1. Reproductive Immunology
Prof. MUDr. Zdenka Ulcova-Gallova, DrSc.
Genetics –Pilsen and Department of Gynecology and Obstetrics of Charles University, Pilsen, Czech Republic

3. Screening for reproductive immunology (RI)
Sperm antibodies
Zona pellucida antibodies
Panel of various antiphospholipid antibodies (eight)
Embryocytotoxic cytokines
Intraacrosomal proteins
Endometrial Ab
Antitrophoblast antibodies
aTSH, aTPO
HLA-I a HLA II
HLA-G
AMH
Inhibin (A, B)
Local CD56+/CD16+
Detailed immunological profile according to basic dg.(CD56/CD16)

4. RI
Men Women Both

5. Immunological origin in unknown/known cause of infertility
Repeated spontaneous abortion or missed abortion
Repeated IVF failure
Patients with autoimmune or oncological diseases
Endometriosis, preeclampsia, PCO, POF, HELLP…..
Andrological factor

6. Zona pellucida antibodies
Sperm antibodies

7. Anti-sperm antibodies (ASA)
History
Incidence in men, women
Methods of measurement
„Treatment“

8. Rich history of ASA
Animals: by Landsteiner and Metchnikoff Metalnikoff
1932 Baskin
Human studies:Rumke 1954, Bronson, Shulman (1991), Dondero, Hendry, Fjallbrandt, Hjort, Isojima, Mori, Koyama,Kutteh, Moghissi, Nakov, Bratanov, Lenzi, Mathur, Ingerslev, Clark,Mettler, Ulcova-Gallova (1985), Bohring, etc.
Multicentric study , comparison of various tests for detection of ASA

9. Origin of ASA in men Damage of blood testis-barrier
Genetic predisposition HLA A-28, B-22

10. Origin of ASA in women Ascendent immunisation
(but other way is possible)

11. Sperm antibodies inhibit from:
Sperm transport
Bloking ZP penetration
Sperm capacitation
Acrosomal reaction
Early fertilization
Postfertilization effect

12. Examination
Serum
Semen
OCM
Peritoneal fluid
Folicular fluid

13. Detection of free ASA Gelatin agglutination tests (GAT-Kibrick)
Tube-Slide Agglutination test
Tray agglutination test (TAT-Friberg)
Sperm-immobilization test(Isojima)
Mixed reaction anti-immunoglobulin tests (MAR-test)
Immunobead technique
ELISA (poor correlation with infertility)
RIA

14. ASA
IgG
IgM
IgA, s IgA
(i IgE)

15. ASA and significance Sperm-agglutinating Sperm-immobilising
Sperm-cytotoxic

16. Prof. Isojima S.
Publication
Science
AJRI,

17. Kibrick test
Shulman,1985

18. Capillary test (Fjallebrandt)
Shulman,1985

19. Sperm-agglutination head to head -TAT

20. Direct /indirect MAR test or immunobead test

21. OCM The Miller-Kurzrok Test) The Sperm Cervical Mucus Contact Test
Kremer test

22. Kremer test

23. Antizonal antibodies (aZP)
Block sperm adherention and sperm penetration into oocyte
Postfertilization effect

24. Zona pellucida Animal models
(Oikawa, Yanagimachi, Kamada, Mori, Aitken, Hasegawa, Koyama, Isojima, Inoue, Yamasaki, Clark, et al.)
Importance of ZP
ZP epitopes

25. ZP
Human ZP X porcine ZP
(Sacco, 1977)

26. AJRI
Ulcova-Gallova Z., Mardesic T.: Does In Vitro Fertilization ( IVF ) Influence the Levels of Sperm and Zona Pellucida ( ZP ) Antibodies in Infertile Women? Amer. J. Reprod. Immunol. 1996, 36, p
Very important!!!

27. ZP antigens Porcine oocytes
ZP1 –ZP4 epitopes (60 kDa, Hasegawa et al. 1995)
Mice –ZP 1- ZP3
Monoclonal antibodies

29. Koyama, Hasegawa, Mochida, Colongos, 2005-2015
Induction of follicular dysfunction by ZP
E.g. ↑aZP in POF
Et al.

30. Examination of aZP antibodies
Passive haemmagglutination tests
ELISA

33. Antiphospholipid antibodies
Interference with membrane phospholipid
1. Prae-conceptionally
2 .Post-conceptionally

34. APA (IgG, IgA, IgM) Ph-L-serine ph-L-glycerol ph-L-inositol
ph-ethanolamine -annexin V
ph-acid
cardiolipin
b2-GP

35. Criteria of APA sy - are not sufficiant for infertility In general:
aPls
Thrombosis (thrombocytopenie?)
Failure in reproduction

36. For instance

37. Analysis of 2965 infertile women (Ulcova-Gallova et al
Analysis of 2965 infertile women (Ulcova-Gallova et al.AJRI, 2005 Aug;54(2):112-7.)
P < 0,001

38. For instance
Immuno-histological detection a-annexin V Ab

39. Červeně: apoptotic.buňky
Zeleně: mrtvé buňky

40. Detection of intracrosomal enzymes
IF (monoclonal antibodies)

42. Immunological tolerance
Mother X foetus

43. Dr. Peter Medawar a prof.Milan Hasek

44. Nidation and implantation

45. Mother -immunotolerance
Barier of decidua
Blockade antibodies
Immunotolerance to foetus antigens
Pregnancy specific proteins (Bohn p.)
HCG, E, P
Anti-idiotype antibodies
Immunosuppressive cells (spec., non-sp.)
Other mechan.

46. Foetus -immunotolerance
Throphoblast bariere of placenta
Sialomucinglycocalix layer
Low expression of MHC
Damping elements:e.g. α FP, PGE, …
C system

47. Ab -detection
Anti-foetus
Anti-placental
Anti-paternal

48. Development of a particular immune response which enhances embryo acceptance ?
Th2
Th2 RESPONSE
ΑΝΤΙΓΟΝΑ ΤΡΟΦΟΒΛΑΣΤΗΣ
HLA-G; CD46; R80k; CD1; ;
«ÔÇ2»
Recognition
by decidual cells
Trophoblast antigens
HLA-G? CD46? R80k? CD1?
γδTcells?
FACILITATION
REACTION
IMMUNOTROPHISM
growth, maturation
of the placenta
Blocking
antibodies
BLOCKAGE
of cytotoxic reactions
Immunoabsorbent placenta
Trophoblast resistant
Local hormone-dependent immunosuppression
to NK cell lysis

49. Rh-incompatibility (ery-C,D,E,c,d,e)
Rh negative mother
Rh positive foetus
Treatment: anti D (to 72 hours)
ABO –incomp.very rare
Keely, Lewin etc.

50. CAVE! - foetus erytroblastosis 1. Anaemia neonatorum
2. Icterus neonati gravis
3. Hydrops fetus universalis

51. 15-20% of infertile couple
Clear immunological cause of infertility or combine with other causes

52. FERTILITY and RI
biologic aging of the hypothalamic-pituitary-ovarian axis in elder women
→changes in endocrinology, immunity, psychology
Tests in RI withe reason

53. Immunological screening for reproductive failure
Sperm antibodies
Zona pellucida antibodies
Panel of various antiphospholipid antibodies (eight)
Embryocytotoxic cytokines
Intraacrosomal proteins
Endometrial Ab
Inhibin (A, B)
Antitrophoblast antibodies
aTSH, aTPO
HLA-I a HLA II
HLA-G
AMH
Local CD56+/CD16+
Detailed immunological profile according to basic dg.(CD56/CD16)

54. Other future in RI
Antigens of Sperm, ZP, beta HCG etc. - contraceptive vaccine
Ways, means of influencing of fertility