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Knockout of Epstein-Barr virus BPLF1 retards B-cell transformation and lymphoma formation in humanized mice.

Whitehurst CB, Li G, Montgomery SA, Montgomery ND, Su L, Pagano JS - MBio (2015)

Bottom Line: EBV also causes aggressive lymphomas in individuals with acquired and innate immune disorders and is strongly associated with diffuse large B-cell lymphomas, classical Hodgkin lymphoma, Burkitt lymphoma, and nasopharyngeal carcinoma (NPC).Typically, EBV initially infects epithelial cells in the oropharynx, followed by a lifelong persistent latent infection in B-cells, which may develop into lymphomas in immunocompromised individuals.Currently, there is no efficacious treatment for EBV, and therapeutic targeting of BPLF1 may lead to a new path to treatment for immunocompromised individuals or transplant recipients infected with EBV.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA cbwhiteh@med.unc.edu.

No MeSH data available.


Related in: MedlinePlus

EBV is detected in mice with tumors. (A) Portions of spleen tissue harvested from mice were disrupted and placed in culture medium. One week later, flasks were examined for GFP (EBV genomes) by immunofluorescence. Top panels are bright-field micrographs, and bottom panels are GFP fluorescence. (B) Detection of EBV genomes in blood of infected mice. Mice were bled weekly throughout the study. The last bleed prior to sacrifice was used to detect EBV genomes by quantitative PCR. Genomes were detected only in mice infected with WT EBV that developed tumors in the spleen. (C) EBV genomes detected in spleens of infected mice. DNA was extracted from splenic tissue at time of harvest, and EBV genome copies were detected with quantitative PCR. Viral genomes were present in all mice presenting with tumors but not in any other mice. EBV genomes were detected in WT2, WT4, WT5, and DUB KO3 (copy numbers 12.7, 99,712, 11.2, and 70.3, respectively) and varied greatly in number likely due to the region utilized for DNA extraction. Some regions of the spleen samples used appeared largely normal and likely resulted in lower genome copy numbers.
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fig5: EBV is detected in mice with tumors. (A) Portions of spleen tissue harvested from mice were disrupted and placed in culture medium. One week later, flasks were examined for GFP (EBV genomes) by immunofluorescence. Top panels are bright-field micrographs, and bottom panels are GFP fluorescence. (B) Detection of EBV genomes in blood of infected mice. Mice were bled weekly throughout the study. The last bleed prior to sacrifice was used to detect EBV genomes by quantitative PCR. Genomes were detected only in mice infected with WT EBV that developed tumors in the spleen. (C) EBV genomes detected in spleens of infected mice. DNA was extracted from splenic tissue at time of harvest, and EBV genome copies were detected with quantitative PCR. Viral genomes were present in all mice presenting with tumors but not in any other mice. EBV genomes were detected in WT2, WT4, WT5, and DUB KO3 (copy numbers 12.7, 99,712, 11.2, and 70.3, respectively) and varied greatly in number likely due to the region utilized for DNA extraction. Some regions of the spleen samples used appeared largely normal and likely resulted in lower genome copy numbers.

Mentions: At the time of harvest, spleen cells from infected mice were cultured to see if EBV could be detected by GFP. Approximately 1 week after plating the spleen cells, samples were examined by immunofluorescence microscopy to detect GFP-positive EBV genomes (Fig. 5A) (bright-field micrographs in upper panels; GFP fluorescence in lower panels). Samples that contained gross tumors were GFP positive, indicating the presence of EBV in the spleen. EBV was only detected in the spleens of animals with spleen tumors (WT2, WT4, WT5, and DUB KO3). Additionally, B-cell lines could only be established from spleens of infected mice that exhibited obvious tumors and presented with GFP-positive genomes. All other cell cultures did not survive.


Knockout of Epstein-Barr virus BPLF1 retards B-cell transformation and lymphoma formation in humanized mice.

Whitehurst CB, Li G, Montgomery SA, Montgomery ND, Su L, Pagano JS - MBio (2015)

EBV is detected in mice with tumors. (A) Portions of spleen tissue harvested from mice were disrupted and placed in culture medium. One week later, flasks were examined for GFP (EBV genomes) by immunofluorescence. Top panels are bright-field micrographs, and bottom panels are GFP fluorescence. (B) Detection of EBV genomes in blood of infected mice. Mice were bled weekly throughout the study. The last bleed prior to sacrifice was used to detect EBV genomes by quantitative PCR. Genomes were detected only in mice infected with WT EBV that developed tumors in the spleen. (C) EBV genomes detected in spleens of infected mice. DNA was extracted from splenic tissue at time of harvest, and EBV genome copies were detected with quantitative PCR. Viral genomes were present in all mice presenting with tumors but not in any other mice. EBV genomes were detected in WT2, WT4, WT5, and DUB KO3 (copy numbers 12.7, 99,712, 11.2, and 70.3, respectively) and varied greatly in number likely due to the region utilized for DNA extraction. Some regions of the spleen samples used appeared largely normal and likely resulted in lower genome copy numbers.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4620474&req=5

fig5: EBV is detected in mice with tumors. (A) Portions of spleen tissue harvested from mice were disrupted and placed in culture medium. One week later, flasks were examined for GFP (EBV genomes) by immunofluorescence. Top panels are bright-field micrographs, and bottom panels are GFP fluorescence. (B) Detection of EBV genomes in blood of infected mice. Mice were bled weekly throughout the study. The last bleed prior to sacrifice was used to detect EBV genomes by quantitative PCR. Genomes were detected only in mice infected with WT EBV that developed tumors in the spleen. (C) EBV genomes detected in spleens of infected mice. DNA was extracted from splenic tissue at time of harvest, and EBV genome copies were detected with quantitative PCR. Viral genomes were present in all mice presenting with tumors but not in any other mice. EBV genomes were detected in WT2, WT4, WT5, and DUB KO3 (copy numbers 12.7, 99,712, 11.2, and 70.3, respectively) and varied greatly in number likely due to the region utilized for DNA extraction. Some regions of the spleen samples used appeared largely normal and likely resulted in lower genome copy numbers.
Mentions: At the time of harvest, spleen cells from infected mice were cultured to see if EBV could be detected by GFP. Approximately 1 week after plating the spleen cells, samples were examined by immunofluorescence microscopy to detect GFP-positive EBV genomes (Fig. 5A) (bright-field micrographs in upper panels; GFP fluorescence in lower panels). Samples that contained gross tumors were GFP positive, indicating the presence of EBV in the spleen. EBV was only detected in the spleens of animals with spleen tumors (WT2, WT4, WT5, and DUB KO3). Additionally, B-cell lines could only be established from spleens of infected mice that exhibited obvious tumors and presented with GFP-positive genomes. All other cell cultures did not survive.

Bottom Line: EBV also causes aggressive lymphomas in individuals with acquired and innate immune disorders and is strongly associated with diffuse large B-cell lymphomas, classical Hodgkin lymphoma, Burkitt lymphoma, and nasopharyngeal carcinoma (NPC).Typically, EBV initially infects epithelial cells in the oropharynx, followed by a lifelong persistent latent infection in B-cells, which may develop into lymphomas in immunocompromised individuals.Currently, there is no efficacious treatment for EBV, and therapeutic targeting of BPLF1 may lead to a new path to treatment for immunocompromised individuals or transplant recipients infected with EBV.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA cbwhiteh@med.unc.edu.

No MeSH data available.


Related in: MedlinePlus