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Heterosubtypic anti-avian H5N1 influenza antibodies in intravenous immunoglobulins from globally separate populations protect against H5N1 infection in cell culture.

Sullivan JS, Selleck PW, Downton T, Boehm I, Axell AM, Ayob Y, Kapitza NM, Dyer W, Fitzgerald A, Walsh B, Lynch GW - J Mol Genet Med (2009)

Bottom Line: Although there were qualitative and quantitative differences in binding and protection between globally different formulations, the heterosubtypic antibody activities for the respective IVIGs were in general quite similar.Of particular note because of the relative geographic proximity to the epicentre of H5N1 and the majority of human infections, was the similarity in the antibody binding responses between IVIGs from the Malayan peninsula, Europe and Australia.They also open a window into a somewhat ill defined arena of antibody immunity, namely heterosubtypic immunity.

View Article: PubMed Central - PubMed

Affiliation: Biosafety, Immunobiology, Global Health and Pandemic Infections Research, Central Clinical School, Faculty of Medicine, The University of Sydney, Camperdown, NSW 2006, Australia.

ABSTRACT
With antigenically novel epidemic and pandemic influenza strains persistently on the horizon it is of fundamental importance that we understand whether heterosubtypic antibodies gained from exposures to circulating human influenzas exist and can protect against emerging novel strains. Our studies of IVIG obtained from an infection-naive population (Australian) enabled us to reveal heterosubtypic influenza antibodies that cross react with H5N1. We now expand those findings for an Australian donor population to include IVIG formulations from a variety of northern hemisphere populations. Examination of IVIGs from European and South East-Asian (Malaysian) blood donor populations further reveal heterosubtypic antibodies to H5N1 in humans from different global regions. Importantly these protect against highly pathogenic avian H5N1 infection in vitro, albeit at low titres of inhibition. Although there were qualitative and quantitative differences in binding and protection between globally different formulations, the heterosubtypic antibody activities for the respective IVIGs were in general quite similar. Of particular note because of the relative geographic proximity to the epicentre of H5N1 and the majority of human infections, was the similarity in the antibody binding responses between IVIGs from the Malayan peninsula, Europe and Australia. These findings highlight the value of employing IVIGs for the study of herd immunity, and particularly heterosubtypic antibody responses to viral antigens such as those conserved between circulating human influenzas and emerging influenza strains such as H5N1. They also open a window into a somewhat ill defined arena of antibody immunity, namely heterosubtypic immunity.

No MeSH data available.


Related in: MedlinePlus

Anti-human influenza binding antibodies in IVIGs: Assays for detection of Ab binding to conformational/ discontinuous influenza epitopes. A. Multiplex assay for anti-influenza IVIG Ab binding showing a comparison of the antibody binding activities of 4 different batches of a single IVIG formulation. IVIG antibody binding to Influenza A: H3N2 A/Shandong/9/1993 and Influenza B: B/Hong Kong/5/1972 antigens displayed on microbeads and performed at the Rules Based Medicine biomarker testing laboratory using multianalyte bead Fluorescence (www.rulesbasedmedicine.com). Binding to herpes simplex virus (HSV), human pappilloma virus (HPV), Influenza B (Infl B) respiratory syncytial virus (RSV) and varicella zoster (V. Zoster) are also included for comparison. The Y-axis values show the amount of IVIG antibody that binds to the respective antigens coated on beads and expressed as the mean ratio of total antibody binding to the respective viral antigens compared to highly-validated RBM internal-reference control standards. B. Microarray slides spotted with viral dilutions (1-0.125mg/ml) of H3N2 (A/Sydney/5/1997), H1N1 (A/Johannesburg/82/1996), tetanus toxoid (Tet Tox) or BSA. IVIG-antibody (D-3) bind robotically arrayed whole inactivated influenza and tetanus toxoid. The BSA is negative. Spots are 10nl and 0.8mm apart. [Note: antigens were bound in duplicate arrays with (top array panel of each slide) and without (bottom array panel on each slide) 0.005% CHAPS. CHAPS was tested as a means of improving liquid flow rates in the Piezorray, but as shown, had no influence on protein binding to the array substrate.]
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Figure 2: Anti-human influenza binding antibodies in IVIGs: Assays for detection of Ab binding to conformational/ discontinuous influenza epitopes. A. Multiplex assay for anti-influenza IVIG Ab binding showing a comparison of the antibody binding activities of 4 different batches of a single IVIG formulation. IVIG antibody binding to Influenza A: H3N2 A/Shandong/9/1993 and Influenza B: B/Hong Kong/5/1972 antigens displayed on microbeads and performed at the Rules Based Medicine biomarker testing laboratory using multianalyte bead Fluorescence (www.rulesbasedmedicine.com). Binding to herpes simplex virus (HSV), human pappilloma virus (HPV), Influenza B (Infl B) respiratory syncytial virus (RSV) and varicella zoster (V. Zoster) are also included for comparison. The Y-axis values show the amount of IVIG antibody that binds to the respective antigens coated on beads and expressed as the mean ratio of total antibody binding to the respective viral antigens compared to highly-validated RBM internal-reference control standards. B. Microarray slides spotted with viral dilutions (1-0.125mg/ml) of H3N2 (A/Sydney/5/1997), H1N1 (A/Johannesburg/82/1996), tetanus toxoid (Tet Tox) or BSA. IVIG-antibody (D-3) bind robotically arrayed whole inactivated influenza and tetanus toxoid. The BSA is negative. Spots are 10nl and 0.8mm apart. [Note: antigens were bound in duplicate arrays with (top array panel of each slide) and without (bottom array panel on each slide) 0.005% CHAPS. CHAPS was tested as a means of improving liquid flow rates in the Piezorray, but as shown, had no influence on protein binding to the array substrate.]

Mentions: IVIGs are highly-purified IgG preparations prepared from large pools of human plasma and the immunoglobulin subtypes in IVIG formulations (i.e., IgG1, IgG2, etc) reflect those of normal plasma. The highly-purified nature of IVIGs is shown in Figure 1 of a protein stained 2D-gel electrophoresis map, and revealed heavy and light chains of these natural antibody preparations and their broad pI properties. In antibody binding studies using multiplex (Rules Based Medicine) and microarray binding assays (Figure 2A and B), there were quantitative and qualitative differences in anti-influenza antibody binding to the antigens of H3N2 influenza A/Shandong/9/93, both between different IVIG preparations (data not shown), and within batches of a single IVIG preparation. Along with quantitative differences in antibody binding of immobilized, H1N1 and H3N2 strains of human influenza (Figure 2B). The relative binding of human antibodies in IVIGs was compared by the multianalyte profiling of antibodies to antigens of a variety of other viruses that commonly infect humans, namely herpes simplex virus (HSV), human papilloma virus (HPV), Influenza B (Infl B), respiratory syncytial virus (RSV) and Varicella Zoster (V. Zoster). As might be predicted the antibody responses against the highly antigenic herpes simplex and influenza A viruses that commonly infect humans predominate over quantitative responses to the other viruses examined.


Heterosubtypic anti-avian H5N1 influenza antibodies in intravenous immunoglobulins from globally separate populations protect against H5N1 infection in cell culture.

Sullivan JS, Selleck PW, Downton T, Boehm I, Axell AM, Ayob Y, Kapitza NM, Dyer W, Fitzgerald A, Walsh B, Lynch GW - J Mol Genet Med (2009)

Anti-human influenza binding antibodies in IVIGs: Assays for detection of Ab binding to conformational/ discontinuous influenza epitopes. A. Multiplex assay for anti-influenza IVIG Ab binding showing a comparison of the antibody binding activities of 4 different batches of a single IVIG formulation. IVIG antibody binding to Influenza A: H3N2 A/Shandong/9/1993 and Influenza B: B/Hong Kong/5/1972 antigens displayed on microbeads and performed at the Rules Based Medicine biomarker testing laboratory using multianalyte bead Fluorescence (www.rulesbasedmedicine.com). Binding to herpes simplex virus (HSV), human pappilloma virus (HPV), Influenza B (Infl B) respiratory syncytial virus (RSV) and varicella zoster (V. Zoster) are also included for comparison. The Y-axis values show the amount of IVIG antibody that binds to the respective antigens coated on beads and expressed as the mean ratio of total antibody binding to the respective viral antigens compared to highly-validated RBM internal-reference control standards. B. Microarray slides spotted with viral dilutions (1-0.125mg/ml) of H3N2 (A/Sydney/5/1997), H1N1 (A/Johannesburg/82/1996), tetanus toxoid (Tet Tox) or BSA. IVIG-antibody (D-3) bind robotically arrayed whole inactivated influenza and tetanus toxoid. The BSA is negative. Spots are 10nl and 0.8mm apart. [Note: antigens were bound in duplicate arrays with (top array panel of each slide) and without (bottom array panel on each slide) 0.005% CHAPS. CHAPS was tested as a means of improving liquid flow rates in the Piezorray, but as shown, had no influence on protein binding to the array substrate.]
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2805843&req=5

Figure 2: Anti-human influenza binding antibodies in IVIGs: Assays for detection of Ab binding to conformational/ discontinuous influenza epitopes. A. Multiplex assay for anti-influenza IVIG Ab binding showing a comparison of the antibody binding activities of 4 different batches of a single IVIG formulation. IVIG antibody binding to Influenza A: H3N2 A/Shandong/9/1993 and Influenza B: B/Hong Kong/5/1972 antigens displayed on microbeads and performed at the Rules Based Medicine biomarker testing laboratory using multianalyte bead Fluorescence (www.rulesbasedmedicine.com). Binding to herpes simplex virus (HSV), human pappilloma virus (HPV), Influenza B (Infl B) respiratory syncytial virus (RSV) and varicella zoster (V. Zoster) are also included for comparison. The Y-axis values show the amount of IVIG antibody that binds to the respective antigens coated on beads and expressed as the mean ratio of total antibody binding to the respective viral antigens compared to highly-validated RBM internal-reference control standards. B. Microarray slides spotted with viral dilutions (1-0.125mg/ml) of H3N2 (A/Sydney/5/1997), H1N1 (A/Johannesburg/82/1996), tetanus toxoid (Tet Tox) or BSA. IVIG-antibody (D-3) bind robotically arrayed whole inactivated influenza and tetanus toxoid. The BSA is negative. Spots are 10nl and 0.8mm apart. [Note: antigens were bound in duplicate arrays with (top array panel of each slide) and without (bottom array panel on each slide) 0.005% CHAPS. CHAPS was tested as a means of improving liquid flow rates in the Piezorray, but as shown, had no influence on protein binding to the array substrate.]
Mentions: IVIGs are highly-purified IgG preparations prepared from large pools of human plasma and the immunoglobulin subtypes in IVIG formulations (i.e., IgG1, IgG2, etc) reflect those of normal plasma. The highly-purified nature of IVIGs is shown in Figure 1 of a protein stained 2D-gel electrophoresis map, and revealed heavy and light chains of these natural antibody preparations and their broad pI properties. In antibody binding studies using multiplex (Rules Based Medicine) and microarray binding assays (Figure 2A and B), there were quantitative and qualitative differences in anti-influenza antibody binding to the antigens of H3N2 influenza A/Shandong/9/93, both between different IVIG preparations (data not shown), and within batches of a single IVIG preparation. Along with quantitative differences in antibody binding of immobilized, H1N1 and H3N2 strains of human influenza (Figure 2B). The relative binding of human antibodies in IVIGs was compared by the multianalyte profiling of antibodies to antigens of a variety of other viruses that commonly infect humans, namely herpes simplex virus (HSV), human papilloma virus (HPV), Influenza B (Infl B), respiratory syncytial virus (RSV) and Varicella Zoster (V. Zoster). As might be predicted the antibody responses against the highly antigenic herpes simplex and influenza A viruses that commonly infect humans predominate over quantitative responses to the other viruses examined.

Bottom Line: Although there were qualitative and quantitative differences in binding and protection between globally different formulations, the heterosubtypic antibody activities for the respective IVIGs were in general quite similar.Of particular note because of the relative geographic proximity to the epicentre of H5N1 and the majority of human infections, was the similarity in the antibody binding responses between IVIGs from the Malayan peninsula, Europe and Australia.They also open a window into a somewhat ill defined arena of antibody immunity, namely heterosubtypic immunity.

View Article: PubMed Central - PubMed

Affiliation: Biosafety, Immunobiology, Global Health and Pandemic Infections Research, Central Clinical School, Faculty of Medicine, The University of Sydney, Camperdown, NSW 2006, Australia.

ABSTRACT
With antigenically novel epidemic and pandemic influenza strains persistently on the horizon it is of fundamental importance that we understand whether heterosubtypic antibodies gained from exposures to circulating human influenzas exist and can protect against emerging novel strains. Our studies of IVIG obtained from an infection-naive population (Australian) enabled us to reveal heterosubtypic influenza antibodies that cross react with H5N1. We now expand those findings for an Australian donor population to include IVIG formulations from a variety of northern hemisphere populations. Examination of IVIGs from European and South East-Asian (Malaysian) blood donor populations further reveal heterosubtypic antibodies to H5N1 in humans from different global regions. Importantly these protect against highly pathogenic avian H5N1 infection in vitro, albeit at low titres of inhibition. Although there were qualitative and quantitative differences in binding and protection between globally different formulations, the heterosubtypic antibody activities for the respective IVIGs were in general quite similar. Of particular note because of the relative geographic proximity to the epicentre of H5N1 and the majority of human infections, was the similarity in the antibody binding responses between IVIGs from the Malayan peninsula, Europe and Australia. These findings highlight the value of employing IVIGs for the study of herd immunity, and particularly heterosubtypic antibody responses to viral antigens such as those conserved between circulating human influenzas and emerging influenza strains such as H5N1. They also open a window into a somewhat ill defined arena of antibody immunity, namely heterosubtypic immunity.

No MeSH data available.


Related in: MedlinePlus