Elsevier

Vaccine

Volume 28, Issue 52, 6 December 2010, Pages 8315-8326
Vaccine

High-throughput automated image analysis of neuroinflammation and neurodegeneration enables quantitative assessment of virus neurovirulence

https://doi.org/10.1016/j.vaccine.2010.07.070Get rights and content

Abstract

Historically, the safety of live attenuated vaccine candidates against neurotropic viruses was assessed by semi-quantitative analysis of virus-induced histopathology in the central nervous system of monkeys. We have developed a high-throughput automated image analysis (AIA) for the quantitative assessment of virus-induced neuroinflammation and neurodegeneration. Evaluation of the results generated by AIA showed that quantitative estimates of lymphocytic infiltration, microglial activation, and neurodegeneration strongly and significantly correlated with results of traditional histopathological scoring. In addition, we show that AIA is a targeted, objective, accurate, and time-efficient approach that provides reliable differentiation of virus neurovirulence. As such, it may become a useful tool in establishing consistent analytical standards across research and development laboratories and regulatory agencies, and may improve the safety evaluation of live virus vaccines. The implementation of this high-throughput AIA will markedly advance many fields of research including virology, neuroinflammation, neuroscience, and vaccinology.

Section snippets

Viruses

LGTV wild-type strain TP21 was received from the Rockefeller Foundation Collection and amplified in Vero cells [31]. The chimeric recombinant TBEV/DEN4Δ30 virus, containing the prM and E genes of the TBEV strain Sofjin and a 30-nucleotide deletion in the 3′ non-coding region of the genome, was originally recovered after transfection of Vero cells with RNA transcripts of its full-length chimeric cDNA genome [32]. The YF 17D vaccine virus was received from Sanofi Pasteur, Inc. (Swiftwater, PA)

Comparison of lymphocytic immunoreactivity and histopathological scores

Two general approaches can be used to evaluate the severity of cellular inflammatory infiltration in the CNS induced by viruses with different neurovirulence. The conventional semi-quantitative approach relies on histopathological analysis of routinely stained (H&E and/or Nissl) sections and assignment of cellular inflammatory infiltration (CII) scores based on the number of perivascular infiltrates and degree of parenchymal infiltration. The representative H&E-stained sections that contained

Acknowledgements

We acknowledge the staff of Bioqual, Inc. (Rockville, MD) and Pathology Associates Division of Charles River Laboratories (Frederick, MD), and Dr. J. Ward, Dr. R. Montali, Lawrence Faucette, Marina Rahman, and Katherine Shea (Comparative Medicine Branch, NIAID, NIH) for their help in conducting the studies. We also thank Drs. J. Taubenberger and S. Whitehead, NIAID, NIH for helpful discussions and critical reading of the manuscript. This work was supported by funds provided by the NIAID

References (33)

  • WHO

    Final report. IABS Scientific Workshop on Neurovirulence Tests for Live Virus Vaccines

    (2005)
  • I. Levenbook et al.

    Neuropathogenesis and neurovirulence of live flaviviral vaccines in monkeys

    J Virol

    (2009)
  • O.A. Maximova et al.

    Neuropathogenesis and neurovirulence of live flaviviral vaccines in monkeys reply

    J Virol

    (2009)
  • N. Nathanson et al.

    Histological studies of the monkey neurovirulence of group B arboviruses. I. A semi-quantitative grading scale

    Am J Epidemiol

    (1965)
  • N. Nathanson et al.

    Histological studies of the monkey neurovirulence of group B arboviruses. II. Selection of indicator centers

    Am J Epidemiol

    (1966)
  • N. Nathanson et al.

    Histological studies of the monkey neurovirulence of group B arboviruses. III. Relative virulence of selected viruses

    Am J Epidemiol

    (1967)
  • Cited by (9)

    • Spleen tyrosine kinase (SYK) blocks autophagic Tau degradation in vitro and in vivo

      2019, Journal of Biological Chemistry
      Citation Excerpt :

      t-SYK levels were also increased in Tg Tau P301S–PBS mice compared with WT littermates and were significantly decreased following chronic SYK inhibition (Fig. 8B). The neuron-specific protein NeuN (45) has been used previously as a marker for neurodegeneration or neuronal loss (46, 47). Neuronal loss revealed by a reduction in NeuN has been described previously in Tg Tau P301S mice (8, 12).

    • Comparative Neuropathology of Major Indian Bluetongue Virus Serotypes in a Neonatal BALB/c Mouse Model

      2018, Journal of Comparative Pathology
      Citation Excerpt :

      Sections (4–5 μm) were stained with haematoxylin and eosin (HE) (Luna, 1968). The histopathological lesions were scored using a modified semiquantitative method (Maximova et al., 2008, 2010): 0, no lesions; 1, minimal lesions; 2, mild lesions; 3, moderate lesions; and 4, severe lesions. This scale was assigned to BTV induced microscopical lesions of brain including: (1) cellular inflammatory infiltration of the brain parenchyma and meninges, (2) vascular endothelial swelling with microhaemorrhage and oedema, and (3) glial cell reaction with ischaemic neuronal degeneration and necrosis.

    • High throughput object-based image analysis of β-amyloid plaques in human and transgenic mouse brain

      2012, Journal of Neuroscience Methods
      Citation Excerpt :

      Such approaches include Acapella™ (Evotec Technologies) Aperio ScanScope XT (Aperio Technologies), BioVision and others (Chubb et al., 2006; Josephs et al., 2008; Willuweit et al., 2009). For example, Aperio, also a whole slide digital based system, has already been implemented for quantitative histopathology in various human and rodent models of CNS disease (Josephs et al., 2008; Krajewska et al., 2010; Maximova et al., 2010). Josephs et al. (2008) reported that amyloid burden in human brain, as assessed by Aperio ScanScope image analysis of whole brain slides, can be measured with sampling in all cortical layers.

    View all citing articles on Scopus
    1

    Tel.: +1 301 594 1616; fax: +1 301 480 5053.

    View full text