The endothelium is an extrahepatic site of synthesis of the seventh component of the complement system

H. Langeggen, M. Pausa, E. Johnson, C. Casarsa, Francesco Tedesco

Research output: Contribution to journalArticle

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Abstract

The level of the terminal complement components secreted by human umbilical vein endothelial cells (HUVEC) was measured by a sensitive ELISA which allows the detection of 30-50 pg/ml of these components. C7 was the only terminal component detected in measurable amounts in the cell supernatant. The mean value was 11 ng/106 cells at 96 h and was slightly higher than that of C3 (9 ng/106 cells). HUVEC and serum C7 analysed by SDS- PAGE and immunoblot exhibited the same electrophoretic mobility. A proportion of C7 secreted by HUVEC was incorporated into the terminal complement complex (TCC) assembled spontaneously in the supernatant of cells cultured in C7- deficient human serum, and was not detected by the standard ELISA for C7 measurement. By adding the amount of C7 present in the TCC to that of free C7, the total amount of the component released by HUVEC was calculated to be approximately 35 ng/106 cells. Further TCC was produced following complement activation of the cell supernatant through the alternative pathway. Synthesis of C7 by HUVEC was confirmed by inhibition experiments in the presence of cycloheximide and by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of C7 mRNA expression. Addition of IL-1α and tumour necrosis factor-alpha to the cell culture stimulated the secretion of C3, but had no effect on the synthesis of C7. By contrast, interferon-gamma had only a marginal effect on the production of C3, but markedly down-regulated the synthesis of C7 as assessed both by ELISA and RT-PCR.

Original languageEnglish
Pages (from-to)69-76
Number of pages8
JournalClinical and Experimental Immunology
Volume121
Issue number1
DOIs
Publication statusPublished - 2000

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Human Umbilical Vein Endothelial Cells
Endothelium
Complement Membrane Attack Complex
Enzyme-Linked Immunosorbent Assay
Reverse Transcriptase Polymerase Chain Reaction
Complement Activation
Cycloheximide
Serum
Interleukin-1
Interferon-gamma
Polyacrylamide Gel Electrophoresis
Cultured Cells
Cell Culture Techniques
Tumor Necrosis Factor-alpha
Messenger RNA

Keywords

  • C7
  • Cytokines
  • Endothelial cells

ASJC Scopus subject areas

  • Immunology

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The endothelium is an extrahepatic site of synthesis of the seventh component of the complement system. / Langeggen, H.; Pausa, M.; Johnson, E.; Casarsa, C.; Tedesco, Francesco.

In: Clinical and Experimental Immunology, Vol. 121, No. 1, 2000, p. 69-76.

Research output: Contribution to journalArticle

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