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

Am J Pathol. 2003 Jul;163(1):69-79. doi: 10.1016/S0002-9440(10)63631-0.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Line
  • Endothelium, Lymphatic / cytology
  • Endothelium, Lymphatic / physiology*
  • Endothelium, Vascular / metabolism
  • Helix-Turn-Helix Motifs
  • Humans
  • In Situ Hybridization
  • Interleukin-33
  • Interleukins
  • Lymph Nodes / cytology
  • Lymph Nodes / metabolism
  • Lymphocytes
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Nucleic Acid Hybridization / methods
  • Palatine Tonsil / cytology
  • Palatine Tonsil / metabolism
  • Peyer's Patches / cytology
  • Peyer's Patches / metabolism
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Venules / cytology
  • Venules / metabolism*


  • IL33 protein, human
  • Interleukin-33
  • Interleukins
  • Nuclear Proteins