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Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules.

ABSTRACT: Lymphocyte homing to secondary lymphoid tissue and lesions of chronic inflammation is directed by multi-step interactions between the circulating cells and the specialized endothelium of high endothelial venules (HEVs). In this study, we used the PCR-based method of suppression subtractive hybridization (SSH) to identify novel HEV genes by comparing freshly purified HEV endothelial cells (HEVECs) with nasal polyp-derived microvascular endothelial cells (PMECs). By this approach, we cloned the first nuclear factor preferentially expressed in HEVECs, designated nuclear factor from HEVs (NF-HEV). Virtual Northern and Western blot analyses showed strong expression of NF-HEV in HEVECs, compared to human umbilical vein endothelial cells (HUVECs) and PMECs. In situ hybridization and immunohistochemistry revealed that NF-HEV mRNA and protein are expressed at high levels and rather selectively by HEVECs in human tonsils, Peyers's patches, and lymph nodes. The NF-HEV protein was found to contain a bipartite nuclear localization signal, and was targeted to the nucleus when ectopically expressed in HUVECs and HeLa cells. Furthermore, endogenous NF-HEV was found in situ to be confined to the nucleus of tonsillar HEVECs. Finally, threading and molecular modeling studies suggested that the amino-terminal part of NF-HEV (aa 1-60) corresponds to a novel homeodomain-like Helix-Turn-Helix (HTH) DNA-binding domain. Similarly to the atypical homeodomain transcription factor Prox-1, which plays a critical role in the induction of the lymphatic endothelium phenotype, NF-HEV may be one of the key nuclear factors that controls the specialized HEV phenotype.

SUBMITTER: Baekkevold ES

PROVIDER: S-EPMC1868188 | biostudies-literature | 2003 Jul

REPOSITORIES: biostudies-literature

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Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules.

Baekkevold Espen S ES Roussigné Myriam M Yamanaka Takeshi T Johansen Finn-Eirik FE Jahnsen Frode L FL Amalric François F Brandtzaeg Per P Erard Monique M Haraldsen Guttorm G Girard Jean-Philippe JP

The American journal of pathology 20030701 1

Lymphocyte homing to secondary lymphoid tissue and lesions of chronic inflammation is directed by multi-step interactions between the circulating cells and the specialized endothelium of high endothelial venules (HEVs). In this study, we used the PCR-based method of suppression subtractive hybridization (SSH) to identify novel HEV genes by comparing freshly purified HEV endothelial cells (HEVECs) with nasal polyp-derived microvascular endothelial cells (PMECs). By this approach, we cloned the fi ...[more]

PMID: 12819012

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Dataset Information

Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules.

Publications

Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules.

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