Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Apr;93(4):830-8.
doi: 10.3382/ps.2013-03511.

Dramatic differences in the response of macrophages from B2 and B19 MHC-defined haplotypes to interferon gamma and polyinosinic:polycytidylic acid stimulation

Maisie E Dawes  1 , Lisa M Griggs, Ellen W Collisson, Worthie E Briles, Yvonne Drechsler
Affiliations

Dramatic differences in the response of macrophages from B2 and B19 MHC-defined haplotypes to interferon gamma and polyinosinic:polycytidylic acid stimulation

Maisie E Dawes et al. Poult Sci. 2014 Apr.

Abstract

The chicken MHC has been associated with disease resistance, though the mechanisms are not understood. The functions of macrophages, critical to both innate and acquired immunity, were compared between the more infectious bronchitis virus-resistant B2 and the more infectious bronchitis virus-susceptible B19 lines. In vivo peripheral blood concentrations of monocytes were similar in B2 or B19 homozygous haplotypes. Peripheral blood-derived macrophages were stimulated with poly I:C, simulating an RNA virus, or IFNγ, a cytokine at the interface of innate and adaptive immunity. Not only did B2-derived peripheral monocytes differentiate into macrophages more readily than the B19 monocytes, but as determined by NO production, macrophages from B2 and B2 on B19 genetic background chicks were also significantly more responsive to either stimulant. In conclusion, the correlation with resistance to illness following viral infection may be directly linked to a more vigorous innate immune response.

Keywords: disease resistance; infectious bronchitis virus; innate immunity; macrophage; monocyte.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Ex vivo differentiation of monocytes prestimulation. Images demonstrate the rate of differentiation of chicken monocytes from homozygous B2 (panel A) and B19 (panel B) chicks at d 2, 4, and 6 d postadherence. Distinct vacuoles, increased numbers of stellate cytoplasmic processes, and decreased nuclear to cytoplasmic ratio are visible on d 4 with B2 cells and on d 6 with B19 cells. Pictures were taken at 20× magnification, inverted, with Nikon Eclipse Ti fluorescence microscope (Nikon Instruments Inc., Melville, NY).
Figure 2
Figure 2
Differentiation of 48 h interferon (IFN)γ-stimulated macrophages. Macrophages cultured for 6 d from B2 and B19 haplotypes were stimulated with IFNγ for 48 h, and pictures were taken with a Nikon Eclipse Ti fluorescence microscope (Nikon Instruments Inc., Melville, NY), inverted at 10× magnification. Macrophage-like features (stellate cytoplasmic processes, vacuolation, and decreased nuclear to cytoplasmic ratio) are more pronounced in B2 cells (A) compared with their B19 counterparts (B). Color version available in the online PDF.
Figure 3
Figure 3
Immunofluorescent labeling of adherent macrophages. Panel A depicts the brightfield image of macrophages. The green fluorescence noted in panel B depicts positive labeling of B2 and B19 chicken cells by monoclonal antibody KUL01-FITC, confirming their monocyte/macrophage cell lineage. Pictures were taken with Nikon Eclipse Ti fluorescence microscope (Nikon Instruments Inc., Melville, NY), inverted at 10× magnification. Color version available in the online PDF.
Figure 4
Figure 4
Kinetics. Nitric oxide release is shown as mean (μg/mL) ± SD by B2 and B19 macrophages at 24, 48, and 72 h poststimulation with poly (I:C); n = 8 (A) and interferon (IFN) γ; n = 6 (B). In the presence of either stimulant, NO production at all 3 time points was significantly lower in B19 than in B2 supernatants (P < 0.01, t-test, paired). Samples were taken at 6 wk of age. Color version available in the online PDF.
Figure 5
Figure 5
A: Nitric oxide release by poly(I:C)-stimulated B2 and B19 macrophages. Cells were stimulated for 48 h with 25 μg/mL poly (I:C). Although not significantly different, basal NO concentration was consistently greater in B19 culture supernatants (10.88 μg/mL) than in B2 supernatants (9.75 μg/mL). Following stimulation, NO production by B2 cells (98.12 μg/mL) was significantly greater than that by B19 macrophages (17.19 μg/mL; P < 0.01, one-way ANOVA). Values are reported as mean with SE, n = 28. B: Nitric oxide release by interferon (IFN)γ-stimulated B2 and B19 macrophages. Cells were stimulated for 48 h with 50 pg/mL of ch-IFNγ. Although not significantly different, basal NO concentration was consistently greater in B19 culture supernatants (11.88 μg/mL) than in B2 supernatants (8.7 μg/mL). Following stimulation, NO production was significantly greater in B2 culture supernatants (92.76 μg/mL) than in B19 culture supernatants (17.9 μg/mL; P < 0.01, one-way ANOVA). Values are reported as mean with SE, n = 21.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Bacon L.D., Hunter D.B., Zhang H.M., Brand K., Etches R. Retrospective evidence that the MHC (B haplotype) of chickens influences genetic resistance to attenuated infectious bronchitis vaccine strains in chickens. Avian Pathol. 2004;33:605–609. 15763730. - PubMed
    1. Banat G.R., Tkalcic S., Dzielawa J.A., Jackwood M.W., Saggese M.D., Yates L., Kopulos R., Briles W.E., Collisson E.W. Association of the chicken MHC B haplotypes with resistance to avian coronavirus. Dev. Comp. Immunol. 2013;39:430–437. 23178407. - PMC - PubMed
    1. Bayyari G.R., Huff W.E., Rath N.C., Balog J.M., Newberry L.A., Villines J.D., Skeeles J.K., Anthony N.B., Nestor K.E. Effect of the genetic selection of turkeys for increased body weight and egg production on immune and physiological responses. Poult. Sci. 1997;76:289–296. 9057208. - PubMed
    1. Bearse G.E., McClary C.F., Miller M.W. The results of eight years’ selection for disease resistance and susceptibility in White Leghorns. Poult. Sci. 1939;18:400–401.
    1. Boockvar K.S., Granger D.L., Poston R.M., Maybodi M., Washington M.K., Hibbs J.B., Jr., Kurlander R.L. Nitric oxide produced during murine listeriosis is protective. Infect. Immun. 1994;62:1089–1100. 7509315. - PMC - PubMed

Publication types

MeSH terms