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从沙特捐赠者分离的B细胞中未发现PEPITEM释放的初步研究结果:对扩大人群研究的启示

Preliminary findings on the absence of PEPITEM release in B cells isolated from Saudi donors: implications for expanded population studies.

作者信息

Alassiri Mohammed, Alanazi Asma, Barhoumi Tlili, Alrfaei Bahauddeen, Alanazi Maisa, Rashid Mamoon, Alhazmi Aiman S, Alasseiri Mohammed, AlMefleh Abdulrahman, Boudjelal Mohammad, Shaibah Hayat, Almuhalhil Khawlah, Mansour Fatmah A, Alehaideb Zeyad, Alghanem Bandar

机构信息

Department of Basic Sciences, College of Science and Health Professions, King Abdullah International Medical Research Center (KAIMRC), King Saud bin Abdulaziz University for Health Sciences (KSAU-HS) Riyadh, KSA.

Department of Pathology and Laboratory Medicine, King Abdulaziz Medical City (KAMC) Riyadh, KSA.

出版信息

Am J Clin Exp Immunol. 2024 Oct 25;13(5):215-225. doi: 10.62347/XNNO3661. eCollection 2024.

Abstract

BACKGROUND

Adiponectin (AQ) plays a role in regulating immune responses. Previous research indicates that B cells can affect T cell transmigration via the adiponectin-induced peptide PEPITEM in Caucasians. This study explores whether this mechanism is also applicable to Saudi populations, considering potential ethnic variations in immune response.

METHODS

We conducted unbiased peptidomic screen on B cells, NK cells, and monocytes isolated from the peripheral blood of male healthy Saudi donors. The cells were stimulated with AQ, and the secretion of PEPITEM and other peptides was assessed using liquid chromatography-mass spectrometry (LC-MS/MS). Flow cytometry was utilized to confirm the purity of isolated cell populations and to verify the expression of adiponectin receptors AR1 and AR2.

RESULTS

PEPITEM was not detected in the supernatants of AQ-stimulated B cells, NK cells, or monocytes. All three cell populations were isolated and purified with high purity, confirmed by flow cytometry showing AR1 and AR2 expression on the surface of these cells. Specifically, less than 47% of B cells expressed ARs, with AR1 at 12% and AR2 at 17%. AQ stimulation increased the number of identified peptides in B cells and monocytes but decreased peptide numbers in NK cells. Dimensionality reduction analysis demonstrated clear segregation of cell types, with strong reproducibility across technical replicates.

CONCLUSION

The inability of B cells to release PEPITEM in response to AQ stimulation is an interesting finding and it needs more confirmatory tests and experiments, however; a hypothesis about the impact of predisposing factors, such as ethnicity could be formulated and tested in the future.

摘要

背景

脂联素(AQ)在调节免疫反应中发挥作用。先前的研究表明,在白种人中,B细胞可通过脂联素诱导的肽PEPITEM影响T细胞迁移。考虑到免疫反应中潜在的种族差异,本研究探讨该机制是否也适用于沙特人群。

方法

我们对从健康沙特男性献血者外周血中分离出的B细胞、自然杀伤细胞(NK细胞)和单核细胞进行了无偏肽组学筛选。用AQ刺激这些细胞,采用液相色谱-质谱联用技术(LC-MS/MS)评估PEPITEM和其他肽的分泌情况。利用流式细胞术确认分离细胞群体的纯度,并验证脂联素受体AR1和AR2的表达。

结果

在AQ刺激的B细胞、NK细胞或单核细胞的上清液中未检测到PEPITEM。通过流式细胞术确认所有三种细胞群体均以高纯度分离和纯化,显示这些细胞表面有AR1和AR2表达。具体而言,不到47%的B细胞表达ARs,其中AR1为12%,AR2为17%。AQ刺激增加了B细胞和单核细胞中鉴定出的肽的数量,但减少了NK细胞中的肽数量。降维分析表明细胞类型明显分离,技术重复间具有很强的可重复性。

结论

B细胞在AQ刺激下不能释放PEPITEM是一个有趣的发现,然而这需要更多的验证性测试和实验;关于诸如种族等易感因素影响的假设未来可以制定并进行测试。

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