OrthoMyxovirus

OrthoMyxovirus
Definition: any of a group of RNA viruses, including the viruses of influenza, parainfluenza, mumps, and Newcastle disease, characteristically causing agglutination of erythrocytes.

Formerly, any of a group of RNA-containing viruses with an affinity for mucins, now included in the families of Orthomyxoviridae and Paramyxoviridae. These viruses include the influenza virus, parainfluenza virus, respiratory syncytial virus, and measles virus.

History

Influenza pandemics have been recognized for several centuries. In the last century, H. influenzae (and other bacteria) were cited as the causal agent. In the 1940's (still pre-tissue culture, primitive immunology, few other viruses known), the characteristic property of haemagglutination was observed, followed by the discovery that the virus could be propagated in embryonated hens eggs (after adaptation) - this made influenza one of the best studied viruses during this period.

More viruses with similar properties were added to the group, until it was split into two families in the 1970s, the Paramyxoviridae and the Orthomyxoviridae:

Orthomyxoviruses Morphology:

Influenza virus particles are highly pleiomorphic (variable), mostly spherical/ovoid, 80-120nm diameter, but many forms occur, including long filamentous particles (up to 300nm long x 200nm diameter). Different strains of virus vary in their tendency to form filaments - this property maps to the matrix protein.

Structure

The outer surface of the particle consists of a lipid envelope from which project prominent glycoprotein spikes (surface proteins) of two types: haemagglutinin (HA), a 135 trimer neuraminidase (NA), a 60 tetramer The inner side of the envelope is lined by the matrix protein. The particles are relative labile (half-life a few hours ),not resistant to drying, etc.

Genome structure
Consists of s/s (-)sense RNA in 8 segments (7 in Influenza C).
The structure of the influenza virus genome is known in great detail because of the tremendous amount of genetic investigation (conventional and molecular) which has been done.

The 5' and 3' terminal sequences of all the genome segments are highly conserved:

Genome structure
The genome segments are packaged into the core. The RNP (RNA + nucleoprotein, N) is in a helical form with the 3 polymerase polypeptides associated with each segment.

Segment:

Polypeptide(s)

Function

1 2 3 4 5 6 7 8

PB2 PB1 PA HA NP NA M1
M2 NS1

Transcriptase: cap binding Transcriptase: elongation Transcriptase: protease activity Haemagglutinin Nucleoprotein: RNA binding; part of
transcriptase complex; nuclear/cytoplasmic transport of vRNA

Neuraminidase: release of virus Matrix protein: major component of virion Integral membrane protein - ion channel
Non-structural: nucleus; effects on cellular
RNA transport, splicing, translation. Antiinterferon protein

NS2

Non-structural: nucleus+cytoplasm,
function unknown

Proteins

The internal antigens (M1 and NP proteins ) are the type-specific proteins (antigens) used to determine if a particular virus is A, B or C. The M1 & NP proteins of all members of each type show cross reactivity.

The external antigens (HA and NA) show more variation and are the subtype and strain-specific antigens. These are used to determine the particular strain of influenza A responsible for an outbreak

HA (hemagglutinin) protein

Hemagglutinin is a lectin that mediates binding of the virus to target cells and entry of the viral genome into the target cell. HA protein is involved in attachment and membrane fusion in the endosome of the infected cell.

NA (neuraminidase) protein

Neuraminidase is an enzyme involved in the release of progeny virus from infected cells, by cleaving sugars (sialic acid =neuraminic acid) that bind the mature viral particles. By late in infection, the sialic acid will have been removed from the infected cell surface by the neuraminidase making it is easier for the progeny virions to diffuse away once they exit the cell. Neuraminidase is also involved in penetration of the mucus layer in the respiratory tract

Host Range: Influenza A viruses

1-Influenza

A viruses infect a wide variety of mammals, including man, horses, pigs, ferrets and birds. 2-The main human pathogen, associated with epidemics and pandemics. 3-There are 16 known haemagglutinin (H) serotypes and 9 known neuraminidase (N) serotypes.

Host Range: Influenza A viruses

Pigs and birds are believed to be particularly important reservoirs, generating pools of genetically/antigenically diverse viruses which get transferred back to the human population via close contact between humans and animals. pig becomes infected with both human and animal A viruses, and that one of the reassortants contains genes for human internal components but a new HA and/or NA segment from the animal virus. If this virus reassortant can infect humans, it will have mainly the same internal components as the current human virus, but new envelope components resulting in little immunity in the population. Influenza A subtypes are therefore classified according to the type of HA and NA protein. It is possible that we do not see such a shift in influenza B because there is no animal reservoir for this virus.

Host Range: Influenza B viruses

Influenza B viruses;
Infect mammals only and Cause disease, but generally not as
severe Influenza B viruses do not have distinguishable serotypes

1.
2. 3.

Host Range: Influenza C viruses

Influenza C viruses;

Infect mammals ,pig . 2. Rarely cause disease. 3. They are genetically and morphologically distinct from A and B types.
1.

Species barrier:

The species which different types of influenza viruses are able to infect are Idetermined by different forms of sialic acid present on the virus glycoproteins. This property depends predominately on the amino acid at position 226 of the haemagglutinin protein: Human viruses: Avian viruses: HA226leu HA226gln
This provides a considerable species barrier between birds and humans which is not easily overcome. However, pigs provide a "mixing pot" - able to be infected by both types of virus & thus allowing the passage of avian viruses to humans

Nomenclature of influenza strains

Type A, B or C/place isolated/strain number /year isolated/virus subtype (A type)

In the case of influenza A, also: HA subtype (H) and NA subtype (N) For example, the three strains for the 2009/2010 vaccine are: A/Brisbane/59/2007 (H1N1)-like A/Brisbane/10/2007 (H3N2)-like B/Brisbane/60/2006 The 2009 H1N1 strain used for the vaccine for the novel, pandemic, 2009 H1N1 influenza is: A/California/07/2009 (H1N1)

Pathogenisis

Typically, influenza is transmitted from infected mammals through the air by coughs or sneezes, creating aerosols containing the virus, and from infected birds through their droppings. Influenza can also be transmitted by saliva, nasal secretions, feces and blood. Infections occur through contact with these bodily fluids or with contaminated surfaces

. Flu viruses can remain infectious for about one week at human body temperature, over 30 days at 0 C (32 F), and indefinitely at very low temperatures. They can be inactivated easily by disinfectants and detergents. The viruses bind to a cell through interactions between its hemagglutinin glycoproteins

Life cycle

Replication:

1-Entry into the cell is facilitated by binding of the HA spikes to mucoproteins containing terminal N-acetyl neuraminic acid ( sialic acid) on the surfaces of epithelial cells
in the lung and throat (Stage 1 )
This interaction can be reversed by polysaccharide cleavage by NA spikes - prevents virus being 'sequestered' by inappropriate cell types

Replication:
2- the particle is engulfed by endocytosis via coated pits into endocytotic vesicles and finally endosomes. - These are acidified by the cell; at about pH 5.0, the HA monomers are cleaved by trypsin-like enzymes in the endosome into the HA1 (upper NH2 portion) and HA2 (trans-membrane COOH portion) polypeptides . This cleavage causes a conformational change in the HA spike which activates the membranefusion function located in HA2 resulting in fusion of the two membranes ( virus envelope & endosomalmembrane) and passage of the nucleocapsid into the cytoplasm.

Replication

Specific nuclear targeting sequences in the NP protein result in translocation of the nucleocapsid into the nucleus.

Genome segments are transcribed by the 3 polymerase polypeptides associated with each genome segment:

in the cell nucleus, the RNA-dependent RNA transcriptase begins transcribing complementary positive-sense vRNA (Steps 3a and b).[29] The vRNA is either exported into the cytoplasm (mRNA) and translated (step 4), or remains in the nucleus . Newly-synthesised viral proteins are either: - secreted through the Golgi apparatus onto the cell surface (in the case of neuraminidase and hemagglutinin, step 5b) or -transported back into the nucleus (NP) to bind vRNA and form new viral genome particles (step 5a).

Other viral proteins have multiple actions in the host cell, including degrading cellular mRNA and using the released nucleotides for vRNA synthesis and also inhibiting translation of host-cell mRNAs.[30]

Negative-sense vRNAs that form the genomes of future viruses, RNA-dependent RNA transcriptase, and other viral proteins are assembled into a virion. Hemagglutinin and neuraminidase molecules cluster into a bulge in the cell membrane. The vRNA and viral core proteins leave the nucleus and enter this membrane protrusion (step 6). The mature virus buds off from the cell in a sphere of host phospholipid membrane, acquiring hemagglutinin and neuraminidase with this membrane coat (step 7). [31] As before, the viruses adhere to the cell through hemagglutinin; the mature viruses detach once their neuraminidase has cleaved sialic acid residues from the host cell.[27] After the release of new influenza virus, the host cell dies ;The cell does not lyse.

DIAGNOSIS

Firm diagnosis :

virus isolation serology. The virus can be isolated from the nose or a throat swab. This is used to infect cells in culture (or eggs).

Hemadsorption may be used to detect infected cells.

Polymerase chain reaction (PCR) test are being developed to detect viral RNA. Recently, rapid tests that can be used in a physician's office have been approved. Provisional diagnosis is often made clinically, based on knowledge of a current outbreak of influenza combined with appropriate clinical symptoms (fever, cough, runny nose, malaise).

Vaccines

Antigenic drift

Antigenic drift is due to mutation. Antibodies to the HA protein are the most important in protection, although those to NA also play a role. Both proteins undergo antigenic drift (i.e. accumulate mutations) and accumulate changes such that an individual immune to the original strain is not immune to the drifted one. Antigenic drift results in sporadic outbreaks and limited epidemics.

Antigenic shift

Antigenic shift is due to reassortment. In the case of influenza A, antigenic shift periodically occurs. Apparently "new" HA and/or NA are found in the circulating viral strains. There is little immunity (particularly if both proteins change, or if new HA is present) and an epidemic/pandemic is seen.

Vaccines
Vaccines are produced by reassortment of egg-adapted strains with strains with the required HA type. Large amounts of virus are then grown in embryonated eggs

(cheap and efficient), purified and formalin inactivated.

The There are two types of vaccine: -TRIVALENT INACTIVATED VACCINE (TIV)

-LIVE, ATTENUATED INFLUENZA VIRUS VACCINE (LAIV)

TRIVALENT INACTIVATED VACCINE (TIV):

It is an inactivated preparation of egg-grown virus is given by injection sub-cutaneously. Only certain formulations of the vaccine are FDA certified for young children. Protection is via IgG antibodies

LIVE, ATTENUATED INFLUENZA VIRUS VACCINE (LAIV) Prepared from egg-grown virus.
It is given nasally and should provide mucosal, humoral and cell-mediated immunity. It is approved for healthy, non-pregnant individuals 2 to 49 years old . should not be given to children under 5 years of age who have possible reactive airways disease (for example, a history of recurrent wheezing). It is contraindicated for children and adolescents on any therapy containing aspirin due to the potential risk of Reye's syndrome since the virus is a live virus.

Both influenza vaccines are

formulated annually using the types and strains of influenza predicted to be the major problems for that year . The vaccines are multivalent, the current ones are trivalent and have two strains of influenza A and one of influenza B. Vaccination needs to be given every year because the most effective strains for the vaccine will change due to drift and/or shift. The vaccines are usually given in the Fall ,once the strains to be used for the influenza season have been determined in the earlier part of the year.

Vaccine currently in clinical trials

A naked DNA vaccine against influenza virus using the nucleoprotein (core) gene of the virus has been developed and is currently in clinical trials. The nucleoprotein is similar across many strains of influenza, unlike envelope or surface proteins which change extensively from one strain to another. Nucleoproteins are therefore not subject to the same humoral immune pressure with antigenic drift as are the surface glycoproteins targeted by conventional vaccines.

Chemotherapy
Zanamivir (Relenza) and Oseltamivir (Tamiflu)

Two neuraminidase inhibitors (Synthetic sialic acid analogues) have been approved by the FDA. They are active against both influenza A and influenza B and can reduce the duration of uncomplicated influenza if taken within two days of the onset of illness.

However, oseltamivir resistance has been seen in some circulating strains recently (the 2009 H1N1 strain is sensitive).

Rimantadine and amantadine. Rimantadine and amantadine block virus entry across the endosome and also interfere with virus release. They may be given as protective agents during an outbreak, especially to those at severe risk and key personnel. Other treatment The best treatments are rest, liquids, anti-febrile agents (not aspirin in the young or adolescent, since Reye's syndrome is a potential problem). Be aware of and treat complications appropriately.