University Women's Hospital, Division of Gynecologicyl Endocrinology and Reproductive Medicine, Inselspital, University of Berne, Effingerstrasse, Berne, Switzerland.
University Institute of Clinical Chemistry, Inselspital, University of Berne, Berne, Switzerland.
Hum Reprod. 2017 Apr 1;32(4):820-831. doi: 10.1093/humrep/dex005.
Are the immune cell profiles and the cytokine concentrations in follicular fluid (FF) and serum at the preovulatory stage different in conventional exogenous gonadotrophin stimulated IVF (c-IVF) compared with natural cycle IVF (NC-IVF)?
The cell counts of CD45+ leucocytes and T cell subpopulations and the cytokine concentrations in FF and serum are different in c-IVF compared to NC-IVF.
FF-derived cells are heterogeneous. Immune cells are involved in intra-ovarian processes and cytokines are required for normal follicular development. Gonadotrophins stimulate the regulatory intrafollicular system and influence the local distribution of immune cells and the intrafollicular release of cytokines. Administration of exogenous gonadotrophins may have a significant effect on this local regulatory system, which then in turn could influence oocyte quality.
STUDY DESIGN, SIZE, DURATION: The study included 105 patients, 69 undergoing c-IVF and 36 undergoing NC-IVF. c-IVF was performed by exogenous ovarian stimulation with hMG and GnRH antagonists.
PARTICIPANTS/MATERIALS, SETTING, METHODS: FF samples were collected from the first dominant follicle in c-IVF without pooling and from single leading preovulatory follicles in NC-IVF. Three different approaches were used to analyze FF samples: (i) microscopic investigation of CD45+ leucocytes, (ii) fluorescence-activated cell sorting to determine CD19+ B cells and CD3+ T cells including T cell subpopulations (CD4+, CD8+), and (iii) evaluation of tumour necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ), interleukins (IL)-2, -6, -8, -10 and vascular endothelial growth factor (VEGF) levels in matched FF and serum samples using the Bio-Plex® platform.
FF obtained from c-IVF contained proportionally more CD45+ leucocytes (P = 0.0384), but fewer CD8+ cytotoxic T cells than FF from NC-IVF. CD3+ T lymphocytes were the most common type of lymphocytes, and the number thereof was comparable in the two study groups. In c-IVF, serum VEGF levels were higher (P = 0.007) than in NC-IVF while FF contained marginally decreased concentrations of IL-8 in c-IVF in comparison to NC-IVF. The cytokine concentration gradient between FF and serum in c-IVF was 10-fold for IL-8 and 8-fold for VEGF and thereby markedly lower than in NC-IVF, where the differences were 32-fold and 30-fold, respectively. Strong positive correlations were determined between FF- IL-10 and FF- VEGF in c-IVF (r = 0.85, P < 0.0001) and in NC-IVF (r = 0.81, P < 0.0001).
N/A.
LIMITATIONS, REASONS FOR CAUTION: The ovulation of NC-IVF follicles was induced by the exogenous administration of hCG, which means that the environment did not fully correspond to the physiological situation.
The differences in the immune profile and the cytokine concentrations in c-IVF and NC-IVF follicles support the hypothesis that conventional ovarian stimulation affects indirectly and heterogeneously the intrafollicular milieu, and thereby possibly affects the oocyte quality and the IVF outcome. However, further studies are needed to confirm our findings and to refine stimulation protocols in the context of optimizing the intrafollicular environment during oocyte maturation.
STUDY FUNDING/COMPETING INTEREST(S): The study was supported by a research grant from IBSA Institut Biochimique SA and MSD Merck Sharp & Dohme GmbH. The authors are clinically involved in low dose mono-follicular stimulation and IVF-therapies, using gonadotrophins from all gonadotrophins distributors on the Swiss market, including Institut Biochimique SA and MSD Merck Sharp & Dohme GmbH.
在常规外源性促性腺激素刺激的体外受精（c-IVF）与自然周期体外受精（NC-IVF）相比，排卵前阶段卵泡液（FF）和血清中的免疫细胞谱和细胞因子浓度是否不同？
与 NC-IVF 相比，c-IVF 中 FF 和血清中的 CD45+白细胞和 T 细胞亚群的细胞计数以及细胞因子浓度不同。
FF 来源的细胞是异质的。免疫细胞参与卵巢内过程，细胞因子是正常卵泡发育所必需的。促性腺激素刺激调节性卵泡内系统，并影响免疫细胞的局部分布和卵泡内细胞因子的释放。外源性促性腺激素的给药可能对这个局部调节系统有显著影响，进而可能影响卵母细胞质量。
研究设计、大小、持续时间：该研究纳入了 105 名患者，其中 69 名接受 c-IVF 治疗，36 名接受 NC-IVF 治疗。c-IVF 通过 hMG 和 GnRH 拮抗剂进行外源性卵巢刺激。
参与者/材料、设置、方法：从 c-IVF 中的第一个主导卵泡中采集 FF 样本，不进行混合，从 NC-IVF 中的单个主导排卵前卵泡中采集 FF 样本。使用三种不同的方法分析 FF 样本：（i）CD45+白细胞的显微镜检查，（ii）荧光激活细胞分选以确定 CD19+B 细胞和 CD3+T 细胞，包括 T 细胞亚群（CD4+、CD8+），以及（iii）使用 Bio-Plex®平台评估匹配的 FF 和血清样本中的肿瘤坏死因子-α（TNF-α）、干扰素-γ（INF-γ）、白细胞介素（IL）-2、-6、-8、-10 和血管内皮生长因子（VEGF）水平。
从 c-IVF 中获得的 FF 中含有比例更高的 CD45+白细胞（P = 0.0384），但含有较少的 CD8+细胞毒性 T 细胞。CD3+T 淋巴细胞是最常见的淋巴细胞类型，两组中的数量相当。在 c-IVF 中，血清 VEGF 水平较高（P = 0.007），而在 NC-IVF 中，FF 中的 IL-8 浓度略有降低。c-IVF 中 FF 与血清之间的细胞因子浓度梯度为 IL-8 的 10 倍，VEGF 的 8 倍，因此明显低于 NC-IVF，其中分别为 32 倍和 30 倍。在 c-IVF 和 NC-IVF 中，均确定了 FF-IL-10 和 FF-VEGF 之间的强正相关（r = 0.85，P < 0.0001）。
无。
局限性、谨慎的原因：NC-IVF 卵泡的排卵是通过 hCG 的外源性给药诱导的，这意味着环境不完全符合生理情况。
c-IVF 和 NC-IVF 卵泡中免疫谱和细胞因子浓度的差异支持以下假设，即常规卵巢刺激会间接且不均匀地影响卵泡内环境，从而可能影响卵母细胞质量和 IVF 结果。然而，需要进一步的研究来证实我们的发现，并在优化卵母细胞成熟过程中的卵泡内环境的背景下改进刺激方案。
研究基金/利益冲突：该研究得到了 IBSA Institut Biochimique SA 和 MSD Merck Sharp & Dohme GmbH 的研究资助。作者在临床上参与了低剂量单卵泡刺激和体外受精治疗，使用了瑞士市场上所有促性腺激素供应商的促性腺激素，包括 Institut Biochimique SA 和 MSD Merck Sharp & Dohme GmbH。
