RU2735999C1 - Method for prediction of placental insufficiency in women with threatening early miscarriage and habitual miscarriage in the anamnesis - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention refers to medicine, namely to obstetrics and gynecology, and can be used for prediction of placental insufficiency in women with threatening early miscarriage and habitual miscarriage in the anamnesis. Method involves 5–12-week gestation in the blood serum of CD178+ monocytes. If its value is 30.3 % and less, placental insufficiency is predicted in the second half of pregnancy.EFFECT: using the invention enables predicting the development of placental insufficiency in the second half of pregnancy with a high degree of accuracy – 98_1 % from early gestation.1 cl, 1 tbl, 3 ex

Description

The invention relates to medicine, namely to obstetrics and gynecology, and allows predicting the development of placental insufficiency in women with threatening early miscarriage and recurrent miscarriage in history.

The relevance of the method is determined both by the high prevalence of placental insufficiency among pregnant women and by the high risk and severity of complications of this pathology for the mother and fetus. According to various world literary sources, placental insufficiency in the population occurs in 30-45% of cases, and among pregnant women with recurrent miscarriage it reaches 75% (Serov V.N. Placental insufficiency // Difficult patient. - 2005. - T. 3, No. 2 - P. 17-20). Placental insufficiency leads to the development of complications of pregnancy, fetal malnutrition and hypoxia, in addition, it is one of the main causes of antenatal fetal death (Afanasyeva N.V., Strizhakov A.N. Pregnancy and childbirth outcomes in fetoplacental insufficiency of varying severity // Questions of gynecology , obstetrics and perinatology. - 2004. - T. 3, No. 2. - S. 7-13).

In connection with the above, an urgent task is to develop methods for predicting the development of placental insufficiency in women with threatening early miscarriage and a history of habitual miscarriage in order to carry out timely preventive and therapeutic measures in order to reduce perinatal complications for both the mother and the child.

A known method for predicting placental insufficiency in the 2nd trimester of pregnancy (Pat. No. 2554827. Russian Federation. A method for predicting placental insufficiency in the 2nd trimester of pregnancy / Botasheva TL, Linde VA, Sargsyan OD, Ermolova NV. , Palieva N.V., Frolov A.A., Tyan Yu.A. // Inventions. Useful models. - 2015. - No. 18). The invention presents a method for predicting placental insufficiency in the 2nd trimester of pregnancy by enzyme-linked immunosorbent assay of blood serum of pregnant women, characterized in that the sex of the fetus is determined by a previously known ultrasound method, then the levels of angiogenic factors and cytokines are determined in the venous blood of pregnant women, and if an increase in the level of interleukin-12 is found to be more 3.2 pg / ml, epidermal growth factor more than 310 pg / ml, a decrease in the level of placenta growth factor less than 40.0 pg / ml in pregnant women carrying male fetuses, and an increase in endothelin-1 more than 0.42 pg / ml, interleukin -1β is more than 17.6 pg / ml, tumor necrosis factor-α is more than 6.5 pg / ml in pregnant women carrying female fetuses, the development of placental insufficiency is predicted in these pregnant women. This method has several disadvantages:

1. It is necessary to carry out a preliminary ultrasound examination to determine the sex of a bearing fetus, which in some cases is difficult or impossible;

2. Prediction is possible only in the 2nd trimester of gestation, since it is impossible to determine the sex of the fetus by ultrasound in early pregnancy;

3. The algorithm for predicting placental insufficiency is not presented in case of difficulty in determining the sex of the fetus;

4. Cost and labor intensity, since the assessment of six immunological indicators, which creates additional difficulties in the interpretation of the results.

A known method for predicting the development of placental insufficiency in the second half of pregnancy in women with the threat of miscarriage of early stages of infectious genesis (Pat. No. 2558795. Russian Federation. A method for predicting the development of placental insufficiency in the second half of pregnancy in women with the threat of early miscarriage of infectious genesis / Malyshkina A. I., Sotnikova N.Yu., Kroshkina N.V., Mozhaeva T.A. // Inventions. Useful models. - 2015. - No. 22), which consists in the fact that women with the threat of miscarriage of infectious genesis in I trimester from the 6th week of pregnancy, determine the content of IL-18 and, if its value is more than 100 pg / ml, predict the development of placental insufficiency in the second half of pregnancy. This method has the following disadvantages:

1. This method is applicable only for women with the threat of interruption of infectious genesis;

2. To perform this method, a preliminary enzyme-linked immunosorbent assay is necessary to assess the infectious status of pregnant women (herpes simplex virus type 1-2; cytomegalovirus; Chlamydia Trachomatis; Mycoplazma Hominis; Ureaplazma Urealyticum);

3. There is no data on the possibility of using this method in the case of detection of other sexually transmitted infections by the method of enzyme immunoassay.

4. Given the need for enzyme immunoassay, the method is costly and time consuming.

The closest to the proposed method is a method for predicting the development of placental insufficiency in the second half of pregnancy (Pat. No. 2600791. Russian Federation. A method for predicting the development of placental insufficiency in the second half of pregnancy / Malyshkina A.I., Sotnikova N.Yu., Kroshkina N.V. ., Batrak N.V. // Inventions. Useful models. - 2016. - No. 30). The method consists in the fact that in the period of 5-12 weeks of gestation, the concentration of DcR3 in the blood serum of women is determined, and if its value is more than 0.87 pg / ml, the development of placental insufficiency is predicted in the second half of pregnancy. This method has the following disadvantages - relatively low indicators of accuracy, sensitivity and specificity. These disadvantages are proposed to be eliminated in the claimed method.

The claimed technical result is achieved by the fact that in the period of 5-12 weeks of gestation in the blood serum of pregnant women with a threatening early miscarriage and habitual miscarriage in the anamnesis, the relative content of CD178 + monocytes is determined and, at its value of 30.3% or less, placental insufficiency is predicted in the second half of pregnancy ...

The technical result is aimed at expanding the arsenal of prognostic tools, increasing the accuracy, sensitivity and specificity of predicting placental insufficiency. The proposed method makes it possible to predict the development of placental insufficiency in women with a threatening early miscarriage and a history of recurrent miscarriage with an accuracy of 98.1%, which makes it possible to carry out timely preventive and therapeutic measures and reduce the level of perinatal morbidity and mortality.

The method is carried out as follows:

1. To 3 ml of heparinized blood from the cubital vein add 3 ml of Medium 199 and isolate the enriched population of mononuclear cells by the standard method of high-speed centrifugation at 1500 rpm for 40 minutes in a density gradient of ficoll-urografin (Bourn A. Isolation of mononuclear cells and granulocytes from human blood and bone marrow // Scand. J. Clin. Invest. - 1968. - Vol.21, No. 97. - P. 77).

2. The suspension of mononuclear cells is washed twice with saline, the cell concentration is adjusted to 1 × 10 ⁶ cells / ml. To 0.1 ml of the resulting suspension was added 20 μl of aHTH-CD178 monoclonal antibodies conjugated with phycoerythrin. The cells are incubated for 30 minutes at room temperature in the dark, washed and fixed in 0.5 ml of CellFix ™ fixative solution (BDBiosciences, Belgium).

3. Determine the relative (percentage) number of CD178 + monocytes in the monocyte gate on a FACScanto flow cytometer (Becton Dickinson, USA). With its value equal to 30.3% or less, the development of placental insufficiency in the second half of pregnancy is predicted.

The novelty of the proposed method lies in the fact that for the first time it is proposed to predict the development of placental insufficiency in pregnant women with threatening early miscarriage and a history of recurrent miscarriage, by determining the content of CD178 + monocytes in the peripheral venous blood.

The CD95-CD178 system is one of the most important inducers of apoptosis, and a disturbance in the expression of one of these molecules may be sufficient to interfere with the processes of apoptosis of the trophoblast and threaten the implantation process or prevent the development of pregnancy. It is known that CD 178 is expressed by monocytes, macrophages, T-lymphocytes, endometrial stroma, and CD95 is mainly expressed by mature T-lymphocytes, as well as in a small number of trophoblastomas (Fas and Fas-L genotype and expression in patients with recurrent pregnancy loss / PCA Banzato, S. Daher, E. Traina et al. // Reprod. Sci. - 2013. - Vol.20, No. 9. - P. 1111-1115).

Due to the interaction of CD95, expressed by trophoblast cells, and CD 178 - stromal cells of the endometrium, apoptosis of stromal cells occurs, which promotes implantation of the ovum (Fas Lon decidual macrophages mediates trophoblast apoptosis: A potential cause of recurrent miscarriage / J. Ding, T. Yin, N. Yan et al. // International Journal of Molecular Medicine. - 2019. - Vol. 43, No. 6. - P. 2376-2386). In addition, monocytes / macrophages expressing CD178 induce apoptosis of endothelial and smooth muscle cells of the spiral arteries of the uterus expressing CD95, which ensures the invasion of the extravillous trophoblast into the muscle layer of the spiral arteries of the uterus, followed by expansion of these vessels and ensuring sufficient blood flow in the placenta of factors (Lnc RNAMEG3 in hi bits trophoblast invasion and trophoblast-mediated VSMC loss in uterine spiral artery remodeling / H. Tao, X. Liu, X. Liu et al. // Mol. Reprod. Dev. - 2019. - Vol. 86, No. 6. - P. 686-695).

A decrease in the expression of CD178 by monocytes in the peripheral blood with threatened early miscarriage may be one of the factors leading to impaired apoptosis, which results in impaired trophoblast invasion and transformation of spiral arteries, which may cause the development of placental insufficiency in women with recurrent miscarriage in history.

Using the proposed method allows with a high degree of accuracy (98.1%) from early gestation to predict the development of placental insufficiency in the second half of pregnancy.

Distinctive features of the claimed method are: determine the relative content of CD178 + monocytes and, if its value is equal to or less than 30.3%, predict the development of placental insufficiency.

The essence of the proposed method is illustrated by the following examples:

Example 1. Patient I., 34 years old, fourth pregnancy, desired, came independently, in a second marriage. She has a history of two early spontaneous miscarriages, one undeveloped early pregnancy. At 6 weeks of pregnancy, she was hospitalized in the gynecological department with complaints of nagging pain in the lower abdomen. With a vaginal examination, the external genital organs are developed correctly, the vagina is narrow, the cervix is conical in shape, up to 4 cm long, the external pharynx is closed, the body of the uterus is slightly larger than normal, rounded, dense, mobile, painless, the discharge is mucous.

Diagnosis: Pregnancy 6 weeks. Threatened miscarriage. Habitual miscarriage.

Prior to the appointment of preserving therapy, a survey was carried out according to the claimed method: the relative content of CD178 + monocytes was 27.7%.

Conclusion: the prognosis is the development of placental insufficiency.

Conservation therapy was carried out with a positive effect.

In the period of 27 weeks of gestation, according to the results of ultrasound and Doppler studies, fetal growth retardation of I degree, impaired uteroplacental circulation of I A degree were revealed.

Diagnosis: Pregnancy 27 weeks. Fetoplacental insufficiency. Fetal growth retardation I degree. Habitual miscarriage.

The forecast for the claimed method was confirmed.

Example 2. Patient E., 23 years old, fifth pregnancy, desired, came independently, in the first marriage. The anamnesis has two spontaneous miscarriages, two non-developing pregnancies in the early stages. At the age of 11 weeks of pregnancy, she was hospitalized in the gynecological department with complaints of nagging pains in the lower abdomen, bloody discharge from the genital tract. With a vaginal examination, the external genital organs are developed correctly, the vagina is narrow, the cervix is conical in shape, up to 4 cm long, the external pharynx is closed, the body of the uterus is enlarged to 11 weeks of gestation, round, dense, mobile, painless, scanty bloody discharge.

Biochemical and ultrasound screening - no pathology.

Diagnosis: Pregnancy 11 weeks. Threatened miscarriage. Habitual miscarriage.

A survey was carried out according to the claimed method: the relative content of CD178 + monocytes was 38.2%.

Conclusion: the prognosis is the absence of placental insufficiency.

Conservation therapy was carried out with a positive effect. Biochemical and ultrasound screening on time - without pathology.

At 39 weeks, the first timely birth through the vaginal birth canal occurred with a female fetus 53 cm long, weighing 3800 g. The Apgar score was 8/9 points.

The forecast for the claimed method was confirmed.

Example 3. Patient M., 25 years old, fourth pregnancy, desired, came independently, in the first marriage. She has a history of three early pregnancies. At the time of 10 weeks of pregnancy, she was hospitalized in the gynecological department with complaints of pulling pains in the lower abdomen. With a vaginal examination, the external genital organs are developed correctly, the vagina is narrow, the cervix is conical in shape, up to 4 cm long, the external pharynx is closed, the body of the uterus is enlarged to 10 weeks of gestation, round, dense, mobile, painless, mucous discharge.

Diagnosis: Pregnancy 10 weeks. Threatened miscarriage. Habitual miscarriage.

A survey was carried out according to the claimed method: the relative content of CD178 + monocytes was 30.3%.

Conclusion: the prognosis is the development of placental insufficiency.

Conservation therapy was carried out with a positive effect.

At 32 weeks of gestation, the results of ultrasound and Doppler studies revealed a growth retardation of the I degree, a violation of the uteroplacental blood circulation of the II degree.

Diagnosis: Pregnancy 32 weeks. Fetoplacental insufficiency. Fetal growth retardation I degree. Habitual miscarriage.

The forecast for the claimed method was confirmed.

In this way, 54 women were examined, the research results are shown in Table 1.

The advantages of the proposed method:

1. High accuracy - 98.1%, sensitivity - 100%) and specificity - 97%.

2. Good reproducibility of the method.

3. Simplicity and availability of the technique.

4. Ease of interpretation of survey results.

5. The method can be used as a screening method for assigning a woman to a risk group for the development of placental insufficiency for the timely appointment of preventive and therapeutic measures in order to reduce perinatal complications for both the mother and the child.

Claims (1)

A method for predicting placental insufficiency in the second half of pregnancy in women with threatened miscarriage and a history of recurrent miscarriage by immunological examination of blood serum at 5-12 weeks of gestation, characterized in that the relative content of CD178 + monocytes is determined and at its values equal to 30.3% and less, predict the development of placental insufficiency.