Tambunan Betty Agustina, Ugrasena I Dewa Gede
Doctoral Program of Medical Science, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.
Department of Clinical Pathology, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.
J Blood Med. 2023 Jan 13;14:7-17. doi: 10.2147/JBM.S393134. eCollection 2023.
Repeated transfusions in thalassemia patients can cause several complications, including alloimmunization and autoimmunization.
This study compares the immune response of T follicular helper (Tfh) lymphocytes expressing T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory domains (TIGIT), programmed cell death-1 (PD-1), and interleukin-21 (IL-21) between patients with allo-auto positive and negative thalassemia before and after hemin administration.
This study used a quasi-experimental pre- and post-test design and was performed between April and November 2021 at the Dr. Soetomo General Academic Hospital in Surabaya, Indonesia. It enrolled 29 patients with allo-auto positive thalassemia and 28 with allo-auto negative, and 9 mL of whole blood (WB) was drawn from each patient. Hemin solution (20 µM) was added to 5 mL of WB, incubated for two hours, processed into peripheral blood mononuclear cells (PBMCs) in RPMI media, and cultured with 5% CO for three days. The 4 mL WB sample was also processed into PBMCs. PBMC cells cultured and without cultured were examined by flow cytometry using a BD FACSCalibur after surface and intracellular staining. Differences in Tfh cells expressing TIGIT, PD-1, and IL-21 between thalassemia groups before and after hemin administration were compared using independent -tests or Mann-Whitney -tests (p < 0.05).
Tfh cell expression did not differ between groups before hemin administration and increased after hemin administration. The increase in Tfh cell expression was higher in the allo-auto positive group. TIGIT and PD-1 expression in Tfh cells did not differ between groups, but TIGIT decreased after hemin administration in contrast to PD-1 result. IL-21 expression in Tfh cells did not differ between groups and did not change after hemin administration.
Hemin affected the expression of Tfh cells in both group thalassemia, but there was no difference of Tfh cell expression between the groups.
地中海贫血患者反复输血可导致多种并发症，包括同种免疫和自身免疫。
本研究比较了给予血红素前后，同种-自身免疫阳性和阴性地中海贫血患者中表达免疫球蛋白和基于免疫受体酪氨酸的抑制域的T细胞免疫受体（TIGIT）、程序性细胞死亡蛋白1（PD-1）和白细胞介素-21（IL-21）的T滤泡辅助（Tfh）淋巴细胞的免疫反应。
本研究采用准实验前后测试设计，于2021年4月至11月在印度尼西亚泗水的苏托莫博士综合学术医院进行。研究纳入了29例同种-自身免疫阳性地中海贫血患者和28例同种-自身免疫阴性患者，从每位患者采集9 mL全血（WB）。将血红素溶液（20 μM）加入5 mL WB中，孵育两小时，在RPMI培养基中处理成外周血单核细胞（PBMC），并在5%二氧化碳条件下培养三天。4 mL WB样本也处理成PBMC。培养和未培养的PBMC细胞在进行表面和细胞内染色后，使用BD FACSCalibur流式细胞仪进行检测。使用独立t检验或曼-惠特尼U检验比较给予血红素前后地中海贫血组中表达TIGIT、PD-1和IL-21的Tfh细胞的差异（p<0.05）。
给予血红素前，各组间Tfh细胞表达无差异，给予血红素后Tfh细胞表达增加。同种-自身免疫阳性组Tfh细胞表达的增加更高。Tfh细胞中TIGIT和PD-1表达在各组间无差异，但与PD-1结果相反，给予血红素后TIGIT表达降低。Tfh细胞中IL-21表达在各组间无差异，给予血红素后也未改变。
血红素影响了两组地中海贫血患者中Tfh细胞的表达，但两组间Tfh细胞表达无差异。
