Proinflammatory cytokine changes in bronchoalveolar lavage fluid cells isolated from pigs infected solely with porcine reproductive and respiratory syndrome virus or co-infected with swine influenza virus

Czyżewska-Dors, Ewelina; Pomorska-Mól, Małgorzata; Dors, Arkadiusz; Pluta, Aneta; Podgórska, Katarzyna; Kwit, Krzysztof; Stasiak, Ewelina; Łukomska, Anna

doi:10.2478/jvetres-2019-0063

Abstract

Introduction

The study evaluated the patterns of local innate immune response in bronchoalveolar lavage fluid (BALF) cells of pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) alone or co-infected with swine influenza virus (SIV).

Material and Methods

The study was performed on 26 seven-week-old pigs in three groups: PRRSV-infected (n = 11), PRRSV and SIV-infected (n = 11), and control (n = 4). BALF was collected post euthanasia at 2 and 4 dpi (three piglets per inoculated group) and at 21 dpi (all remaining pigs). Expression of IFN-α, IFN-γ, IL-1β, IL-6, IL-8, and IL-10 mRNA was quantified in BALF cells. PRRSV RNA was quantified in BALF samples using a commercial real-time RT-PCR kit.

Results

The three cytokines IFN-α, IFN-γ, and IL-1β presented significant expression changes in all experimental pigs. In PRRSV-infected animals IL-8 also did, but in co-infected subjects IL-6 and IL-10 were the additional upregulated cytokines. The highest number of differentially expressed genes was observed at 4 dpi, and significant differences in cytokine gene expression did not occur between the experimental groups at any other time point. The mean PRRSV load in the BALF of PRRSV-infected pigs was higher than that of co-infected pigs at each time point, having statistical significance only at 4 dpi.

Conclusion

The results of the study indicate that infection with PRRSV alone as well as with SIV interferes with innate and adaptive immune response in the infected host. They also showed that co-infection demonstrates additive effects on IL-6 and IL-10 mRNA expression levels.

References

Barbé F., Atanasova K., Van Reeth K.: Cytokines and acute phase proteins associated with acute swine influenza infection in pigs. Vet J 2011, 187, 48–53.
Search in Google Scholar Back to article
Choi Y.K., Goyal S.M., Joo H.S.: Retrospective analysis of etiologic agents associated with respiratory diseases in pigs. Can Vet J 2003, 44, 735–737.
Search in Google Scholar Back to article
Czyżewska-Dors E., Dors A., Kwit K., Stasiak E., Pomorska-Mól M.: Pig lung immune cytokine response to the swine influenza virus and the Actinobacillus pleuropneumoniae infection. J Vet Res 2017, 61, 259–265.
Search in Google Scholar Back to article
Díaz I., Darwich L., Pappaterra G., Pujols J., Mateu E.: Immune responses of pigs after experimental infection with a European strain of porcine reproductive and respiratory syndrome virus. J Gen Virol 2005, 86, 1943–1951.
Search in Google Scholar Back to article
Dobrescu I., Levast B., Lai K., Delgado-Ortega M., Walker S., Banman S., Townsend H., Simon G., Zhou Y., Gerdts V., Meurens F.: In vitro and ex vivo analyses of co-infections with swine influenza and porcine reproductive and respiratory syndrome viruses. Vet Microbiol 2014, 169, 18–32.
Search in Google Scholar Back to article
Duvigneau J.C., Hartl R.T., Groiss S., Gemeiner M.: Quantitative simultaneous multiplex real-time PCR for the detection of porcine cytokines. J Immunol Methods 2005, 306, 16–27.
Search in Google Scholar Back to article
Dwivedi V., Manickam C., Binjawadagi B., Linhares D., Murtaugh M.P., Renukaradhya G.J.: Evaluation of immune responses to porcine reproductive and respiratory syndrome virus in pigs during early stage of infection under farm conditions. Virol J 2012, 9, 45.
Search in Google Scholar Back to article
Fablet C., Marois-Crehan C., Simon G., Grasland B., Jestin A., Kobisch M., Madec F., Rose N.: Infectious agents associated with respiratory diseases in 125 farrow-to-finish pig herds: a cross-sectional study. Vet Microbiol 2012, 157, 152–163.
Search in Google Scholar Back to article
García-Nicolás O., Rosales R.S., Pallarés F.J., Risco D., Quereda J.J., Graham S.P., Frossard J.P., Morgan S.B., Steinbach F., Drew T.W., Strickland T.S., Salguero F.J.: Comparative analysis of cytokine transcript profiles within mediastinal lymph node compartments of pigs after infection with porcine reproductive and respiratory syndrome genotype 1 strains differing in pathogenicity. Vet Res 2015, 46, 34.
Search in Google Scholar Back to article
Gimeno M., Darwich L., Diaz I., de la Torre E., Pujols J., Martin M., Inumaru S., Cano E., Domingo M., Montoya M., Mateu E.: Cytokine profiles and phenotype regulation of antigen presenting cells by genotype-I porcine reproductive and respiratory syndrome virus isolates. Vet Res 2011, 42, 9.
Search in Google Scholar Back to article
Gómez-Laguna J., Salguero F.J., Barranco I., Pallarés F.J., Rodríguez-Gómez I.M., Bernabé A., Carrasco L.: Cytokine expression by macrophages in the lung of pigs infected with the porcine reproductive and respiratory syndrome virus. J Comp Pathol 2010, 142, 51–60.
Search in Google Scholar Back to article
Guo B., Lager K.M., Henningson J.N., Miller L.C., Schlink S.N., Kappes M.A., Kehrli M.E. Jr, Brockmeier S.L., Nicholson T.L., Yang H.C., Faaberg K.S.: Experimental infection of United States swine with a Chinese highly pathogenic strain of porcine reproductive and respiratory syndrome virus. Virology 2013, 435, 372–384.
Search in Google Scholar Back to article
Halbur P.G., Paul P.S., Frey M.L., Landgraf J., Eernisse K., Meng X.J., Lum M.A., Andrews J.J., Rathje J.A.: Comparison of the pathogenicity of two US porcine reproductive and respiratory syndrome virus isolates with that of the Lelystad virus. Vet Pathol 1995, 32, 648–660.
Search in Google Scholar Back to article
Huang C., Zhang Q., Feng W.H.: Regulation and evasion of antiviral immune responses by porcine reproductive and respiratory syndrome virus. Virus Res 2015, 202, 101–111.
Search in Google Scholar Back to article
Khatri M., Dwivedi V., Krakowka S., Manickam C., Ali A., Wang L.: Swine influenza H1N1 virus induces acute inflammatory immune responses in pig lungs: a potential animal model for human H1N1 influenza virus. J Virol 2010, 84, 11210–11218.
Search in Google Scholar Back to article
Kowalczyk A., Pomorska-Mól M., Kwit K., Pejsak Z., Rachubik J., Markowska-Daniel I.: Cytokine and chemokine mRNA expression profiles in BALF cells isolated from pigs single infected or co-infected with swine influenza virus and Bordetella bronchiseptica Vet Microbiol 2014, 170, 206–212.
Search in Google Scholar Back to article
Liu Y., Du Y., Wang H., Du L., Feng W.H.: Porcine reproductive and respiratory syndrome virus (PRRSV) up-regulates IL-8 expression through TAK-1/JNK/AP-1 pathways. Virology 2017, 506, 64–72.
Search in Google Scholar Back to article
Loving C.L., Brockmeier S.L., Vincent A.L., Palmer M.V., Sacco R.E., Nicholson T.L.: Influenza virus coinfection with Bordetella bronchiseptica enhances bacterial colonization and host responses exacerbating pulmonary lesions. Microb Pathog 2010, 49, 237–245.
Search in Google Scholar Back to article
Meier W.A., Galeota J., Osorio F.A., Husmann R.J., Schnitzlein W.M., Zuckermann F.A.: Gradual development of the interferon-γ response of swine to porcine reproductive and respiratory syndrome virus infection or vaccination. Virology 2003, 309, 18–31.
Search in Google Scholar Back to article
Meurens F., Berri M., Auray G., Melo S., Levast B., Virlogeux-Payant I., Chevaleyre C., Gerdts V., Salmon H.: Early immune response following Salmonella enterica subspecies enterica serovar Typhimurium infection in porcine jejunal gut loops. Vet Res 2009, 40, 5.
Search in Google Scholar Back to article
Miguel J.C., Chen J., Van Alstine W.G., Johnson R.W.: Expression of inflammatory cytokines and Toll-like receptors in the brain and respiratory tract of pigs infected with porcine reproductive and respiratory syndrome virus. Vet Immunol Immunopathol 2010, 135, 314–319.
Search in Google Scholar Back to article
Moore K.W., de Waal Malefyt R., Coffman R.L., O’Garra A.: Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 2001, 19, 683–765.
Search in Google Scholar Back to article
Pfaffl M.W.: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001, 29, 2002–2007.
Search in Google Scholar Back to article
Pomorska-Mól M., Markowska-Daniel I., Kwit K., Czyżewska E., Dors A., Rachubik J., Pejsak Z.: Immune and inflammatory response in pigs during acute influenza caused by H1N1 swine influenza virus. Arch Virol 2014, 159, 2605–2614.
Search in Google Scholar Back to article
Rola-Łuszczak M., Materniak-Kornas M., Pluta A., Podgórska K., Nielsen J., Stadejek T., Kuźmak J.: Transcriptional profiles of PBMCs from pigs infected with three genetically diverse porcine reproductive and respiratory syndrome virus strains. Mol Biol Rep 2018, 45, 675–688.
Search in Google Scholar Back to article
Rossow K.D.: Porcine reproductive and respiratory syndrome. Vet Pathol 1998, 35, 1–20.
Search in Google Scholar Back to article
Solano G.I., Segalés J., Collins J.E., Molitor T.W., Pijoan C.: Porcine reproductive and respiratory syndrome virus (PRRSV) interaction with Haemophilus parasuis Vet Microbiol 1997, 55, 247–257.
Search in Google Scholar Back to article
Thanawongnuwech R., Brown G.B., Halbur P.G., Roth J.A., Royer R.L., Thacker B.J.: Pathogenesis of porcine reproductive and respiratory syndrome virus-induced increase in susceptibility to Streptococcus suis infection. Vet Pathol 2000, 37, 143–152.
Search in Google Scholar Back to article
Theofilopoulos A.N., Baccala R., Beutler B., Kono D.H.: Type I interferons (α/β) in immunity and autoimmunity. Annu Rev Immunol 2005, 23, 307–336.
Search in Google Scholar Back to article
Tu P.Y., Tsai P.C., Lin Y.H., Liu P.C., Chang H.L., Kuo T.Y., Chung W.B.: Expression profile of Toll-like receptor mRNA in pigs co-infected with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2. Res Vet Sci 2015, 98, 134–141.
Search in Google Scholar Back to article
Zuckermann F.A., Garcia E.A., Luque I.D., Christopher-Hennings J., Doster A., Brito M., Osorio F.: Assessment of the efficacy of commercial porcine reproductive and respiratory syndrome virus (PRRSV) vaccines based on measurement of serologic response, frequency of gamma-IFN-producing cells and virological parameters of protection upon challenge. Vet Microbiol 2007, 123, 69–85.
Search in Google Scholar Back to article