Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Human SAA and/or HDL proteins compared in this study †

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Sequencing, Recombinant

Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Binding of SAA preparations to influenza A virus (IAV)—Binding of SAAs to the Phil82 strain of IAV was tested by ELISA as described in Section 2 (“Materials And Methods”) using an anti‐serum amyloid A (anti‐SAA1) antibody (panel A). Binding of SAA1 was also tested in presence of maltose (panel B) and using an anti‐DDK antibody for detection (SAA1 has a DDK tail) (panel C). HDL binding to IAV was tested as well (panel D). * = P < 0.05 and ** = P < 0.01 compared to control. Results represent mean ± semfor 3 to 5 experiments

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Binding Assay, Virus, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Neutralization of influenza A virus (IAV) by serum SAA, but not recombinant, SAA preparations—The indicated strains of IAV were pre‐incubated with SAA and or HDL preparations and then these were used to infection MDCK cell monolayers as described in Section 2 (“Materials And Methods”). Serum SAA from Abcam (panel A), SAA complexed with HDL (SAAHDL; panel B), serum HDL (panel C), and two recombinant serum amyloid A (SAA1) preparations were compared (panel D; HDL and SAAHDL included for comparison). * = P < 0.05 and ** = P < 0.01 and *** = < 0.001 compared to control. Results represent mean ± semfor 3 to 5 experiments

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Neutralization, Virus, Recombinant, Incubation, Infection, Comparison, Control

Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Effects of TLR2 blocking antibodies, pertussis toxin (PT), and wortmannin of neutrophil responses to Apo‐serum amyloid A (Apo‐SAA1)

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Blocking Assay, Control, Virus

Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Stimulation of neutrophil hydrogen peroxide production by SAA and or HDL—Neutrophil H2O2generation was measured through decline in scopoletin fluorescence as described in Section 2 (“Materials And Methods”). Influenza A virus (IAV; Phil82 strain) was pre‐incubated with the indicated concentrations of SAA and/or HDL preparations and added to suspensions of neutrophils at time 0. Results of unstimulated cells (PBS) are included for comparison. Instances where serum amyloid A (SAA1) proteins increased IAV‐induced H2O2production are noted by * or ** in in legend, where * = P < 0.05 and ** = P < 0.01 compared to control. Results represent mean ± semfor 3 to 5 experiments

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Fluorescence, Virus, Incubation, Comparison, Control

Journal: Journal of leukocyte biology

Article Title: Effects of serum amyloid protein A on influenza A virus replication and viral interactions with neutrophils

doi: 10.1002/JLB.4AB0220-116RR

Figure Lengend Snippet: Effect of SAA and or HDL on neutrophil IL‐8 production or caspase activity—In panel A, IL‐8 production by neutrophils was measured by ELISA as described in Section 2 (“Materials And Methods”). ** = P < 0.01 compared to control. Results represent mean ± semfor 3 to 5 experiments. Panel B shows IL‐8 production in response to influenza A virus (IAV) alone or IAV combined with Apo‐serum amyloid A (Apo‐SAA1). Panel C shows caspase 3 activation by IAV alone or IAV combined with Apo‐SAA1, SAAHDL (SAA complexed with HDL), SAA1, or HDL. * = P < 0.05 compared to results with neutrophils in media alone. Results are mean ± semof 4 experiments

Article Snippet: TABLE 1 Protein (amino acids included) Source Sequence Description Designation used in this paper Serum amyloid A (SAA1) (19‐94) Human serum(Abcam) RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDS No N‐terminal signal peptide, no C‐terminus Serum SAA SAAHDL Human Serum (EMD Millipore) Serum Amyloid A Protein‐Rich‐High‐Density Lipoprotein (262 μg/ml SAA, 1.508 μg/ml HDL) No N‐terminal signal peptide, With HDL SAAHDL Apo‐SAA1 (19‐122) Recombinant E. coli (Peprotech) MRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY No N‐ terminal signal peptide ApoSAA1 SAA1 (1‐122) Recombinant HEK cells (Origene) MKLLTGLVFCSLVLGVSSRSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY Full length (contains signal peptide) SAA1 HDL Human serum (Sigma‐Aldrich) HDL Open in a separate window † Human apolipoprotein serum amyloid A‐1 gene codes for a 122 amino acid nonglycosylated polypeptide including an 18 amino acid N‐terminal sequence.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Control, Virus, Activation Assay