Conformational changes in influenza virus haemagglutinin and its monomer detected by monoclonal antibodies

doi:10.1016/0264-410X(85)90099-4

Vaccine

Volume 3, Issue 3, September 1985, Pages 175-181

https://doi.org/10.1016/0264-410X(85)90099-4 Get rights and content

Abstract

Exposure of influenza virus haemagglutinin to pH 5 results in conformational changes occurring in the molecule which are accompanied by antigenic modifications. Furthermore, isolated haemagglutinin (HA) at a concentration of 0.1 nM undergoes dissociation from the trimeric to a monomeric form when exposed to pH 5. Whether present on intact virus or as the isolated monomer, each form of haemagglutinin from pH 5 exhibits similar alterations in antigenic characteristics. These forms of HA show modifications in the antigenic sites located in the hinge (site C), tip (site B) and subunit interface (site D) regions. Whereas binding of monoclonal antibodies recognizing the tip and interface is abrogated or diminished, binding of antibodies to the hinge region is greatly enhanced following exposure of virus or the monomeric form of HA to pH 5.

References (34)

T. Maeda et al.
Activation of influenza virus by acidic media causes haemolysis and fusion of erythrocytes
FEBS Lett.
(1980)
J. Lenard et al.
pH-dependent haemolysis by influenza virus, Semliki Forest virus and Sendai virus
Virology
(1981)
S.C. Lazarowitz et al.
Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the haemagglutinin polypeptide
Virology
(1975)
R.T.C. Huang et al.
Influenza viruses cause haemolysis and fusion of cells
Virology
(1981)
A. Scheid et al.
Two disulfide linked polypeptide chains constitute the active F protein of paramyxoviruses
Virology
(1977)
R.G. Webster et al.
Determination of the number of nonoverlapping antigenic areas on Hong Kong (H3N2) influenza virus haemagglutinin with monoclonal antibodies and the selection of variants with potential epidemiological significance
Virology
(1980)
R.G. Webster et al.
Changes in the antigenicity of the haemagglutinin molecule of H3 influenza viruses at acidic pH
Virology
(1983)
W.G. Laver et al.
Antigenic drift in type A influenza viruses: sequence differences in the haemagglutinin of Hong Kong (H3N2) variants selected with monoclonal hybridoma antibodies
Virology
(1979)
S.E. Newton et al.
Sequence of the haemagglutinin gene of influenza virus A/Memphis/1/71 and previously uncharacterized monoclonal antibody-derived variants
Virology
(1983)
P.L. Ey et al.
Isolation of pure IgG₁, IgG_2a and IgG_2b immunoglobulins from mouse serum using protein A-Sepharose
Immunochemistry
(1978)

H. Kida et al.

Biological activity of monoclonal antibodies to operationally defined antigenic regions to the haemagglutinin molecule of A/Seal/Massachusetts/1/80 (H7N7) influenza virus

Virology

(1982)

M.A. Bothwell et al.

A sedimentation equilbrium method for determining molecular weights of proteins with a tabletop high speed air turbine centrifuge

J. Biol. Chem.

(1978)

T. Maeda et al.

Interactions of influenza virus haemagglutinin with target membrane lipids is a key step in virus-induced hemolysis and fusion at pH 5.2

A. Yoshimura et al.

Infectious cell entry mechanism of influenza virus

J. Virol.

(1982)

A. Helenius et al.

On the entry of Semliki Forest virus into BHK-21 cells

J. Cell. Biol.

(1980)

J.M. White et al.

Cell fusion by Semliki Forest, influenza and vesicular stomatitis viruses

J. Cell. Biol.

(1981)

J.J. Skehel et al.

Studies on the primary structure of the influenza virus hemagglutinin

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Citation Excerpt :
Therefore, HA proteins can be incubated in media at varying pH, reneutralized, incubated with trypsin, and then resolved by SDS-PAGE to determine the exposure pH at which HA degradation occurs [43,44]. Conformation-specific monoclonal antibodies and flow cytometry can also be used to measure the pH at which the HA protein is activated [45–50]. HA activation pH values typically fall within a range of pH 4.8–6.2 (reviewed in [21,22]).
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A subtype specific ELISA using purified hemagglutinin (HA) from influenza A H1N1 and H3N2 was developed to detect antibodies present in swine previously exposed to either H1N1 or H3N2 influenza viruses. The HA was extracted using the detergent octylglucoside followed by ion exchange chromatography. All HA preparations were free of contaminating nucleoprotein and matrix protein contamination. Monospecific swine anti-H1N1 and swine anti-H3N2 sera were used to demonstrate the subtype specificity of the assay. Monospecific rabbit anti-H1N1 or H3N2 was used to sterically block crossreacting determinants and thus enhance assay specificity. A linear relationship between single dilution point ELISA and the hemagglutination inhibition (HI) test was established. This enabled the accurate estimation of HI titer from ELISA. Further refinement of this ELISA based HI estimation system could allow it to replace the current HI procedures in instances where identification at the subtype level of specificity is acceptable. The substantial specificity requirements associated with the detection of strain specific antibody would still necessitate the use of the HI procedure.
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The enhancement of hormone activity by antibodies has been known for many years; however, investigation into the molecular basis of the phenomenon has only recently begun. A number of mechanisms for this enhancement, including “buffering” or slow release, bivalency and Fc region, and conformational and receptor “restriction” effects, have been documented or proposed. The availability of panels of monoclonal antibodies of distinct combining site specificity have aided in these studies and contributed substantially to our understanding of hormone-receptor interactions.
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Conformational changes in influenza virus haemagglutinin and its monomer detected by monoclonal antibodies

Abstract

FEBS Lett.

Virology

Virology

Virology

Virology

Virology

Virology

Virology

Virology

Immunochemistry

Virology

J. Biol. Chem.

Interactions of influenza virus haemagglutinin with target membrane lipids is a key step in virus-induced hemolysis and fusion at pH 5.2

Infectious cell entry mechanism of influenza virus

J. Virol.

On the entry of Semliki Forest virus into BHK-21 cells

J. Cell. Biol.

Cell fusion by Semliki Forest, influenza and vesicular stomatitis viruses

J. Cell. Biol.

Studies on the primary structure of the influenza virus hemagglutinin