Astroviridae Pathogenesis

Geoff Potts BIO 437 4/11/07

Introduction
Discovered in 1975 after an outbreak in children Characteristics
Linear, naked, +ssRNA genome of about 7.5k nt 30 nm star-like isometric capsid of 3 protein subunits Ribosomal shifting of overlapping genome

Pathogenesis and Epidemiology

Infects many mammals and birds with severe infection with 8 mild to moderate human strains Source is contaminated water, sewage, or hands and enters through the fecal-oral route Infects intestinal mucosa
Causes mild gastroenteritis and acute watery diarrhea Causes death by dehydration mostly in young and elderly

Starry Starry Night

Genome
Genome codes for three genes, ORF1a containing the proteases, ORF1b containing the RNA-dep RNA polymerase, and ORF2 containing the capsid
Replication and assembly in the cytoplasm with subgenomic RNA expression ORF1a and ORF1b are nonstructural while ORF2 is structural
ORF2 codes for at least three capsid proteins that are cleaved by proteases before assembly ORF1a/b overlap and ribosomal frameshifting occurs in order to translate the both genes

Human Strain 1 is most common worldwide and cDNA is infectious when capped

Pathogenesis
Entry intro intestinal epithelium by means of endocytosis
Seems to bind homologous PVR/CD155 (Polio Virus Receptor) upon entry along with homologous viral protein E1 in pH dependent fusion (Similar to Sindbis Virus)

ORF1a codes for nsp1a and accumulates in the nucleus and leads to apoptosis and viral spread Damage to intestinal epithelium causes loss of water from cells and watery diarrhea In turkey model and the immunocompromised, adaptive immunity plays no role while innate macrophage Nitric Oxide seems to limit replication
In humans, antibodies seem to prevent reinfection of the same viral strain

Overall Course of Infection

Latent period of 1 to 4 days during viral replication in intestines Symptomatic phase for about 2-4 days with high antigen levels in feces and blood
Person-to-person spread occurs most during this period

After 7-9 days viral load drops and symptoms subside

Major Symptoms
Diarrhea Headache Nausea Low-Grade fever Abdominal pain Sometimes vomiting Can lead to dehydration

Diagnosis

Antibody based with heavy virus shedding in feces

ELISA Immunofluorescence Titer (Often not helpful as a majority of population is exposed)

RT-PCR (Reverse Transcriptase PCR)

Using RT and Taq polymerase from Thermus aquaticus

For research purposes EM and cell cultures can be used but not useful for medical purposes

Diarrheal Infectious Agents

Viruses cause watery diarrhea
Rotavirus, Calcivirus, Astrovirus often co-infected in winter peak months Adenovirus all year round

Bacteria
E. coli, Shigella, Salmonella, V. cholerae, C. difficile

Parasites
Giardia, Amebic dysentery, Cryptosporidium
Giardia cysts are resistant to chlorination

Prevention/Treatment
Wash hands, cook food thoroughly, probiotics like yogurt, and proper sanitation infrastructure Lay off the alcohol or Starbucks
Though if you are in Mexico drink the beer and not the water to prevent gastroenteritis in the first place

Oral Rehydration Therapy (WHO formula)

Electrolytes, ions, sugars, and water Kids always love the Pedialyte Teens love the Gatorade

Pepto-Bismol eases symptoms in adults

Some evidence says that the non-aspirin salicylate may still lead to Reyes syndrome in children during viral infection

ORT Plan

Conclusion
Characteristics
Naked, linear ss+RNA in star-like capsid

Pathogenesis
Mild gastroenteritis course that infects mammals and birds

Diagnosis
RT-PCR and ELISA

Infectious agents of gastroenteritis

Diverse range of bacteria, viruses, and protozoans

Prevention/Treatment
Good personal sanitation, avoid person-person or common source spread, political infrastructure, avoidance of diuretics, oral rehydration therapy

Future Directions
Better studies on molecular pathogenesis in humans rather than animal cell cultures or models Improving awareness of dehydration due to diarrhea in developed world by promoting ORT treatment and preventing death in children Looking into drugs to decrease severity and length of symptoms Measuring efficacy and cost of vaccination

Questions to know
Name the leading causes of viral gastroenteritis in children Name three ways to diagnose astrovirus infection Name three symptoms of astrovirus gastroenteritis Name the basic characteristics of the astrovirus genome

References
Bhattacharya, R., et al. (2006). Molecular epidemiology of human astrovirus infections in Kolkata, India. Infection, Genetics and Evolution. 6:425-435. Koci, M.D., Kelley, L.A, Larsen, D., Schultz-Cherry, S. (2004). Astrovirus-Induced Synthesis of Nitric Oxide Contributes to Virus Control during Infection. Journal of Virology. 78: 1564-1574. Krishna, N.K. (2005). Identification of Structural Domains involved in Astrovirus Capsid Biology. Viral Immunol. 18:17-26. Marshall, J.A., Bruggink, L.D., Sturge, K., Subasinghe, N., Tan, A., Hogg, G.G. Molecular features of astrovirus associated with gastroenteritis outbreak in an aged-care centre. European Journal of Clinical Microbiology & Infectious Diseases. 26:67-71. Tang, Y., Murgia, M.V., Ward, L., Saif, Y.M. Pathogenicity of Turkey Astroviruses in Turkey Embryos and Poults. Avian Diseases. 50:526-531. Zhang, Z., et al. (2006). Quantitation of human astrovirus by real-time reverse transcription-polymerase chain reaction to examine correlation with clinical illness. Journal of Virological Methods. 134:190-196.

Questions/Comments/Pra ise?