DataSheet1_HLA variations in patients with diffuse large B-cell lymphoma and association with disease risk and prognosis: a case-control study.docx

IntroductionHuman leukocyte antigen (HLA) polymorphisms have been associated with the development of various autoimmune diseases, as well as malignant neoplasms. Non-Hodgkin lymphomas (NHLs) are a heterogenous group of lymphoid malignancies in which a genetic substrate has been established and is deemed to play a crucial role in disease pathogenesis. This study aimed to identify whether variations in the HLA gene region were associated with diffuse large B-cell lymphoma (DLBCL) risk and prognosis.
MethodsWe defined HLA class I (HLA-A, HLA-B, HLA-C) and class II (HLA-DRB1, HLA-DQB1) alleles in 60 patients with DLBCL and compared the results to those found by 236 healthy adult donors from the bone marrow bank of Northern Greece. HLA typing was performed by two molecular methods, Sequence - Specific Oligonucleotide HLA typing (SSO) and Sequence - Specific Primer HLA typing (SSP), from white blood cells recovered from peripheral blood. The phenotypic frequencies of HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 between patients and controls were compared with the 2-sided Fisher’s exact test. Results with p-value <0.05 were considered statistically significant. Odds Ratios with 95% Confidence Intervals were calculated to further strengthen the results. The 2-sided Fisher’s exact test was also applied to alleles found only in one of the two groups, while the odds ratios together with the confidence intervals were corrected with Haldane-Anscombe method.
ResultsAmong the studied HLA polymorphisms, the frequency HLA-C*12 allele was significantly lower in patients with DLBCL compared with control subjects (6.7% vs. 34.7%, OR = 0.16, 95% CI: 0.04–0.44). Frequency of HLA-B*39 was significantly lower in patients with DLBCL compared with controls, but due to the low frequency of this polymorphism in the studied population and small sample size, determinations regarding the significance of this findings were limited. Survival analysis revealed that the presence of HLA-C*12 was not associated with improved or worsened overall and progression-free survival. No statistically significant associations were observed in the phenotypic frequencies of HLA-A, HLA-DQB1, HLA-DRB1 and the rest of HLA-B alleles between the control and DLBCL groups.
DiscussionCollectively, our results provide valuable insight regarding the role of HLA variations on DLBCL risk. Further studies are required to consolidate our findings and ascertain the clinical implications of these genetic variations on DLBCL management and prognosis.

引言 人类白细胞抗原（Human leukocyte antigen，HLA）多态性与多种自身免疫性疾病及恶性肿瘤的发生发展密切相关。非霍奇金淋巴瘤（Non-Hodgkin lymphomas，NHLs）作为一类异质性淋巴系统恶性肿瘤，其遗传易感基础已被证实，并被认为在疾病发病机制中发挥关键作用。本研究旨在探讨HLA基因区域的变异是否与弥漫大B细胞淋巴瘤（diffuse large B-cell lymphoma，DLBCL）的发病风险及预后存在关联。
方法 本研究对60例DLBCL患者的HLA I类（HLA-A、HLA-B、HLA-C）及II类（HLA-DRB1、HLA-DQB1）等位基因进行分型，并将结果与希腊北部骨髓库的236名健康成人供者的分型结果进行对照。从外周血分离白细胞后，采用两种分子生物学方法完成HLA分型：序列特异性寡核苷酸探针HLA分型（Sequence-Specific Oligonucleotide，SSO）与序列特异性引物HLA分型（Sequence-Specific Primer，SSP）。对比患者组与对照组间HLA-A、HLA-B、HLA-C、HLA-DRB1及HLA-DQB1的表型频率时，采用双侧Fisher确切概率法（2-sided Fisher’s exact test）进行统计分析，以P值<0.05作为差异具有统计学意义的判定标准。同时计算比值比（Odds Ratios，OR）及其95%置信区间（Confidence Intervals，CI）以进一步强化研究结果的可信度。针对仅在两组中某一组检出的等位基因，同样采用双侧Fisher确切概率法进行分析，并采用Haldane-Anscombe法对比值比及其置信区间进行校正。
结果 在本次研究纳入的HLA多态性位点中，DLBCL患者组的HLA-C*12等位基因频率显著低于对照组（6.7% vs. 34.7%，OR=0.16，95%CI：0.04~0.44）。DLBCL患者组的HLA-B*39基因频率同样低于对照组，但由于该多态性在研究人群中频率较低且样本量有限，该结果的统计学显著性判定存在一定局限性。生存分析结果显示，HLA-C*12的携带与否与患者的总生存期及无进展生存期均无显著关联。对照组与DLBCL患者组间，HLA-A、HLA-DQB1、HLA-DRB1及其他HLA-B等位基因的表型频率均未观察到统计学意义上的显著差异。
讨论 综合来看，本研究结果为HLA变异在DLBCL发病风险中的作用提供了具有价值的参考依据。未来仍需开展更大样本量的研究以验证本研究结果，并明确此类遗传变异在DLBCL诊疗及预后评估中的临床意义。