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Transgenic expression of the chemokine receptor encoded by human herpesvirus 8 induces an angioproliferative disease resembling Kaposi's sarcoma.

Yang TY, Chen SC, Leach MW, Manfra D, Homey B, Wiekowski M, Sullivan L, Jenh CH, Narula SK, Chensue SW, Lira SA - J. Exp. Med. (2000)

Bottom Line: Human herpesvirus 8 (HHV8, also known as Kaposi's sarcoma [KS]-associated herpesvirus) has been implicated as an etiologic agent for KS, an angiogenic tumor composed of endothelial, inflammatory, and spindle cells.These lesions are characterized by a spectrum of changes ranging from erythematous maculae to vascular tumors, by the presence of spindle and inflammatory cells, and by expression of vGPCR, CD34, and vascular endothelial growth factor.We conclude that vGPCR contributes to the development of the angioproliferative lesions observed in these mice and suggest that this chemokine receptor may play a role in the pathogenesis of KS in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.

ABSTRACT
Human herpesvirus 8 (HHV8, also known as Kaposi's sarcoma [KS]-associated herpesvirus) has been implicated as an etiologic agent for KS, an angiogenic tumor composed of endothelial, inflammatory, and spindle cells. Here, we report that transgenic mice expressing the HHV8-encoded chemokine receptor (viral G protein-coupled receptor) within hematopoietic cells develop angioproliferative lesions in multiple organs that morphologically resemble KS lesions. These lesions are characterized by a spectrum of changes ranging from erythematous maculae to vascular tumors, by the presence of spindle and inflammatory cells, and by expression of vGPCR, CD34, and vascular endothelial growth factor. We conclude that vGPCR contributes to the development of the angioproliferative lesions observed in these mice and suggest that this chemokine receptor may play a role in the pathogenesis of KS in humans.

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Detection of the expression of vGPCR and VEGF within angioproliferative lesions of CDVG mice. (A) RT-PCR analysis of vGPCR expression within ears and muscle from animals in two separate transgenic lines of CDVG mice, 19 and 122. Samples obtained from tissues with gross lesions (+) and without lesions (−) were used. Amplification of a segment of G3PDH was used as an internal control for the reactions. (B) Magnetically sorted CD2+ cells found within tumors express vGPCR. Positive (+) and neg–ative (−) controls for this reaction were thymi from transgenic and control mice, respectively. No signal was observed in CD2 samples not reverse transcribed, indicating that the signal was derived from transgenic transcripts rather than genomic DNA. (C) In situ hybridization analysis of the expression of vGPCR and VEGF within the muscle of CDVG mice, dark field. The silver grains are only associated with infiltrating cells in the lesion. No signals found in the muscle cells (indicated by white *). Inset, higher magnification of the boxed area, bright field.
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Figure 5: Detection of the expression of vGPCR and VEGF within angioproliferative lesions of CDVG mice. (A) RT-PCR analysis of vGPCR expression within ears and muscle from animals in two separate transgenic lines of CDVG mice, 19 and 122. Samples obtained from tissues with gross lesions (+) and without lesions (−) were used. Amplification of a segment of G3PDH was used as an internal control for the reactions. (B) Magnetically sorted CD2+ cells found within tumors express vGPCR. Positive (+) and neg–ative (−) controls for this reaction were thymi from transgenic and control mice, respectively. No signal was observed in CD2 samples not reverse transcribed, indicating that the signal was derived from transgenic transcripts rather than genomic DNA. (C) In situ hybridization analysis of the expression of vGPCR and VEGF within the muscle of CDVG mice, dark field. The silver grains are only associated with infiltrating cells in the lesion. No signals found in the muscle cells (indicated by white *). Inset, higher magnification of the boxed area, bright field.

Mentions: To investigate whether the vascular changes observed in the CDVG mice were associated with the expression of vGPCR, we performed RT-PCR and in situ hybridization studies. Transgene transcripts were detected in samples with macroscopic lesions (ear, muscle) but were not found in transgenic tissue samples that lacked lesions (Fig. 5 A and Fig. 5 C, left panel). As the transgene expression was directed by CD2 regulatory elements, we investigated whether CD2+ cells could be found within the lesions. To this end, we isolated CD2+ cells from the tumors using magnetic sorting. Next, we investigated if these CD2+ cells expressed the transgene by RT-PCR analysis. As shown in Fig. 5 B, transgene expression could be detected by RT-PCR in these CD2+ cells. To investigate the distribution of the vGPCR-expressing cells, we performed in situ hybridization. Interestingly, similar to human KS 18, vGPCR expression can be detected in a subset of cells within the lesion (Fig. 5 C, left panel). Morphologically, most of these cells were either round or oval- shaped; we only rarely detected expression in spindle-shaped cells.


Transgenic expression of the chemokine receptor encoded by human herpesvirus 8 induces an angioproliferative disease resembling Kaposi's sarcoma.

Yang TY, Chen SC, Leach MW, Manfra D, Homey B, Wiekowski M, Sullivan L, Jenh CH, Narula SK, Chensue SW, Lira SA - J. Exp. Med. (2000)

Detection of the expression of vGPCR and VEGF within angioproliferative lesions of CDVG mice. (A) RT-PCR analysis of vGPCR expression within ears and muscle from animals in two separate transgenic lines of CDVG mice, 19 and 122. Samples obtained from tissues with gross lesions (+) and without lesions (−) were used. Amplification of a segment of G3PDH was used as an internal control for the reactions. (B) Magnetically sorted CD2+ cells found within tumors express vGPCR. Positive (+) and neg–ative (−) controls for this reaction were thymi from transgenic and control mice, respectively. No signal was observed in CD2 samples not reverse transcribed, indicating that the signal was derived from transgenic transcripts rather than genomic DNA. (C) In situ hybridization analysis of the expression of vGPCR and VEGF within the muscle of CDVG mice, dark field. The silver grains are only associated with infiltrating cells in the lesion. No signals found in the muscle cells (indicated by white *). Inset, higher magnification of the boxed area, bright field.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195818&req=5

Figure 5: Detection of the expression of vGPCR and VEGF within angioproliferative lesions of CDVG mice. (A) RT-PCR analysis of vGPCR expression within ears and muscle from animals in two separate transgenic lines of CDVG mice, 19 and 122. Samples obtained from tissues with gross lesions (+) and without lesions (−) were used. Amplification of a segment of G3PDH was used as an internal control for the reactions. (B) Magnetically sorted CD2+ cells found within tumors express vGPCR. Positive (+) and neg–ative (−) controls for this reaction were thymi from transgenic and control mice, respectively. No signal was observed in CD2 samples not reverse transcribed, indicating that the signal was derived from transgenic transcripts rather than genomic DNA. (C) In situ hybridization analysis of the expression of vGPCR and VEGF within the muscle of CDVG mice, dark field. The silver grains are only associated with infiltrating cells in the lesion. No signals found in the muscle cells (indicated by white *). Inset, higher magnification of the boxed area, bright field.
Mentions: To investigate whether the vascular changes observed in the CDVG mice were associated with the expression of vGPCR, we performed RT-PCR and in situ hybridization studies. Transgene transcripts were detected in samples with macroscopic lesions (ear, muscle) but were not found in transgenic tissue samples that lacked lesions (Fig. 5 A and Fig. 5 C, left panel). As the transgene expression was directed by CD2 regulatory elements, we investigated whether CD2+ cells could be found within the lesions. To this end, we isolated CD2+ cells from the tumors using magnetic sorting. Next, we investigated if these CD2+ cells expressed the transgene by RT-PCR analysis. As shown in Fig. 5 B, transgene expression could be detected by RT-PCR in these CD2+ cells. To investigate the distribution of the vGPCR-expressing cells, we performed in situ hybridization. Interestingly, similar to human KS 18, vGPCR expression can be detected in a subset of cells within the lesion (Fig. 5 C, left panel). Morphologically, most of these cells were either round or oval- shaped; we only rarely detected expression in spindle-shaped cells.

Bottom Line: Human herpesvirus 8 (HHV8, also known as Kaposi's sarcoma [KS]-associated herpesvirus) has been implicated as an etiologic agent for KS, an angiogenic tumor composed of endothelial, inflammatory, and spindle cells.These lesions are characterized by a spectrum of changes ranging from erythematous maculae to vascular tumors, by the presence of spindle and inflammatory cells, and by expression of vGPCR, CD34, and vascular endothelial growth factor.We conclude that vGPCR contributes to the development of the angioproliferative lesions observed in these mice and suggest that this chemokine receptor may play a role in the pathogenesis of KS in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.

ABSTRACT
Human herpesvirus 8 (HHV8, also known as Kaposi's sarcoma [KS]-associated herpesvirus) has been implicated as an etiologic agent for KS, an angiogenic tumor composed of endothelial, inflammatory, and spindle cells. Here, we report that transgenic mice expressing the HHV8-encoded chemokine receptor (viral G protein-coupled receptor) within hematopoietic cells develop angioproliferative lesions in multiple organs that morphologically resemble KS lesions. These lesions are characterized by a spectrum of changes ranging from erythematous maculae to vascular tumors, by the presence of spindle and inflammatory cells, and by expression of vGPCR, CD34, and vascular endothelial growth factor. We conclude that vGPCR contributes to the development of the angioproliferative lesions observed in these mice and suggest that this chemokine receptor may play a role in the pathogenesis of KS in humans.

Show MeSH
Related in: MedlinePlus