Fig 1.
Efficient transmission of seasonal H1N1 and H3N2 influenza viruses to naïve animals under short or periodic exposures.
(A) Schematic of experimental procedure. Shaded gray bars depict exposure times. Three to four donor ferrets were infected with A/CA/07/2009 (H1N1) (B-D) or A/Perth/16/2009 (H3N2) (E-G) and a naïve recipient ferret was placed in the adjacent cage at 24 hour post-infection for 7 (B and E) or 2 (C and F) continuous days, or 8 hours a day for 5 consecutive days (D and G). Nasal washes were collected from all ferrets on the indicated days and each bar indicates an individual ferret. Limit of detection is indicated by dashed line. Viral titers for donor animals in part G have previously been published in [45].
Fig 2.
Pre-existing heterosubtypic immunity blocks H3N2 transmission but not H1N1pdm09 transmission.
(A) Schematic of experimental procedure, gray bars depict 2-day exposure time. (B) All donor and recipient animals were infected with A/Perth/16/2009 (H3N2) virus 60 days prior to H1N1pdm09 (A/CA/07/2009) transmission. Donor animals are denoted as H3-H1 INF, and recipient animals are ‘H3-imm’. (C) All donor and recipient animals were infected with A/CA/07/2009 (H1N1pdm09) virus 84 days prior to H3N2 (Perth/16/2009) transmission. Donor animals are denoted as H1-H3 INF, and recipient animals are ‘H1-imm’. The contribution of donor and recipient immunity was tested separately for A/Perth/16/2009 (H3N2) transmission. Donors only (D) or recipient only (E) had pre-existing immunity to A/CA/07/09 H1N1pdm09. Donor animals are denoted as H1-H3 INF or H3N2 INF (no prior immunity), and recipient animals are either ‘naïve’ to indicate no prior immunity or ‘H1-imm’. Each bar represents an individual animal. Limit of detection is denoted by a dashed line.
Fig 3.
Neutralizing antibody titers against H1N1 and H3N2 viruses.
We examined the neutralizing antibody titers against A/CA/07/09 (H1N1pdm09; red lines) and A/Perth/16/09 (H3N2; green lines) from ferrets depicted in Fig 2B and 2C. Sera from animals from Fig 2C; H1N1pdm09 primary infection followed by H3N2 transmission are depicted in panel A. Sera from animals from Fig 2B, H3N2 primary infection followed by H1N1pdm09 transmission, are depicted in panel B. At the indicated times, serum was collected and neutralizing antibodies against both H1N1pdm09 (red lines) and H3N2 (green lines) were determined by microneutralization assay. Donor infected animals are denoted in solid circles and recipient animals are denoted by open circles. Each point is an individual animals and trend lines for donor (solid) or recipient (dashed) animals are shown. Limit of detection is indicated by the dashed line.
Fig 4.
Innate immunity does not block H3N2 transmission.
(A) Schematic of experimental procedure, gray bars depict 2-day exposure time. (B) Recipient animals were infected with B/Brisbane/60/2008 (IBV) virus 67 days prior to H3N2 (A/Perth/16/2009) transmission. Donor animals are denoted as H3 INF, and recipient animals are ‘IBV-imm’. Each bar represents an individual animal. Limit of detection is denoted by a dashed line.