NK cells not only in endometrium but also in ovulatory cervical mucus in patients with decreased fertility.

OBJECTIVE: An increased number of NK cells is associated with autoimmune disorder and is known to play a role in infertility. The aim of our research was to monitor the density of NK cells CD56+ and CD16+ in ovulatory cervical mucus (OCM) and in endometrium in infertile women as well as in connection with the actual status of antibodies against phospholipids, sperm and HHV-6 antibodies. TYPE OF STUDY: Original aticle. SETTING: Genetika - Plzeň. METHODS: Seventy-two randomly selected women aged 20-39 (mean age: 32.3) years old resulted in fifty-seven patients with repeated unexplained miscarriages, and fifteen fertile healthy women. The hormonal status was studied including ovulation, the humoral autoimmune responses to eight phospholipids, trombophilia, karyotyping, hysteroscopy, and endometrium immunohistology. Patients were without any clinical and laboratory symptoms of vaginitis at the time of OCM sampling and endometrium study. In one patient antiphospholipid syndrome was present, and in one woman diabetes mellitus was identified. Uterine NK cells CD56+ , CD16+ and NK cells in OCM were identified by immunocytochemistry, antiphospholipid antiboides by ELISA. We used indirect MAR-test for study of local spermagglutinating antibodies in OCM. Indirect immunofluorescent method was used for detection of serum and OCM IgM, IgG antibodies against HHV-6 levels at the time of ovulation. RESULTS: We found both high density of NK cells CD56+ and CD16+ in OCM and in endometrium in only two infertile women with repeated abortions. NK cells in OCM were missing in other samples of patients. The prevalence of high density of NK cells CD56+ in the endometrium was seen in twenty three (40%), NK cells CD16+ in eleven (19%), NK cells 56+ and NK cells 16+ together in eight (14%). Levels of serum and OCM IgG against HHV-6 in all examined patients were not elevated, no cervical sperm antibodies were found. CONCLUSION: We compared density of NK cells CD56+ and CD16+ in OCM and secretory endometrium in all infertile patients. Our results show that cell mucosal activity in the cervical area at the time of ovulation in two infertile patients was evident. We excluded the abnormal number of NK cells owing to local and general viral infection (HHV-6). But our question still remains - are cervical NK cells fixed or still migrating from endometrium into OCM? New research is planned.

[Results of the treatment in selected infertile patients with high density of endometrial NK cells CD56+ and CD16+ Second part]. / Výsledky lécby u vybraných infertilních pacientek s vysokou denzitou endometriálních NK bunek CD56+ a CD16+ Druhá cást.

INTRODUCTION: This statements follows the first part of our publication entitled Screening of endometrial NK cells in selected infertile patients (First part - Methods and current results), where we dealt with methodology and endometrial findings of the natural killer cells CD16+ and CD56+. These cells are among the most important in preimplantaion, and in implantation period, in early pregnancy too, but can also negatively influence above mentioned processes. OBJECTIVE: We focused on the immunomodulatory treatment with intramuscular immunoglobulins in 21 infertile patients with a high density of endometrial NK cells CD16+ and CD56+. DESIGN: Original work-prospective study. SETTING: Genetics-Pilsen. METHODS: From the originally investigated 57 women aged 25-41 (average 34) years with their history of 3-9 abortions, we target on a high selective subgroup of 21 infertile patients with pathological density of endometrial NK cells CD56+ and CD16+. RESULTS: Twelve patients underwent immunomodulatory treatment with intramuscular immunoglobulin in time of positive HCG and went on the therapeutic scheme, nine infertile women continued with IVF-ET but did not achieve HCG positivity despite of initial treatment with immunoglobulins. Of the 12 treated patients, seven of them gave a birth to healthy babies, as to the next five pregnancies - one was extrauterine pregnancy, one genetically defective fetus, three pregnancies go on successfully.Nine women from the IVF program did not get pregnat. CONCLUSION: Immunomodulatory treatment with immunoglobulins also influences endometrial NK cells CD56+ and CD16+ associating with an overproduction of embryocytoxic cytokines. Treatment of our patients with the pathological endometrial density of NK cells must be solved strictly individually.

[Screening of endometrial NK cells in selected infertile patients First part - Methods and current results]. / Screeningové vysetrení endometriálních NK bunek u vybraných infertilních pacientek 1. cást - metodika a prubezné výsledky.

INTRODUCTION: Generaly, natural killer cells (NK cells) are among the most important cells of our immune defense system. They are present in the blood, decidua but also in secretory endometrium. OBJECTIVE: We investigate an association between high density of NK cells CD56+ and NK cells CD16+ into blood and secretory endometrium in patients with up to now unexplained recurrent miscarriage. At the same time, we focused on diagnosis of antiphospholipid syndrome, congenital trombophilic factors, stress factor, and lenght of hormonal contraception before conception. DESIGN: Original work-prospective study. SETTING: Genetics-Pilsen. METHODS: We investigated 59 patients aged 25-41 (average 34) years with their history of 3-9 abortions. We monitored their blood cells by flow cytometry and endometrium obtained by hysteroscopy performed on the 22nd-24th day of the menstrual cycle. We concentrated on the cellular immunity focused on the identification of lymphocytes CD56+ and CD16+. ELISA method was used for identification of antiphospholipid antibodies. RESULTS: Our study goes on, we are publishing our preliminary results. We found a high density of endometrial NK cells CD56+ in 41/57, NK cells CD16+ in 40/57, both NK cells CD56+ and CD16+ in 36/57 patients. In 13 women, we identified only sporadically these cells in the secretory endometrium. NK cells CD56+ and NK cells CD16+ in the blood were all normal number in all our patients. Three of them have a homozygot form of Leiden mutation, and 18/57 primary antiphospholipid syndrome, 12 women have university education, four of them, medical doctors, have regular night services in the hospital. CONCLUSION: Overproduction of endometrial NK cells is associated with increased local activity of embryocytotoxic cytokines that may negatively affect pregnancy. Treatment of patients with pathological immunological findings must be solved individually, as we will show soon in part two of our study.

[Organ-specific antibodies in children with idiopathic inflammatory bowel disease and their use in clinical practice]. / Orgánove specifické protilátky u detí s idiopatickými strevními zánety a jejich uzití v klinické praxi.

BACKGROUND: Ethiopathogenesis of the idiopathic inflammatory bowel disease has not been yet fully explained. Several abnormalities of the humoral immunity supporting the concept of autoimmune character of the disease have been identified. The aim of our study was to characterise occurrence of the organ specific antibodies against the intestinal goblet cells and against acinar cells of the exocrine pancreatic tissue and to evaluate their significance for the diagnostics of the idiopathic inflammatory bowel disease. METHODS AND RESULTS: 69 children were included in the study. The group consisted of 20 patients with idiopathic proctocolitis (11 boys and 9 girls, 6 to 18 years old, average age 15.5) and 14 patients with Crohn's disease (9 boys and 5 girls, 5 to 18 years old, average age 14.7). Control group included 35 children (20 boys and 15 girls, average age 14.7). In patients of the idiopathic proctocolitis group, antibodies against the intestinal goblet cells were assayed by indirect immunofluorescence method in 55%. In patients with Crohn's disease, antibodies against acinar cells of the exocrine pancreatic tissue were found in 64.2%. Differences in manifestation of antibodies against acinar cells of the exocrine pancreatic tissue in Crohn's disease were statistically significant (p = 0.001). Statistically significant (p = 0.01) was also the difference of levels of antibodies against the intestinal goblet cells in patients with idiopathic proctocolitis when compared to patients with Crohn's disease. Statistically significant difference (p = 0.01) was found in levels of antibodies against acinar cells of the exocrine pancreatic tissue in Crohns disease and antibodies against the intestinal goblet cells in patients with idiopathic proctocolitis. CONCLUSION: Testing on presence of specific antibodies against acinar cells of the exocrine pancreatic tissue and against intestinal goblet cells is a valuable tool for the diagnostics of the idiopathic inflammatory bowel disease in childhood and adolescence.

[Does therapeutic membrane plasmapheresis increase protein synthesis?]. / Urychluje lécebná membránová plazmaferéza proteosyntézu?

The objective of the investigation was to assess whether therapeutic membrane plasmapheresis accelerates protein synthesis. To this end pseudouridine (PSI), a modified nucleoside was investigated which provides information on the tRNA turnover and thus indirectly also on protein synthesis. The authors made 10 plasmapheresis on a A 2008 PF monitor with Plasmaflux P2 filters which they use to exchange 1 plasma volume of the patients. Laboratory indicators were investigated one day before plasmapheresis, on the day of plasmapheresis and during its course, and on the 1st, 2nd and possibly 3rd day after plasmapheresis. They revealed that the clearance of the plasma filter for PSI (0.41 +/- 0.04 ml/s, arithmetical mean +/- SEM) did not differ significantly from the filtration rate (0.49 +/- 0.01 ml/s, p = 0.15). As compared with the initial examination (0.49 +/- 0.06 ml/s) on the first day after plasmapheresis as a result of reduced glomerular filtration rate the renal clearance of PSI was reduced (0.33 +/- 0.05, p less than 0.01). PSI serum concentrations were therefore expressed as the serum PSI/serum creatinine ratio. This ratio was, as compared with the initial examination (80.4 +/- 4.8 nmol/mumol), raised midway during the procedure (100.8 +/- 8.6, p much less than 0.05) and after its termination (132.3 +/- 6.1, p much less than 0.01). The increase was not due to disintegration of cells or dietary factors. The rise of the serum PSI/serum creatinine ratio was due to a more rapid tRNA turnover and thus provided evidence that therapeutic membrane plasmapheresis accelerates protein synthesis.