Method of human blood serum analysis for soluble form of antigen cd50 dimer

FIELD: medicine.

SUBSTANCE: invention refers to a method of human blood serum analysis for a soluble form of antigen CD50 dimer involving the use of CD50-specific monoclonal antibodies whereat a tray reaction includes tetramethyl benzidine as a substratum.

EFFECT: invention provides detecting the soluble form of antigen CD50 dimer in human blood serum.

1 cl, 2 ex

 

The invention relates to the field of biology and medicine, in particular to molecular immunology, and can be used for differential diagnosis of small-cell lung cancer.

At the present time is characterized by more than 240 membrane differencirovannyh antigens of the cells of the immune system, are classified in the International workshops. For more than 40 membrane protein antigens of nature demonstrated the existence of a soluble form. These proteins belong to different families and superstratum. Effector action of soluble membrane antigens of cells of the immune system depends on whether there is protein in Monomeric or oligomeric form, that is, from the peculiarities of structural organization.

Known methods for determining the soluble forms of some of the antigens of human blood based on enzyme-linked immunosorbent assay. In particular, the developed method for the evaluation of serum content of soluble CD38 antigen (R. Mallone, Ferrua s, Morra M. et al. Characterization of a CD38-like 78-kilodalton soluble protein released from cell lines derived from patients with X-linked agammaglobulinemia // J. Clin. Invest., 1998, Vol.101, No. 12, P.2821-2830), and enzyme-linked immunosorbent way of identifying its dimeric form (Egorova N, Kurnikov GU, Novikov V.V. Serum content of total and oligomeric fractions of soluble CD54 and CD38 antigens with urogenital chlamydiosis // Mater. Ross. scientific-practical use. proc. "Central issues of infection control", S.-Petersburg, 2004, s). Also known methods for the determination of soluble forms of adhesion antigens in the serum of a person belonging to different families of proteins - immunoglobulins, integrins, selectins (M.A. del Pozo, Pulido R., Munoz C. Regulation of ICAM-3 (CD50) membrane expression on human neutrophils through a proteolytic shedding mechanism // Eur. J. Immunol., 1994, Vol.24 (11), P.2586-2594; Tsujisaki m, Imai K., Hirata H. et al. Detection of circulating intercellular adhesion molecule-1 antigen in malignant diseases // Clin. Exp. Immunol., 1991, Vol.85 (1), P.3-8). Some of these methods are used with the use of specific monoclonal antibodies series IR (Baryshnikov, A., Tonevitsky A.G. Monoclonal antibodies, Moscow, 1997, 212 C.), purification by ammonium sulfate and rivanol is generally accepted methods (Primal, Immunological methods. - M.: Medicine, 1987, p.390-413). To prepare the substrate solution is usually used citrate buffer and tetramethylbenzidine, and for washing unreacted components - phosphate-saline with tween (SDF-T) (e.g., Egorov A.M., Osipov A.P., Dzantiev B.B., Gavrilova E.M. Theory and practice of immunoassay. - M.: Higher. HQ., 1991,288 C.).

The object of the invention is a method allowing to detect in the serum of a person a new form of soluble membrane protein CD50, the contents of which, as studies have shown, correl the plans with different morphological variants of lung cancer (squamous non-squamous cell lung cancer, squamous cell orogovevshi lung cancer, glandular cell lung cancer, small cell lung cancer, a mixed form of lung cancer, malignant carcinoid). The molecular mass of the protein detected corresponds to a soluble form of the dimer CD50 antigen.

According to available information, any information about the identified protein and its relationship with morphological variants of lung cancer is not present.

Thus, obtained when using the present invention the technical result consists in the detection of a soluble form of the dimer CD50 antigen in the serum of a person.

This technical result in the implementation of the invention is achieved by the fact that wells contribute used as CD50-specific monoclonal antibodies monoclonal antibodies IR-60, diluted in the ratio 1:700 0,85% NaCl in a volume of 100 μl, stand the tablet in a humid chamber at 42°C for 2 hours, wash off unbound antibodies 4-5 times SDF-T (pH 7,2-7,6), 25-fold concentrate which consists of 250 g NaCl, 25 g NaHPO4·12H2O, 100 g of urea, 25 ml of tween-80) was dissolved in 1 l of distilled water, bring in the first row of wells 100 ál of positive control serum, diluted SDF-T in the following ratios: whole serum, 1:1, 1:3, 1:7, 1:15, 1:31, 1:63, 1:127, in 3 holes contribute SDF-T about what JaME 100 μl, in the other hole making analyzed serum in a volume of 100 μl, stand the tablet in a humid chamber at a temperature of 18-25°C for 18-24 hours, wash off unbound components of the reaction SDF-T 4-5 times, make monoclonal antibodies IR-60, associated with the enzyme horseradish peroxidase, diluted SDF-T in the ratio of 1:500 in a volume of 100 μl, stand the tablet for 1 hour at a temperature of 42°C, washed unreacted components of the reaction SDF-T 5-6 times, make a substrate solution in a volume of 100 μl consisting of 10% tetramethylbenzidine dissolved in 10 ml of citrate buffer solution (pH 4.0)containing 0.05% hydrogen peroxide and consisting of 5 g of citric acid, 5 g sodium citrate translesanas dissolved in 1 l of distilled water, stand the tablet in a dark place at a temperature of 18-25°C for 15-20 min, make a 10% solution of sulfuric acid in a volume of 50 μl, measure the optical density of the colored product peroxidase activity at a wavelength of 450 nm, and translate the values of optical density in arbitrary units using a calibration curve constructed on the basis of rascaroli positive control serum.

The method for determining a soluble form of dimer CD50 antigen in the serum of human blood as follows.

For the synthesis of conjugate "Monoclonal ant the body IR-60-horseradish peroxidase" periodate method used CD50-specific monoclonal antibodies IR-60. To a solution containing 5 mg of horseradish peroxidase (RZ=3,0) in 1 ml of N2Oh, add 0.2 ml of a freshly prepared 0.02 M solution of periodate sodium NaJO4and stirred for 20 minutes at 20°C in a dark chamber. The resulting solution cialiswhat against 0.001 M Na-acetate buffer (pH 4,4) for 24 hours at 4°C. To the solution was added 10 mg of antibody, dissolved in 2 ml of 0.2 M Na-carbonate buffer (pH 9,6). The solution is stirred for 2 hours at a temperature of 20°C in a dark cell, add 0.1 ml of a freshly prepared aqueous solution NaHB4(4 mg/ml), and stirred for two hours at 4°C. the resulting conjugate is precipitated with a saturated solution (NN4)2SO4and then cialiswhat against saline. To stabilize the conjugate add an equal volume of glycerol and stored at -20°C.

The optimal concentration of monoclonal antibodies IR-60 for inscription on the tablet is determined by measuring the optical density at different dilutions from 1:100 to 1:1500. The maximum difference between experimental and baseline values was observed at a dilution of monoclonal antibodies IR-60 1:700. The most effective concentration of monoclonal antibodies IR-60, conjugated with horseradish peroxidase, also determined using dilution of the conjugate 1:100 to 1:1500. The most effective was is the giving of the conjugate 1:500.

Example 1. In wells contribute purified monoclonal antibodies IR-60, diluted in the ratio 1:700 0,85%NaCl solution, in a volume of 100 μl.

Tablet incubated in a humid chamber at 42°C for 2 hours. Unbound antibodies are washed 4 times SDF-I.

In the first row of wells contribute 100 ál of positive control serum, diluted SDF-T in the following ratios: whole serum, 1:1, 1:3, 1:7, 1:15, 1:31, 1:63; 1:127.

In 3 holes make 100 ál of SDF-T for control of background reactions.

In the other hole making 100 μl of test sera, diluted SDF-T in 1:1 ratio.

Tablet incubated in a humid chamber at a temperature of 18°C for 18 hours and washed 4 times SDF-I.

Then make 100 ál of conjugate solution "Monoclonal antibodies ICA-60-horseradish peroxidase", divorced SDF-T in the ratio of 1:500.

The tablet is maintained at a temperature of 42°C for 1 hour and washed 5 times SDF-I.

Introduce 100 μl of substrate solution, which is used as a solution of tetramethylbenzidine.

Tablet stand for 15 minutes at a temperature of 18°C in a dark place.

Add 100 ál of 10% sulfuric acid solution and immediately determine the optical density of the colored product at a wavelength of 450 nm using a photometer, Multiskan EX. Translate units of optical density in the base unit (U/ml) using a calibration curve built on the basis of rascaroli positive control serum. For 1000 U/ml take the value of optical density corresponding to the working dilution of the analyzed serum samples. The content of soluble forms of the dimer CD50 antigen in the analyzed serum was 153 U/ml.

Example 2. In wells subjected to UV radiation, making purified monoclonal antibodies IR-60, diluted in the ratio 1:700 0,85% NaCl in a volume of 100 μl.

Tablet incubated in a humid chamber at 42°C for 2 hours. Unbound antibodies are washed 5 times SDF-I.

In the first row of wells contribute 100 ál of positive control serum, diluted SDF-T in the following ratios: whole serum, 1:1, 1:3, 1:7, 1:15, 1:31, 1:63; 1:127.

In 3 holes make 100 ál of SDF-T for control of background reactions.

In the other hole making 100 μl of test sera, diluted SDF-T in 1:1 ratio.

Tablet stand in a moist chamber at 22 ░ C for 21 hours and washed 5 times SDF-I.

Then make 100 ál of conjugate solution "Monoclonal antibodies ICA-60-horseradish peroxidase", divorced SDF-T in the ratio of 1:500.

The tablet is maintained at a temperature of 42°C for 1 hour and washed 6 times SDF-I.

Introduce 100 μl of substrate solution, as to the th use a solution of tetramethylbenzidine in 10 ml of citrate buffer solution (pH 4.0), containing 0.05% hydrogen peroxide.

Tablet stand for 15 minutes at a temperature of 18°C in a dark place.

Add 100 ál of 10% sulfuric acid solution and immediately determine the optical density of the colored product at a wavelength of 450 nm using a photometer, Multiskan EX. Translate units of optical density in arbitrary units (U/ml) using a calibration curve constructed on the basis of rascaroli positive control serum. For 1000 U/ml take the value of optical density corresponding to the working dilution of the analyzed serum samples. The content of soluble forms of the dimer CD50 antigen in the analyzed serum was 53 U/ml.

When determining the soluble form of the dimer CD50 antigen in serum 85 healthy donors was shown that the content of the norm is 65.8±1,6 U/ml. Serum samples were obtained from 94 patients with lung cancer with different morphological variants of the tumor (squamous non-squamous cell lung cancer, squamous cell orogovevshi lung cancer, glandular cell lung cancer, small cell lung cancer, a mixed form of lung cancer, malignant carcinoid) at all stages of tumor.

The content of soluble oligomeric fractions CD50 by small cell variant of the tumor, amounting to 112.4±9,9 /ml, showed a significant 1.7-fold (p=0.001) exceeding the level of this antigen in the group of healthy donors and all forms of cancer of the lung: 2.15 times the level squamous non-squamous cancer, equal 52,2±2,6 U/ml, to 2.29 times higher than squamous cell orogovevshih lung cancer, equal 48,9±6,9 U/ml, 2.68 times higher than the level of the mixed lung cancer, equal 41,9±4,9 U/ml. Indicators antigen in squamous cell cancers and mixed form tumors was significantly lower level of healthy donors (p<0,05).

Thus, the determination of the soluble form of the dimer CD50 antigen lawfully be used for differential diagnosis of small-cell lung cancer and other variants of this nosology as additional auxiliary test.

The method for determining a soluble form of dimer CD50 antigen in the serum of a person, consisting in the fact that wells contribute used as CD50-specific monoclonal antibodies monoclonal antibodies IR-60, diluted in the ratio 1:700 0,85%NaCl solution, in a volume of 100 μl, stand the tablet in a humid chamber at 42°C for 2 h, washed unbound antibodies 4-5 times with phosphate-saline (SDF-T), (pH 7,2-7,6), 25-fold concentrate which consists of 250 g NaCl, 25 g NaHPO4·12H2O, 100 g of urea, 8 ml of Triton X100 and 25 ml of tween-80, RAS is voennih in 1 l of distilled water, make the first row of wells 100 ál of positive control serum, diluted SDF-T in the following ratios: whole serum, 1:1, 1:3,1:7, 1:15, 1:31, 1:63, 1:127, in 3 holes contribute SDF-T in a volume of 100 μl in the other hole making analyzed serum in a volume of 100 μl, stand the tablet in a humid chamber at a temperature of 18-25°C for 18-24 h, washed unbound components of the reaction SDF-T 4-5 times, make monoclonal antibodies IR-60, associated with the enzyme horseradish peroxidase, diluted SDF-T in the ratio of 1:500 in a volume of 100 μl, stand the tablet for 1 h at a temperature of 42°C, washed unreacted components of the reaction SDF-T 5-6 times, make a substrate solution in a volume of 100 μl consisting of 10%tetramethylbenzidine dissolved in 10 ml of citrate buffer solution (pH 4.0)containing 0.05% hydrogen peroxide and consisting of 5 g of citric acid, 5 g sodium citrate translesanas dissolved in 1 l of distilled water, stand the tablet in a dark place at a temperature of 18-25°C for 15-20 min, make a 10%solution of sulfuric acid in a volume of 50 μl, measure the optical density of the colored product peroxidase activity at a wavelength of 450 nm, and translate the values of optical density in arbitrary units using a calibration curve constructed on the basis of rascaroli will put the school control sera.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention refers to medicine and veterinary science, namely to microbiology and immunology. The method involves cultivation of Brucella abortus I-206 L-strain on a dense nutrient medium for L-brucellas at 37°C for 3-5 days. Thereafter, a microbial mass is washed of with buffered 0.9% normal saline of pH 7.2±0.2 and inactivated by adding 2.5% formalin. A bacterial suspension is kept for one day at 37°C with specific sterility being controlled. The bacterial suspension is reduced to concentration 4.5·1010-5·10 m.c. in ml with buffered 0.9% normal saline (pH 7.2±0.2) and thermally treated on a water bath at 100°C for 45-50 minutes. The suspension is centrifuged at 7000 rpm for 50-60 minutes; the supernatant is separated and frozen-dried. The method allows producing the preparation exhibiting high specificity and activity, available for preparing based immunobiological preparations and a test-system for human and animal blood serum examination for L-brucella antibodies.

EFFECT: method is technological, accessible, does not require using expensive devices and equipment.

2 ex

FIELD: medicine.

SUBSTANCE: substance of the invention involves a method of detecting an antibody in a sample being tested containing a body fluid of a specified mammal where said antibody is a biological marker of a disease state or propensity for a disease where the method involves (a) contacting said sample being tested with a set of various amounts of an antigen specific to said antibody, (b) evaluating specific binding of said antibody and said antigen, (c) drawing a diagram or calculating a curve of said specific binding to the amount of the antigen for each amount of the antigen used at the stage (a), and (d) stating the presence or absence of said disease state or propensity for the disease by the specific binding of said antibody and said antigen for each individual antigen concentration used where the presence of said disease state or propensy for the disease is detected by screening a titration curve starting with the stage (c) for observing an S-shaped or sigmoid curve.

EFFECT: higher sensitivity of the diagnostic technique.

23 cl, 7 ex, 6 tbl, 15 dwg

FIELD: medicine.

SUBSTANCE: method for typing lepromatous leprosy consists in evaluating neutrophilic granulocyte (phagocytes) recovered from peripheral blood, incubated with lepromin; a chemoluminescence (CL) level is specified, and maximum intensity lepromin activated CL enables to type lepromatous leprosy.

EFFECT: method provides higher accuracy and information value of typing lepromatous leprosy.

1 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to infectious disease, and can be used for prediction of antiretroviral therapy efficiency in case of HIV-infection. For this purpose by method of solid-phase immunoenzyme assay level of cytokines is determined. If indices of soluble tumour necrosis factor alpha receptor protein 75 are from 4335.48 to 6001.86 pg/ml, soluble tumour necrosis factor alpha receptor protein 55 - from 768.72 to 1323.87 pg/ml and soluble interleukin-6 receptor from 1770.77 to 3800.31 pg/ml, favourable clinic course of HIV-infection after 1-3 months since beginning of antiretroviral therapy is predicted.

EFFECT: method ensures increase of accuracy of antiretroviral therapy efficiency prediction due to selection of certain immunological criteria.

1 dwg, 7 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: offered is a method for selection of a contingent indicated for diphtheria prophylactic immunisation: vaccination or revaccination. Venous blood serum of a person being tested and a reference venous blood serum sample are examined for levels of specific IgG to diphtheria antitoxin, IgGTDA and IgGRDA respectively. Saliva of the person being tested and a reference saliva sample are examined for levels of specific secretory IgA to diphtheria dialysate antigen, slgATDDA and slgARDDA respectively. The derived values slgATDDA and slgARDDA are compared, and the produced values are also compared thereby. Provided IgGTDA≤IgGRDA and slgATDDA≤slgARDDA, diphtheria prophylactic immunisation is considered to be indicated.

EFFECT: method presents more reliable determination of indications for diphtheria prophylactic immunisation.

FIELD: medicine.

SUBSTANCE: admission laboratory blood plasma examination in the patients with acute calculous cholecystitis is added with evaluating a cholecystokinin level both preoperative one, and on the 9th day following cholecystectomy, and if the examination shows decreasing concentration more than in 2 times, function-type Oddi sphincter dyssynergia is predicted.

EFFECT: invention enables early prediction of developing function-type Oddi sphincter dyssynergia following cholecystectomy.

2 ex, 5 dwg

FIELD: medicine.

SUBSTANCE: in newborn babies blood is sampled from umbilical artery and concentration of IL-6, IL-8 and NO2 in umbilical blood serum is determined by ELISA. After that diagnostic index (DI) is calculated by formula and if DI is lower than 0, it is concluded that laboratory indices of perinatal affection of nervous system in newborn babies are absent, and if DI is higher than 0, it is concluded that laboratory indices of perinatal affection of nervous system in newborn babies are present.

EFFECT: application of the method makes it possible to diagnose perinatal affection of nervous system in a newborn baby at pre-clinic stage.

3 ex

FIELD: medicine.

SUBSTANCE: in order to diagnose reproductive disorders in men, concentration of cytokines IL-8, IL-10 and IL-1RA in blood serum is determined by ELISA, after which diagnostic index DI is calculated. If DI is higher than 0, early reproductive disorders in men are diagnosed, if DI is lower than 0, it is concluded that laboratory indices of reproductive disorders in men are absent.

EFFECT: application of the method makes it possible to increase accuracy of diagnostics of early impaired fertility in men with normal indices of spermogram and to take decision about necessity of carrying out therapeutic correction of detected disorders in due time.

2 ex

FIELD: medicine.

SUBSTANCE: claimed is method of diagnosing phenotype of multiple medication resistance. Unicellular suspensions are prepared from tumour tissue and comparison cultures and fixed, after which incubation with primary and secondary antibodies and double washing are carried out. D values of statistical test of Kholmogorov-Smirnov are calculated and on the basis of obtained data determined is coefficient of level of expression of multiple medication resistance markers, showing phenotype expression. Coefficient of expression level makes it possible to point out three degrees of its expression: low, medium and high. On the basis of coefficient it is possible to determine tumour sensitivity to chemical preparations.

EFFECT: method makes it possible to increase accuracy of determining multiple medication resistance markers and carry out analysis in short terms.

2 dwg

FIELD: medicine.

SUBSTANCE: method includes introduction of sample into sample hole of the device, the device being adjusted for delivering said sample into lateral flow matrix, which has multitude of varieties of IgE antigens, immobilised in respective positions in result window. Method additionally includes possibility of sample movement along lateral flow matrix through immobilised multitude of IgE antigen varieties into the second position lower on flow from the first position, introduction of liquid buffer into lateral flow matrix for mobilisation of labeled reagent, which is adapted for binding aHTH-IgE-antibody and is dried in lateral flow matrix in position higher on flow from introduction of filtered sample into lateral flow matrix, and possibility of movement of labeled reagent, mobilised by liquid buffer, along lateral flow matrix through multitude of immobilised varieties of antigens IgE into position lower on flow from the first position.

EFFECT: simplification of process.

46 cl, 1 ex, 1 tbl, 8 dwg

FIELD: medicine, ophthalmology.

SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.

EFFECT: higher accuracy of prediction.

2 ex

FIELD: medicine, medicinal microbiology.

SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.

EFFECT: improved assay method.

3 tbl, 3 ex

FIELD: medicine, biology.

SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.

EFFECT: improved an valuable properties of nutrient medium.

3 ex

FIELD: medicine, cardiology.

SUBSTANCE: in peripheral blood one should detect the level of CD95(+) and CD16(+) neutrophilic granulocytes and at combination of increased level of CD95(+) neutrophilic granulocytes by 4 times and more and CD16(+) neutrophilic granulocytes by 0.6 times against the norm with ECG signs of myocardial infarction one should predict lethal result of large-focal myocardial infarction.

EFFECT: higher accuracy of prediction.

FIELD: medicine, parasitology.

SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.

EFFECT: higher efficiency and accuracy of diagnostics.

1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.

EFFECT: higher accuracy of detection.

FIELD: medicine, immunology.

SUBSTANCE: one should carry out reaction of blast-transformation, detect proliferation of T-lymphocytes activated with antibodies to CD3 in the presence of interleukin-7 (ACT IL-7) and in the presence of interleukin-7 and dexametazone (ACT IL-7 D), calculate the index for dexametazone action as the ratio of ACT IL-7 to ACT IL-7 D, moreover, the value of dexametazone action index being above 1.2 indicates increased production of cytokins that suppress T-lymphocytes in neonatals. The method enables to detect functional defect of immune system that characterizes neonatal period.

EFFECT: higher efficiency of detection.

2 ex

FIELD: medicine.

SUBSTANCE: method involves measuring forced exhalation volume per 1 s (FEV1) in l, full right ventricle evacuation time (RVE) in ms and angiotensin II value (AII) in ng/l. Discriminant relationship is built as D=0.504·RVE+3.038·FEV1 - 2.0·AII. D being less than 83.88, pulmonary hypertension occurrence is predicted within 1 year. D being equal to or greater than 83.88, no pulmonary hypertension is predicted to occur.

EFFECT: enhanced accuracy of prediction.

FIELD: medicine, medicinal immunology.

SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.

EFFECT: improved method for assay.

5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.

EFFECT: high accuracy of diagnosis.

Up!