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

Czech version

Authors: Z. Ulčová-Gallová ^1,2; P. Mukenšnabl ³; M. Haschová ⁴; M. Pešek ⁵; P. Chaloupka ⁵; P. Lošan ¹; K. Bibková ¹; Z. Mičanová ¹; J. Cibulka ¹; M. Švecová ⁶
Authors‘ workplace: Reprodukční imunologie, Genetika – Plzeň, přednosta MUDr. P. Lošan ¹; Gynekologicko-porodnická klinika LF UK a FN, Plzeň, přednosta doc. MUDr. Z. Novotný, CSc. ²; Šiklův anatomicko-patologický ústav LF UK, Plzeň, přednosta prof. MUDr. M. Michal ³; Oddělení klinické biochemie a imunologie, Nemocnice Privamed, Plzeň, přednosta Ing. Z. Jeřábek ⁴; Gynekologicko-porodnické oddělení, Nemocnice Privamed, Plzeň, přednosta MUDr. Z. Knaizl ⁵; Oddělení mikrobiologie a virologie, FN Plzeň, přednostka MUDr. M. Švecová ⁶
Published in: Ceska Gynekol 2019; 84(3): 184-189
Category: Original Article

Overview

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.

Keywords:

endometrium – infertility – NK cells – ovulatory cervical mucus – HHV-6

Sources

1. Bulmer, JN., Morrison, L., Longfellow, M., et al. Granulated lymphocytes in human endometrium: histochemical and imunohistochemical studies. Hum Reprod, 1991, 6, p. 791–798.

2. Crha, I., Ventruba, P., Žáková, J., et al. Děložní mikrobiom jako faktor receptivity endometria. Čes Gynek, 2019, 84(1), s. 49–54.

3. Coulam, CB., Bilal, M., Salazar Garcia, MD., et al. Prevalence of HHV-6 in endometrium from women with recurrent implantation failure. Am J Reprod Immunol, 2018, 80(1):e12862. doi: 10.1111/aji.12862. Epub 2018 Apr 17.

4. Coulam, C. Recurrent Implantation Failure. 15th International Symposium for Immunology of Reproduction, Varna, Bulgaria, 2018, June 15–17, abstr.p. 17.

5. Dvořan, M., Vodička, J., Dostál, J., et al. Implantace a diagnostika receptivity endometria. Čes Gynek, 2018, 83(4), s. 291–299.

6. Eggert-Kruse, W., Botz, I., Pohl, S., et al. Antimicrobial activity of human cervical mucus. Hum Reprod, 2000, 15, p. 778–784.

7. Hansen, LK., Becher, N., Glavind, J., et al. The cervical mucus plug inhibits, but does not block, the passage of ascending bacteria from vagina during pregnancy. Acta Obstet Gynecol Scand, 2014, 93, p. 102–108.

8. Kuon, RJ., Vomstein, K., Weber, M., et al. The „killer cell story“ in recurrent miscarriage: Association between activated peripheral lymphocytes and uterine natural killer cells. J Reprod Immunol, 2017, 119, p. 9–14.

9. Kwak-Kim, J., Bao, S., Lee, SK., et al. Immunological modes of pregnancy loss: inflammation, immune effectors, and stress. Am J Reprod Immunol, 2014, 72, p. 129–140.

10. Mirmonsef, P., Gilbert, D., Zariffard, MR., et al. Effect of commensal bacteria on innate immune responses in the female genital tract. Am J Reprod Immunol, 2011, 65, p. 190–195.

11. Seshadri, S., Sunkara, SK. Natural killer cells in female infertility and recurrent miscarriage: a systematic review and metaanalysis. Hum Reprod Update, 2014, 20, p. 429–438.

12. Tang, AW., Alfirevic, Z., Quenby, S. Natural killer cells and pregnancy outcomes in women with recurrent miscarriages and infertility: a systemic review. Hum Reprod, 2011, 26, p. 1971–1980.

13. Ulcova-Gallova, Z. Immunological and physicochemical properties of cervical ovulatory mucus. J Reprod Immunol, 2010, 86(2), p. 115–121.

14. Ulcova-Gallova, Z. Ten-years experience with antispermatozoal activity in ovulatory cervical mucus and local hydrocortisone treatment. Am J Reprod Immunol, 1997, 38(3), p. 231–234.

15. Ulcova-Gallova, Z., Pesek, M., Mukensnabl, P., et al. [Screening of endometrial NK cells in selected infertile patients. First part -⁠ Methods and current results.] Ces Gynek, 2017, 82(5), p. 366 –371.

16. Vassiliadou, N., Searle, RF., Bulmer, JN. Elevated expression of activation molecules by decidual lymphocytes in women suffering spontaneous early pregnancy loss. Hum Reprod, 1999, 14(5), p. 1194–2000.