Chemokines and their receptors have been implicated in cancer biology. The CXCL12/CXCR4 axis is essential for the homing and retention of hematopoietic stem cells in bone marrow niches, and has a significant role in neonatal development. It is also implicated in multiple facets of cancer biology including metastasis, angiogenesis/neo-vasculogenesis, and immune cell trafficking at the tumor microenvironment (TME). Immunohistochemistry (IHC) is an ideal method for investigating involvement of CXCL12 in the TME. Three antibodies were evaluated here for their suitability to stain CXCL12. Both D8G6H and K15C gave apparent specific staining in both lymphoid and tumor tissue, but with converse staining patterns. D8G6H stained cells in the parafollicular zone whereas K15C showed staining of lymphoid cells in the interfollicular zone of tonsil tissue. Using a cell line with high CXCL12 expression, TOV21G, as a positive control, it was found that D8G6H gave strong staining of TOV21G cells whereas no staining was observed with K15C indicating that D8G6H specifically stains CXCL12. Significant staining of CXCL12 in the ovarian TME using tissue microarray was observed using D8G6H. These data demonstrate the importance of antibody characterization for IHC applications, and provide further evidence for the involvement of CXCL12 in ovarian cancer biology.

中文翻译：

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趋化因子CXCL12免疫组织化学染色抗体的表征和验证。

趋化因子及其受体与癌症生物学有关。CXCL12 / CXCR4轴对于骨髓小生境中的造血干细胞的归巢和保留至关重要，并且在新生儿发育中具有重要作用。它还涉及癌症生物学的多个方面，包括转移，血管生成/新血管生成以及肿瘤微环境（TME）处的免疫细胞运输。免疫组织化学（IHC）是研究CXCL12是否参与TME的理想方法。在此评估了三种抗体对CXCL12染色的适用性。D8G6H和K15C都在淋巴样组织和肿瘤组织中均给出了明显的特异性染色，但具有相反的染色模式。D8G6H在卵泡旁区域染色细胞，而K15C在扁桃体组织的卵泡间区域染色淋巴样细胞。使用具有高CXCL12表达的细胞系TOV21G作为阳性对照，发现D8G6H对TOV21G细胞产生强染色，而未观察到用K15C染色，表明D8G6H特异性染色CXCL12。使用D8G6H，使用组织微阵列观察到卵巢TME中CXCL12的显着染色。这些数据证明了抗体表征对于IHC应用的重要性，并为CXCL12参与卵巢癌生物学提供了进一步的证据。