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. 2009 Sep 15;88(5):640-5.
doi: 10.1097/TP.0b013e3181b242a7.

TNF-alpha signaling is not required for in vivo transcriptional reactivation of latent murine cytomegalovirus

Zheng Zhang  1 , Zhigao Li, Shixian Yan, Xueqiong Wang, Michael Abecassis
Affiliations

TNF-alpha signaling is not required for in vivo transcriptional reactivation of latent murine cytomegalovirus

Zheng Zhang et al. Transplantation. .

Abstract

Background: Reactivation of cytomegalovirus (CMV) is frequently observed in recipients of solid organs and bone marrow transplants and is associated with increased risk of acute and chronic allograft rejection, opportunistic infection, graft failure, and patient mortality. The molecular mechanisms by which reactivation occurs are not well understood. Previous studies have suggested that tumor necrosis factor (TNF)-alpha, which is induced by allogeneic transplantation, may have a role in reactivation of CMV through activation of nuclear factor kappa-light-chain-enhancer of activated B cells and subsequent transcriptional reactivation of immediate early (ie) gene expression.

Methods and results: We have tested the role of TNF-alpha in the reactivation of CMV directly by testing whether TNF-alpha is required to initiate transcription of ie gene expression in a murine model of allogeneic transplantation of kidneys latently infected with mouse CMV.

Conclusions: Our studies show that although TNF-alpha seems to be sufficient, it is not required for initiating transcription of ie gene expression in this model, suggesting that both TNF-alpha-dependent and -independent pathways play an important role in the reactivation of latent CMV infection.

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Conflict of interest statement

The authors have no conflicts of interest to disclose.

Figures

Fig. 1
Fig. 1. TNF-α RNA is derived from both donor and recipient cells in kidney allografts
RNAs from control and transplanted kidneys of latently infected TNF-α-sufficient BALB/c mice transplanted into TNF-sufficient C57BL/6 mice treated with control antibody (n=7) or TNF-deficient (n=5) recipients treated with anti-TNF-α were analyzed for TNF-α expression by real time PCR. Expression was normalized to HPRT. The fold change for each mouse was calculated as the ratio of the transplanted kidney to the contralateral control as described in the methods. Results shown are the average fold change plus standard error. *, p < 0.001.
Fig. 2
Fig. 2. Role of TNF–α in transcriptional reactivation of ie-1 expression
RT-PCR and Southern blot hybridization analysis of RNA extracted from kidneys of six MCMV (Smith strain) latently infected BALB/c mice transplanted into allogeneic C57BL/6 recipient mice treated with control antibody (A) or into TNF–α-deficient mice treated with anti-TNF–α (B). Pairs of contralateral control (C) and transplanted (G) kidneys from each mouse are shown grouped together. ie-1 RNA was amplified from the cDNA with ie-1 specific primers CH16 and CH17 and detected by hybridization with CH15 (6) as previously described (15). ie-1 DNA and RNA are distinguishable by size. R, RNA size marker; D, DNA size marker. Fig. 2C and 2D. In a separate experiment, ie-1 and ie-3 are induced in 3/3 and 1/3 WT mice (control antibody) respectively, compared to 2/4 and 1/4 respectively in TNF-α KO mice treated with anti-TNF-α antibody.
Fig. 3
Fig. 3. Acute infection of Δm157 virus in TNFR1/TNFR2-deficient mice is similar to that of Smith virus in wild type BALB/c mice
Organs from infected mice (n=3) were homogenized and analyzed for the presence of infectious virus by plaque assay 4 days after infection. WT, BALB/c mice infected with Smith strain virus; TNFR-/-, B6;129S-Tnfrsflatm1ImxTnfrsf1btm1Imx mice infected with Δm157 virus. Results shown are the average plus standard deviation.
Fig. 4
Fig. 4. Δm157 virus establishes latency in TNFR1/TNFR2-deficient mice
(A) Nested PCR analysis of MCMV ie-1 DNA in lungs and kidneys of Δm157-infected B6;129S-Tnfrsflatm1ImxTnfrsf1btm1Imx mice. Lanes: L, nested PCR analysis of lung DNA from a Smith-infected BALB/c mouse; 1,2,3, nested PCR analysis of DNA isolated from lungs and kidneys of three Δm157 latent B6;129S-Tnfrsflatm1ImxTnfrsf1btm1Imx mice; N, no template control. (B) Titers of reactivated virus in supernatants derived from spleen explants of Smith infected BALB/c mice (WT) or Δm157-infected B6;129S-Tnfrsflatm1ImxTnfrsf1btm1Imx (TNFR-/-) mice. Results shown are the average titers of explants from three mice analyzed in duplicate plus standard deviation.
Fig. 5
Fig. 5. TNFR signaling in donor kidneys is not required for transcriptional reactivation of ie gene expression
RNA extracted from control (C) and transplanted (G) kidneys of eight Δm157 latently infected B6;129S-Tnfrsflatm1ImxTnfrsf1btm1Imx mice transplanted into allogeneic BALB/c recipients was analyzed by RT-PCR and Southern blot hybridization for expression of ie-1 RNA. R, RNA size marker; D, DNA size marker.
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References

    1. Razonable RR, Paya CV. Beta-Herpesviruses in transplantation. Reviews in Medical Microbiology. 2002;13:163.
    1. Reddehase MJ, Simon CO, Seckert CK, Lemmermann N, Grzimek NK. Murine model of cytomegalovirus latency and reactivation. Curr Top Microbiol Immunol. 2008;325:315. - PubMed
    1. Kurz SK, Rapp M, Steffens HP, Grzimek NK, Schmalz S, Reddehase MJ. Focal transcriptional activity of murine cytomegalovirus during latency in the lungs. J Virol. 1999;73(1):482. - PMC - PubMed
    1. Simon CO, Seckert CK, Dreis D, Reddehase MJ, Grzimek NK. Role for tumor necrosis factor alpha in murine cytomegalovirus transcriptional reactivation in latently infected lungs. J Virol. 2005;79(1):326. - PMC - PubMed
    1. Grzimek NK, Dreis D, Schmalz S, Reddehase MJ. Random, asynchronous, and asymmetric transcriptional activity of enhancer-flanking major immediate-early genes ie1/3 and ie2 during murine cytomegalovirus latency in the lungs. J Virol. 2001;75(6):2692. - PMC - PubMed

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