A novel P-selectin glycoprotein ligand-1 monoclonal antibody recognizes an epitope within the tyrosine sulfate motif of human PSGL-1 and blocks recognition of both P- and L-selectin
A novel P-selectin glycoprotein ligand-1 monoclonal antibody recognizes an epitope within the tyrosine sulfate motif of human PSGL-1 and blocks recognition of both P- and L-selectin
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DOI：
10.1182/blood.v91.1.154.154_154_164
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发表时间：
1998-01-01
期刊：
影响因子：
20.3
通讯作者：
Kansas, GS
中科院分区：
文献类型：
--
作者：
Snapp, KR;Ding, H;Kansas, GS
p -选择素和p -选择素糖蛋白配体-1 （PSGL-1）之间的相互作用介导了炎症部位内皮细胞管腔表面白细胞的最早“滚动”。先前，PSGL-1已被证明是中性粒细胞与p -选择素相互作用的主要媒介，但关于PSGL-1介导p -选择素与其他白细胞亚群相互作用的能力的研究产生了不同的和相互矛盾的结果，产生了一种新的针对人PSGL-1的IgG单克隆抗体（MoAb），该MoAb的特异性通过流式细胞分析和Western blotting对转染人PSGL-1的细胞进行了证实。这个新开发的MoAb， KPL1，抑制表达p -选择素的COS细胞与HL60细胞、中性粒细胞或淋巴细胞之间的相互作用。此外，在生理条件下的流动实验中，KPL1完全抑制了p -选择素与纯化CD4 T细胞或中性粒细胞之间的相互作用，但对T细胞或中性粒细胞与e -选择素的相互作用没有影响。此外，KPL1阻断了转染l -选择素的淋巴样细胞与表达PSGL-1的COS细胞之间的相互作用。KPL1表位被定位到PSGL-1一致的酪氨酸硫酸化基序中的一个位点，先前被证明是与p -选择素相互作用的必要条件，现在被证明是与l -选择素相互作用的必要条件，并且与PL1功能阻断抗PSGL-1 MoAb鉴定的表位不同。正常白细胞的双色流式细胞术显示，自然杀伤（NK）细胞（CD16(+)）、单核细胞、CD4和CD8 T细胞以及α / β和γ / δ T细胞均呈PSGL-1阳性，而B细胞则表达低水平的KPL1表位。这种低水平的KPL1染色也在生发中心的免疫组织学上观察到，没有检测到KPL1染色，而t细胞区域（滤泡间区）KPL1阳性。有趣的是，原位浆细胞和白细胞介素-6依赖性骨髓瘤细胞系是KPL1(+)，因此，PSGL-1基本上在所有的血液中性粒细胞、NK细胞、B细胞、T细胞和单核细胞上表达。B细胞分化过程中酪氨酸硫酸化的变化可能影响B细胞与P-选择素和l -选择素相互作用的能力。(C) 1998年由美国血液病学会出版。
Interactions between P-selectin and P-selectin glycoprotein ligand-l (PSGL-1) mediate the earliest "rolling" of leukocytes on the lumenal surface of endothelial cells at sites of inflammation. Previously, PSGL-1 has been shown to be the primary mediator of interactions between neutrophils and P-selectin, but studies on the ability of PSGL-1 to mediate interactions between P-selectin and other subsets of leukocytes have yielded variable and conflicting results, A novel IgG monoclonal antibody (MoAb) to human PSGL-1 was generated, and the specificity of this MoAb was confirmed by both flow cytometric analysis and Western blotting of cells transfected with human PSGL-1. This newly developed MoAb, KPL1, inhibited interactions between P-selectin expressing COS cells and either HL60 cells, neutrophils, or lymphocytes. Furthermore, KPL1 completely inhibited interactions between P-selectin and either purified CD4 T cells or neutrophils in a flow assay under physiological conditions, but had no effect on interactions of T cells or neutrophils with E-selectin. In addition, KPL1 blocked interactions between lymphoid cells transfected with L-selectin and COS cells expressing PSGL-1. The KPL1 epitope was mapped to a site within a consensus tyrosine sulfation motif of PSGL-1, previously shown to be essential for interaction with P-selectin and now shown to be essential for interaction with L-selectin, and to be distinct from the epitope identified by the PL1 function blocking anti-PSGL-1 MoAb. Two-color flow cytometry of normal leukocytes showed that while natural killer (NK) cells (CD16(+)), monocytes, CD4 and CD8 T cells, and alpha/beta and gamma/delta T cells were uniformly positive for PSGL-1, B cells expressed low levels of the KPL1 epitope. This low level of KPL1 staining was also observed immunohistologically in germinal centers, which had no detectable KPL1 staining, whereas T-cell areas (interfollicular region) were positive for KPL1. Interestingly, plasma cells in situ and interleukin-6-dependent myeloma cell lines were KPL1(+), Thus, PSGL-1 is expressed on essentially ail blood neutrophils, NK cells, B cells, T cells, and monocytes. Variation in tyrosine sulfation during B-cell differentiation may affect the ability of B cells to interact with P- and L-selectin. (C) 1998 by The American Society of Hematology.