Francesca Massenzio

Abnormal pattern of lymphocyte subpopulations in the endometrium of infertile women with chronic endometritis.

Problem  Endometrial lymphocytes play a critical role in endometrial receptivity. This study aimed at evaluating the variations induced by chronic endometritis (CE) on endometrial lymphocyte subsets. We compared the results in infertile women diagnosed with CE with those in unexplained infertile women without any sign of CE.

ORIGINAL ARTICLE: Abnormal Pattern of Lymphocyte Subpopulations in the Endometrium of Infertile Women with Chronic Endometritis

Problem  Endometrial lymphocytes play a critical role in endometrial receptivity. This study aimed at evaluating the variations induced by chronic endometritis (CE) on endometrial lymphocyte subsets. We compared the results in infertile women diagnosed with CE with those in unexplained infertile women without any sign of CE.

Reduced percentage of natural killer cells associated with impaired cytokine network in the secretory endometrium of infertile women with polycystic ovary syndrome

OBJECTIVE To evaluate lymphocyte subset distribution in the secretory endometrium from infertile patients with polycystic ovary syndrome (PCOS), and the expression of the cytokines known to play a role in determining the endometrial lymphocyte pattern. DESIGN Experimental clinical study. SETTING Outpatient clinic in a university hospital. PATIENT(S) Twenty-eight patients with PCOS (PCOS group) and 6 fertile patients (control group). INTERVENTION(S) On days 22-26 of a spontaneous cycle, subjects underwent endometrial biopsies. MAIN OUTCOMES MEASURE(S) In 19 of 28 patients with PCOS and 6 controls with a late secretory endometrium, the percentage and phenotype of lymphocyte subsets were analyzed by flow cytometry. In the late secretory endometrium of 11 patients with PCOS and 3 controls, the expression of interleukins 15 and 18 and of chemokine ligand 10 was also analysed by polymerase chain reaction. RESULT(S) In patients with PCOS the percentage of CD56+/CD16- and of CD56bright/CD16- cells was significantly lower (median [confidence interval]: 38% [31%-52.7%] vs. 63.7% [57.7%-69%] and 17.4% [8%-41.6%] vs. 52% [43%-60%], respectively), whereas the percentage of CD3+ was significantly higher (45% [33.3%-64%] vs. 26.1% [21%-32%]) as compared with controls. Accordingly, polymerase chain reaction analysis revealed a significantly lower expression of interleukins 15 and 18 and of chemokine ligand 10 in patients with PCOS than in controls. CONCLUSION(S) Results demonstrated an abnormal percentage of endometrial lymphocyte subsets, associated with an impaired cytokine network in patients with PCOS. This could explain the poor reproductive potential in these patients.

Preliminary evidence for high anti-PLAC1 antibody levels in infertile patients with repeated unexplained implantation failure

OBJECTIVE Placenta-specific1 (PLAC1) is a trophoblast-specific gene encoding for a protein that is highly expressed in human placenta, on the surface of the syncytiotrophoblast. PLAC1 was found to elicit spontaneous antibody responses in cancer patients. We aimed to determine the levels of anti-PLAC1 antibodies in infertile women with a history of unexplained repeated implantation failure after IVF cycles as compared to fertile women. STUDY DESIGN An observational case-control clinical study. MAIN OUTCOME MEASURE(S) Two groups of patients were analysed in two different experimental settings: 21 infertile women and 81 control patients were enrolled in the first group, 16 infertile women and 67 fertile controls in the second group. Anti-PLAC1 antibody levels and ranking were analysed by ELISA test. RESULTS In both groups of infertile patients enrolled, optical densities (OD) from ELISA test ranked significantly higher than those of controls (0.27 ± 0.2 vs. 0.13 ± 0.1 respectively; p = 0.0009 in the first group), (0.62 ± 0.38 vs. 0.39 ± 0.35 respectively; p = 0.0044 in the second experiment). In the first group about one case in four (29%) had OD levels above the 95th percentile (0.337) for healthy controls (p = 0.005). In the second experiment 4 out of 16 cases (25%) had OD levels above the 95th percentile (0.878) for healthy controls (p = 0.023). CONCLUSIONS Anti-PLAC1 antibodies could represent a biomarker associated with infertility and with high probability of repeated implantation failure after ovarian stimulation and IVF-ET, greatly improving the diagnostic work up of infertile couples.

Human monocyte-derived dendritic cells exposed to hyperthermia show a distinct gene expression profile and selective upregulation of IGFBP6

Fever plays a role in activating innate immunity while its relevance in activating adaptive immunity is less clear. Even brief exposure to elevated temperatures significantly impacts on the immunostimulatory capacity of dendritic cells (DCs), but the consequences on immune response remain unclear. To address this issue, we analyzed the gene expression profiles of normal human monocyte-derived DCs from nine healthy adults subjected either to fever-like thermal conditions (39°C) or to normal temperature (37°C) for 180 minutes. Exposure of DCs to 39°C caused upregulation of 43 genes and downregulation of 24 genes. Functionally, the up/downregulated genes are involved in post-translational modification, protein folding, cell death and survival, and cellular movement. Notably, when compared to monocytes, DCs differentially upregulated transcription of the secreted protein IGFBP-6, not previously known to be specifically linked to hyperthermia. Exposure of DCs to 39°C induced apoptosis/necrosis and resulted in accumulation of IGFBP-6 in the conditioned medium at 48 h. IGFBP-6 may have a functional role in the hyperthermic response as it induced chemotaxis of monocytes and T lymphocytes, but not of B lymphocytes. Thus, temperature regulates complex biological DC functions that most likely contribute to their ability to induce an efficient adaptive immune response.Fever plays a role in activating innate immunity while its relevance in activating adaptive immunity is less clear. Even brief exposure to elevated temperatures significantly impacts on the immunostimulatory capacity of dendritic cells (DCs), but the consequences on immune response remain unclear. To address this issue, we analyzed the gene expression profiles of normal human monocyte-derived DCs from nine healthy adults subjected either to fever-like thermal conditions (39°C) or to normal temperature (37°C) for 180 minutes. Exposure of DCs to 39°C caused upregulation of 43 genes and downregulation of 24 genes. Functionally, the up/downregulated genes are involved in post-translational modification, protein folding, cell death and survival, and cellular movement. Notably, when compared to monocytes, DCs differentially upregulated transcription of the secreted protein IGFBP-6, not previously known to be specifically linked to hyperthermia. Exposure of DCs to 39°C induced apoptosis/necrosis and resulted in accumulation of IGFBP-6 in the conditioned medium at 48 h. IGFBP-6 may have a functional role in the hyperthermic response as it induced chemotaxis of monocytes and T lymphocytes, but not of B lymphocytes. Thus, temperature regulates complex biological DC functions that most likely contribute to their ability to induce an efficient adaptive immune response.

PLAC1 immunization does not induce infertility in mice

AIM Placenta specific 1 (PLAC1) is a protein rarely expressed in normal cells, except it is important for placental development, with a possible role in the establishment of the mother-fetus interface. The gene is also highly active in a wide variety of cancers and therefore, immunization with PLAC1 peptides could possibly be part of future immunotherapeutic strategies. We investigated whether vaccination against PLAC1 could induce infertility. MATERIALS & METHODS We inoculated female mice with PLAC1 peptides, put them in mating, measured antibody response (ELISA assay) and checked, in immunohistochemistry, binding of the induced antibodies to the native antigen. RESULTS We demonstrated that mice consistently develop antibody responses. We also demonstrated that female mice, after being inoculated with the PLAC1 peptide mix, do became pregnant and can give birth to normal infants. CONCLUSION PLAC1 antigens as a specific anti-cancer vaccine could induce anti-PLAC1 antibodies which do not necessarily cause infertility.

ORIGINAL ARTICLE: Abnormal Pattern of Lymphocyte Subpopulations in the Endometrium of Infertile Women with Chronic Endometritis: UNK CELLS IN WOMEN WITH CHRONIC ENDOMETRITIS

Problem  Endometrial lymphocytes play a critical role in endometrial receptivity. This study aimed at evaluating the variations induced by chronic endometritis (CE) on endometrial lymphocyte subsets. We compared the results in infertile women diagnosed with CE with those in unexplained infertile women without any sign of CE.