The influence of different cultivating conditions on polymorphonuclear leukocyte apoptotic process in vitro, II: Ultrastructural characteristics of PMN populations incubated with proteinase 3 anti-neutrophil autoantibodies

L. Guejes, N. Zurgil, I. Stambler, Mordechai Deutsch, B. Gilburd, Y. Shoenfeld

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The present study shows the effects of proteinase 3 antineutrophil cytoplasmic autoantibodies (PR3 ANCA) on polymorphonuclear leukocytes (PMN) apoptotic processes in vitro. The results are part of a generalized morphological analysis of 3 identical experiments on the influence of different cultivating conditions on the apoptotic processes. As controls, the authors use the results on spontaneous PMN apoptosis (Guejes L, Zurgil N, Deutsch M, Gilburd B, Shoenfeld Y. Ultrastruct Pathol. 2003;27: 23-32) and PMN populations incubated with normal human IgG. Interaction of PR3 ANCA with the target antigen proteinase 3 (PR3) is one of the crucial pathogenic factors in Wegener granulomatosis (systemic autoimmune vasculitis). Following 40 min and 12 h incubation, PMN populations were evaluated by light microscopy, transmission electron microscopy, and immunogold electron microscopy. Twelve-hour cultures, either control or incubated with PR3 ANCA, contained different cell forms ranging from normal cells to cells at the final stages of apoptosis. Neutrophils at the state of complete manifestation of apoptotic phenotype were analyzed and compared. Three morphologically distinct apoptotic cell lines were characteristic for all PMN populations studied, regardless of cultivating conditions. As in spontaneous apoptosis, these cell lines are code-named "first," "second," and "third." The present study has shown, firstly, that in the presence of PR3 ANCA, all 3 apoptotic lines were modified or altered. Secondly, the modifications or alterations of apoptotic cell lines effected by PR3 ANCA are specific for each cell line: the "first" line is characterized by intensification and modification of activation; the "second" by vacuolized cell forms; and the "third" by pronounced lytic alterations of the nuclei, while the cytoplasm is fully identical to that of control cell lines.

Original languageEnglish
Pages (from-to)37-51
Number of pages15
JournalUltrastructural Pathology
Volume29
Issue number1
DOIs
StatePublished - Jan 2005

Bibliographical note

Funding Information:
This research was supported by the Horowitz Foundation.

Keywords

  • Apoptosis
  • In vitro cultures
  • Polymorphonuclear leukocytes (PMN)
  • Proteinase 3 (PR3)
  • Proteinase 3 anti-neutrophil cytoplasmic auto antibodies (PR3 ANCA)

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