional units, from 31.5 to 90.5 u units, KSAI indicators in patients with terminal stage (III) do not significantly differ from those for

generalized stage (II),

Example 1, Sick M., 35 years old. The diagnosis is systemic scleroderma (SJS) of a chronic course, stage I, stage II activity. Raynaud's syndrome, scleroderma polyarthritis, flexion contractures of the joints of the hands, hyperpigmentation of the skin, calcification of soft tissues. The duration of the disease is 7 years. Onset of the disease with pain and stiffness in small joints

hands followed by Raynaud’s syndrome, skin lesions. Objectively: Raynaud’s syndrome, knee and ankle joint configuration, flexion contractures of small joints of the hands, hyperpigmentation of the skin of the face and neck, calcification of soft tissues along the spine, in the area of knee joints, years, Osteoporosis on the roentgenograms of bones and feet, narrowing of the joints between the joints .

X-ray of the knee joints: against the background of the femur in soft tissues, multiple petrified shadows, In total.Ai. blood acceleration of ESR up to 30 mm / h, albumin - 38.1%, γ-globulins - 35.7%,

RF in the 1/320 latex test, TJ - lymphocytes - 6.7%, Tyi - lymphocytes - 10.1%, ASL - 8%, KSAI-11.8 conventional units, which corresponds to stage 1 SSD.

Example 2. Sick Sh., 48 years old. Diagnosis: SJS of chronic course, first stage, II degree of activity. Induracil and hyperpigmentation of the skin, Raynaud's syndrome, polyarthralgia. The duration of the disease is 8 years. The onset of the disease with the syndrome

Raynaud followed by damage to the skin, joints. An objective examination of the patient revealed a dense swelling of the hands, face, induction and hyperpigmentation of the skin of the extremities, a pouch symptom, trophic

impaired fingers, marked Raynaud's syndrome is noted. On the roentgenogram of the hands - periarticular osteoporosis. On the ECG - sinus arrhythmias. B - total an.blood: ESR acceleration up to 30 mm / hour,

CEC in serum - 2.2 units. OD, ASL - 10%, 1J - lymphocytes -5.3%, Tjfi-lymphocytes - 7.8%, KSAI - 24.2 conventional units, which corresponds to stage I SSD.

Example 3. Sick A., 33 years old. Diagnosis. Chronic respiratory disease, stage II, stage I activity. Dense edema, induction and hyperpigmentation of the skin, Raynaud's syndrome, polyarthralgia, initial manifestations of damage to the gastrointestinal tract. Disease duration 4 years. Onset of the disease with Raynaud's syndrome. Objectively: dense swelling of the hands, induction and hyperpigmentation of the skin of the limbs, abdomen, face, severe Raynaud's syndrome. Gastrointestinal fluoroscopy - gastroptosis, gastritis vein, spastic deformity of the large intestine. On the ECG, dystrophic changes in the myocardium. In total blood, an acceleration of ESR was not observed (14 mm / h), and CEC in serum was 1.3 units. OD, IJ - suppressors-4.2%, ty-helpers-7.7%, ASL-11%, KSAI-34.0 srvc. This value of KSAI is a reflection of the onset of immunological generalization of the disease, preceding clinical generalization and corresponds to stage I SSD.

PRI me R 4. Patient K., 54 years old. Diagnosis: SJS chronic course, stage II, stage I activity. Diffuse pneumofibrosis, cardiosclerosis, esophagitis, hyperpigmentation of the skin, Raynaud's syndrome. The duration of the disease is 23 years. Objectively: the condition of a patient of moderate severity, cyanosis of the lips, acrocyanosis, shortness of breath. Marked Raynaud's syndrome, hyperpigmentation of the skin of the face, and a pouch symptom are noted. On the chest radiograph - diffuse pneumofibrosis. On an ECG - sinus bradycardi. Hypertrophy of the left ventricle, overload of the myocardium of the left atrium. In the general blood test: ESR acceleration to 25 mm / May, CEC-2.2 units, OD, Tj suppressors - 4.2%, b, - helpers - 5.4%, ASL - 18%, KSAI - 79, 4 conventional units, which corresponds to stage II SSD.

Example 5. Sick X., 42 years old. Diagnosis: chronic SJS, terminal stage (III), activity of the III degree. Severe vascular and trophic disorders (Raynaud's syndrome, multiple persistent ulcerations at the fingertips, hair loss, weight loss by 15 kg), skin lesions (induction and atrophy), scleroderma polyarthritis, diffuse pneumofibrosis, pulmonary insufficiency of the third degree, decompensated pulmonary heart - NK II B - III Art. The patient died due to increasing pulmonary heart disease.

The duration of the disease is 15 years. Onset of the disease with Raynaud's syndrome. Objectively: the patient's condition is extremely serious. Diffuse cyanosis, growing shortness of breath, swollen cervical veins, swelling on the legs, the liver protrudes 4 cm from the edge of the costal arch. Marked Raynaud's syndrome, cyanosis of the fingers with ulceration and dystrophic changes, cyanosis of the toes, induction and hyperpigmentation of the skin of the face and limbs, and a symptom of pouch are noted. On the chest x-ray diffuse pievmofibroz. On an ECG sinus tachycardi, ventricular extrasystoles, blockade of the right bundle branch block. Hypertrophy of both ventricles of the left atrium with dystrophic changes and myocardial overload. In total an. blood - ESR acceleration up to 42 mm / hour, CEC - 2.8 units. OP, RF in the 1/120 latex test, T-suppressors - 4.7%. Tu - helpers - 4.4%. ASL - 16%. KSAI - 77.4 conventional units, which corresponds to the expanded stage of the disease (II and III),

This example was not included in the table. However, an immunological examination of this case showed that the severity of immunological changes, as well as the digital value of the CSI, do not go beyond the indicators of the generalized stage, which confirms the priority of physical wear of the affected organs in the pathogenesis of the development of the terminal stage of the disease.

The positive effect of using the proposed method, in comparison with the prototype, is to increase the accuracy, which is confirmed by the indicators of all groups of patients who do not overlap, allowing us to follow the development of the stages, showing its reliability. In addition, the method provides early diagnosis and prediction of the transition of the initial stage and the expanded stage of the SJS,

Early diagnosis is possible subject to the specificity of the method. T ak, with a resolution of d «100 (conventional units), where

the ratio

T; x T} v

ASL reacting with the same skin antigen in the group of patients with chronic eczema, its digital value is 5.55 ± 0.5 conventional units. at M ± 2 st from 2.6 to 8.4 conventional units against 16.9 ± 0.9 conventional units at M ± 2 (7 from 8.7 to 25.1 conventional units in patients with stage I of SJS (, 01). The acceleration of the method and the reduction of labor intensity, in comparison with the prototype, is ensured by a simultaneous study of 10 people, study time 24 hours (prototype 5-7 days) The method is non-invasive and can be performed on an outpatient basis, while the prototype provides for endoscopic, radiological and other comprehensive examinations requiring in-patient observation.

Claims (1)

SUMMARY OF THE INVENTION A method for diagnosing stages of development of systemic scleroderma, including a blood test, characterized in that, in order to increase accuracy, ensure early diagnosis, speed up the method and reduce labor intensity, the amount of antigen-binding lymphocytes reacting with skin antigen, 7 suppressors and T helper cells. the ratio of antigen-binding lymphocytes to the product of TJ and T is calculated in arbitrary units, and a value of 11.8 to 24.2 is used to diagnose stage I of the disease, from 34.0 to 79.5, stage II of systemic scleroderma is diagnosed. KSAI indicators in healthy individuals and patients with SJS depending on the stage of the disease