Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Nov;87(21):11908-11.
doi: 10.1128/JVI.01620-13. Epub 2013 Aug 21.

Latency locus complements MicroRNA 155 deficiency in vivo

Sang-Hoon Sin  1 , Yong Baek Kim, Dirk P Dittmer
Affiliations

Latency locus complements MicroRNA 155 deficiency in vivo

Sang-Hoon Sin et al. J Virol. 2013 Nov.

Abstract

MicroRNA-155 (miR-155) is expressed in many cancers. It also executes evolutionary conserved functions in normal B cell development. We show that the Kaposi's sarcoma-associated herpesvirus (KSHV) latency locus, which contains an ortholog of miR-155, miR-K12-11, complements B cell deficiencies in miR-155 knockout mice. Germinal center (GC) formation was rescued in spleen, lymph node, and Peyer's patches. Immunoglobulin levels were restored. This demonstrates that KSHV can complement the normal, physiological function of miR-155.

PubMed Disclaimer

Figures

Fig 1
Fig 1
Generation of KSHV latency locus transgenic mice without miR-155. (A) Sequence of human and mouse miR-155. Overlaid is the deep sequencing coverage at each nucleotide position. This represents a new feature in mirBase version 19, which integrates sequence coverage from multiple, previously published RNAseq experiments. Underlined are identical residues. (B) Genotyping was done using qPCR, and PCR products were visualized via the LabChip system (Caliper). WT, KSHV latency locus transgenic mouse; het, KSHV latency locus transgene+/− miR-155+/− mouse; KO, miR-155−/− mouse. Shown is a representative of 3 independent experiments. (C) Expression of mature miR-155 was confirmed by qPCR-based TaqMan assay (catalog no. 4427975; Life Technologies). A dotted or solid line represents an miR-155−/− or an miR-155+/+ KSHV latency locus transgenic mouse. (D) Quantitative analysis of miR-155. The expression level is expressed as percentage of U6 in log10 scale on the vertical axis and genotype for KSHV transgene on the vertical axis. The miR-155 genotype is shown on top of each subpanel. Shown are individual data points in dark gray and box-whisker plots in gray. The line indicated median expression. The limit of detection was 0.001% of U6 levels.
Fig 2
Fig 2
Rescue of miR-155 deficiency-induced phenotypes by the KSHV latency locus. Lymphocytes from KSHV latency locus transgenic mice on the miR-155ko background (n = 6) were subjected to flow cytometry analysis. miR-155ko mice (n = 7) were used as a control. The percentage of GC B cells (CD19+ CD38− CD95+) was plotted from PP (A), spleen (B), and mLN (C). *, P ≤ 0.05. (C) KSHV latency locus transgenic mice on the miR-155ko background (n = 6) and miR-155ko mice (n = 6) were immunized intraperitoneally (i.p.) with 200 μg NP-CGG (Biosearch Technologies) per mouse. Percentage of SP GC B cells (CD19+ CD38− CD95+) was determined on day 28 by flow cytometry. *, P ≤ 0.05. All flow cytometry data were acquired on CyAn (Beckman Coulter) and analyzed using FlowJo (version 7.6.5; Tree Star). Two-tailed Student's t tests were used for statistical analysis. General architecture of PP revealed by H&E staining was shown in the transgenic (D) and wild-type (E) mice on the miR-155ko background. White arrows indicate GC area. PNA+ GC B cells (black arrows) were shown in the transgenic (F) and wild-type (G) mice on the miR-155ko background. Formalin-fixed, paraffin-embedded tissue sections were stained with PNA (Vector Laboratories). Black arrows indicate PNA+ GC B cells. Magnification, ×40. TG means KSHV latency transgenic mice on the miR-155ko background.
Fig 3
Fig 3
KSHV latency locus complements reduced immunoglobulin levels in miR-155ko mice. KSHV latency locus transgenic mice with the miR-155ko background (n = 7) and miR-155ko mice (n = 6) were immunized i.p. with 100 μg alum-precipitated NP-KLH and boosted on day 21. Serum was collected before immunization and on day 25. Levels of non-NP-specific (resting) and NP-specific Ig isotypes were determined using unlabeled anti-mouse Ig antibodies (Southern Biotech) or NP34-bovine serum albumin (Biosearch Technologies). Alkaline phosphatase-labeled antibodies and p-nitrophenyl phosphate were used for detection (all from Southern Biotech). (A) Levels of resting non-NP-specific immunoglobulins of the KSHV transgenic × miR-155ko mice (black dots, n = 7) and miR-155ko (gray triangles, n = 6) were plotted, respectively. (B) Levels of NP-specific immunoglobulins of the KSHV transgenic × miR-155ko mice (black dots, n = 7) and miR-155ko (gray triangles, n = 6) were analyzed 5 days after the first boost with NP-KLH and plotted. *, P ≤ 0.05.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Gottwein E, Corcoran DL, Mukherjee N, Skalsky RL, Hafner M, Nusbaum JD, Shamulailatpam P, Love CL, Dave SS, Tuschl T, Ohler U, Cullen BR. 2011. Viral microRNA targetome of KSHV-infected primary effusion lymphoma cell lines. Cell Host Microbe 10:515–526 - PMC - PubMed
    1. Haecker I, Gay LA, Yang Y, Hu J, Morse AM, McIntyre LM, Renne R. 2012. Ago HITS-CLIP expands understanding of Kaposi's sarcoma-associated herpesvirus miRNA function in primary effusion lymphomas. PLoS Pathog. 8:e1002884.10.1371/journal.ppat.1002884 - DOI - PMC - PubMed
    1. Dittmer D, Lagunoff M, Renne R, Staskus K, Haase A, Ganem D. 1998. A cluster of latently expressed genes in Kaposi's sarcoma-associated herpesvirus. J. Virol. 72:8309–8315 - PMC - PubMed
    1. Fakhari FD, Dittmer DP. 2002. Charting latency transcripts in Kaposi's sarcoma-associated herpesvirus by whole-genome real-time quantitative PCR. J. Virol. 76:6213–6223 - PMC - PubMed
    1. O'Hara AJ, Vahrson W, Dittmer DP. 2008. Gene alteration and precursor and mature microRNA transcription changes contribute to the miRNA signature of primary effusion lymphoma. Blood 111:2347–2353 - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources