Limits...
Polymerase chain reaction selects a novel disintegrin proteinase from CD40-activated germinal center dendritic cells.

Mueller CG, Rissoan MC, Salinas B, Ait-Yahia S, Ravel O, Bridon JM, Briere F, Lebecque S, Liu YJ - J. Exp. Med. (1997)

Bottom Line: Dendritic cell precursors or immature dendritic cells express no or low levels of the message.It is induced to high levels upon spontaneous or CD40-dependent maturation and in a mixed lymphocyte reaction.In situ hybridization showed distinct expression of this gene in the germinal center.

View Article: PubMed Central - PubMed

Affiliation: Schering-Plough Laboratory for Immunological Research, 69571 Dardilly, France.

ABSTRACT
To identify genes expressed by a specific subset of dendritic cells found in vivo a polymerase chain reaction-based cDNA subtraction technique was applied to the recently described germinal center dendritic cells. A novel member of the disintegrin metalloproteinase family was cloned which comprises a not typical zinc-chelating catalytic site most similar to a bacterial metalloproteinase. Dendritic cell precursors or immature dendritic cells express no or low levels of the message. It is induced to high levels upon spontaneous or CD40-dependent maturation and in a mixed lymphocyte reaction. In situ hybridization showed distinct expression of this gene in the germinal center. This, together with the findings that certain disintegrin metalloproteinases regulate the activity of tumor necrosis factor alpha and that metalloproteinases have also been implicated in FasL processing, suggest that this novel molecule may play an important role in dendritic cell function and their interactions with germinal center T cells.

Show MeSH

Related in: MedlinePlus

Decysin is highly expressed in mature DCs. (A) 28 cycle PCR on GCDCs and CD11c+ blood DCs immediately after cell sorting, after 24 h  incubation in complete medium and after CD40 stimulation for 24 h in complete medium. Day 12 harvested CD34 stem cell– and monocyte-derived  DC were analyzed for decysin before and after CD40 ligation on CD40L-transfected mouse L cells for the indicated time. (B) PCR on stem cell–generated DCs before and after coculture with alloreactive total naive T cells for 12 and 24 h. β actin was amplified with 28 cycles, decysin, CD83, and IL-2  with 35 cycles.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2199019&req=5

Figure 4: Decysin is highly expressed in mature DCs. (A) 28 cycle PCR on GCDCs and CD11c+ blood DCs immediately after cell sorting, after 24 h incubation in complete medium and after CD40 stimulation for 24 h in complete medium. Day 12 harvested CD34 stem cell– and monocyte-derived DC were analyzed for decysin before and after CD40 ligation on CD40L-transfected mouse L cells for the indicated time. (B) PCR on stem cell–generated DCs before and after coculture with alloreactive total naive T cells for 12 and 24 h. β actin was amplified with 28 cycles, decysin, CD83, and IL-2 with 35 cycles.

Mentions: Since decysin was identified in CD40-stimulated GCDCs, we wondered whether the metalloproteinase might be expressed at high levels in CD40-activated DCs. Indeed, 28 cycle PCR coupled to reverse transcribed RNA (RT-PCR) on freshly isolated GCDCs failed to detect decysin (Fig. 4 A). Its expression is induced by spontaneous maturation in culture (4) and increases in response to CD40 activation. Similarly, in another ex vivo isolate, blood CD11c+ DCs do not contain detectable decysin mRNA, but maturation in culture (23) and more importantly CD40 activation result in decysin induction. In vitro generated DCs from CD34 progenitor cells or monocytes rapidly synthesize the message in response to CD40 ligation and a mixed lymphocyte reaction (B) results in decysin expression together with CD83. Thus, the novel metalloproteinase represents a DC maturation marker synthesised in response to T cell signals.


Polymerase chain reaction selects a novel disintegrin proteinase from CD40-activated germinal center dendritic cells.

Mueller CG, Rissoan MC, Salinas B, Ait-Yahia S, Ravel O, Bridon JM, Briere F, Lebecque S, Liu YJ - J. Exp. Med. (1997)

Decysin is highly expressed in mature DCs. (A) 28 cycle PCR on GCDCs and CD11c+ blood DCs immediately after cell sorting, after 24 h  incubation in complete medium and after CD40 stimulation for 24 h in complete medium. Day 12 harvested CD34 stem cell– and monocyte-derived  DC were analyzed for decysin before and after CD40 ligation on CD40L-transfected mouse L cells for the indicated time. (B) PCR on stem cell–generated DCs before and after coculture with alloreactive total naive T cells for 12 and 24 h. β actin was amplified with 28 cycles, decysin, CD83, and IL-2  with 35 cycles.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199019&req=5

Figure 4: Decysin is highly expressed in mature DCs. (A) 28 cycle PCR on GCDCs and CD11c+ blood DCs immediately after cell sorting, after 24 h incubation in complete medium and after CD40 stimulation for 24 h in complete medium. Day 12 harvested CD34 stem cell– and monocyte-derived DC were analyzed for decysin before and after CD40 ligation on CD40L-transfected mouse L cells for the indicated time. (B) PCR on stem cell–generated DCs before and after coculture with alloreactive total naive T cells for 12 and 24 h. β actin was amplified with 28 cycles, decysin, CD83, and IL-2 with 35 cycles.
Mentions: Since decysin was identified in CD40-stimulated GCDCs, we wondered whether the metalloproteinase might be expressed at high levels in CD40-activated DCs. Indeed, 28 cycle PCR coupled to reverse transcribed RNA (RT-PCR) on freshly isolated GCDCs failed to detect decysin (Fig. 4 A). Its expression is induced by spontaneous maturation in culture (4) and increases in response to CD40 activation. Similarly, in another ex vivo isolate, blood CD11c+ DCs do not contain detectable decysin mRNA, but maturation in culture (23) and more importantly CD40 activation result in decysin induction. In vitro generated DCs from CD34 progenitor cells or monocytes rapidly synthesize the message in response to CD40 ligation and a mixed lymphocyte reaction (B) results in decysin expression together with CD83. Thus, the novel metalloproteinase represents a DC maturation marker synthesised in response to T cell signals.

Bottom Line: Dendritic cell precursors or immature dendritic cells express no or low levels of the message.It is induced to high levels upon spontaneous or CD40-dependent maturation and in a mixed lymphocyte reaction.In situ hybridization showed distinct expression of this gene in the germinal center.

View Article: PubMed Central - PubMed

Affiliation: Schering-Plough Laboratory for Immunological Research, 69571 Dardilly, France.

ABSTRACT
To identify genes expressed by a specific subset of dendritic cells found in vivo a polymerase chain reaction-based cDNA subtraction technique was applied to the recently described germinal center dendritic cells. A novel member of the disintegrin metalloproteinase family was cloned which comprises a not typical zinc-chelating catalytic site most similar to a bacterial metalloproteinase. Dendritic cell precursors or immature dendritic cells express no or low levels of the message. It is induced to high levels upon spontaneous or CD40-dependent maturation and in a mixed lymphocyte reaction. In situ hybridization showed distinct expression of this gene in the germinal center. This, together with the findings that certain disintegrin metalloproteinases regulate the activity of tumor necrosis factor alpha and that metalloproteinases have also been implicated in FasL processing, suggest that this novel molecule may play an important role in dendritic cell function and their interactions with germinal center T cells.

Show MeSH
Related in: MedlinePlus