H7 Australian Avian Influenza Supplemental Material

H7 Australian Avian Influenza Supplemental Materia

H7 Australian Avian Influenza Sequences

H7 Australian Avian Influenza Sequence

Intranasal vaccination with γ−Flu provides superior protection to heterotypic virus challenge.

<p>Groups of 10 BALB/c mice were either mock treated (A) or vaccinated with γ−A/PC (3.2×10⁶ PFU equivalents) intravenously (B) or intranasally (C). Mice were challenged intranasally after 3 weeks with a lethal dose (6×10² PFU) of A/PR8 and weight recorded daily for 21 days. Survival (D) of mice mock treated, or vaccinated i.n., i.v., i.p., or s.c. and challenged as for (A–C) and monitored for 21 days.</p

H5N1 infectivity and viral genetic loads in lung and brain.

<p>NOTE. Viral infectivity and relative viral genetic loads are expressed as log₁₀ TCID₅₀/g and log₁₀ U per 20 ng of extracted RNA (geometric mean±s.d. of triplicate reactions), respectively, where 1 unit (1 U) of viral RNA is arbitrarily defined as the number of RNA molecules which, when reverse transcribed and subjected to real-time PCR, produced a C_T value of 38.</p>*<p>Day post-challenge.</p>†<p>Undetectable (<10^3.2 TCID₅₀/g (infectivity) or <1 U per 20 ng of extracted RNA (genetic load)).</p

Intranasal vaccination with γ−Flu (γ−A/PR8[H1N1]) protects against H5N1 challenge.

<p>Groups of 10 BALB/c mice were either mock treated (A) or vaccinated with γ−A/PR8 (B). Mice were challenged 4 weeks later with 3 MID₅₀ of A/Vietnam/1203/2004[H5N1] intranasally and weight recorded daily for 21 days.</p

Cross-reactive cytotoxic T cell responses induced by γ-Flu.

<p>10-week-old BALB/c mice were either infected or vaccinated with live A/PR8, γ-A/PR8, live A/PC, or γ-A/PC. Six days later, splenocytes from these mice were tested for their killing activity against mock, A/PC-, A/PR8-, A/JAP-infected, and NPP-labelled P815 targets. Data represent % specific lysis after 6 h assay time at an effector to target cell ratio of 120∶1.</p

Phylogenetic relationship of the polymerase protein of the Mapputta Group viruses and other selected orthobunyaviruses.

<p>Relationship was inferred by Bayesian analysis of the protein sequence alignment. A WAG model of aa substitution with gamma+invariant site heterogeneity was used. Numbers represent Bayesian posterior probabilities (Maximum Likelihood Bootstrap values). Tomato spotted wilt virus (TSWV) has been included as an out-group. Tree is drawn to scale measured in substitutions/site as indicated by the scale bar.</p

Phylogenetic relationship of the M segment polyprotein of the Mapputta Group viruses and other selected orthobunyaviruses.

<p>Relationship was inferred by Bayesian analysis of the protein sequence alignment. A WAG model of aa substitution with gamma+invariant site heterogeneity was used. Numbers represent Bayesian posterior probabilities (Maximum Likelihood Bootstrap values). Tomato spotted wilt virus (TSWV) has been included as an out-group. Tree is drawn to scale measured in substitutions/site as indicated by the scale bar.</p

Genome terminal sequences of MAPV, MPKV and BUCV compared with the orthobunyavirus type species, BUNV.

<p>MAPV and BUCV have the universally conserved complementary terminal 11 bases (in bold) typically observed in orthobunyaviruses, including a non-canonical base pairing at position 9 (<u>underlined</u>). MPKV differs from the consensus sequence with an additional non-canonical base pairing at position 8 (<u>underlined</u>).</p

Cross neutralisation of viruses of the Mapputta group.

<p>Cross neutralisation of viruses of the Mapputta group.</p