Data Sheet 1_Incomplete B-cell reconstitution in ART-treated people living with HIV is associated with EBV-linked lymphoma progression.xlsx

BackgroundDespite the widespread use of antiretroviral therapy (ART), people living with HIV continue to exhibit persistent immune alterations. Among the most affected cell populations are B lymphocytes, which show disruptions in differentiation, class-switch recombination, and the development of immunological memory. However, the relationship between these abnormalities, Epstein–Barr virus (EBV) coinfection, and clinical outcomes across different stages of HIV disease remains poorly understood.
MethodsWe analyzed 272 patients living with HIV from Hospital Universitario San Ignacio (Bogotá, Colombia), divided in four groups: clinical stage (S1, S2, S3 and lymphoma), clinical categories: AIDS-defining diseases, non-AIDS-defining diseases, and coinfections (CI), time on ART (<1 year, >1 year); and EBV coinfection. B lymphocyte subpopulations and immunoglobulin isotypes were quantified (next-generation flow cytometry). Cytokines (multiplex assays) and EBV viral load (quantitative PCR) were measured.
ResultsHIV progression was observed, associated with B cell compartment remodeling, characterized by depletion of naïve and memory B cells (smIgA1–2 and smIgG1–4) and an increase in immature/transitional cells and alterations in smIgM with isotype switching. These variations correlated negatively with clinical stage (ρ up to –0·43). The cytokine profile showed a persistent inflammatory signature (MIP-1β, G-CSF, FLT-3L, and IL-3). EBV coinfection intensified this phenotype, being associated with elevated levels of IL-6, IL-10, IL-15, TNF-α, and sCD40L, and with greater loss of memory cell subpopulations. Patients with AIDS-defining diseases and lymphomas exhibited the most profound alteration. Even after prolonged ART (>1 year), B cell reconstitution remained incomplete and biased toward immature phenotypes.
ConclusionsThis study shows that recovery of the B cell compartment under ART is incomplete and functionally unbalanced. Humoral memory loss, bias toward immature phenotypes, and persistent inflammation create a state of dysregulation that is exacerbated by EBV coinfection, especially in advanced clinical stages. These immunological defects provide a basis for the pathogenesis of non-AIDS-defining diseases and Epstein-Barr-associated lymphomas. Our findings support the need to integrate advanced immunological assessments, including standardized flow cytometry, as a complement to CD4+ count and viral load, in order to more accurately characterize immune competence and anticipate clinical risks in people living with HIV.

【研究背景】尽管抗逆转录病毒疗法（antiretroviral therapy, ART）已在临床广泛应用，但HIV感染者仍持续存在免疫功能异常。其中受影响最为显著的细胞群为B淋巴细胞（B lymphocytes），其分化、类别转换重组及免疫记忆发育过程均出现紊乱。然而，上述免疫异常、爱泼斯坦-巴尔病毒（Epstein–Barr virus, EBV）合并感染与HIV不同疾病阶段的临床结局之间的关联，目前仍未得到充分阐明。
【研究方法】本研究纳入来自哥伦比亚波哥大圣伊格纳西奥大学医院的272名HIV感染者，按以下维度分组：临床分期（S1、S2、S3及淋巴瘤组）、临床类别（艾滋病定义性疾病、非艾滋病定义性疾病及合并感染组）、抗逆转录病毒治疗时长（<1年、>1年）以及EBV合并感染情况。采用下一代流式细胞术（next-generation flow cytometry）定量检测B淋巴细胞亚群与免疫球蛋白同种型；通过多重检测（multiplex assays）测定细胞因子水平，采用定量PCR（quantitative PCR）检测EBV病毒载量。
【研究结果】随着HIV疾病进展，B细胞库发生重塑，具体表现为初始B细胞与记忆B细胞（smIgA1-2及smIgG1-4）耗竭，未成熟/过渡型B细胞比例升高，同时smIgM伴随类别转换出现异常。上述变化与临床分期呈显著负相关（相关系数ρ最高可达-0.43）。细胞因子谱呈现持续的炎症特征，涉及MIP-1β、G-CSF、FLT-3L及IL-3。EBV合并感染会加剧这一异常表型，与IL-6、IL-10、IL-15、TNF-α及sCD40L水平升高以及记忆细胞亚群大量丢失相关。合并艾滋病定义性疾病与淋巴瘤的患者，其免疫异常程度最为显著。即使接受了长期抗逆转录病毒治疗（>1年），患者的B细胞重建仍不完全，且偏向未成熟细胞表型。
【研究结论】本研究证实，接受抗逆转录病毒治疗后，HIV感染者的B细胞库恢复并不完全，且功能失衡。体液记忆丢失、偏向未成熟细胞表型以及持续炎症共同导致免疫失调状态，而EBV合并感染会进一步加重这一状态，尤其在疾病晚期临床阶段。上述免疫缺陷为非艾滋病定义性疾病及爱泼斯坦-巴尔病毒相关淋巴瘤的发病机制提供了理论依据。本研究结果提示，需将包括标准化流式细胞术在内的先进免疫学评估作为CD4+细胞计数及病毒载量检测的补充手段，从而更准确地评估HIV感染者的免疫功能状态，并预判其临床风险。