Department of Pharmacology, Experimental Therapy, and Toxicology, Institute of Experimental and Clinical Pharmacology and Pharmacogenomik and ICePhA, University of Tuebingen, Tuebingen, Germany.
Children's Hospital and Interdisciplinary Center for Infectious Diseases, University of Tuebingen, Tuebingen, Germany.
Immun Inflamm Dis. 2021 Mar;9(1):210-222. doi: 10.1002/iid3.380. Epub 2020 Nov 23.
Chemokine receptors and their corresponding ligands are key players of immunity by regulation of immune cell differentiation and migration. CXCR1 is a high-affinity receptor for CXCL8. Differential expression of CXCR1 is associated with a variety of human pathologies including cancer and inflammatory diseases. While various studies have highlighted the importance of CXCR1-mediated CXCL8-sensing for neutrophil trafficking and function, its role in B-cell responses remains unsolved. Therefore, our aim was to investigate innate and adaptive antibody responses in CXCR1-deficient mice.
Cell populations of the spleen and the peritoneal cavity were identified and quantified via flow cytometry. To investigate thymus-independent (TI) and thymus-dependent (TD) antibody responses, mice were immunized intraperitoneally with TNP-Ficoll, Pneumovax23, and TNP-Chicken Gamma Globulin. Mice were bled before as well as 7 and 14 days after vaccination to collect serum. Serum antibody levels overtime were analyzed according to their specificity by enzyme-linked immunosorbent assay. B-1 cell functionality was examined by IL-5/IL-5Rα-dependent stimulation of peritoneal and splenic cells in vitro. To analyze CXCR1/2-expression, CD19 splenocytes were enriched by magnetic-activated cell sorting before isolation of total RNA contents, followed by reverse transcription and real-time polymerase chain reaction.
The distribution of natural B-1 cell populations was disturbed in the absence of CXCR1, while their responsiveness towards TI antigens and in vitro stimulation remained functional. Besides, CXCR1-deficiency was accompanied by increased frequencies of follicular B-2 cells in the spleen. Interestingly, these mice produced elevated levels of antigen-specific IgG upon TD immunization and harbored a significantly enlarged proportion of CXCR5-expressing T helper (H) cells. CXCR1-expression was detectable in CD19 splenocytes derived from wild-type, but not CXCR1-deficient mice.
Our data demonstrate a previously unknown relevance of CXCR1 for the production of specific IgG in response to vaccination. These findings identify CXCR1 as a promising candidate for future studies on the regulation of adaptive antibody responses.
趋化因子受体及其相应的配体通过调节免疫细胞的分化和迁移，是免疫的关键因素。CXCR1 是 CXCL8 的高亲和力受体。CXCR1 的差异表达与多种人类疾病有关，包括癌症和炎症性疾病。虽然各种研究强调了 CXCR1 介导的 CXCL8 感应对中性粒细胞迁移和功能的重要性，但它在 B 细胞反应中的作用仍未解决。因此，我们的目的是研究 CXCR1 缺陷小鼠的先天和适应性抗体反应。
通过流式细胞术鉴定和定量脾和腹腔细胞群。为了研究胸腺非依赖性（TI）和胸腺依赖性（TD）抗体反应，将小鼠用 TNP-Ficoll、Pneumovax23 和 TNP-鸡γ球蛋白经腹腔免疫接种。在接种前以及接种后 7 天和 14 天采血以收集血清。根据酶联免疫吸附试验的特异性分析血清抗体水平随时间的变化。通过体外刺激腹腔和脾细胞，IL-5/IL-5Rα 依赖性检测 B-1 细胞功能。为了分析 CXCR1/2 的表达，用磁性激活细胞分选法富集 CD19 脾细胞，然后分离总 RNA 含量，进行逆转录和实时聚合酶链反应。
缺乏 CXCR1 时，天然 B-1 细胞群的分布受到干扰，但其对 TI 抗原的反应性和体外刺激仍然具有功能。此外，CXCR1 缺陷小鼠脾脏中滤泡 B-2 细胞的频率增加。有趣的是，这些小鼠在 TD 免疫接种后产生了更高水平的抗原特异性 IgG，并具有显著增加的 CXCR5 表达的辅助性 T（H）细胞比例。从野生型但不是 CXCR1 缺陷型小鼠的 CD19 脾细胞中可检测到 CXCR1 的表达。
我们的数据表明，CXCR1 对疫苗接种后特异性 IgG 的产生具有先前未知的相关性。这些发现将 CXCR1 确定为未来调节适应性抗体反应的研究的有希望的候选物。
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