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Influenza virus vaccine based on the conserved hemagglutinin stalk domain.

Steel J, Lowen AC, Wang TT, Yondola M, Gao Q, Haye K, García-Sastre A, Palese P - MBio (2010)

Bottom Line: The strain specificity of vaccines presently in use mirrors the exquisite specificity of the neutralizing antibodies that they induce, that is, antibodies which bind to the highly variable globular head domain of hemagglutinin (HA).Furthermore, the headless HA vaccine provided full protection against death and partial protection against disease following lethal viral challenge.Our results suggest that the response induced by headless HA vaccines is sufficiently potent to warrant their further development toward a universal influenza virus vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, New York, USA.

ABSTRACT
Although highly effective in the general population when well matched to circulating influenza virus strains, current influenza vaccines are limited in their utility due to the narrow breadth of protection they provide. The strain specificity of vaccines presently in use mirrors the exquisite specificity of the neutralizing antibodies that they induce, that is, antibodies which bind to the highly variable globular head domain of hemagglutinin (HA). Herein, we describe the construction of a novel immunogen comprising the conserved influenza HA stalk domain and lacking the globular head. Vaccination of mice with this headless HA construct elicited immune sera with broader reactivity than those obtained from mice immunized with a full-length HA. Furthermore, the headless HA vaccine provided full protection against death and partial protection against disease following lethal viral challenge. Our results suggest that the response induced by headless HA vaccines is sufficiently potent to warrant their further development toward a universal influenza virus vaccine.

No MeSH data available.


Related in: MedlinePlus

Antisera from mice vaccinated with the PR8 4G headless HA show broad cross-reactivity by ELISA. The vaccine groups from which sera are derived are identified at the top of each column, and the ELISA substrate used is indicated to the right of each row. The colors used are consistent throughout the figure, with sera from Gag-only-vaccinated mice indicated in black, HK68 4G-vaccinated mice indicated in green, PR8 4G-vaccinated mice indicated in blue, and full-length-PR8-HA-vaccinated mice indicated in red. Each mouse is represented by a unique symbol that is the same in each panel. A rabbit antiserum raised against whole PR8 virus is shown in gray with open triangles, and a serum sample taken from a naive mouse is shown in gray with open squares. The reactivities of mouse sera to whole PR8 virus (A), purified recombinant A/New Caledonia/20/1999 HA protein (B), purified recombinant A/California/04/2009 HA (C), purified recombinant A/Singapore/1/1957 HA (D), purified recombinant A/Viet Nam/ 1203/2004 HA (E), and purified recombinant A/Hong Kong/ 1/1968 HA (F) are shown.
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f6: Antisera from mice vaccinated with the PR8 4G headless HA show broad cross-reactivity by ELISA. The vaccine groups from which sera are derived are identified at the top of each column, and the ELISA substrate used is indicated to the right of each row. The colors used are consistent throughout the figure, with sera from Gag-only-vaccinated mice indicated in black, HK68 4G-vaccinated mice indicated in green, PR8 4G-vaccinated mice indicated in blue, and full-length-PR8-HA-vaccinated mice indicated in red. Each mouse is represented by a unique symbol that is the same in each panel. A rabbit antiserum raised against whole PR8 virus is shown in gray with open triangles, and a serum sample taken from a naive mouse is shown in gray with open squares. The reactivities of mouse sera to whole PR8 virus (A), purified recombinant A/New Caledonia/20/1999 HA protein (B), purified recombinant A/California/04/2009 HA (C), purified recombinant A/Singapore/1/1957 HA (D), purified recombinant A/Viet Nam/ 1203/2004 HA (E), and purified recombinant A/Hong Kong/ 1/1968 HA (F) are shown.

Mentions: By ELISA, prechallenge sera were tested against a panel of HA substrates in order to evaluate the breadth of reactivity (Fig. 6). Against concentrated PR8 virion (Fig. 6A), Gag-only and HK68 4G antisera showed only a low level of background activity at the lowest dilution (1:50), while sera from the animals vaccinated with PR8 full-length HA gave a positive signal at a 1:6,250 dilution. Antisera against the PR8 4G headless HA were less potent than those against the full-length HA but reacted positively at a 1:50 dilution. When tested against recombinant HA proteins derived from a recent seasonal H1N1 influenza virus (A/New Caledonia/20/1999) (Fig. 6B) and a 2009 pandemic H1N1 influenza virus (A/California/04/2009) (Fig. 6C), Gag-only and HK68 4G antisera were negative, while sera from mice that received the PR8 full-length HA were either negative or showed a low level of reactivity. On these heterologous H1 substrates, the PR8 4G antisera showed the highest levels of reactivity, with the sera from two of the five mice in particular demonstrating high titers. Similar results were seen with recombinant HA proteins of the H2 and H5 subtypes: against the A/Singapore/1/1957 (H2N2) and A/Viet Nam/1203/2004 (H5N1) HAs, sera derived from PR8 4G-vaccinated mice showed moderate to high activity, while sera from the remaining groups (including those receiving the full-length HA) were largely negative (Fig. 6D and E). Finally, against the H3 subtype HA of A/Hong Kong/1/1968 (H3N2), only the HK68 4G antisera produced a positive signal (Fig. 6F). Thus, overall, sera obtained from mice vaccinated with the headless PR8 HA showed greater activity against heterologous strains than did sera from animals vaccinated with PR8 full-length HA. While serum titers of PR8 4G-vaccinated mice appeared to be higher against the heterologous HA proteins than against the homologous PR8 virus, a direct comparison should not be made due to the differing substrates used (purified HA versus whole virus). Within the PR8 4G group, the sera from two mice in particular consistently showed relatively high titers by ELISA. These serologic findings correlated with the protection data in that these same two mice were fully protected from disease while their three remaining counterparts each exhibited some weight loss after challenge (data not shown).


Influenza virus vaccine based on the conserved hemagglutinin stalk domain.

Steel J, Lowen AC, Wang TT, Yondola M, Gao Q, Haye K, García-Sastre A, Palese P - MBio (2010)

Antisera from mice vaccinated with the PR8 4G headless HA show broad cross-reactivity by ELISA. The vaccine groups from which sera are derived are identified at the top of each column, and the ELISA substrate used is indicated to the right of each row. The colors used are consistent throughout the figure, with sera from Gag-only-vaccinated mice indicated in black, HK68 4G-vaccinated mice indicated in green, PR8 4G-vaccinated mice indicated in blue, and full-length-PR8-HA-vaccinated mice indicated in red. Each mouse is represented by a unique symbol that is the same in each panel. A rabbit antiserum raised against whole PR8 virus is shown in gray with open triangles, and a serum sample taken from a naive mouse is shown in gray with open squares. The reactivities of mouse sera to whole PR8 virus (A), purified recombinant A/New Caledonia/20/1999 HA protein (B), purified recombinant A/California/04/2009 HA (C), purified recombinant A/Singapore/1/1957 HA (D), purified recombinant A/Viet Nam/ 1203/2004 HA (E), and purified recombinant A/Hong Kong/ 1/1968 HA (F) are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2912658&req=5

f6: Antisera from mice vaccinated with the PR8 4G headless HA show broad cross-reactivity by ELISA. The vaccine groups from which sera are derived are identified at the top of each column, and the ELISA substrate used is indicated to the right of each row. The colors used are consistent throughout the figure, with sera from Gag-only-vaccinated mice indicated in black, HK68 4G-vaccinated mice indicated in green, PR8 4G-vaccinated mice indicated in blue, and full-length-PR8-HA-vaccinated mice indicated in red. Each mouse is represented by a unique symbol that is the same in each panel. A rabbit antiserum raised against whole PR8 virus is shown in gray with open triangles, and a serum sample taken from a naive mouse is shown in gray with open squares. The reactivities of mouse sera to whole PR8 virus (A), purified recombinant A/New Caledonia/20/1999 HA protein (B), purified recombinant A/California/04/2009 HA (C), purified recombinant A/Singapore/1/1957 HA (D), purified recombinant A/Viet Nam/ 1203/2004 HA (E), and purified recombinant A/Hong Kong/ 1/1968 HA (F) are shown.
Mentions: By ELISA, prechallenge sera were tested against a panel of HA substrates in order to evaluate the breadth of reactivity (Fig. 6). Against concentrated PR8 virion (Fig. 6A), Gag-only and HK68 4G antisera showed only a low level of background activity at the lowest dilution (1:50), while sera from the animals vaccinated with PR8 full-length HA gave a positive signal at a 1:6,250 dilution. Antisera against the PR8 4G headless HA were less potent than those against the full-length HA but reacted positively at a 1:50 dilution. When tested against recombinant HA proteins derived from a recent seasonal H1N1 influenza virus (A/New Caledonia/20/1999) (Fig. 6B) and a 2009 pandemic H1N1 influenza virus (A/California/04/2009) (Fig. 6C), Gag-only and HK68 4G antisera were negative, while sera from mice that received the PR8 full-length HA were either negative or showed a low level of reactivity. On these heterologous H1 substrates, the PR8 4G antisera showed the highest levels of reactivity, with the sera from two of the five mice in particular demonstrating high titers. Similar results were seen with recombinant HA proteins of the H2 and H5 subtypes: against the A/Singapore/1/1957 (H2N2) and A/Viet Nam/1203/2004 (H5N1) HAs, sera derived from PR8 4G-vaccinated mice showed moderate to high activity, while sera from the remaining groups (including those receiving the full-length HA) were largely negative (Fig. 6D and E). Finally, against the H3 subtype HA of A/Hong Kong/1/1968 (H3N2), only the HK68 4G antisera produced a positive signal (Fig. 6F). Thus, overall, sera obtained from mice vaccinated with the headless PR8 HA showed greater activity against heterologous strains than did sera from animals vaccinated with PR8 full-length HA. While serum titers of PR8 4G-vaccinated mice appeared to be higher against the heterologous HA proteins than against the homologous PR8 virus, a direct comparison should not be made due to the differing substrates used (purified HA versus whole virus). Within the PR8 4G group, the sera from two mice in particular consistently showed relatively high titers by ELISA. These serologic findings correlated with the protection data in that these same two mice were fully protected from disease while their three remaining counterparts each exhibited some weight loss after challenge (data not shown).

Bottom Line: The strain specificity of vaccines presently in use mirrors the exquisite specificity of the neutralizing antibodies that they induce, that is, antibodies which bind to the highly variable globular head domain of hemagglutinin (HA).Furthermore, the headless HA vaccine provided full protection against death and partial protection against disease following lethal viral challenge.Our results suggest that the response induced by headless HA vaccines is sufficiently potent to warrant their further development toward a universal influenza virus vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, New York, USA.

ABSTRACT
Although highly effective in the general population when well matched to circulating influenza virus strains, current influenza vaccines are limited in their utility due to the narrow breadth of protection they provide. The strain specificity of vaccines presently in use mirrors the exquisite specificity of the neutralizing antibodies that they induce, that is, antibodies which bind to the highly variable globular head domain of hemagglutinin (HA). Herein, we describe the construction of a novel immunogen comprising the conserved influenza HA stalk domain and lacking the globular head. Vaccination of mice with this headless HA construct elicited immune sera with broader reactivity than those obtained from mice immunized with a full-length HA. Furthermore, the headless HA vaccine provided full protection against death and partial protection against disease following lethal viral challenge. Our results suggest that the response induced by headless HA vaccines is sufficiently potent to warrant their further development toward a universal influenza virus vaccine.

No MeSH data available.


Related in: MedlinePlus