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

IF 1.4 Q4 IMMUNOLOGY American journal of clinical and experimental immunology Pub Date : 2024-10-25 eCollection Date: 2024-01-01 DOI:10.62347/XNNO3661
Mohammed Alassiri, Asma Alanazi, Tlili Barhoumi, Bahauddeen Alrfaei, Maisa Alanazi, Mamoon Rashid, Aiman S Alhazmi, Mohammed Alasseiri, Abdulrahman AlMefleh, Mohammad Boudjelal, Hayat Shaibah, Khawlah Almuhalhil, Fatmah A Mansour, Zeyad Alehaideb, Bandar Alghanem
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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.

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关于从沙特捐献者体内分离出的 B 细胞不释放 PEPITEM 的初步发现:对扩大人群研究的影响。
背景:脂肪连接素(AQ)在调节免疫反应中发挥作用。以前的研究表明,在白种人中,B 细胞可通过脂肪连接素诱导的多肽 PEPITEM 影响 T 细胞的迁移。考虑到免疫反应中潜在的种族差异,本研究探讨了这一机制是否也适用于沙特人:我们对从沙特男性健康捐献者外周血中分离出的 B 细胞、NK 细胞和单核细胞进行了无偏见的肽组筛选。用 AQ 刺激细胞,并使用液相色谱-质谱联用仪(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的纯度,并验证脂肪素受体 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均未检测到PEPITEM。流式细胞仪显示这些细胞表面均有 AR1 和 AR2 表达,从而证实了这三种细胞群的分离和纯化纯度都很高。具体来说,只有不到 47% 的 B 细胞表达 ARs,其中 AR1 为 12%,AR2 为 17%。AQ刺激增加了B细胞和单核细胞中识别出的肽的数量,但减少了NK细胞中肽的数量。降维分析显示了细胞类型的明显分离,并且在不同技术重复中具有很强的可重复性:B细胞在AQ刺激下不能释放PEPITEM是一个有趣的发现,需要更多的确认测试和实验。
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