Anti-endometrial and anti-endothelial auto-antibodies in women with endometriosis

The present study investigates the specificity of anti-endometrialantibodies present in serum of women with endometriosis. A twocolour indirect immuno-histochemical method was used, so thatthe antigenic reactivity of endogenous immunoglobulins was blockedand specific anti-endometrial antibodies were readily distinguishable.Serum was collected from women with endometriosis, before andafter 6 months of treatment, and from women without the disease.The reactivity of serum with uterine and ectopic endometriumfrom women with and without the disease was studied. The frequencyof anti-endometrial antibodies in the serum of women with endometriosiswas higher (P < 0.001) than in control sera. Most antibodiesspecifically reacted with glands in ectopic and uterine endometrium.Antibody reactivity was strongest with endometrium from controlwomen, compared with uterine and ectopic endometrium from womenwith endometriosis (P < 0.01). A proportion of sera containinganti-endometrial antibodies also reacted with vascular endothelium.Binding was strongest to vessels in uterine and ectopic endometriumfrom women with endometriosis compared to endometrium from womenwithout the disease (P < 0.01). The presence of anti-endometrialantibodies was associated with infertility.     

Immunology: Bone marrow-derived cell populations in uterine and ectopic endometrium

Uterine endometrium contains numerous bone marrow-derived cells.The spectrum of cell types is different from that of any othertissue, and the differences in endometrium from women with endometriosismay reflect a different endometrial phenotype in these women.The cell types of bone marrow origin found in ectopic endometriummay indicate the degree of differentiation of the tissue. Itwas found that, in normal endometrium, the CD45⁺ cell populationcomprised T cells, macrophages, CD56⁺ large granular lymphocytes,some CD16⁺ cells and a few B cells. Changes in these cell populationsduring the menstrual cycle were similar in endometrium fromboth controls and patients with endometriosis, and resembledthat reported previously by others. In ectopic endometrium,the frequency of CD45⁺ cells remained within the same rangeas that of uterine endometrium but without any obvious patternof change during the menstrual cycle. CD56⁺ large granular lymphocytes,an immune cell type characteristic of uterine endometrium, werealso found in ectopic endometrium. Our results indicate thatectopic endometrium, as well as comprising both glandular andstromal cells, contains bone marrow-derived cell populationssimilar to those of uterine endometrium. This suggests thatthe same processes of cell migration and/or differentiationoccur in ectopic and uterine endometrium.     

Anti-endometrial antibodies in women measured by an enzyme-linked immunosorbent assay

An enzyme-linked immunosorbent assay (ELISA) was developed tomeasure anti-endometrial antibody concentrations in the serumof women with endometriosis. Pooled cytosolic protein extractsfrom the endometrial gland cells of 10 women were used as anantigen source. Serum samples were obtained from women withendometriosis before (n = 51) and after 6 months treatment withdanazol or nafarelin (n = 30). Control sera came from womenwith a normal pelvis at laparoscopy, performed for sterilization(n = 23) or the investigation of pain and/or infertility (n= 22), 13 women with Rokitansky syndrome, and 10 umbilical cordbloods and adult males. There were no significant differencesin serum anti-endometrial antibody concentrations before andafter treatment, or between women with endometriosis and withoutendometriosis. Concentrations were lower in male and cord bloodserum than in female's serum (P < 0.0001). We conclude thatthe ELISA is not a useful diagnostic tool for endometriosisunless more specific antigens can be isolated.     

Peritoneal fluid cytokines and the relationship with endometriosis and pain

It is generally accepted that the current scoring system forendometriosis has little correlation with clinical symptomssuch as pain, and therefore we may deduce that either endometriosisdoes not cause pain, or that the current scoring system doesnot indicate the biological activity of the disease. Pain mayoccur because the presence of endometriosis produces an intraperitonealinflammatory response, and several studies have shown that thecytokine content of peritoneal fluid differs between women withand without endometriosis. We studied the relationship betweentumour necrosis factor a (TNFa), platelet-derived growth factor(PDGF), interleukin (IL)-6, IL-4 and TNF (a and P) activityin peritoneal fluid and the clinical history of pain and infertility.TNFa concentrations were increased in peritoneal fluid of womenwith endometriosis and of infertile women; PDGF concentrationswere increased in peritoneal fluid of parous women; EL-6 wasincreased in peritoneal fluid of women with adhesions; IL-4was absent from peritoneal fluid. PDGF and IL-6 concentrationswere cycle related, with the highest amounts in the menstrualand proliferative phases respectively. We failed to demonstrateany association between concentrations of cytokines in vitroand pain symptoms or severity of endometriosis.     

Expression of intermediate filament in endometrial glands changes with the onset of pregnancy and in endometriosis.

Appropriate endometrial differentiation is believed to be a prerequisite for pregnancy success. This study investigates the expression of two intermediate filament proteins, cytokeratin and vimentin, in human endometrium and first trimester decidua and in ectopic endometrium from women with endometriosis. Stromal elements, including vascular endothelial cells, were consistently vimentin-positive and cytokeratin-negative. Surface and glandular epithelial cells of human endometrium co-expressed vimentin and cytokeratin during all stages of the menstrual cycle, but failed to express vimentin after the onset of pregnancy. This suggests that intermediate filaments, and especially vimentin, may have a role to play in the proliferation and/or differentiation of the endometrial glands during decidualization. Ectopic endometrium showed a staining pattern similar to normal endometrium.     

Vitrification can modify embryo cleavage stage after warming. Should we change endometrial preparation?

Purpose

Studies have shown that embryo metabolism and cell cleavage after warming vitrified embryos is faster than after thawing frozen embryos. We study vitrified embryo transfer (VET) results depending on the developmental stage of warmed embryos and the duration of progesterone treatment before embryo transfer.

Methods

We designed a prospective study, patients were randomized in two groups, starting progesterone three (D + 3) or four days (D + 4) before embryo transfer. We recruited 88 patients with embryos vitrified on day 3.

Results

We didn’t find statitistical differences in pregnancy rate when we transferred embryos in D + 3 vs D + 4 (38.2 % vs 40.5 % p ≥ 0.05). The day after warming, 54.6 % of embryos had developed to morula or early blastocyst, 32.4 % to cleavage stage and 13 % didn’t cleave. Transfers were with morula/blastocysts stage embryos (52.1 %; n:37), cleavage stage embryos (18.3 %; n:13) or mixed (29.6 %; n:21). Implantation rate was significantly higher in morula/blastocyst stage than in cleavage stage or mixed transfers (44 %, 22 % and 16.3 %; p = 0.011). Pregnancy and implantation rates were significantly higher in morula/blastocyst transfers on D + 4 than on D + 3 (68.7 % and 64.7 % vs 33.3 %, and 33.3 %, p = 0.033 and p = 0.034).

Conclusions

Our findings suggest that a majority of embryos will develop to morula/blastocyst stage after warming. VET results with morula/blastocysts, and after four days of progesterone supplementation, are better than with cleavage stage embryos.