Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Nov 15;112(10):4227-34.
doi: 10.1182/blood-2008-04-151498. Epub 2008 Aug 29.

Rhesus macaque rhadinovirus-associated non-Hodgkin lymphoma: animal model for KSHV-associated malignancies

Beata U Orzechowska  1 , Michael F Powers, Jerald Sprague, He Li, Bonnie Yen, Robert P Searles, Michael K Axthelm, Scott W Wong
Affiliations

Rhesus macaque rhadinovirus-associated non-Hodgkin lymphoma: animal model for KSHV-associated malignancies

Beata U Orzechowska et al. Blood. .

Abstract

Rhesus macaque rhadinovirus (RRV) is closely related to Kaposi sarcoma-associated herpesvirus (KSHV) and is associated with the development of B-cell hyperplasia and persistent lymphadenopathy resembling multicentric Castleman disease in rhesus macaques (RMs) coinfected with simian immunodeficiency virus (SIV). Here we investigated whether RMs experimentally infected with SIV and RRV can develop other disease manifestations observed in HIV- and KSHV-infected patients. As reported earlier, inoculation of SIV-infected RMs with RRV results in persistent RRV infection, whereas immunocompetent animals infected with RRV exhibit viremia 2 weeks after infection, followed by a period of no virus detection until they are subsequently made immunodeficient by SIV infection. A subset of animals developed abnormal cellular proliferations characterized as extranodal lymphoma and a proliferative mesenchymal lesion. In situ hybridization and immunohistochemistry analysis indicate RRV is present in both malignancies, and DNA microarray analysis detected viral interleukin-6 (vIL-6) and viral FLICE-like inhibitory protein (vFLIP) transcripts. Reverse-transcriptase polymerase chain reaction analysis confirmed vIL-6 and vFLIP expression, and that of RRV open reading frames 72 and 73, homologs of KSHV open reading frames shown to be expressed in primary effusion lymphoma. These data support the utility of the RRV-/SIV-infected RM as an excellent animal model to investigate KSHV-like pathogenesis.

PubMed Disclaimer

Figures

Figure 1
Figure 1
RRV load in peripheral blood compartment of experimentally inoculated RMs. Real-time PCR analysis of viral DNA loads using DNA isolated from PBMCs and RRV-specific primers from each cohort of animals. (A) Cohort1. (B) Cohort 2A and 2B. (C) Cohort 3. (D) Cohort 4.
Figure 2
Figure 2
Lymphoma and retroperitoneal fibromatosis (RF) in RRV-infected RMs. (A) Hematoxylin and eosin stain of lymphoma isolated at biopsy from the breast of animal 19185 (original magnification ×630). (B) Combined in situ hybridization of lymphoma with RRV cosmid probe (purple) and immunohistochemistry with mouse anti-CD20 (brown) and anti-CD3+ (gray) demonstrates that RRV is present in CD20+ cells comprising the lymphoma (original magnification ×630). (C) Same as panel B, except cosmid vector control replaces the RRV cosmid probe. (D) Hematoxylin and eosin stain of lymphoma isolated from liver of animal 19286 during necropsy (original magnification ×200). (E) Same as panel B, except RRV cosmid probe detected by immunofluorescence (purple) with no evidence of CD20 staining. (F) Similar to panel C; this is the control for panel E. (G) Hematoxylin and eosin stain of RF attached to the stomach of animal 18483 (original magnification ×400). (H) Combined in situ hybridization with RRV cosmid probe (purple) and immunohistochemistry with anti-CD20 (brown) and anti-CD3 (gray) shows that RRV is not present in the CD20+ or CD3+ cells (original magnification ×1000). Arrows point to RRV-positive cells that have spindle-like morphology. (I) Identical to panel H, except vector control replaces the RRV cosmid (original magnification ×630).
Figure 3
Figure 3
Analysis of RRV gene expression in lymphoma of animal 19185. RNA isolated from lymphoma of animal 19185 was analyzed by reverse transcriptase (RT)-PCR to confirm expression of vIL-6 and vFLIP and to analyze expression of ORFs 72 and 73. Lanes 1 and 5 indicate 100 base pair ladder; lane 2, RNA from lymphoma amplified with vIL-6-specific primers; lane 3, same as lane 2, minus RT; lanes 4 and 7, GADPH-specific primers as control for RNA; lane 6, no template control; lane 8, ORF-73–specific primers; lane 9, ORF-72–specific primers; and lane 10, ORF-71–specific primers.
Figure 4
Figure 4
Detection of vIL-6 in RRV-associated lymphomas, RF, and MCD. (A) Lymphoma from RM 19185 was stained with murine monoclonal antibodies specific for RRV vIL-6 and CD20. (C) Lymphoma from animal 19286 was stained with murine monoclonal antibodies specific for RRV vIL-6 and goat antihuman IgM, and nuclei. (B,D) Isotype controls (original magnification ×630). (A) vIL-6 (green) and CD20 (red). (B) Isotype controls for vIL-6 and CD20. (C) vIL-6 (green) and IgM (red), and nuclei (blue). (D) Isotype controls for vIL-6 and IgM. (E) RF from animal 18483 was stained for vIL-6 (green), vimentin (red), and nuclei (blue). Original magnification ×630. (F) MCD lesion from animal 19455 was stained for vIL-6 (green), CD20 (red), and nuclei (blue). Original magnification ×630.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Chang Y, Cesarman E, Pessin MS, et al. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science. 1994;265:1865–1869. - PubMed
    1. Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. N Engl J Med. 1995;332:1186–1191. - PubMed
    1. Soulier J, Grollet L, Oksenhendler E, et al. Kaposi's sarcoma-associated herpesvirus-like DNA sequences in multicentric Castleman's disease. Blood. 1995;86:1276–1280. - PubMed
    1. Oksenhendler E, Boulanger E, Galicier L, et al. High incidence of Kaposi sarcoma-associated herpesvirus-related non-Hodgkin lymphoma in patients with HIV infection and multicentric Castleman disease. Blood. 2002;99:2331–2336. - PubMed
    1. Russo JJ, Bohenzky RA, Chien MC, et al. Nucleotide sequence of the Kaposi sarcoma-associated herpesvirus (HHV8). Proc Natl Acad Sci U S A. 1996;93:14862–14867. - PMC - PubMed

Publication types

MeSH terms