pregnant woman holding her belly and yellow flower

Dr. Ioannis Toliopoulos, PhD, Post Doc, Molecular Biologist, Physiologist, Immunologist in Reproductive Medicine

The huge issue of infertility rises day by day in national and international level, especially nowadays where civilization and technology has advanced gradually. On the other hand, all this evolution has provoked serious issues in terms of fertility in humans. The parameters that concern this issue are many such as genetics, but more important nowadays are considered to be exogenic and endogenic factors that influence the genetic material in husband and wife. In these cases, in a large percentage the wife’s immune system seems to be affected majorly and as a result her reproductive system. In addition, the major issue in cases of infertility, RSAs (rapid spontaneous abortions, and IVF failures, has to be the male factor, which is the sperm and has to be excluded so that the wife can’t be accused for infertility. The parameters that should be tested for sperm are sperm number, vitality, motility, and DNA fragmentation, where the diagnostic and therapeutic approach is estimated individually. Moreover, the diagnostic evaluation and the therapeutic approach must be consulted by specialized scientists. With the exclusion of sperm as an infertility reason, the wife does not get prosecuted somatically and psychologically.

On the other hand, biological specific markers as NK cells, NK activation, as well as embryotoxicity (ETA) influence in high levels woman’s fertility and the evolution of pregnancy until the first trimester. The results below are an important step of immune regulation of couples that face infertility, RSAs and IVF failures.

1) Couple (woman 38 years old, husband 42 years old) that had been in IVF procedure 6 times since 2009, DNA fragmentation of husband was measured 25 % and in wife the markers that were tested were MHTFR 677 C>T (heterozygous in thrombophilia, AMH and inhibin B (ovary reservation panel) were normal, while the immunological factors and NK activation and embryotoxicity were not normal. Specifically, biological therapy was given to the husband, while in wife intralipid, baby aspirin and IVIg were administered for certain time. After 6 months within 2013, the wife was pregnant and the couple has a baby today.

2) Couple (woman 33 years old, husband 39 years old) had 3 RSAs in 4th and 5th week of gestation.  DNA fragmentation was tested for husband, which was normal (9%). Wife was diagnosed for Hashimoto’s thyroiditis with high levels of anti-TPO and anti-TG. Also, levels and NK activation were high and abnormal respectively, while embryotoxicity was negative. The therapeutic approach was intralipid, baby aspirin and cortisol and the lady had a successful pregnancy 4 months after and today the couple has a baby.

3) Couple (woman 44 years old, husband 46 years old) faced infertility and last 10 years and one failed IVF attempt. DNA fragmentation of the husband was checked which was 23%. In wife, AMH and Inhibin B were tested which were out of normal range, and NK levels were normal, but NK activation was high, while embryotoxicity was negative.  For the sperm, biological therapy was applied, while in wife biological supplements were administered for the regulation of the phyco-neuro-endocrine-immune system, and intralipid 2 weeks before the embryo transfer in IVF attempt with donor egg. The woman became pregnant and has a healthy baby today.

4) Couple (woman 31 years old, husband 34 years old) with infertility issue the last 4 years. The wife was diagnosed with one salpynx after an ectopic pregnancy from the past. DNA fragmentation of the husband was normal, while wife was diagnosed for heterozygous thombophilia in mutations GP IIb/IIIa A1/A2 and PAI-I 4G/5G, as well as increased NK activation. The markers of egg quality were normal as well as the levels of NK cells and embryotoxicity. Intralipid was administered and baby aspirin and pregnancy was detected after 8 months and today this couple has a child.

5) Couple (woman 32 years old, husband 43 years old) with 5 IVF failed attempts.  DNA fragmentation was tested which was 43%, which seemed that the sperm was the main factor for the failed pregnancy. In wife, markers of egg quality were normal, but NK levels and NK activation were high.  Embyotoxicity was negative.  Biological therapy for the sperm was given for 1.5 month and the DNA fragmentation was tested again and was 25%, which means that husband has more chances to participate in successful pregnancy. Also, intralipid was administered and IVIg to wife for the next 3 months and still trying naturally without IVF because of the age of wife. This case here is stated in order to stress the importance of the diagnostic tests before a couple proceeds to IVF procedure.

6) Woman 51 years old, who was in menopause period three years ago and husband (same age with good sperm), was tested for NK levels that had been increased as well as their activation. She went on intralipid therapy, she became pregnant with donor egg, but she didn’t complete the therapy. In year 2013 (she received 2 doses before and 1 after conception), where she had spontaneous abortion. Later, she went under IVF again in summer 2014, she went on IVF with better preparation this time with intralipid and after the pregnancy test she is pregnant today with twins.

7) Woman 41 years old without salpinx (removed) and husband 45 years old without sperm problems. She has one child, one RSA in week 20. She performed 2 IVFs in 2012 and 2013 with embyo transfer, where both failed without conception. After a period of time, she performed the NK tests, and she received intralipid therapy (3 doses) and nowadays, she is pregnant.


Unexplained infertility of couple                                  Result=Pregancy with a baby
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 30 years old ΝΚ subpopulation= normal 2 Doses 6 months
Without child ΝΚ activity=↑

Unexplained infertility for over 2 years. Wife with no certain problem after complete check up as well as the husband. Three sperm inseminations were performed with no result. After the immunological diagnosis the therapeutic approach (before IVF trials), pregnancy was achieved and the couple has a female baby they year 2013.


Rapid Spontaneous Abortions (RSAs)                                  Result=STILL ON THE ATTEMPT
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 30 years old ΝΚ levels = ↑ 1 Dose 2 months
Without child ΝΚ activity=

Two RSAs before laboratorial diagnosis. Wife without a certain problem after complete check up as well as the husband. After the diagnosis and the therapeutic approach (1 dose only was administered so therapy was incomplete because she stopped for her own reasons), pegnancy was achieved naturally, but she had an RSA again at the 10th week.  They tried with IVF attempts and immunological vaccinations in monthly doses and they had another pregnancy and another RSA the 8th week as a result.  Since then, the couple is still trying for conception until today.


Unexplained infertility of couple                                                                Result=Pregnancy
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 27 years old ΝΚ subpopulation= normal 1 Dose 13 months
Without child ΝΚ activity= normal

Unexplained infertility the last 3 years. Wife and husband had no certain problem after complete check up. Three sperm insemination were performed without the desired result. After the immunologic diagnostic and treatment, pregnancy was achieved naturally without any intervention one month before where the couple was determined to start the IVF process.


Unexplained infertility of couple                                             Result=Divorce of the couple
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 33 years old ΝΚ subpopulation= ↑ 2 Doses NEVER
Without child ΝΚ activity= ↑
ETA= normal

This couple had infertility problem 4 years ago. Woman with endometriosis went under surgery 2 times, while the husband had no problem. Two therapies were performed, but she decided on her own to go to different gynecologist in order to get in 2 sperm inseminations and 3 IVFs with no results at all. The final result was the break of the marriage.


Unexplained infertility of couple                                               Result=Pregnancy with twins
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 31 years old ΝΚ subpopulation= normal 1 Dose 2 months
Without child ΝΚ activity= normal

This couple had unexplained infertility the last 4 years. Wife and husband had no certain problem after complete check up. Two stimulations of ovaries were performed with no sperm insemination and without certain result. After the immunological diagnosis and therapeutic approach pregnancy with twins was achieved with ovary stimulation (2 months before the scheduled attempt for IVF). Today, the couple is happy with the 2 female twins who were born in 2013.


Rapid Spontaneous Abortion (RSA)                                     Result=STILL ON THE ATTEMPT
AGE DIAGNOSIS April  2012 THERAPY Pregnancy time
Woman 31 years old ΝΚ subpopulation= normal NONE
Without child ΝΚ activity= normal

Three RSAs were reported for the lady before specific diagnosis. The lady had no problems and complete check up (only low AMH). The husband’s DNA sperm fragmentation was normal. She did not follow the individual therapy for her own reasons and since there no pregnancy was reported neither naturally or with IVF attempt. The couple continuous their IVF attempts.


Rapid Spontaneous Abortion (RSA)                                          Result=PREGNANCY with baby
AGE DIAGNOSIS January  2012 THERAPY Pregnancy time
Woman 33 years old ΝΚ subpopulation= normal First dose 3 months
Without child ΝΚ activity= normal

Two RSAs were reported two years ago before the diagnosis. The wife and husband had no diagnostic problem. After the 2 RSAs, the vaccine for blocking factors was performed from no specialized IVF center and there was no pregnancy detection for half a year. After the diagnosis and therapeutic approach with stimulating ovulation and natural contact, the couple today has a new born baby.


Unexplained infertility of couple                                     Result=STILL ON THE ATTEMPT
ρ AGE DIAGNOSIS June  2013 THERAPY Pregnancy time
Woman 36 years old ΝΚ subpopulation= normal
Without child ΝΚ activity=
ETA=Weak Positive

There was unexplained infertility for the last 8 years. Seven IVF attempts were performed and one only was successful, which led to RSA because of cervix insufficiency. After the diagnosis, the wife did not follow the suggested treatment (her personal choice). She proceeded to embryo transfer with frozen embryos, but again failed to be pregnant. Then she followed possibly the vaccine protocol without result. The couple is still trying even today for results.


Rapid Spontaneous Abortion (RSA)                                                   Result=PREGNANCY
AGE DIAGNOSIS January 2012 THERAPY Pregnancy time
Woman 36 years old ΝΚ subpopulation= ↑ 2 doses 1 month
Without child ΝΚ activity= normal

There were 2 RSAs the last year. Wife and husband without any serious problem. She followed the suggested therapy and after natural contact she became pregnant and she is in the 6th month.

All the above results are very satisfactory. On the other hand, they offer the couple exceptional diagnostic result, which investigates the reasons for infertility, RSAs, and failed IVFs and they don’t depend on the age, but rather in the regulation of the immune system. In other words, every couple should perform the diagnostic procedures consulted only from specialized doctors who will safe money for them and from many stressors.  The specialized scientists can guide the couple to follow the targeted diagnostic and therapeutic strategies in order to increase the chances of the couple to reach successful pregnancy, and to avoid the redundant test that may lead to confusion. By this way, the couple will know their reproductive issues, they won’t be tricked, they will know what to do at the right time and the will create their own chances to bring the next generations in life with no fears and no superstitious behaviours.,


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