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T-Cell tropism of simian varicella virus during primary infection.

Ouwendijk WJ, Mahalingam R, de Swart RL, Haagmans BL, van Amerongen G, Getu S, Gilden D, Osterhaus AD, Verjans GM - PLoS Pathog. (2013)

Bottom Line: Except for pneumonitis, pathology produced by SVV-EGFP was less compared to SVV-wt.In ganglia, SVV was found primarily in neurons and occasionally in memory T-cells adjacent to neurons.In conclusion, the data suggest the role of memory T-cells in disseminating SVV to its target organs during primary infection of its natural and immunocompetent host.

View Article: PubMed Central - PubMed

Affiliation: Department of Viroscience, Erasmus MC, Rotterdam, the Netherlands.

ABSTRACT
Varicella-zoster virus (VZV) causes varicella, establishes a life-long latent infection of ganglia and reactivates to cause herpes zoster. The cell types that transport VZV from the respiratory tract to skin and ganglia during primary infection are unknown. Clinical, pathological, virological and immunological features of simian varicella virus (SVV) infection of non-human primates parallel those of primary VZV infection in humans. To identify the host cell types involved in virus dissemination and pathology, we infected African green monkeys intratracheally with recombinant SVV expressing enhanced green fluorescent protein (SVV-EGFP) and with wild-type SVV (SVV-wt) as a control. The SVV-infected cell types and virus kinetics were determined by flow cytometry and immunohistochemistry, and virus culture and SVV-specific real-time PCR, respectively. All monkeys developed fever and skin rash. Except for pneumonitis, pathology produced by SVV-EGFP was less compared to SVV-wt. In lungs, SVV infected alveolar myeloid cells and T-cells. During viremia the virus preferentially infected memory T-cells, initially central memory T-cells and subsequently effector memory T-cells. In early non-vesicular stages of varicella, SVV was seen mainly in perivascular skin infiltrates composed of macrophages, dendritic cells, dendrocytes and memory T-cells, implicating hematogenous spread. In ganglia, SVV was found primarily in neurons and occasionally in memory T-cells adjacent to neurons. In conclusion, the data suggest the role of memory T-cells in disseminating SVV to its target organs during primary infection of its natural and immunocompetent host.

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SVV infects predominantly memory T-cells in blood after infection in African green monkeys.(A) Average SVV DNA load in PBMC of SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys determined by SVV-specific real-time qPCR. (B) PBMC from SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys were analyzed for infectious SVV by co-cultivation with BSC-1 cells. (C) PBMC from SVV-EGFP−infected monkeys were analyzed for EGFP expression by flow cytometry. (D) EGFP expression in PBMC subsets from SVV-EGFP−infected monkeys. Data are given as percentage of EGFPpos cells within each lymphocyte subset relative to the total number of PBMC, as determined by flow cytometry. Lymphocyte subsets were defined by differential expression of the following markers: T-cells = CD3posCD16neg cells, B-cells = CD20posMHC-IIposcells, natural killer (NK) cells = CD3negCD16pos cells, dendritic cells (DC) = CD3negCD14negCD16negCD20negCD14negMHC-IIpos cells, and monocytes = CD3negCD14posMHC-IIpos cells. (E and F) Percentage of EGFPpos cells among each T-cell subset relative to the number of CD8bright, CD8dim and CD4pos T-cells (E) and in naive, central memory and effector memory T-cells (F) from SVV-EGFP−infected monkeys as evaluated by flow cytometry. In all panels, data are means ± SEM.
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ppat-1003368-g004: SVV infects predominantly memory T-cells in blood after infection in African green monkeys.(A) Average SVV DNA load in PBMC of SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys determined by SVV-specific real-time qPCR. (B) PBMC from SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys were analyzed for infectious SVV by co-cultivation with BSC-1 cells. (C) PBMC from SVV-EGFP−infected monkeys were analyzed for EGFP expression by flow cytometry. (D) EGFP expression in PBMC subsets from SVV-EGFP−infected monkeys. Data are given as percentage of EGFPpos cells within each lymphocyte subset relative to the total number of PBMC, as determined by flow cytometry. Lymphocyte subsets were defined by differential expression of the following markers: T-cells = CD3posCD16neg cells, B-cells = CD20posMHC-IIposcells, natural killer (NK) cells = CD3negCD16pos cells, dendritic cells (DC) = CD3negCD14negCD16negCD20negCD14negMHC-IIpos cells, and monocytes = CD3negCD14posMHC-IIpos cells. (E and F) Percentage of EGFPpos cells among each T-cell subset relative to the number of CD8bright, CD8dim and CD4pos T-cells (E) and in naive, central memory and effector memory T-cells (F) from SVV-EGFP−infected monkeys as evaluated by flow cytometry. In all panels, data are means ± SEM.

Mentions: To determine the kinetics of virus infection and identify the blood lymphocyte subsets infected during the viremic phase of varicella, peripheral blood mononuclear cells (PBMC) isolated at multiple dpi from SVV-infected monkeys were analyzed. SVV DNA was detected in PBMC from 2 dpi until necropsy (Fig. 4A). Viral DNA load in PBMC peaked at 7 dpi and was higher in SVV-wt− compared to SVV-EGFP−infected monkeys (Fig. 4A). Infectious virus was isolated from PBMC of both SVV-EGFP− and SVV-wt−infected monkeys until 9 and 11 dpi (Fig. 4B). EGFPpos lymphocytes were detected from 5 to 11 dpi, peaking at 7 dpi (Fig. 4C). Together, the data indicate that the kinetics of viral DNA load and infectious virus titer represent the temporal change in the number of circulating SVV-infected lymphocytes, but not in the replication of SVV in blood lymphocytes during viremia. The rapid loss of SVV-infected lymphocytes from the circulation could be caused by virus-induced apoptosis [26] or, alternatively, infected lymphocytes may be cleared from the circulation by the SVV-specific adaptive immune response [14], [21], [27].


T-Cell tropism of simian varicella virus during primary infection.

Ouwendijk WJ, Mahalingam R, de Swart RL, Haagmans BL, van Amerongen G, Getu S, Gilden D, Osterhaus AD, Verjans GM - PLoS Pathog. (2013)

SVV infects predominantly memory T-cells in blood after infection in African green monkeys.(A) Average SVV DNA load in PBMC of SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys determined by SVV-specific real-time qPCR. (B) PBMC from SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys were analyzed for infectious SVV by co-cultivation with BSC-1 cells. (C) PBMC from SVV-EGFP−infected monkeys were analyzed for EGFP expression by flow cytometry. (D) EGFP expression in PBMC subsets from SVV-EGFP−infected monkeys. Data are given as percentage of EGFPpos cells within each lymphocyte subset relative to the total number of PBMC, as determined by flow cytometry. Lymphocyte subsets were defined by differential expression of the following markers: T-cells = CD3posCD16neg cells, B-cells = CD20posMHC-IIposcells, natural killer (NK) cells = CD3negCD16pos cells, dendritic cells (DC) = CD3negCD14negCD16negCD20negCD14negMHC-IIpos cells, and monocytes = CD3negCD14posMHC-IIpos cells. (E and F) Percentage of EGFPpos cells among each T-cell subset relative to the number of CD8bright, CD8dim and CD4pos T-cells (E) and in naive, central memory and effector memory T-cells (F) from SVV-EGFP−infected monkeys as evaluated by flow cytometry. In all panels, data are means ± SEM.
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ppat-1003368-g004: SVV infects predominantly memory T-cells in blood after infection in African green monkeys.(A) Average SVV DNA load in PBMC of SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys determined by SVV-specific real-time qPCR. (B) PBMC from SVV-wt− (closed squares) and SVV-EGFP− (open squares) infected monkeys were analyzed for infectious SVV by co-cultivation with BSC-1 cells. (C) PBMC from SVV-EGFP−infected monkeys were analyzed for EGFP expression by flow cytometry. (D) EGFP expression in PBMC subsets from SVV-EGFP−infected monkeys. Data are given as percentage of EGFPpos cells within each lymphocyte subset relative to the total number of PBMC, as determined by flow cytometry. Lymphocyte subsets were defined by differential expression of the following markers: T-cells = CD3posCD16neg cells, B-cells = CD20posMHC-IIposcells, natural killer (NK) cells = CD3negCD16pos cells, dendritic cells (DC) = CD3negCD14negCD16negCD20negCD14negMHC-IIpos cells, and monocytes = CD3negCD14posMHC-IIpos cells. (E and F) Percentage of EGFPpos cells among each T-cell subset relative to the number of CD8bright, CD8dim and CD4pos T-cells (E) and in naive, central memory and effector memory T-cells (F) from SVV-EGFP−infected monkeys as evaluated by flow cytometry. In all panels, data are means ± SEM.
Mentions: To determine the kinetics of virus infection and identify the blood lymphocyte subsets infected during the viremic phase of varicella, peripheral blood mononuclear cells (PBMC) isolated at multiple dpi from SVV-infected monkeys were analyzed. SVV DNA was detected in PBMC from 2 dpi until necropsy (Fig. 4A). Viral DNA load in PBMC peaked at 7 dpi and was higher in SVV-wt− compared to SVV-EGFP−infected monkeys (Fig. 4A). Infectious virus was isolated from PBMC of both SVV-EGFP− and SVV-wt−infected monkeys until 9 and 11 dpi (Fig. 4B). EGFPpos lymphocytes were detected from 5 to 11 dpi, peaking at 7 dpi (Fig. 4C). Together, the data indicate that the kinetics of viral DNA load and infectious virus titer represent the temporal change in the number of circulating SVV-infected lymphocytes, but not in the replication of SVV in blood lymphocytes during viremia. The rapid loss of SVV-infected lymphocytes from the circulation could be caused by virus-induced apoptosis [26] or, alternatively, infected lymphocytes may be cleared from the circulation by the SVV-specific adaptive immune response [14], [21], [27].

Bottom Line: Except for pneumonitis, pathology produced by SVV-EGFP was less compared to SVV-wt.In ganglia, SVV was found primarily in neurons and occasionally in memory T-cells adjacent to neurons.In conclusion, the data suggest the role of memory T-cells in disseminating SVV to its target organs during primary infection of its natural and immunocompetent host.

View Article: PubMed Central - PubMed

Affiliation: Department of Viroscience, Erasmus MC, Rotterdam, the Netherlands.

ABSTRACT
Varicella-zoster virus (VZV) causes varicella, establishes a life-long latent infection of ganglia and reactivates to cause herpes zoster. The cell types that transport VZV from the respiratory tract to skin and ganglia during primary infection are unknown. Clinical, pathological, virological and immunological features of simian varicella virus (SVV) infection of non-human primates parallel those of primary VZV infection in humans. To identify the host cell types involved in virus dissemination and pathology, we infected African green monkeys intratracheally with recombinant SVV expressing enhanced green fluorescent protein (SVV-EGFP) and with wild-type SVV (SVV-wt) as a control. The SVV-infected cell types and virus kinetics were determined by flow cytometry and immunohistochemistry, and virus culture and SVV-specific real-time PCR, respectively. All monkeys developed fever and skin rash. Except for pneumonitis, pathology produced by SVV-EGFP was less compared to SVV-wt. In lungs, SVV infected alveolar myeloid cells and T-cells. During viremia the virus preferentially infected memory T-cells, initially central memory T-cells and subsequently effector memory T-cells. In early non-vesicular stages of varicella, SVV was seen mainly in perivascular skin infiltrates composed of macrophages, dendritic cells, dendrocytes and memory T-cells, implicating hematogenous spread. In ganglia, SVV was found primarily in neurons and occasionally in memory T-cells adjacent to neurons. In conclusion, the data suggest the role of memory T-cells in disseminating SVV to its target organs during primary infection of its natural and immunocompetent host.

Show MeSH
Related in: MedlinePlus