Interferon- Protects the Female Reproductive Tract from Viral and Bacterial Infection
Ka Yee Fung1,*,
Niamh E. Mangan1,*,
Jay C. Horvat2,
Jemma R. Mayall2,
Sebastian A. Stifter1,
Nicole De Weerd1,
Laila C. Roisman1,3,
Sarah A. Robertson4,
John E. Schjenken4,
Caroline E. Gargett7,8,
Hong P. T. Nguyen7,
Daniel J. Carr9,
Philip M. Hansbro2, and
Paul J. Hertzog1,
1 Centre for Innate Immunity and Infectious Diseases, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia.
2 Centre for Asthma and Respiratory Disease and Hunter Medical Research Institute, The University of Newcastle, Newcastle, New South Wales, Australia.
3 Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
4 Robinson Institute and School of Paediatrics and Reproductive Health, University of Adelaide, South Australia, Australia.
5 Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
6 Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Australia.
7 The Ritchie Centre, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia.
8 Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia.
9 Department of Ophthalmology and Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
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Fig. 1. Interferon- signals through the type I IFN receptor but is not induced by TLR ligands nor regulated by IRFs. (A and B) BMDMs from WT, Ifnar1–/–, and Ifnar2–/– C57BL/6 mice were stimulated with recombinant mouse Ifn-α1, Ifn-β, or Ifn- (0.1 μg/ml) for 3 hours. (A) Irf-7 and (B) 2'5'oas expression was measured by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR). Data are expressed as mean ± SEM (error bars) of at least three independent experiments. (C) BMDMs from C57BL/6 WT mice were treated with a range of TLR ligands or transfected with Poly (I:C) and Poly (dA:dT) for 3 hours at 37°C. Ifn-β, Il-6, and Ifn- were measured by qRT-PCR. Data are expressed as mean ± SEM of at least three independent experiments. (D) Luciferase reporter plasmids containing Ifn-α, Ifn-β, p125, or Ifn- were cotransfected with empty vector or IRF-3, IRF-7, or IRF-5 expression vectors into HEK293 cells. Data are expressed as mean ± SEM. All values are means of at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (unpaired Student's t test).|
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Fig. 3. Ifn-–/– mice are more susceptible to HSV-2 vaginal infection. (A) Isg15, (B) Irf-7, and (C) 2'5'oas expression between WT and Ifn-–/– C57BL/6 mice was determined by qRT-PCR. The values represent means ± SEM (error bars) of four individual mice. (D to F and H) Mice pretreated with medroxyprogesterone acetate (Depo-Ralovera, Pfizer) at day –5 were infected with HSV-2 (D, E, H) at a level of 2400 or (F) 24 pfu per mouse on day 0. (D) Representative images demonstrating overt genital lesions, redness, and swelling in HSV-2–infected Ifn-–/– mice at day 7 pi; these qualities are absent in C57BL/6 WT mice. (E)Clinical scores of WT and Ifn-–/– C57BL/6 mice during the 7-day course of infection. Data are means ± SEM of five individual mice and are representative of at least three separate experiments. (F and G) HSV-2 titers (pfu) from vaginal tissue of WT and Ifn-–/– C57BL/6 mice infected with (F) 2400 and (G) 24 pfu, respectively, at day 3 pi were determined by titration of clarified vaginal tissue samples on Vero cell monolayers by plaque assay. Data are expressed as mean ± SEM of five individual mice. (H) HSV-2 titers from homogenates of vaginal tissue, spinal cord, and brain stem of infected WT and Ifn-–/– C57BL/6 mice at day 7 pi were determined as in (F) and (G). Data are expressed as mean ± SEM of five individual mice. *P < 0.05; **P < 0.01 (unpaired Student's t test).|