Virology lecture 4 Flashcards
Influenza virus and HIV in detail as they ilustrate many principles of virology
Basica influenza virus structure
1 - -ve ssRNA, 8 segments
2 - nucleocapsid has helical symmetry. envelope with 2 prominant spikes. one is haemagglutinin (HA) the other is neuraminidase (NA) = glycoproteins
3 - 3rd viral protein in envelope = M2. an ion channel.
4 - M1 protein under envelope
5 - NP (nucleoprotein) associated with RNA genome within the nucleocapsid
6 - PB1, PB2 and PA polypeptides also in virion core which collectively make the RNA dependent RNA polymerase for -ve to +ve ssRNA.
7 - variable but spherical shape in culture. in fresh clinical isolates however it can be filamentous.
8 - an exception to the -ve ssRNA rule in that it replicates in the nucleus.
Influenza classification?
1 - orthomyxovirus family
2 - 3 types (A,B and C). A = most disease in man.
3 - 2 methods of classification - type, country of isolation, isolate number, year. ie A/PR/8/34 = type A, Puerto rico, isolate 8, 1934.
4 - second classification mechanism - spcific HA and NA proteins, H1N1 = H1 HA and N1 NA
the 8 influenza RNA segments, the protein encoded and its function
1 - PB2 - RNA pol
2 - PB1 - RNA pol
3 - PA - RNA pol
4 - HA - binds sialic acid
5 - NP (nucleoprotein) - encapsidates the RNA genome
6 - NA - surface glycoprotein - cleaves sialic acid fro infected cell surface and virions.
7 - M1 and M2 (matrix proteins) - M1 interacts with envelpoe and nucleocapsid, M2 is an ion channel
8 - NS1 and NS2 (non-structural) - NS1 = evasion of innate immunity
how many types of HA and NA in influenza A? natural reservoirs?
1 - 17 HA types (1-17) and 9 NA types (1-9)
2 - birds are the natural reservoirs for most subtypes (all bar H17 but a few can infect man (H1,2,3,5 and N1,2,8) , pigs (H1 and 3, N1 and 2), horses (H1 and 7, N7 and 8), specific fruit bats (H17) and seals (H4 and 7, N7).
influenza patterns of transmission and death
1 - highly seasonal, epidemics during winter due to close proximity as an aerosol with an envelope.
2 - even in an ordinary year it causes 10-15,000 deaths in the UK
influenza entry pattern
HA binds sialic acid, endocytosis, acidification of endosome, confromational change of HA, membrane fusion, nucleocapsid released into the cytosol then transported to nucleus for replication.
HA structure
1 - trimer. each monomer include a HA1 and a HA2 - cleaved from HA0.
2 - HA1 is the globular head with sialic acid binding site
3 - HA2 is the stalk connecting the head to the envelope
mechanism of influenza membrane fusion during entry. drugs?
1 - N terminus of HA2 has a hydrophobic peptide called the fusion peptide. at neutrla pH its buried on acidification HA1 moves to expose it. it inserts in to the endosome membrane, destabilises it and promotes fusion. hence fusion is pH dependent.
- amantadine and rimantidine raise the pH to inhibit entry.
define epidemic
επί (epi), meaning “upon or above” and δήμος (demos), meaning “people”) occurs when new cases of a certain disease, in a given human population, and during a given period, substantially exceed what is expected based on recent experience.
define pandemic
pandemic comes from the Greek pandemos meaning “pertaining to all people”. The Greek word pan means “all” and the Greek word demos means “people”. Denoting a disease affecting or attacking the population of an extensive region, country, continent, global; extensively epidemic
which influenza mRNAs produce more than one protein? how? can they be used to replicate the genome?
1 -segment 7 for M1 and M2, segment 8 for NS1 and NS2. alternative splicing.
viral mRNA from RNA-dep RNA pol are not complete copies so cant make up the genome. viral RNA pol makes +ve from the -ve then -ve from that - go to templates for more mRNA or packaged
how does new influenza escape its host cell? drugs?
Neuraminidase cleaves sialic residues from proteins to stop it getting stuck to the outside of the cell. it also removes sialic acid residues from HA and NA on virions to prevent their aggregation.
tamiflue and relenza inhibit NA.
how does influenza cause repeated infection?
antigenic drift and shift. also new viruses from animal reservoirs ie H5N1 bird flu and H1N1 swine flu.
antigenic drift in influenza?
1 - neutralising antibodies bind HA, causes natural selection for aa mutations to evades these. this gradual accumilation of changes = drift.
H3 found in man in 1968 is very different to the H3 strain circulated in 2003. huge numbers of aa changes.
antigenic shift in influenza?
1 - acquisition of a completely new HA from another influenza virus.
2 - more than one influenza type infects a cell, reassortment during packaging of new virions produces a new virus. often this is of no consequence, one combination is dangerous - if 7 genes are fro the huma virus it’ll replicate well, add the avian gene for HA and it’ll evade any existing immunity. this causes pandemics.
date of influenza pandemics and genotype.
1 - 1918 - H1N1 - devastating pandemic w/ 40-50 million dead.
2 - 1957 - H2N2
3 - 1968 - H3N2 - HA changes more rapidly as its critical to reinfection, the NA antibody is less important.
4 - 1977 - H1N1 - reemergence from a 1930s circulation
5 - 2009 - H1N1 - derived from mexican swine. low 0.03% mortality rate vs the 2.5% of the 1918 strain
why is the H5N1 influenza virus of concern?
1 - highly virulent, severe epidemics in birds, spread to other species inc man. 60% mortality in man. several hundered deaths but no evidence of human to human transmission yet.
2- factors influencing its adaptation to man-
a - better binding
b - better replication in cells
c - better escape from innate immune system
d - better transmission between humans
difference between human influenza and avian in HA binding to sialic acid?
1 - human preferentially binds sialic acid with a alpha2’-6’ link to the terminal galactose, avian to a alpha2’-6’ link. one amino acid mutation in the HA structure can change the preference allowing adaptation!
what influences the ability of influenza to replicate in human cells?
the aa in the PB2 subunit of the RNA pol. is the aa E627 is present it replicates well in avian but not human, change to K627 and this switches.
how does the influenza virus resist host immunity?
NS1 protein confers resistance to interferon mediated inhibition. avian influenza version works well in birds but less efficient in humans but it could adapt to increase its virulence in humans.
HIV first investigated and why? year identified? close relative?
no infected in 1990 and 1996
1 - 1981 - CDC spotted unusual cluster of opportunistic infections - pneumonitis carinii, disseminated HCMV, mucosal candida and chronic HSV.
2 - retrovirus identified in paris in 1983
3 - 1990 - 307,000 AIDS cases, 9 million HIV.
4 - 1996 - 28million infected, 5.8million AIDS cases and over 75% of these were dead.
outcome of HIV?
death to infection ratio in 2010?
1 - DEATH
2 - for every 2 death, 3 more were infected so the pandemic is spreading. 7000 new infections a day, 42% in 15-24yo.
HIV family and general structure
1 - retrovirus, lentivirus subfamily. more complex genome than standard retrovirus so slow infection.
2 - +ve ssRNA, 5’ can 3’ poly A tail. but doesnt act as mRNA.
3 - diploid genome (2 sets)
4 - genome contains a tRNA to prime reverse transcription. dsDNA - provirus.
HIV capsid and envelope structure
1 - cone like capsid, composed of gag24 protein.
2 - lipid envelope with gp120 and gp41 proteins from gp160 precursor. gp120 is eavily glycosylated to provide a glycan shield to make it harder for antibodies to target. quite variable env proteins.
3 - p24 gag more conserved than env protein so target of antigen in diagnostic kits.
