RU2568904C1 - Method for prediction of clinical outcome of pseudotuberculosis infection in children - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention can be used to predict the unfavourable outcomes of pseudotuberculosis infection (PTI) in children. That is ensured by measuring the IgA and CD38+ concentration in the acute period of pseudotuberculosis infection in the children and compared to immunological values in the children suffered PTI infection and recovered completely with no unfavourable consequences in the period of the long-term case follow-up (reference group). If the CD38+ count increases to 54.07±0.60% with no statistically significant IgA changes in the acute period of PTI infection as compared to the reference group of children, gastrointestinal diseases are predicted. If the CD38+ concentration increases to 53.89±1.79% alongside a decrease of the IgA count to 142.58±20.14 mg/ml in the acute period of PTI infection, allergic syndrome is predicted to be developed in the case follow-up period.

EFFECT: using the presented method enables considering the pathological process developing in the case follow-up period as shown by the varying values in the acute period of the disease.

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Description

The invention relates to medicine, namely to infectious immunology, and can be used to predict adverse outcomes of pseudotuberculosis (PST) infection in children.

In Russia, PST infection is registered everywhere; the highest incidence rates are characteristic of the Siberian region. In addition to Russia, PST infection is endemic to Finland, Japan and other countries [E.A. Voskresenskaya, O.A. Burgasov. Methods of genetic typing of Iersinia pseudotuberculosis. Infectious diseases, 2011. - T. 9. - No. 2. - S. 81-88]. Among patients with PST, 70-75% are children under the age of 14 years [G.P. Somov, V.I. Pokrovsky, N.N. Besednova // Pseudotuberculosis. M .: Medicine, 2001. - S. 256]. In a number of cases, complications arise after an transmitted PST infection [A.P. Pomogaeva et al. // Clinical and immunological features of pseudotuberculosis in children. Bulletin of Siberian medicine, 2006. - No. 4. - S. 103-110]. Yersiniosis can become chronic in 6-35% of cases [Yersiniosis in children: etiology, epidemiology, clinical features, diagnosis and treatment tactics / I.V. Shestakova, N.D. Yushchuk // Attending physician No. 11, 2009. - Found on the Internet https://www.lvrach.ru/2009/l1/11184197/].

Based on the foregoing, the problem of predicting the outcome of pseudotuberculosis infection is relevant in terms of timely prevention of the development of the pathological process.

Several methods for predicting the outcome of the course of yersiniosis are known. To predict the development of the infectious process, indicators of T and B lymphocytes, completeness of phagocytosis, IgA and IgM levels are used [O.V. Popova et al. Th1 / Th2 response in various variants of the course of yersiniosis infection // Medical Immunology. - 2005. - Volume 7. - No. 2-3. - S. 120-121; V.F. Uchaikin et al. Yersiniosis in children // Geotar-Media Publishing Group. - Moscow. - 2005. - S. 73-77].

The proposed methods can predict the severity of the disease, but do not allow to predict the long-term consequences of the transferred infection process.

It is proposed to predict a prolonged course of infection on the basis of studying the immune status of patients in the acute phase of the disease according to indicators of CD8 +, CD20 +, CD25 + and CD95 +, the level of IgE, IFN-γ in the blood [G.F. Zheleznikova et al. Immunological criteria for the protracted course of yersiniosis infection in children. - Medical immunology. - 2001. - Volume 3. - No. 2. - S. 215-223). The method allows to predict a protracted course of yersiniosis infection, however, this method does not allow to determine the possibility of complications in the long-term period after a previous PST infection.

As a prototype, a method for predicting the course of yersiniosis in children based on clinical and immunological data in the initial period of the disease was chosen [patent RU No. 2422828 from 06/27/2011]. The proposed method allows to prevent the chronic and protracted course of yersiniosis infection, but does not make it possible to predict the formation of adverse effects in the long-term follow-up of children undergoing PST infection.

The objective of the present invention is to predict possible adverse effects after a pseudotuberculosis infection in children based on the study of immunological parameters in the acute period of the disease.

The technical result is achieved in that the population of pathogens of pseudotuberculosis is heterogeneous, it contains bacteria cultivated and uncultivated in laboratory conditions, and the concentration of uncultured bacteria significantly exceeds the number of cultivated ones. This is primarily due to the fact that the human body is an unfavorable environment for the reproduction of these bacteria. Yersinia pseudotuberculosis in high concentrations accumulates in the refrigerator at + 4 ° C. I.P. Yudin under the concept of uncultured bacteria means "viable" potentially, but under other conditions [I.P. Yudin. Modern approaches to assessing the viability of bacteria with a focus on the phenomenon of unculture / Annals of Mechnicov Institute, 2007, N 3. - P. 8-16]. Uncultured bacteria in the body are in an inactive state and therefore an adequate immune response is not formed on them. They are excreted from the body as a foreign substance. Therefore, by the intensity of the immune response, one can judge the quantitative ratio of cultured and uncultured bacteria in the patient's body in the stage of an acute infectious process. It is uncultured bacteria that can affect the development of adverse long-term outcomes after an infection.

The technical result is simple, not requiring large material costs and is based on the determination in the acute period of the disease in children of two immunological parameters: IgA and CD38 +. By changing these indicators in the acute period of the disease, it is possible to judge the development of the pathological process during the follow-up of children who have had a PST infection. The prevalence of uncultured bacteria in the population during the period of the disease leads to a change in these immunological parameters and in the long term these bacteria go into the cultivated state, causing the development of gastrointestinal diseases, or allergic syndrome develops in patients. With an increase in the number of CD38 + and the absence of statistically significant changes in IgA in the acute period of the disease compared with the control group of children (children who did not observe complications caused by this infection during the long-term follow-up after PST infection), the occurrence of gastrointestinal diseases was noted . An increase in the concentration of CD38 + and a decrease in the amount of IgA in the acute period of the disease compared with the control group made it possible to predict the development of an allergic syndrome during the follow-up period. The noted changes in immunological parameters are associated with the heterogeneity of the microbial population of Yersinia pseudotuberculosis, which contains cultured and uncultured bacteria. The concentration of uncultured bacteria significantly exceeded the number of cultivated bacteria, since the optimum temperature conditions for the reproduction of these bacteria is a temperature of + 4 ° C. At higher temperatures, a significant part of the bacteria goes into an uncultured state. The development of the pathological process in children during the follow-up period is connected with this. It has been established that uncultured bacteria under certain conditions can go into a cultivated state.

When developing the proposed method, a non-standard approach to the evaluation of immunological data was used. Usually, immunological data are compared with the immune status of a healthy person. The immunological data obtained by us were compared with immunological parameters in children who underwent PST infection, which ended in complete recovery without the formation of adverse effects during a long-term follow-up.

The technical result is achieved by the fact that according to the invention, in the acute period of pseudotuberculosis infection, the concentration of IgA and CD38 + in children is determined and compared with immunological parameters in children who underwent PST infection, which ended in complete recovery without the formation of adverse effects during long-term follow-up (control group), with an increase in the number of CD38 + to 54.07 ± 0.60% and the absence of statistically significant changes in IgA in the acute period of PST infection compared with The control group of children predicts the occurrence of gastrointestinal diseases, and with an increase in the concentration of CD38 + to 53.89 ± 1.79% and a decrease in the amount of IgA to 142.58 ± 20.14 mg / ml in the acute period of PST infection, development is predicted during the follow-up period allergic syndrome.

The proposed method is as follows: to identify defects and establish the degree of their severity in the cellular, humoral links of the immune system and the system of factors of nonspecific defense of the body, a complex of immunological tests of level 1 and 2 is used. All immunological parameters are evaluated in heparinized (25 PIECES / ml) venous blood. Blood sampling is carried out in the morning on an empty stomach at the same time in the initial period of the disease (up to 10 days). The following methods are used: Immunophenotyping of peripheral blood lymphocytes using an expanded panel of monoclonal antibodies to differentiation antigens (CD7 +, CD71 +, CD54 +, CD50 +, HLADR, CD3 +, CD4 +, CD8 +, CD16 +, CD11b +, CD20 +, CD23 +, CD38 +); determination of the concentration of serum immunoglobulins of class A, M, G (IgA, IgM, IgG) by the method of radial diffusion in a gel according to Mancini.

Subpopulations of lymphocytes are determined by flow cytometry. Lymphocytes are isolated from heparinized blood by centrifugation on a ficoll-verographin gradient. Contribute to centrifuge tubes in a volume of 50 μl. 5 μl of the tested monoclonal antibody (Sorbent LLP, Moscow) was added to the cells, incubated for 30-40 minutes, 150 ml of Hanks solution was added to the mixture and centrifuged for 5 minutes. Remove supernatant. To the pellet of the washed cells add 50 μl of F (ab) 2 - fragments of sheep antibodies labeled with FITC. Cells are suspended for 30 minutes at a temperature of + 4 ° C, then washed twice. The analysis is carried out on a flow laser cytofluorimeter FACSCAN firm "Becton Dickinson", USA. Blood for research is taken in the acute period of the infectious process (up to 10 days from the onset of the disease).

Evaluation of the results is carried out as follows: In the acute period of pseudotuberculosis infection, the concentration of IgA and CD38 + in children is determined and compared with immunological parameters in children who underwent PST infection, which ended in complete recovery without the formation of adverse effects during prolonged follow-up (control group), with an increase in the number of CD38 + to 54.07 ± 0.60% and the absence of statistically significant changes in IgA in the acute period of PST infection compared with the control group of children predict the occurrence of gastrointestinal diseases, and with an increase in the concentration of CD38 + to 53.89 ± 1.79% and a decrease in the number of IgA to 142.58 ± 20.14 mg / ml in the acute period of PST infection, they are predicted during the follow-up period development of an allergic syndrome.

Examples of practical use of the proposed method.

Example 1

Patient V., 9 years old, medical history No. 1548 Diagnosis: Pseudotuberculosis, scarlet fever-like form, moderate. He became acutely ill, with a temperature increase of up to 38.5 ° С, weakness, lethargy, complaints of abdominal pain, the appearance of a small-pointed rash with thickening in the area of natural folds. In organs and systems without features. There is mild hyperemia in the throat, no raids. The tongue is raspberry. The abdomen is normal in shape, painless, the liver extends 2.0 cm from under the edge of the costal arch, the spleen is not palpable. Liquid stool 1-2 times a day, without pathological impurities. Urine is a normal color. Based on the totality of clinical and medical history data, the diagnosis was made: Pseudotuberculosis, confirmed serologically (in RPHA the titer of antibodies is 1: 400).

Epidanamosis: an indication of the use of fresh cabbage and carrot salads in the school cafeteria. The patient denies the presence of allergic reactions to drugs and products. The patient receives a standard course of treatment, including the appointment of etiotropic drugs, pathogenetic and symptomatic therapy. Discharged on the 24th day of the disease with recovery.

Immunological examination was carried out on the 7th day of illness. Immunograms: CD38 + - 55.2%, IgA - 138.2 mg / ml. In the control group, these indicators were 44.80 ± 2.66% and 243.92 ± 19.54 mg / ml, respectively. There was an increase in CD38 + to 55.2% and a decrease in IgA to 138.2 mg / ml. The child is predicted to develop allergic syndrome during the follow-up period.

During follow-up observation after 1, 3 and 6 months, asthenization phenomena persisted, and complaints for the first time revealed allergy manifestations in the form of urticaria for citrus fruits, sneezing for the smell of fish. The patient was examined by an allergist. Diagnosed with Food Allergy.

Example 2

Patient B., 10 years old, medical history No. 2435 Diagnosis: Pseudotuberculosis, scarlet fever-like form, moderate. Was admitted to the infectious diseases hospital on the 2nd day of illness with a diagnosis of Scarlet fever. She got sick acutely, with fever up to febrile numbers, sore throat when swallowing, pain in the extremities, a rash of a small-pointed nature on the trunk, extremities - on the inner surface of the thighs, arms, feet, palms with a thickening in the natural folds, above the joints, in the places of bends petechiae are noted. The face is pasty, the eyelids are hyperemic. The conjunctiva is hyperemic, the sclera is injected. The tongue is papillary, cleansed of plaque, Tonsils 2 degrees, white gaps in the gaps, are removed, the surface does not bleed after removal, there is no swelling. The abdomen is normal in shape, painless, the liver extends 2.0 cm from under the edge of the costal arch, the edge of the spleen is palpated. The chair is decorated. Urine is a normal color. Based on the totality of clinical and anamnestic data, the diagnosis was made: Pseudotuberculosis, confirmed serologically (in RPGA antibody titer 1: 200).

Epidanamnesis: the patient used a salad of fresh cabbage and carrots (vegetables were stored in the cellar), lives in a private house. The patient denies the presence of allergic reactions to drugs and products. A standard course of treatment was carried out, including the administration of etiotropic drugs, pathogenetic and symptomatic therapy. Discharged on the 18th day of illness with recovery.

Immunological examination of the patient was carried out on the 5th day of illness. Immunograms: CD38 + 54.6%, IgA 126.8 mg / ml. In the control group, these indicators were 44.80 ± 2.66% and 243.92 ± 19.54 mg / ml, respectively. A marked increase in CD38 + to 546% and a decrease in IgA to 126.8 mg / ml. The child is predicted to develop allergic syndrome during the follow-up period.

During follow-up observation after 1, 3, and 6 months, asthenization phenomena persisted, the first identified complaints of allergy manifestations in the form of urticaria on chocolate, sneezing on the smell of garlic. Tearing, nasal congestion in the spring. The patient was examined by an allergist. Diagnosed with "Spring hay fever."

Example 3

Patient N., 7 years old, medical history No. 2846 Diagnosis: Pseudotuberculosis, scarlet fever-like form, moderate. Was admitted to the infectious diseases hospital on the 2nd day of the disease in a state of moderate severity. She fell ill acutely, with a temperature increase of up to 40.0 ° C, a rash of a small-pointed nature on the trunk, limbs - along the inner surface of the thighs, arms, feet, hands and feet - is hyperemic and swollen. Tonsils of 1 degree, hyperemic, no plaque. The tongue is overlaid. The abdomen is normal in shape, soft, painless, the liver extends 1.5 cm from under the edge of the costal arch, the edge of the spleen is palpated. The chair is decorated. Urine is a normal color. Based on the totality of clinical and anamnestic data, the diagnosis was made: Pseudotuberculosis, confirmed serologically (in RPGA antibody titer 1: 200). Epidanamnesis: the patient ate salads of fresh cabbage and carrots stored in the refrigerator at + 4 ° C. The patient received a standard course of treatment, including the appointment of etiotropic drugs, pathogenetic and symptomatic. Discharged on the 16th day of illness with recovery.

Immunological examination was carried out on the 6th day of illness. Immunograms: CD38 + 54.6%, IgA 234.2 mg / ml. In the control group, these indicators were 44.80 ± 2.66% and 243.92 ± 19.54 mg / ml, respectively. A marked increase in CD38 + to 54.6% and the absence of statistically significant changes in IgA compared with the control group. The patient is predicted to develop a disease of the gastrointestinal tract after PST infection.

During follow-up observation after 1, 3 and 6 months, complaints appeared about abdominal pain after physical exertion, intake of fatty foods. Periodically nausea, stool disorders. Ultrasound of the abdominal organs was performed. The patient was examined by a gastroenterologist. Diagnosed with DZHVP hypotonic type.

The child confirmed the prognosis of the development of gastrointestinal pathology during the follow-up period.

The revealed criteria allow predicting the development of adverse long-term consequences already in the acute period of PST infection and thereby optimizing the follow-up of children with the involvement of specialized specialists (gastroenterologist and allergist) with the necessary preventive measures.

A method for predicting the outcome of pseudotuberculosis infection in children during the follow-up period was developed on the basis of the Tyumen Infectious Diseases Hospital. Under observation were 34 children aged 3 to 15 years. The results of immunological studies in various groups of children with yersiniosis infection are presented in table 1, where, depending on the outcome of the disease, all children are divided into 3 groups: the first group - children whose disease of PST infection ended in complete recovery and there were no complications in the dispensary period - 12 person (control group); the second group - children who, after undergoing PST, developed gastrointestinal diseases (gastritis, gastroduodenitis, biliary dyskinesia, dyspancreatism); the third group - children who, after PST, developed an allergic syndrome in the form of atopic dermatitis, aggravation of food allergies, hay fever. The proposed method allows to save material resources by 1.47 million rubles. when conducting 100 predictions of an adverse outcome of PST infection.

Notes:

- Group 1 - a control group of children in whom during the period of long-term follow-up observation there were no complications associated with PST infection;

- Group 2 - the appearance in the dispensary period of gastrointestinal diseases;

- Group 3 - the appearance in the dispensary period of allergic diseases;

- n is the number of children examined.

Claims (1)

A method for predicting the outcome of pseudotuberculosis infection (PST) in children, characterized in that the concentration of IgA and CD38 + in children is determined in the acute period of pseudotuberculosis infection and compared with immunological parameters in children who underwent PST infection, which ended in complete recovery without the formation of adverse effects during a long period follow-up (control group), with an increase in the number of CD38 + to 54.07 ± 0.60% and the absence of statistically significant changes in IgA in the acute period of PST in In comparison with the control group of children, the projections predict the occurrence of gastrointestinal diseases, and with an increase in the concentration of CD38 + to 53.89 ± 1.79% and a decrease in the amount of IgA to 142.58 ± 20.14 mg / ml in the acute period of PST infection, they are predicted period of follow-up observation of the development of allergic syndrome.