Besteht ein Effekt des Immuntherapeutikums LeukoNorm® auf die Steigerung der Implantationsfähigkeit im humanen Endometrium?

Zusammenfassung

Einleitung: Selbst nach künstlicher Befruchtung betragen die Schwangerschaftsraten bei einem Transfer von 2 teilungsfähigen Embryonen maximal 30–50 %. Dies kann am ehesten mit einer eingeschränkten endometrialen Implantation erklärt werden. Ob eine Immuntherapie wie mit LeukoNorm® zu einer Veränderung der endometrialen Immunzellen und Funktionsmarker führen kann, soll deshalb auf molekularbiologischer Ebene eruiert werden. Methoden: Bei 17 Frauen mit rezidivierendem Implantationsversagen wurden LH-datierte endometriale Biopsien (LH+7/8) in 2 konsekutiven Zyklen einmal ohne und einmal direkt nach einer LeukoNorm®-Therapie gewonnen. Immunhistochemisch wurden endometriale Immunzellen, molekularbiologische Implantationsmarker, insbesondere Immunmodulatoren, Glukosetransporter und Adhäsionsfaktoren, die zum größten Teil bei Implantationsversagern als dysreguliert beschrieben wurden, mittels Multi-Probe RNAse Protection Assay untersucht. Ergebnisse: Immunhistochemisch zeigten die endometrialen CD45+- und CD56+-Immunzellen keine signifikanten Zellzahlunterschiede nach einer LeukoNorm®-Behandlung. Molekularbiologisch wiesen auch die untersuchten endometrialen Funktionsmarker keine signifikanten Unterschiede (p = 0,49) auf, bei jedoch deutlichem Variationskoeffizient von 0,84 (β3-Integrin), 0,33 (Osteopontin), 0,64 (GLUT1), 0,50 (GLUT3), 0,66 (LIF), 0,26 (TGFb-1), 0,61 (IL-1b) und 0,50 (M‐CSF). Diskussion: Durch die Behandlung mit LeukoNorm® konnte anhand der untersuchten Parameter kein direkter Effekt auf die endometriale Funktion nachgewiesen werden. Eine mögliche Beeinflussung anderer Parameter kann jedoch nicht sicher ausgeschlossen werden.

Abstract

Objective: Even with artificial reproduction techniques, the implantation rates are only around 30–50 % per transfer of two viable embryos. These low implantation rates are most likely due to impaired endometrial receptivity. We therefore examined the potential effects of the immune therapeutic agent LeukoNorm® at the molecular level, and investigated both immune cells and known implantation factors in endometrial tissue. Methods: LH (LH+7/8) dated biopsies were taken from 17 women with recurrent implantation failure prior to and after the systemic administration of LeukoNorm®. Endometrial immune cells were examined immunohistochemically and individual implantation markers, such as immunmodulators, glucose transporters and adhesion factors, all of which are known to be dysregulated in women with implantation failure, were examined by RNAse protection assay to detect RNA expression changes. Results: Immunohistochemical analysis of CD45+ and CD 56+ cells did not show significant changes in cell numbers after LeukoNorm® treatment. Furthermore, the RNA expression of the examined implantation factors also did not show significant changes, but analysis demonstrated large variation coefficients of 0.84 (β3-integrin), 0.33 (osteopontin), 0.64 (GLUT1), 0.5 (GLUT3), 0.66 (LIF), 0,26 (TGFb-1), 0.61 (IL-1b) and 0.5 (M-CSF). Conclusions: Looking at specific implanation markers, we were not able to demonstrate a direct impact on endometrial function by LeukoNorm® therapy. Nevertheless, a potential effect on other parameters cannot be excluded.

Implantationsversagen - Immuntherapeutikum - Genexpression des Endometriums

implantation failure - immune therapeutic agent - endometrial gene expression

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