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United States Biological
May 2009 Issue 1| Subscribe                                                                                               www.usbio.net
Newsletter

Influenza A :
virion visualized by TEMInfluenza A and B are the two types of influenza viruses that cause epidemic human disease. Influenza A viruses are further categorized into subtypes on the basis of two surface antigens: hemagglutinin and neuraminidase. Since 1977, influenza A (H1N1) viruses influenza A (H3N2) viruses have been in global circulation. United States Biological can supply you with the highest-quality antibodies and antigens to study this ongoing threat. USBio also carries a variety of antigens and antibodies to Influenza B, as well as to other infectious diseases.


Antibodies to H1N1:

virion visualized by TEM

Cat # Antibody to: Type: Apps:
I7650-20 Influenza A, Puerto Rico 8/34 (H1N1) Pab E, N
I7650-81 Influenza A, USSR (H1N1) Pab IF, IH
I7650-19B Influenza A, H1N1, Nucleoprotein Mab IC
I7650-17F Influenza A, H1N1, Hemagglutinin (HA) Mab E, IH, IF
I7650-10B Influenza A H1N1 Mab IF
I7650-78B Influenza A, USSR (H1N1) Pab E, IF, B, IH
I7650-17F Influenza A, H1N1, Hemagglutinin Mab E, IH, IF
I7650-05 Influenza A, USSR (H1N1) (FITC) Pab IH, IC, IF
I7650-05E Influenza A, USSR (H1N1) Pab E, B, IC
I7650-09 Influenza A, H1N1, (FITC) Pab E, IF

This month's Special:
10% off selected H1N1 Antibodies

I7650-10 Anti- Influenza A, H1N1
Recognizes Influenza A, H1N1. Does not crossreact with Influenza B, Parainfluenza or RSV. Does not react to HEp-2 cells. Suitable for use in Immunohistochemistry, Immunofluorescence and Western Blot. Other applications have not been tested.

I7650-17 Anti- Influenza A, H1N1, Hemagglutinin (HA)
Recognizes Influenza virus type A (hemagglutinin). Reacts with hemagglutinin of H1 serotype. Suitable for use in ELISA, Immunofluorescence, Immunocytochemistry, and Hemagglutinin Inhibition. Other applications have not been tested.

I7650-10A Anti- Influenza A, USSR (H1N1) (Biotin)
Specific to Influenza A, H1N1 by IHA. May crossreact with chicken cellular proteins. Does not crossreact Influenza B, RSV, Parainfluenza 1-3, Adenovirus or HEp-2 cells. Suitable for use in Fluorescence Microscopy, Immunofluorescence, Immunohistochemistry, and Hemaglutination Inhibition. Other applications not tested.

Hatching TimeInfluenza virus is a single-stranded RNA virus, segmented, ~100nm diameter, and enveloped. Strains are described by geographic origin, strain number, year of isolation and hemagglutination (H) and neuraminidase (N) antigens. It is an acute viral disease of the upper respiratory tract characterized by fever, headache, chills, myalgia, weakness, runny nose and sore throat. Cough can be severe. Nausea and vomiting are uncommon.


Influenza A, H1N1
Swine Flu and You

Influenza Virus

With the recent outbreak of Influenza A/Mexico/09 (H1N1) aka Swine flu A/Mexico/09 (H1N1) initially in Mexico, in the US, and around the world, it may be useful to summarize the features of the influenza virus and its avenues for vaccination and treatment.

Structure

The influenza A virus particle is ~100nm in diameter and usually roughly spherical in shape that is coated in a lipid bilayer derived from the plasma membrane of its host. The influenza virus is composed of a viral envelope containing two main types of glycoprotein, wrapped around a central core. Studded over the surface of the virion are the two glycoproteins, hemagglutinin (H or HA) and neuraminidase (N or NA). HA is a lectin that is involved in binding of the virus to target cells and entry of the viral genome into the target cell, while NA is involved in the release of the virus from infected cells, by cleaving sugars that bind the mature viral particles.

The central core contains the viral RNA genome and other viral proteins that package and protect this RNA. Most viruses contain a single piece of nucleic acid, but influenza virus is different in that it contains eight pieces of negative sense RNA. The eight Influenza A RNA molecules code for:
1. The HA gene. It encodes the hemagglutinin. 3 distinct hemagglutinins, H1, H2, and H3) are found in human infections; 13 others have been found in animal flu viruses.
2. The NA gene. It encodes the neuraminidase. 2 different neuraminidases (N1 and N2) have been found in human viruses; 7 others in other animals.
3. The NP gene encodes the nucleoprotein. Influenza A, B, and C viruses have different nucleoproteins.
4. The M gene encodes two proteins: a matrix protein (M1) and an ion channel (M2) spanning the lipid bilayer.
5. The NS gene encodes two different non-structural proteins. These are found in the cytoplasm of the infected cell but not within the virion itself.
6. – 8. One RNA molecule (PA, PB1, PB2) for each of the 3 subunits of the RNA polymerase.

Life Cycle

Influenza infection and replication is a multi-step process: first the virus has to bind to and enter the cell, then deliver its genome to a site where it can produce new copies of viral proteins and RNA, assemble these components into new viral particles and finally exit the host cell.

Influenza viruses bind through Hemagglutinin (HA) onto sialic acid sugars on the surfaces of epithelial cells; typically in the nose, throat and lungs of mammals and intestines of birds. The virus is imported into the cell by endocytosis. The acidic conditions in the endosome cause the M2 ion channel to allow protons to pass through the viral envelope and acidify the core of the virus. This causes the virus core to disassemble and to release the viral RNA and core proteins. One of the anti-viral drugs on the market, amantadine, blocks the action of the M2 ion channel, thus preventing infection. Since 2006, the Center for Disease Control found rates of amantadine resistance to be much higher than in previous seasons. For some strains and in some areas the resistance has reached 100% signaling the need for additional anti-viral treatments.

After their release into the nuclease, the core proteins and vRNA and the RNA-dependent RNA polymerase begin transcribing complementary positive-sense vRNA. The vRNA is either exported into the cytoplasm and translated, or remains in the nucleus. Newly synthesized viral proteins are either secreted through the Golgi apparatus onto the cell surface (in the case of NA and HA) or transported back into the nucleus to bind vRNA and form new viral genome particles.

Negative-sense vRNAs that form the genomes of future viruses, RNA-dependent RNA polymerase, and other viral proteins are assembled into a virion. The mature virus buds off from the cell. The mature viruses detach once their neuraminidase has cleaved sialic acid residues from the host cell. Drugs that inhibit neuraminidase, such as zanamavir (Relenza) and oseltamivir (Tamiflu), prevent the release of new infectious viruses and halt viral replication. After the release of new influenza viruses, the host cell dies.

Influenza Strains A, B, C

There are three types of influenza viruses: A, B and C. Influenza A and B viruses cause seasonal epidemics of disease almost every winter in the United States. Influenza type C infections cause a mild respiratory illness and are not thought to cause epidemics.
Influenza A viruses are divided into subtypes based on two proteins on the surface of the virus: the hemagglutinin (H) and the neuraminidase (N). There are 16 different hemagglutinin subtypes and 9 different neuraminidase subtypes, Influenza A viruses can be further broken down into different strains. The current subtypes of influenza A viruses found in people are A/ (H1N1), A/ (H3N2) A/(H5N1) (or bird flu, found predominantly in Asia). Influenza A can also affect a variety of animal species.

Influenza B viruses are not divided into subtypes. Influenza B viruses also can be further broken down into different strains; these strains are found only in human.
Most cases of the flu, especially those that occur in epidemics or pandemics, are caused by the influenza A virus, which can affect a variety of animal species (ducks, chickens, pigs, whales, horses and seals).


Antigenic Drift and Shift

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. This is one of the main reasons why people can get the flu more than one time. In most years, one or two of the three virus strains in the influenza vaccine are updated to keep up with the changes in the circulating flu viruses. People, who want to be protected from flu, need to get a flu shot every year.
The separation of the genome into eight separate segments of vRNA allows mixing or reassortment of vRNAs if more than one type of influenza virus infects a single cell. The resulting rapid change in viral genetics produces antigenic shifts, which are sudden changes from one antigen to another. These sudden large changes allow the virus to infect new host species and quickly overcome protective immunity. This is important in the emergence of pandemics.

A description of antigenic shift is as follows: Pigs can be infected with both human and avian influenza viruses in addition to swine influenza viruses. Infected pigs get symptoms similar to humans, such as cough, fever and runny nose. Because pigs are susceptible to avian, human and swine influenza viruses, they potentially may be infected with influenza viruses from different species (e.g., ducks and humans) at the same time. If this happens, it is possible for the genes of these viruses to mix and create a new virus. For example, if a pig were infected with a human influenza virus and an avian influenza virus at the same time, the viruses could mix (reassort) and produce a new virus that had most of the genes from the human virus, but a hemagglutinin and/or neuraminidase from the avian virus. The resulting new virus would likely be able to infect humans and spread from person to person, but it would have surface proteins (hemagglutinin and/or neuraminidase) not previously seen in influenza viruses that infect humans. This type of major change in the influenza A viruses is known as antigenic shift.

The 2009 H1N1 flu virus is a mix of antigens and consists of North American swine influenza viruses, North American avian influenza viruses, human influenza viruses and swine influenza viruses found in both Asia and Europe.

Pandemics

Antigenic shift results when a new influenza A subtype to which most people have little or no immune protection infects humans. If this new virus causes illness in people and can be transmitted easily from person to person, a pandemic can occur. In a normal influenza season, the greatest mortality is typically seen in the most susceptible populations; in infants and the elderly. A pandemic can be recognized because of the greater mortality of adults in mid-life (25-50 age range). Because adults in mid-life do not have as robust an immune systems that young adults have and because they have not lived long enough to have seen a similar flu strain, and develop antibodies such as the older age groups, they often become the hardest hit segment of the population.

Influenza was responsible for the most devastating plague in human history: the “Spanish flu” that swept around the world in 1918 killing 675,000 people in the U.S. and an estimated 20-50 million people worldwide. The timeline for the 1918 breakout involved an initial illness that occurred in the spring of 1918. Over the summer, the flu outbreak lessened, which is typical for influenza. The exception to this was for military personnel who lived in close quarters who continued to fall ill. In September of 1918, when flu typically makes a comeback in the northern hemisphere because of the change in weather and humidity, a second wave infected all segments of the population. This was the most serious outbreak. Again, in the late spring of 1919, a third wave took place. Pandemics can be comprised of a complex series of outbreaks over many population groups worldwide.

 

virion visualized by TEMSwine Flu Virus

Influenza A Antigens
from Allantoic Fluid:

These products have been treated in a manner consistent with methods of inactivation using 0.005% merthiolate. Generally accepted good laboratory practice appropriate to microbiological/viral safe handling practices and techniques is required for use.

Influenza A, Taiwan (H1N1)
Suitable for use in serological studies of influenza A virus, or as an immunogen for antibody production. Has been tested with anti-influenza A monoclonal antibodies.
~90% pure.

Influenza A, Beijing (H1N1)
Suitable for use in Serological studies of influenza A virus, immunogen for antibody production. Has been tested with anti-influenza A monoclonal antibodies.

~90% pure.

Influenza A, Caledonia (H1N1)
Recombinant Influenza A was harvested from Allantoic fluid of 10-day old embryonated eggs, inoculated with influenza strain A/New Caledonia/20/99 IVR 116.

~90% pure.

Influenza A, Taiwan (H1N1)
Recombinant Influenza A was harvested from Allantoic fluid of 10-day old embryonated eggs, inoculated with the influenza strain- Taiwan/1/86.
~90% pure.

other strains available


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