RU2712627C1 - Method for infertility prediction in females with hypothalamic syndrome - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to obstetrics and gynecology, and can be used for prediction of infertility in females of reproductive age with hypothalamic syndrome. For this purpose, in peripheral blood and aspirate from uterine cavity in women of reproductive age with hypothalamic syndrome and body weight index greater than or equal to 30, lymphocyte subpopulations values are determined CD3⁺CD19^-, CD3⁺CD4⁺, CD3⁺CD8⁺, CD3^-CD19⁺, CD3^-CD(16⁺56⁺) and immunoregulatory index – CD4⁺/CD8⁺ depending on the menstrual cycle, namely on 5th and 21st days. If the lymphocyte subpopulations are on 5th day of the menstrual cycle in the peripheral blood CD3⁺CD19^- from 1.08 to 1.19×10⁹/l; CD3⁺CD4⁺ from 0.51 to 0.55×10⁹/l; CD3⁺CD8⁺ from 0.35 to 0.40×10⁹/l; CD3^-CD19⁺ from 0.38 to 0.47×10⁹/l; CD3^-CD(16⁺56⁺) from 0.37 to 0.43×10⁹/l immunoregulatory index – CD4⁺/CD8⁺ 1.45–1.37, on 21st day of menstrual cycle in peripheral blood CD3⁺CD19^- from 1.20 to 1.51×10⁹/l; CD3⁺CD4⁺ from 0.56 to 0.65×10⁹/l; CD3⁺CD8⁺ from 0.41 to 0.45×10⁹/l; CD3^-CD19⁺ from 0.38 to 0.46×10⁹/l; CD3^-CD(16⁺56⁺) from 0.38 to 0.44×10⁹/l immunoregulatory index – CD4⁺/CD8⁺ 1.36–1.22, in an endometrial aspirate on 5th day of the menstrual cycle CD3⁺CD19^- from 1.1 to 1.22×10⁹/l; CD3⁺CD4⁺ from 0.54 to 0.58×10⁹/l; CD3⁺CD8⁺ from 0.36 to 0.42×10⁹/l; CD3^-CD19⁺ from 0.40 to 0.49×10⁹/l; CD3^-CD(16^-56⁺) from 0.45 to 0.54×10⁹/l; immunoregulatory index – CD4⁺/CD8⁺ 1.59–1.21, on 21st day of the menstrual cycle in the endometrial aspirate CD3⁺CD19^- from 1.22 to 1.53×10⁹/l; CD3⁺CD4⁺ from 0.54 to 0.57×10⁹/l; CD3⁺CD8⁺ from 0.43 to 0.47×10⁹/l; CD3^-CD19⁺ from 0.49 to 0.55×10⁹/l; CD3^-CD(16^-56⁺) from 0.39 to 0.46×10⁹/l immunoregulatory index – CD4⁺/CD8⁺ 1.34–1.42 predict infertility in females with hypothalamic syndrome.

EFFECT: using this method enables predicting infertility in females of reproductive age with hypothalamic syndrome and body weight index greater than or equal to 30 by evaluating immune responses at the systemic and local level of the female body.

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Description

The invention relates to clinical medicine, in particular to the specialty of obstetrics and gynecology, and for the search for new opportunities for predicting infertility in women of reproductive age with hypothalamic syndrome and body mass index greater than or equal to 30.

Currently, one of the main factors leading to a decrease in the birth rate is the deterioration of the reproductive health of women. Infertility is a major medical and social problem not only in Russia, but also in Europe and America. In recent years, there has been a steady increase in neuroendocrine pathology. Up to 18-25% of women of reproductive age suffer from hypothalamic syndrome. In most cases, without treatment, the hypothalamic syndrome transforms into a neuroendocrine syndrome, against which 25-72% of women develop infertility (Kulakov V.I., Prilepskaya V.N., Radzinsky V.E. // Outpatient Clinic Manual assistance in obstetrics and gynecology. M.: GEOTAR-Media, - 2007. - S. 849-855). In this regard, it is necessary to search for new methods for predicting reproductive disorders in women with hypothalamic syndrome.

The well-known "Method for predicting the development of reproductive disorders" (RF Patent No. 2495431, published 10.10.2013 g), characterized in that women examine the cervical discharge on the tenth day of the menstrual cycle, as the parameters determine the levels of interleukin 1 beta and interferon gamma at the same time. When the content of interleukin 1 beta is more than 38.8 ± 3.8 pg / ml and the simultaneous content of interferon gamma is more than 8.1 ± 0.5 pg / ml, the formation of reproductive disorders is predicted.

The disadvantage of this method is that the study is conducted in women with sexually transmitted infections, which already implies the presence of an inflammatory process at the level of the cervical canal, and interleukin 1 beta and interferon gamma were used to characterize local immunity in the inflammatory process, emphasizing the infectious nature of reproductive disorders functions.

A known method for predicting reproductive function in girls with hypothalamic syndrome of puberty (RF Patent No. 2438136, published December 27, 2011). The essence of the method lies in the fact that before and after treatment in menstrual blood determine the level of glycodelin. When the glycodelin value is 11.12 ng / ml or more after treatment, pathogenetic therapy is assessed as effective.

The disadvantage of this method is that the marker used for prediction is a marker of the protein-forming function of the endometrium and does not reflect the state of cellular immunity in the endometrium. In addition, the study is carried out in menstrual blood.

A known method for predicting reproductive disorders in women with overweight and obesity (RF Patent No. 2617491, published 04/25/2017). The essence of the proposed method lies in the fact that early prediction is achieved by determining the activity of NF-kB in aspirate from the uterine cavity, with an unfavorable prognosis for pregnancy - NF-kB> 6.86 pg / ml.

The disadvantage of this method is that it determines the modulating processes of apoptosis, angiogenesis, differentiation and proliferation at the endometrial level, but does not reflect the state of cellular immunity.

A known method for predicting the onset of pregnancy in women with tubal-peritoneal infertility in the program of in vitro fertilization / RF Patent 2523401 Published July 20, 2014 /

The essence of the method lies in the fact that before the IVF program in the patient in the luteal phase of the cycle, an endometrial biopsy is performed and the relative content of CD95 + macrophages in the leukocyte endometrial infiltrate is determined. With a relative CD95 + macrophage content of 48.8% or more, pregnancy is predicted, and with a value of less than 48.8%, pregnancy is not expected.

The disadvantage of this method is that only CD95 + macrophages are determined in the endometrial biopsy and in the IVF program, without the possibility of a natural pregnancy. There is no comprehensive assessment of the state of immunity at the systemic and local levels.

Known methods for predicting and mechanisms for the development of infertility in women: Kulakov V.I., Prilepskaya V.N., Radzinsky V.E. // Guidelines for outpatient care in obstetrics and gynecology. M .: GEOTAR-Media, - 2007. - S. 849-855 / Krutova V.A., B.G. Ermoshenko A.M. Chulkova, Possibilities for predicting the development of female infertility // Kuban Scientific Medical Bulletin - 2009 - No. 9 (114) - P. 73-80 /. Zhukovets I.V., Leshchenko O.Ya., Atalyan A.V. The main mechanisms of primary infertility in women of reproductive age with hypothalamic dysfunction in the puberty Acta Biomedica scietifica - 2017, - Volume 2, - No. 5, Part 1. - P. 15-19 /;

The disadvantage of these studies is that they do not reflect the state of immune responses at the systemic and local levels in women with reproductive disorders.

A known method for predicting reproductive disorders in women of reproductive age with hypothalamic dysfunction / RF Patent No. 2617247, published 04.24.2017 /. The essence of the method is that early prediction is achieved by simultaneously evaluating pro- and anti-inflammatory cytokines (TNF-α and IL-10) in aspirate from the uterine cavity. With an unfavorable prognosis for pregnancy, TNF-α> 20.2 pg / ml, IL-10 <4.78 pg / ml, with an unfavorable prognosis for gestation with TNF-α> 18.9-20.1 pg / ml, IL -10 <6.06-4.79 pg / ml.

The disadvantage of this method is the determination of one pro and one anti-inflammatory interleukins at the local level, on the 21st day of the menstrual cycle, which does not allow for a comprehensive assessment of all parts of the immune system, both at the local and systemic levels and in a timely manner (with the onset of the menstrual cycle) preventive measures to the stage of pregnancy planning.

It is known that the normal development and functioning of the reproductive system is carried out with the close interaction of the immune and endocrine systems. Immune system disorders are associated with reproductive dysfunction. Cells of the immune system are involved in the regulation of folliculogenesis, ovulation, in the formation and regression of the corpus luteum. If estrogen production is disturbed in a woman’s body, an increase in the number of CD8 lymphocytes and low CD4 / CD8 is recorded. The role of immune factors in the functioning of the female reproductive system / N.O. Motovilova, I.Yu. Kogan, Areg A. Totolyan // Ural Medical Journal. - 2010. - No. 4. - S. 117-121). There is evidence that estradiol and progesterone regulate the activity of cytolytic T cells (CD3, CD8), a decrease in which is noted during the secretory phase. It was proved that immune cells are present everywhere in the reproductive tract, and the number of macrophages, B and T cells in the endometrium changes during the menstrual cycle (Gadieva F.G. Relationship of the immune and endocrine systems in women of reproductive age // Obstetrics and Gynecology - 2001. - No. 1. - S. 11-13).

The present invention is aimed at earlier prediction of infertility in women of reproductive age with hypothalamic syndrome.

The technical result is achieved by the fact that in the peripheral blood and aspirate from the uterus in women of reproductive age with hypothalamic syndrome and a body mass index greater than or equal to 30, lymphocyte subpopulations CD3 ⁺ CD19 ^- , CD3 ⁺ CD4 ⁺ , CD3 ⁺ CD8 ⁺ , CD3 ^{- are} determined CD19 ⁺ , CD3 ^- CD (16 ⁺ 56 ⁺ ) and immunoregulatory index - CD4 ⁺ / CD8 ⁺ depending on the day of the menstrual cycle, namely on days 5 and 21, and with the lymphocyte subpopulations on the 5th day of the menstrual cycle in the peripheral blood CD3 ⁺ CD19 ^- from 1.08 to 1.19 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.51 to 0.55 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.35 to 0.40 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.38 to 0.47 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) from 0.37 to 0.43 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.45-1.37, on the 21st day of the menstrual cycle in the peripheral blood CD3 ⁺ CD19 ^- from 1.20 to 1.51 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.56 to 0.65 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.41 to 0.45 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.38 to 0.46 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) from 0.38 to 0.44 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.36-1.22, in endometrial aspirate on day 5 of the CD3 menstrual cycle ⁺ CD19 ^- from 1.1 to 1.22 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.54 to 0.58 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.36 to 0.42 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.40 to 0.49 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) from 0.45 to 0.54 × 10 ⁹ / l, immunoregulatory index - CD4 ⁺ / CD8 ^- - 1, 59-1.21, on the 21st day of the menstrual cycle in endometrial aspirate CD3 ⁺ CD19 ^- from 1.22 to 1.53 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.54 to 0.57 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.43 to 0.47 × 10 ⁹ / L, CD3 ^- CD19 ⁺ from 0.49 to 0.55 × 10 ⁹ / L, CD3 ^- CD (16 ^- 56 ⁺ ) from 0.39 to 0.46 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.34-1.42 predict infertility in women with hypothalamic syndrome.

To assess the cellular immunity link at the system level, the subpopulation composition of peripheral blood lymphocytes is determined CD3 ⁺ , CD4 ⁺ , CD8 ⁺ , CD16 ⁺ , CD19 ⁺ , CD56 ⁺ . To do this, blood was drawn from a vein using a disposable syringe into 2 test tubes with a volume of 10 ml into a dry sterile and sterile test tube containing heparin at a final concentration of 25 PIECES in 1 ml of blood on the 5th and 21st day of the menstrual cycle. Peripheral blood from a vein was taken on an empty stomach from 8 to 10 in the morning. Within 1.5 hours at a temperature of 37 ° C, the biological material was delivered to the laboratory.

Whole blood was stained for 30 minutes. in the dark using monoclonal antibodies CD3, CD4, CD8, CD (16 + 56), CD19 fluorochrome-labeled FITC, PE and PerCP / Cy5 (Becton Dickinson, DAKO) to detect immunocompetent cells. The following combinations of monoclonal antibodies were used to determine target cell populations.

1) Peripheral blood:

• TgG1 / IgG2a / CD45 - to control the non-specific binding of lymphocytes to anti-human antibodies and the selection of lymphocytes;

• CD3 / CD19 / CD45 - to determine the relative content of B-lymphocytes [CD3 ^- CD19 ⁺ ] and T-limocytes [CD3 ⁺ CD19 ^- ];

• CD3 / CD4 / CD45 - to determine the relative number of T-helpers [CD3 ⁺ CD4 ⁺ ];

• CD3 / CD8 / CD45 - to determine the relative number of cytogenous T-lymphocytes [CD3 ⁺ CD8 ⁺ ];

• CD3 / CD (16 + 56) / CD45 - to determine the relative number of natural killers [CD3 ^- CD (16 + 56) ⁺ ].

After staining peripheral blood samples, red blood cells were lysed using a hypotonic lysing solution (DAKO Easy Lyse). For this, 700 μl of the prepared solution was added to each tube, thoroughly mixed and incubated for 10-12 minutes. in the dark at room temperature. Then, stained mononuclear cells were washed from lysed red blood cells and unbound antibodies by two successive centrifuges with Hanks solution for 5 minutes. at 300 g (1000 rpm.). The results were taken into account using an Epics flow cytometer (Beckman Coulter, USA) using standard protocols. In addition, an immunoregulatory index (IRI, CD4 + / CD8 +) was calculated. Studies were performed within 6 hours after blood sampling.

Evaluation of the state of local cellular immunity in patients with hypothalamic syndrome and a body mass index greater than or equal to 30 was carried out in endometrial tissue with subsequent immunohistochemical examination. Endometrial tissue was obtained on days 5 and 21 of the menstrual cycle by aspiration biopsy using Pipel (C.C.D., France). Up to 3 ml of the obtained tissue was placed in a test tube with 100 IU of heparin sodium in phosphate-buffered saline (PBS). The aspirate was subsequently mechanically ground and homogenized by passing through a decreasing volume syringe with needles of smaller diameter. Then, a fraction of mononuclear cells was isolated from the obtained endometrial homogenate: the homogenate was carefully layered on 2 ml of Ficoll-Paque, Amersham, (1.077 g / l), and then centrifuged for 40 min. at 450 g (1500 rpm.). After centrifugation in a density gradient, the opalescent ring of mononuclear cells was gently transferred to a dry clean tube using a dispenser and further washed with FSB (centrifugation for 15 minutes at 450 g (1500 rpm). Endometrial mononuclear cells were stained for 30 minutes in the dark with monoclonal antibodies CD3, CD4, CD8, CD (16 + 56), CD19, CD45, fluorochrome-labeled FITC, PE and PerCP / Cy5 (Becton Dickinson, DAKO) for detection of immunocompetent cells. The following combinations of monoclonal antibodies were used to determine target cell populations:

2) Endometrial aspirate:

- IgG1 / IgG2a / CD45 - to control the non-specific binding of lymphocytes to anti-human antibodies and the allocation of lymphocytes;

- CD3 / CD4 / CD45 - to determine the relative number of T-helpers [CD3 ⁺ CD4 ⁺ ];

- CD3 / CD8 / CD45 - to determine the relative number of cytotoxic T-lymphocytes [CD3 ⁺ CD8 ⁺ ];

• CD3 / CD19 / CD45 - to determine the relative content of B-lymphocytes [CD3 ^- CD19 ⁺ ] and T-limocytes [CD3 ⁺ CD19 ^- ];

- CD3 / CD (16 ⁺ 56 ⁺ ) / CD45. - to determine the relative amount of natural killers [CD3 ^- CD (16 ⁺ 56 ⁺ )], [CD3 ^- CD (16 ^- 56 ⁺ )];

The results were taken into account using a Beckman Coulter Epics flow cytometer. (USA) using standard protocols. In addition, an immunoregulatory index (IRI, CD4 ⁺ / CD8 ⁺ ) was calculated.

This method allows you to predict infertility before planning a pregnancy and timely correction of the identified violations. Also, the proposed method provides the possibility of a screening examination of patients with hypothalamic syndrome and a body mass index of 30 or more, namely, to predict reproductive disorders, timely correction, which will assess the risk, accelerate the implementation of the generative function and reduce the material costs of therapy.

New features of the claimed method are:

- prediction of infertility before planning pregnancy;

- assessment of the subpopulation composition of lymphocytes in the endometrium (local level) and peripheral blood (systemic level) simultaneously;

- determination of the cellular link of immunity at the systemic and local levels by determining the subpopulation composition of lymphocytes is carried out on the 5th and 21st day of the cycle;

- determination of the cellular link of immunity at the systemic and local levels by determining the subpopulation composition of lymphocytes is carried out in patients with hypothalamic syndrome and a body mass index greater than or equal to 30;

- venous blood (systemic level of regulation of immunity) and endometrial tissue (local level of regulation of immunity) are used as biological material.

The proposed method is based on a new direction, obtained by laboratory research on changes in the content of cellular immunity indicators at the systemic and local levels, namely the determination of the subpopulation composition of peripheral blood lymphocytes CD3 ⁺ , CD4 ⁺ , CD8 ⁺ , CD16 ⁺ , CD19 ⁺ , CD56 ⁺ in women of reproductive age with hypothalamic syndrome, body mass index greater than or equal to 30 on the 5th and 21st day of the menstrual cycle compared with women of reproductive age who did not have disorders in the hypothalamic-pituitary system topic (relatively healthy).

For the first time, it was found that in women of reproductive age with hypothalamic syndrome in the peripheral blood, on the 5th day of the menstrual cycle, there is an increase in lymphocyte subpopulations CD3 ⁺ CD19 ^- , CD3 ⁺ CD4 ⁺ , CD3 ⁺ CD8 ⁺ , CD3 ^- CD19 ⁺ , CD3 ^- CD (16 ⁺ 56 ⁺ ) in the peripheral blood on the 5th and 21st day of the cycle, with an increase in performance by the 21st day of the menstrual cycle. In addition, with an increase in the weight of patients, the concentrations of subpopulations of these lymphocytes in the blood also increase. In the endometrium, the tendency is to increase the subpopulations of lymphocytes CD3 ⁺ CD19 ^- , CD3 ⁺ CD4 ⁺ , CD3 ⁺ CD8 ⁺ , CD3 ^- CD19 ⁺ , CD3 ^- CD (16 ^- 56 ⁺ ). NK cells (CD3 ^- CD (16 ^- 56 ⁺ ) in the endometrium increase to the greatest extent, CD3 ⁺ CD4 ⁺ slightly decrease, against the background of an increase in CD3 ⁺ CD8 ⁺ .

The study was conducted in 64 women of reproductive age with hypothalamic syndrome and 30 conditionally healthy women from 2010 to 2017 on the basis of women's consultation and women's consultation No. 1 of GBUZ JSC City Polyclinic No. 8 named after N.I. Pirogov. "

When assessing the levels of pro-inflammatory cytokines on the 5th day of the menstrual cycle in the peripheral blood and endometrial aspirate, a significant increase in CD3 ⁺ CD19 ^is noted ^- in relation to the control (p <0.05), also CD3 ⁺ CD4 ⁺ (p <0.01) CD3 ^- CD19 ⁺ (p <0.01), CD3 ^- CD (16 ^- 56 ⁺ ) (p <0.05). The level of CD3 ^- CD (16 ^- 56 ⁺ ) in the endometrium significantly increased in relation to the control (p <0.05). The described trend is reflected in table 1, 2, 3 and Fig., Where:

FIG. 1 Dynamics of a subpopulation of lymphocytes in peripheral blood with hypothalamic syndrome in women on the 5th day of the menstrual cycle;

FIG. 2 Dynamics of a subpopulation of lymphocytes in peripheral blood with hypothalamic syndrome in women on the 21st day of the menstrual cycle;

FIG. 3 Dynamics of a subpopulation of lymphocytes in aspirate from the uterine cavity in case of hypothalamic syndrome in women on the 5th day of the menstrual cycle

FIG. 4 Dynamics of a subpopulation of lymphocytes in an aspirate from the uterine cavity in case of hypothalamic syndrome in women on the 21st day of the menstrual cycle

Case studies:

Example 1

Patient K., 25 years old,

Complaints at the time of the examination: irregular menstruation, absence of pregnancy for 3 years with regular sex without contraception,

History: menstruation from 12 years, 4-6 days, after 30-35 days, from 15 years - after 50-65 days, irregular. In reproductive age, an obstetrician-gynecologist did not consult a doctor. In the puberty period, I applied to the children's gynecologist at the Children's City Polyclinic No. 1 JSC. After the examination, the diagnosis was made: Hypothalamic dysfunction of the puberty period. Secondary oligomenorrhea.

Sexual life since 19 years in marriage, without contraception. There was no pregnancy. Objectively: BMI 33.5 kg / m ² , pearl-colored stria in the thighs and buttocks, mammary glands. Clinical diagnosis: Secondary oligomenorrhea. Obesity 1 tbsp. Complicated gynecological history (dysfunction of the hypothalamus in the puberty). The examination was performed on the 5th day of the menstrual cycle in the peripheral blood: CD3 ⁺ CD19 ^- - 1.15 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.53 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.37 × 10 ⁹ / l, CD3 ^- CD19 ⁺ - 0.41 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56) - 0.39 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.43, 21 days of the menstrual cycle: CD3 ⁺ CD19 ^- 1.44 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.58 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.43 × 10 ⁹ / L, CD3 ^- CD19 ⁺ - 0.42 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) - 0.43 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.34. In the endometrium, on the 5th day of the menstrual cycle, the content of lymphocyte subpopulations was: CD3 ⁺ CD19 ^- - 1.19 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.55 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.39 × 10 ⁹ / l, CD3 ^- CD19 ⁺ - 0.45 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) - 0.49 × 10 ⁹ / l, immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.41, on the 21st day of the menstrual cycle, CD3 ⁺ CD19 ^- - 1.42 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ - 0.56 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ - 0.45 × 10 ⁹ / l, CD3 ^- CD19 ⁺ - 0.52 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) 0.42 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.33 predict infertility.

After examination, the patient underwent pathogenetic therapy for 5 months with recommendations on the use of barrier methods of contraception. 7 months after treatment, pregnancy occurred. Ended in the birth of a full-term baby.

Example 2

Patient S., 23 years old.

Complaints at the time of the examination: does not show.

History: menstruation from 11 years, 5-7 days, after 55-80 days, irregular. Regarding menstrual irregularities, she received combined oral contraceptives for 3 years. Sexual life since 20 years. Before taking combined oral contraceptives, she was not protected from pregnancy. Pregnancy did not occur. In the puberty period, I applied to the children's gynecologist at the Children's City Polyclinic No. 1 JSC. After the examination, the diagnosis was made: Hypothalamic dysfunction of the puberty period. Primary oligomenorrhea.

Objectively: BMI - 34.7 kg / m ² , nacreous stria in the thighs and buttocks, mammary glands. Clinical diagnosis: Primary oligomenorrhea. Complicated gynecological history (dysfunction of the hypothalamus in the puberty).

The patient is recommended to cancel combined oral contraceptives. 2 months later, after the abolition of combined oral contraceptives, a survey was conducted. The content of lymphocyte subpopulations in the peripheral blood on the 5th day of the menstrual cycle was: CD3 ⁺ CD19 ^- - 1.16 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ - 0.54 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ - 0, 38 × 10 ⁹ / l, CD3 ^- CD19 ⁺ - 0.42 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) - 0.40 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1, 42, on the 21st day of the menstrual cycle: CD3 ⁺ CD19 ^- 1.45 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.57 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.43 × 10 ⁹ / L, CD3 ^- CD19 ⁺ - 0.42 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) - 0.44 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.32. In the endometrium, on the 5th day of the menstrual cycle, the content of lymphocyte subpopulations was: CD3 ⁺ CD19 ^- - 1.18 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.55 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.39 × 10 ⁹ / l, CD3 ^- CD19 ⁺ - 0.44 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) - 0.47 × 10 ⁹ / l, immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.41, on the 21st day of the menstrual cycle, CD3 ⁺ CD19 ^- - 1.43 × 10 ⁹ / L, CD3 ⁺ CD4 ⁺ - 0.57 × 10 ⁹ / L, CD3 ⁺ CD8 ⁺ - 0.45 × 10 ⁹ / L, CD3 ^- CD19 ⁺ - 0.50 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) from 0.45 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ - 1.35 predict infertility.

After the examination, pathogenetic therapy was prescribed for 6 months. One month after treatment, pregnancy occurred. Ended in the birth of a full-term baby.

The proposed method achieves early prognosis of infertility, i.e. before pregnancy planning, which allows timely correction of the revealed violations and risk assessment and accelerate the implementation of the generative function and reduce money for therapy.

Claims (1)

A method for predicting infertility in women with hypothalamic syndrome, namely, that in the peripheral blood and aspirate from the uterine cavity in women of reproductive age with hypothalamic syndrome and a body mass index greater than or equal to 30, lymphocyte subpopulations CD3 ⁺ CD19 ^- , CD3 ⁺ CD4 ^{+ are} determined , CD3 ⁺ CD8 ⁺ , CD3 ^- CD19 ⁺ , CD3 ^- CD (16 ⁺ 56 ⁺ ) and the immunoregulatory index - CD4 ⁺ / CD8 ⁺ depending on the day of the menstrual cycle, namely on days 5 and 21, and when the subpopulations of lymphocytes are 5th day of the menstrual cycle in the peripheral blood and CD3 ⁺ CD19 ^- from 1.08 to 1.19 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.51 to 0.55 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.35 to 0.40 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.38 to 0.47 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) from 0.37 to 0.43 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ 1.45-1.37, on the 21st day of the menstrual cycle in the peripheral blood CD3 ⁺ CD19 ^- from 1.20 to 1.51 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.56 to 0.65 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.41 to 0.45 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.38 to 0.46 × 10 ⁹ / l, CD3 ^- CD (16 ⁺ 56 ⁺ ) from 0.38 to 0.44 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ 1.36-1.22, in the endometrial aspirate on day 5 of the menstrual cycle CD3 ⁺ CD19 ^- from 1.1 to 1.22 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.54 to 0.58 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.36 to 0.42 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.40 to 0.49 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) from 0.45 to 0.54 × 10 ⁹ / l, immunoregulatory index - CD4 ⁺ / CD8 ⁺ 1.59-1.21, on the 21st day of the menstrual cycle in endometrial aspirate CD3 ⁺ CD19 ^- from 1.22 to 1.53 × 10 ⁹ / l, CD3 ⁺ CD4 ⁺ from 0.54 to 0.57 × 10 ⁹ / l, CD3 ⁺ CD8 ⁺ from 0.43 to 0.47 × 10 ⁹ / l, CD3 ^- CD19 ⁺ from 0.49 to 0.55 × 10 ⁹ / l, CD3 ^- CD (16 ^- 56 ⁺ ) from 0.39 to 0.46 × 10 ⁹ / l immunoregulatory index - CD4 ⁺ / CD8 ⁺ 1.34-1.42 predict infertility in women with hypothalamic syndrome.

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