Morphologic changes in viral infections

1. Morphological Changes in Viral Infections Shridhan Patil 25 June 2012

2. "The Father of Modern Vaccines." Nobel laureate John Franklin Enders • Known for polio viruses •
Classified viruses on the basis of morphologic effects on cells.

3. Virus NOT considered as cells • Small, obligate intracellular parasites • Strucure – DNA/RNA and protein
coat • Mobile genetic elements • Virion - Function
4. Classification • Size and shape • Chemical composition • Structure of genome • Modes of replication
5. Cytopathic effect(CPE) • Morphological changes in cells caused by viral infections; the responsible virus is
said to be cytopathogenic • Degree of CPE depends on o Virus and host type o Multiplicity of infection
(MOI) o Some viruses do not cause CPE – Interference/ hemadsorption • Application- Virus identification,
unknown virus
6. How virus infects • Live inside cells- avoid immune response • Attachment receptors- DOCKING •
PENETRATION- cytoplasm • BIOSYNTHESIS of viral components • MATURATION • RELEASE
7. Sampling • What types of cases): stool, tissue, specimens are • Gastrointestinal tract saliva, brain biopsy,
collected to diagnose? infections: cerebrospinal fluid stool and rectal swabs • Respiratory tract • Genital
infections: infections: • Vesicular rash: vesicle vesicle fluid or Nasal fluid, skin Swab and bronchial Scrapings
washings, throat • Urinary tract infections: and nasal swabs, sputum • Maculopapular rash: urine throat, stool,
• Eye infections: throat and rectal swabs • Bloodborne infections: and blood • conjunctival • CNS
(encephalitis and swab/scraping meningitis
8. Techniques of virus study • Direct 1. Electron microscopy/ Immuno-electroon microscopy 2. Light
microscopy 3. Ag detection, molecular methods etc.. • Indirect 1. Cell culture 2. Egg pocks on chorio-
allantoic membrane 3. Animal disease/death
9. • Find suitable host – primary, semi-continuous, continuous cells • Host grown as MONOLAYER on a solid
surface • Infect cell with virus of interest- ONE VIRION/cell • Circle of death- easily counted and quantified
10. Cytopathic effects • Clinical signs, symptoms- morphologic- physiologic- biochemical- immunologic
effects of viruses • Unfixed, unstained smears- 10x- some CPE directly observed • Translucent cells-
condenser down- iris diaphragm partly closed • Inclusion bodies fixation and staining
11. • Requires experience to use CPE as diagnostic tool • Virus may not conform to norm for its family • CPE
best viewed by daily observation of cultures admixed with low MOI (<0.1) • Normal cell ageing to be
distinguished • Rate of CPE appearance- characteristic • General rule : Slow virus- 4-5 days • Rapid – 1-2
days for CPE to appear
12. Types 1) Total destruction o Of all monolayer- most severe o Cells shrink rapidly, become pyknotic o
Detach from glass within 72 hrs o Typical of ENTEROVIRUSES 20x objective within 24 hrs  Bovine fetal
spleen cells
13. 2) Subtotal Bovine fetal spleen cells 2 destruction days • Some cells postinfection dead/detached
Rhabdovirus • Togavirus • Some picornavirus • Paramyxovirus 20x objective
14. 3) Focal degeneration • Herpesvirus, Poxvirus • Localised foci of infection • D/t cell to cell virus trasnfer
rather than extracellularly • Enlarged, rounded, refractile cells Bovine fetal spleen cells 2 days postinfection,
Herpesvirus • Strangling of cytoplasm Black arrows - cell rounding in a focal pattern. • Cell fusion Blue
arrows -cytoplasmic stranding. 20x objective . IMP: view at a low MOI
15. 4) Swelling and clumping • Adenovirus • Cells greatly enlarge and clump together in ‘grape-like’ clusters
Bovine fetal spleen cells 4 days postinfection with a high MOI of the bovine adenovirus, an Adenovirus,
showing cell rounding and small amounts of clumping. 10X objective
16. 5) Foamy degeneratioon • d/t large/numerous cytoplasmic vacuoles • Retrovirus • Paramyxovirus •
Flavivirus • Difficult to see without Giemsa-stained bovine fetal spleen cells 1 day staining Flavivirus, showing
vacuoles (arrow). 40X objective
17. 6) Cell fusion • Syncytium/polykaryon • Plasma membranes of 4 or more cells fuse • Enlarged cell with 4
or more nuclei • Easily seen when stained • To be distinguished from cell Giemsa-stained bovine fetal
clumping/clustering spleen cells 4 days • Paramyxovirus postinfection with the • Herpesvirus bovine
respiratory syncytial virus, a Paramyxovirus, showing syncytia (arrows) and faint basophilic cytoplasmic
inclusion bodies (dashed arrows). 20x objective
18. 7) Inclusion bodies • Areas of altered cell staining • CANNOT be seen in live cell cultures • Single or
multiple- depends on virus • Large/small • Round/irregular • Intranuclear/intracytoplasmic •
Eosinophilic/basophilic • Chromatin margination • Indicate areas of viral protein synthesis or nucleic acid
synthesis or virion assembly • Some cases no virus is present Herpesvirus, 40x. Viral scarring
19. Cytoplasmic stranding Adenovirus, 20x. Intranuclear inclusion Rough-edged nuclear inclusion bodies
20. Poxvirus, showing pink eosinophilic cytoplasmic Feline nasal turbinate primary cell culture infected
inclusion bodies (arrows) and cell swelling near the with feline herpes virus-1, a Herpesvirus, showing top of
the field, 20x. chromatin margination (dark ring around the edge of the nucleus).
21. CPE of Common Viruses • MEASLES o dilated skin vessels, edema o Lymph nodes, lungs o Warthin-
Finkeldey cells
22. • MUMPS o Parotitis o Orchitis o Pancreas o Rarely encephalitis Mumps parotitis Enlarged glands, C/S
reddish brown, Glistening. Macrophage, lymphocyte infiltration. Oedematous
23. • HERPES SIMPLEX o Fever blisters o Gingivostomatitis o Genital herpes o Herpes epithelial keratitis o
Herpes stromal keratitis o Cowdry type A inclusion bodies • Pink to purple intranuclear inclusions Glassy
intranuclear herpes simplex inclusion bodies