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. 2009 Mar;83(5):2226-36.
doi: 10.1128/JVI.01655-08. Epub 2008 Dec 10.

Role for CCR5 in dissemination of vaccinia virus in vivo

Ramtin Rahbar  1 , Thomas T Murooka, Eleanor N Fish
Affiliations

Role for CCR5 in dissemination of vaccinia virus in vivo

Ramtin Rahbar et al. J Virol. 2009 Mar.

Abstract

In an earlier report, we provided evidence that expression of CCR5 by primary human T cells renders them permissive for vaccinia virus (VACV) replication. This may represent a mechanism for dissemination throughout the lymphatic system. To test this hypothesis, wild-type CCR5(+/+) and CCR5 null mice were challenged with VACV by intranasal inoculation. In time course studies using different infective doses of VACV, we identified viral replication in the lungs of both CCR5(+/+) and CCR5(-/-) mice, yet there were diminished viral loads in the spleens and brains of CCR5(-/-) mice compared with CCR5(+/+) mice. Moreover, in association with VACV infection, we provide evidence for CD4+ and CD8+ T-cell as well as CD11c+ and F4/80+ cell infiltration into the lungs of CCR5(+/+) but not CCR5(-/-) mice, and we show that the CCR5-expressing T cells harbor virus. We demonstrate that this CCR5 dependence is VACV specific, since CCR5(-/-) mice are as susceptible to intranasal influenza virus (A/WSN/33) infection as CCR5(+/+) mice. In a final series of experiments, we provide evidence that adoptive transfer of CCR5(+/+) bone marrow leukocytes into CCR5(-/-) mice restores VACV permissiveness, with evidence of lung and spleen infection. Taken together, our data suggest a novel role for CCR5 in VACV dissemination in vivo.

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Figures

FIG. 1.
FIG. 1.
CCR5−/− mice are less susceptible to VACV infection. Groups of female mice age 6 to 8 weeks were either mock infected (PBS) or infected with 104, 105, or 106 PFU of VACV by intranasal inoculation. (A) Body weight was measured daily, and values are recorded as the mean percent weight loss at the indicated time point compared to that of uninfected control mice ± standard error. 104 PFU, n = 10; 105 PFU, n = 12 until day 8 and n = 7 thereafter; 106 PFU, n = 12 until day 3 and n = 7 thereafter. CCR5+/+ mock infected, ▴; CCR5+/+ VACV infected, ⧫; CCR5−/− mock infected, •; CCR5−/− VACV infected, ░⃞. (B) Viral titers were measured in lungs, spleens, and brains of CCR5+/+ (▪) and CCR5−/− (formula image) mice (n = 10) at the indicated times p.i. with 104 PFU of VACV, as described in Materials and Methods.(C) Lungs of mock-infected or VACV-infected (104 PFU) CCR5+/+ and CCR5−/− mice were harvested on day 7 p.i., fixed in 2% paraformaldehyde, and embedded in paraffin, and 6-μm-thick histological sections were prepared and stained with hematoxylin-eosin. The dotted line indicates the lower level of detection for VACV. *, P < 0.05. ND, not detected.
FIG. 2.
FIG. 2.
Intranasal inoculation with VACV leads to an increase in BAL fluid viral titer and an influx of CD4+ and CD8+ T cells into the lungs of CCR5+/+but not CCR5−/− mice. Groups of female mice (n = 10), age 6 to 8 weeks, were infected with 104 PFU of EGFP-VACV by intranasal inoculation. Mice were euthanized at the indicated times p.i. and their BAL fluid collected as described in Materials and Methods. (A) Viable cell counts in the BAL fluid of CCR5+/+ (▪) and CCR5−/− (▒) were determined by trypan blue exclusion. Values are means ± standard errors and are representative of two independent experiments. (B) Viral titers in BAL fluid of CCR5+/+ (▪)and CCR5−/− (░⃞)mice were measured at the indicated times p.i. The dotted line indicates the lower level of detection for VACV. (C and E) CD4+ (C) and CD8+ (E) T-cell counts were determined by flow cytometry, using PE-Cy5-conjugated anti CD4/CD8 and APC-conjugated CD3 antibodies. ▪, total CD4+/CD8+ cell counts; ░⃞, VACV-infected CD4/CD8+ T cells as determined by quantitation of EGFP-positive cells as described in Materials and Methods. (D and F) The data are partitioned as VACV-infected CD4+ (D) and CD8+ (F) T cells in the BAL fluid of CCR5+/+ and CCR5−/− mice, recorded as the percent positive ± standard errors; results are representative of two independent experiments. ▒, CD4+ CCR5−/CD8+ CCR5− T-cell count in the BAL fluid from CCR5+/+ mice; □, CD4+ CCR5+/CD8+ CCR5+ T-cell count in the BAL fluid from CCR5+/+ mice; ▧, CD4+ CCR5−/CD8+ CCR5− T-cell count in the BAL fluid from CCR5−/− mice. *, P < 0.05; **, P < 0.01.
FIG. 3.
FIG. 3.
VACV infection of CCR5+/+ but not CCR5−/− mice influences the T-cell responses in secondary lymphoid organs. Groups of female mice (n = 10), age 6 to 8 weeks, were infected with 104 PFU of EGFP-VACV by intranasal inoculation. Mice were euthanized at the indicated times p.i., and CD3+ CD4+ and CD3+ CD8+ T-cell counts in the spleens (A and B) and mLNs (C and D) were determined by flow cytometry, using PE-Cy5-conjugated anti CD4/CD8 and APC-conjugated CD3 antibodies. ▪, total CCR5+/+ CD4+/CD8+; ░⃞, total CCR5−/− CD4+/CD8+. Data are shown as mean cell counts ± standard errors and are representative of two independent experiments. *, P < 0.05; **, P < 0.01.
FIG. 4.
FIG. 4.
VACV infection modulates CD11c+ DC and F4/80+ macrophage populations in the mLNs and spleens of CCR5+/+ but not CCR5−/− mice. CD11c+ and F4/80+ cell infiltration into the mLNs (A and C) and spleens (B and D) of EGFP-VACV-infected mice (104 PFU) was determined by flow cytometry, using APC-conjugated anti-CD11c and PE-Cy5-conjugated anti-F4/80 antibodies. Data are presented as total cell number infiltrate of CD11c+/F4/80cells. ▪, total CCR5+/+ CD11c+/F4/80+; ░⃞, total CCR5−/− CD11c+/F4/80+. Data are shown as mean cell counts ± standard errors and are representative of two independent experiments. *, P < 0.05 **, P < 0.01.
FIG. 5.
FIG. 5.
CCR5−/− mice are permissive for influenza A/WSN/33 virus infection. (A) Groups of female mice (n = 6), age 6 to 8 weeks, were infected with 0.1 HAU influenza A/WSN/33 virus and body weight measured daily. Body weight loss was expressed as the mean percent weight loss of animals at the indicated time points compared to that of controls ± standard errors. ▪, CCR5+/+; ░⃞, CCR5−/−. (B) Lungs of mock- and influenza A/WSN/33 virus-infected CCR5+/+ and CCR5−/− mice harvested on days 3 and 6 p.i. were fixed in 2% paraformaldehyde and embedded in paraffin, and 6-μm-thick histological sections were prepared and stained with hematoxylin and eosin.
FIG. 6.
FIG. 6.
Adoptive transfer of CCR5+/+ leukocytes into CCR5−/− mice restores permissiveness to VACV infection. Hematopoietic leukocytes from CCR5+/+ mice were isolated and transplanted into irradiated CCR5+/+ and CCR5−/− mice as described in Materials and Methods. After 2 weeks, transplanted mice were either mock or EGFP-VACV infected (104 PFU). (A) At the indicated times p.i., mice were euthanized and VACV titers in lungs, BAL fluid, and spleens of recipient CCR5+/+ (▪)and CCR5−/− (░⃞) mice were determined. The data are representative of two independent experiments. The dotted line indicates the lower level of detection for VACV. ND, not detected. (B to M) Single-cell suspensions from the BAL fluid, spleens, and mLNs of recipient CCR5+/+ (▪) and CCR5−/− (░⃞) mice were obtained and stained with PE-Cy5-conjugated anti-CD4/CD8/F480 and APC-conjugated anti-CD3/CD11c antibodies at the indicated time points. Values are the mean CD4+ (B, D, and F) and CD8+ (C, E, and G) T-cell, CD11c+ cell (H, I, and J), and F4/80+ cell (K, L, and M) counts ± standard errors from two independent experiments. ░⃞, total VACV-infected CCR5+/+ CD4+/CD8+ cells; (□), total VACV-infected CCR5−/− CD4+/CD8+/CD11c+/F4/80+cells. *, P < 0.05; **, P < 0.01.
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References

    1. Alcami, A., J. A. Symons, P. D. Collins, T. J. Williams, and G. L. Smith. 1998. Blockade of chemokine activity by a soluble chemokine binding protein from vaccinia virus. J. Immunol. 160624-633. - PubMed
    1. Andres, P. G., P. L. Beck, E. Mizoguchi, A. Mizoguchi, A. K. Bhan, T. Dawson, W. A. Kuziel, N. Maeda, R. P. MacDermott, D. K. Podolsky, and H. C. Reinecker. 2000. Mice with a selective deletion of the CC chemokine receptors 5 or 2 are protected from dextran sodium sulfate-mediated colitis: lack of CC chemokine receptor 5 expression results in a NK1.1+ lymphocyte-associated Th2-type immune response in the intestine. J. Immunol. 1646303-6312. - PubMed
    1. Baird, A. M., R. M. Gerstein, and L. J. Berg. 1999. The role of cytokine receptor signaling in lymphocyte development. Curr. Opin. Immunol. 11157-166. - PubMed
    1. Bartlett, N. W., K. Buttigieg, S. V. Kotenko, and G. L. Smith. 2005. Murine interferon lambdas (type III interferons) exhibit potent antiviral activity in vivo in a poxvirus infection model. J. Gen. Virol. 861589-1596. - PubMed
    1. Breman, J. G., and D. A. Henderson. 2002. Diagnosis and management of smallpox. N. Engl. J. Med. 3461300-1308. - PubMed

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