Infection of cynomolgus macaques (Macaca fascicularis) and rhesus macaques (Macaca mulatta) with different wild-type measles viruses

Lemon

Ludlow

et al. 2010

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The global increase in measles vaccination has resulted in a significant reduction of measles mortality. The standard route of administration for the live-attenuated measles virus (MV) vaccine is subcutaneous injection, although alternative needle-free routes, including aerosol delivery, are under investigation. In vitro, attenuated MV has a much wider tropism than clinical isolates, as it can use both CD46 and CD150 as cellular receptors. To compare the in vivo tropism of attenuated and pathogenic MV, we infected cynomolgus macaques with pathogenic or attenuated recombinant MV expressing enhanced green fluorescent protein (GFP) (strains IC323 and Edmonston, respectively) via the intratracheal or aerosol route. Surprisingly, viral loads and cellular tropism in the lungs were similar for the two viruses regardless of the route of administration, and CD11c-positive cells were identified as the major target population. However, only the pathogenic MV caused significant viremia, which resulted in massive virus replication in B and T lymphocytes in lymphoid tissues and viral dissemination to the skin and the submucosa of respiratory epithelia. Attenuated MV was rarely detected in lymphoid tissues, and when it was, only in isolated infected cells. Following aerosol inhalation, attenuated MV was detected at early time points in the upper respiratory tract, suggesting local virus replication. This contrasts with pathogenic MV, which invaded the upper respiratory tract only after the onset of viremia. This study shows that despite in vitro differences, attenuated and pathogenic MV show highly similar in vivo tropism in the lungs. However, systemic spread of attenuated MV is restricted.Measles virus (MV) is one of the most contagious human viruses and is transmitted via aerosols or by direct contact with contaminated respiratory secretions. Clinical symptoms appear approximately 2 weeks after infection and include fever, rash, cough, coryza, and conjunctivitis (20). Measles is associated with immunosuppression, resulting in increased susceptibility to opportunistic infections. While significant progress has been made in global control programs, 164,000 deaths were attributed to measles in 2008 (46).MV was first isolated in cell culture in 1954 (16). This Edmonston wild-type MV strain was passaged multiple times in primary human kidney and amnion cells and adapted to eggs and chicken embryo fibroblasts to produce the live-attenuated Edmonston-B vaccine virus (15), which was later replaced by the more attenuated MV strains (Edmonston-Zagreb, Moraten, and Schwarz) (34). These vaccines have been shown to be safe and effective, and high coverage in two-dose regimens has successfully interrupted endemic MV transmission in large geographic areas (6).For many years, laboratory-adapted MV-Edmonston strains were used as the prototype virus and were shown to display a wide cellular tropism in vitro. The virus efficiently infected epithelial cells, which were considered the target cells for primary MV infection in vivo (22). In 1993,...

Section: In Vitrosupporting

confidence: 57%

“…Isolation of MV was performed on human B-LCL using an infectious-center test as previously described (14). Virus isolations were monitored by UV microscopy for EGFP fluorescence after cocultivation with B-LCL for 3 to 6 days.…”

Section: Cells and Virusesmentioning

confidence: 99%

In VivoTropism of Attenuated and Pathogenic Measles Virus Expressing Green Fluorescent Protein in Macaques

Lemon

Ludlow

et al. 2010

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“…Isolation of MV was performed on Vero cells using an infectious-center test as previously described (19,20). Virus isolations were monitored for cytopathic effect and/or fluorescence on day 6 after isolation.…”

Section: Methodsmentioning

confidence: 99%

Live-Attenuated Measles Virus Vaccine Targets Dendritic Cells and Macrophages in Muscle of Nonhuman Primates

Rennick

Carsillo

et al. 2015

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Although live-attenuated measles virus (MV) vaccines have been used successfully for over 50 years, the target cells that sustain virus replication in vivo are still unknown. We generated a reverse genetics system for the live-attenuated MV vaccine strain Edmonston-Zagreb ( IMPORTANCE Even though MV strain Edmonston-Zagreb has long been used as a live-attenuated vaccine (LAV) to protect against measles, nothing is known about the primary cells in which the virus replicates in vivo.This is vital information given the push to move toward needle-free routes of vaccination, since vaccine virus replication is essential for vaccination efficacy. We have generated a number of recombinant MV strains expressing enhanced green fluorescent protein. The virus that best mimicked the nonrecombinant vaccine virus was formulated according to protocols for production of commercial vaccine virus batches, and was subsequently used to assess viral tropism in nonhuman primates. The virus primarily replicated in professional antigen-presenting cells, which may explain why this LAV is so immunogenic and efficacious.

“…Cells were counted using a hemocytometer and used directly for flow cytometry and virus isolation. Isolation of CDV was performed on Vero-dogSLAM (VDS) cells (4) using an infectious center test as previously described (28). Virus isolations were monitored by UV microscopy for EGFP fluorescence after cocultivation with VDS for 3 to 6 days, and results were expressed as number of virus-infected cells/10 6 total cells.…”

Section: Methodsmentioning

confidence: 99%

“…After being thawed, samples were vortexed, the swab was removed, and the remaining transport medium was used for virus isolation. Isolation of CDV was performed on VDS cells using an infectious center test as previously described (28). The isolations were screened for EGFP fluorescence at days 3 and 7 posttitration, and results are expressed as TCID 50 /ml.…”

Section: Bronchoalveolar Lavage (Bal)mentioning

confidence: 99%

Measles Vaccination of Nonhuman Primates Provides Partial Protection against Infection with Canine Distemper Virus

Ludlow

Verburgh

et al. 2014

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Measles virus (MV) is being considered for global eradication, which would likely reduce compliance with MV vaccination. As a result, children will grow up without MV-specific immunity, creating a potential niche for closely related animal morbilliviruses such as canine distemper virus (CDV). Natural CDV infection causing clinical signs has never been reported in humans, but recent outbreaks in captive macaques have shown that CDV can cause disease in primates. We studied the virulence and tropism of recombinant CDV expressing enhanced green fluorescent protein in naive and measles-vaccinated cynomolgus macaques. In naive animals CDV caused viremia and fever and predominantly infected CD150 ؉ lymphocytes and dendritic cells. Virus was reisolated from the upper and lower respiratory tracts, but infection of epithelial or neuronal cells was not detectable at the time points examined, and the infections were self-limiting. This demonstrates that CDV readily infects nonhuman primates but suggests that additional mutations are necessary to achieve full virulence in nonnatural hosts. Partial protection against CDV was observed in measles-vaccinated macaques, as demonstrated by accelerated control of virus replication and limited shedding from the upper respiratory tract. While neither CDV infection nor MV vaccination induced detectable cross-reactive neutralizing antibodies, MV-specific neutralizing antibody levels of MV-vaccinated macaques were boosted by CDV challenge infection, suggesting that cross-reactive VN epitopes exist. Rapid increases in white blood cell counts in MV-vaccinated macaques following CDV challenge suggested that cross-reactive cellular immune responses were also present. This study demonstrates that zoonotic morbillivirus infections can be controlled by measles vaccination. IMPORTANCEThroughout history viral zoonoses have had a substantial impact on human health. Given the drive toward global eradication of measles, it is essential to understand the zoonotic potential of animal morbilliviruses. Morbilliviruses are thought to have evolved from a common ancestral virus that jumped species and adapted to new hosts. Recently, canine distemper virus (CDV), a morbillivirus normally restricted to carnivores, caused disease outbreaks in nonhuman primates. Here, we report that experimental CDV infection of monkeys resulted in fever and leukopenia. The virus replicated to high levels in lymphocytes but did not spread to epithelial cells or the central nervous system. Importantly, like measles virus in macaques, the infections were selflimiting. In measles-vaccinated macaques CDV was cleared more rapidly, resulting in limited virus shedding from the upper respiratory tract. These studies demonstrate that although CDV can readily infect primates, measles immunity is protective, and CDV infection is self-limiting.