Publication: A human in vitro granuloma model for the investigation of multinucleated giant cell and granuloma formation
Authors
Seitzer, U. ; Haas, H. ; Gerdes, J.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
A method for the in vitro generation of
granulomas and its use in the analysis of the human
granulomatous response is summarized. As a target for
the cellular response L3 larvae of Nippostrongylus
brasiliensis are coincubated with human mononuclear
blood cells, and within seven to fourteen days the
development of blood monocytes to mature macrophages
and to epithelioid cells and multinucleated giant cells
(MGC) as typical constituents of granulomas clustered
around the nematode is observed. The following review
describes the uses and applications of this model for
phenotyping, functional, formation and modulating
studies of granulomas and MGCs, taking into account its
unique features compared to other in vitro models.
With respect to MGC formation, procedures are
described and examples are given which allow the
phenotyping of these cells using immunofluorescence
and immunohistological techniques. In addition, the
potential of this model for illuminating functional
aspects of MGC is described applying an isolation
protocol for MGC and a subsequent reverse-transcriptase
polymerase chain reaction method for the analysis of
single cells. Moreover, the significance and relevance of
using this granuloma model is discussed in the follow up
analysis of in vivo findings of interleukin-6 expression in
MGC of granulomas of patients with sarcoidosis. These in vivo results implicated a role for interleukin-6 in
granuloma and MGC development. The in vitro
granuloma model was used to investigate potential
modulatory effects of this cytokine by analysing the cell
numbers and the number of MGC per in vitro
granuloma, the size of the MGC formed, the fusion
index and the morphology of the in vitro granuloma. The
results demonstrated significant modulatory effects of
interleukin-6 on the cell number per in vitro granuloma
and on the morphology of the cells involved.
Conceivably, elevated interleukin-6 levels may modulate
granuloma formation with respect to the number of cells
involved and in influencing distinct cell populations involved in granuloma formation.
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