Hostname: page-component-8448b6f56d-m8qmq Total loading time: 0 Render date: 2024-04-18T06:54:27.826Z Has data issue: false hasContentIssue false

Parasite-specific proliferative responses of chicken spleen cells upon in vitro stimulation with Eimeria tenella antigen

Published online by Cambridge University Press:  09 November 2018

Eva Wattrang*
Affiliation:
Department of Microbiology, National Veterinary Institute, SE-751 89 Uppsala, Sweden
Per Thebo
Affiliation:
Department of Microbiology, National Veterinary Institute, SE-751 89 Uppsala, Sweden
Osama Ibrahim
Affiliation:
Department of Microbiology, National Veterinary Institute, SE-751 89 Uppsala, Sweden
Tina Sørensen Dalgaard
Affiliation:
Department of Animal Science, Aarhus University, Blichers Allé 20, P.O. box 50, DK-8830, Tjele, Denmark
Anna Lundén
Affiliation:
Department of Microbiology, National Veterinary Institute, SE-751 89 Uppsala, Sweden
*
Author for correspondence: Eva Wattrang, E-mail: eva.wattrang@sva.se

Abstract

This study aimed to set up methodology to monitor parasite-specific T-cell activation in vitro using Eimeria tenella-infected chickens. A sonicated E. tenella sporozoite protein preparation was used for the activation of chicken spleen cell cultures. Proliferation assessed by 3H-thymidin incorporation or blast transformation of T-cells assessed by immunofluorescence labelling and flow cytometry were used as read-outs for activation. Results showed that E. tenella-specific proliferation was detected in cultures of spleen cells collected in a ‘window’ between 8 and 14 days after primary infection. However, due to high variation in proliferative responses between individuals and to high background proliferation, large numbers of observations were needed to obtain significant results. Moreover, the outcome was not improved by increasing the infection dose to chickens or by depletion of T-cell receptor (TCR) γ/δ expressing cells from cultures. An E. tenella-specific blast transformation response was observed for TCRα/β expressing cells within the same ‘window’, confirming the identity of the responding cells as classic T-cells. Thus, it is possible to study the kinetics of E. tenella-specific T-cell responses in vitro. However, more in-depth phenotypic identification of the responding T-cells could improve the methodology.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anonymous (1971). Manual of Veterinary Parasitological Laboratory Techniques. London: Ministry of Agriculture, Fisheries and Food.Google Scholar
Blake, DP and Tomley, FM (2014) Securing poultry production from the ever-present Eimeria challenge. Trends in Parasitology 30, 1219.Google Scholar
Blake, DP, Hesketh, P, Archer, A, Carroll, F, Shirley, MW and Smith, AL (2005) The influence of immunizing dose size and schedule on immunity to subsequent challenge with antigenically distinct strains of Eimeria maxima. Avian Pathology 34, 489494.Google Scholar
Braukmann, M, Methner, U and Berndt, A (2015) Avian CD25( + ) gamma/delta (γδ) T cells after Salmonella exposure. Veterinary Immunology and Immunopathology 168, 1418.Google Scholar
Breed, DG, Dorrestein, J and Vermeulen, AN (1996) Immunity to Eimeria tenella in chickens: phenotypical and functional changes in peripheral blood T-cell subsets. Avian Diseases 40, 3748.Google Scholar
Breed, DG, Dorrestein, J, Schetters, TP, Waart, LV, Rijke, E and Vermeulen, AN (1997a) Peripheral blood lymphocytes from Eimeria tenella infected chickens produce gamma-interferon after stimulation in vitro. Parasite Immunology 19, 127135.Google Scholar
Breed, DG, Schetters, TP, Verhoeven, NA and Vermeulen, AN (1997b) Characterization of phenotype related responsiveness of peripheral blood lymphocytes from Eimeria tenella infected chickens. Parasite Immunology 19, 563569.Google Scholar
Bumstead, JM, Bumstead, N, Rothwell, L and Tomley, FM (1995) Comparison of immune responses in inbred lines of chickens to Eimeria maxima and Eimeria tenella. Parasitology 111(Pt 2), 143151.Google Scholar
Chapman, HD and Shirley, MW (2003) The Houghton strain of Eimeria tenella: a review of the type strain selected for genome sequencing. Avian Pathology 32, 115127.Google Scholar
Chapman, HD, Barta, JR, Blake, D, Gruber, A, Jenkins, M, Smith, NC, Suo, X and Tomley, FM (2013) A selective review of advances in coccidiosis research. Advances in Parasitology 83, 93171.Google Scholar
Dalgaard, TS, Norup, LR, Rubbenstroth, D, Wattrang, E and Juul-Madsen, HR (2010) Flow cytometric assessment of antigen-specific proliferation in peripheral chicken T cells by CFSE dilution. Veterinary Immunology and Immunopathology 138, 8594.Google Scholar
Gaines, H and Biberfeld, G (2000) Measurement of lymphoproliferation at the single-cell level by flow cytometry. Methods in Molecular Biology 134, 243255.Google Scholar
Haug, A, Thebo, P and Mattsson, JG (2007) A simplified protocol for molecular identification of Eimeria species in field samples. Veterinary Parasitology 146, 3545.Google Scholar
Hermosilla, C, Burger, HJ and Zahner, H (1999) T cell responses in calves to a primary Eimeria bovis infection: phenotypical and functional changes. Veterinary Parasitology 84, 4964.Google Scholar
Hoek, A, Rutten, VP, Kool, J, Arkesteijn, GJ, Bouwstra, RJ, Van Rhijn, I and Koets, AP (2009) Subpopulations of bovine WC1( + ) gammadelta T cells rather than CD4( + )CD25(high) Foxp3( + ) T cells act as immune regulatory cells ex vivo. Veterinary Research 40, 6.Google Scholar
Huang, X, Zou, J, Xu, H, Ding, Y, Yin, G, Liu, X and Suo, X (2011) Transgenic Eimeria tenella expressing enhanced yellow fluorescent protein targeted to different cellular compartments stimulated dichotomic immune responses in chickens. Journal of Immunology 187, 35953602.Google Scholar
Johnson, J and Reid, WM (1970) Anticoccidial drugs: lesion scoring techniques in battery and floor-pen experiments with chickens. Experimental Parasitology 28, 3036.Google Scholar
Joyner, LP and Norton, CC (1973) The immunity arising from continuous low-level infection with Eimeria tenella. Parasitology 67, 333340.Google Scholar
Kruisbeek, AM, Shevach, E and Thornton, AM (2004) Proliferative assays for T cell function. Current Protocols in Immunology, Chapter 3, Unit 3 12, pp. 1–20. doi: 10.1002/0471142735.im0312s60.Google Scholar
Leathem, WD and Burns, WC (1967) Effects of the immune chicken on the endogenous stages of Eimeria tenella. Journal of Parasitology 53, 180185.Google Scholar
Leathem, WD and Burns, WC (1968) Duration of acquired immunity of the chicken to Eimeria tenella infection. Journal of Parasitology 54, 227232.Google Scholar
Lillehoj, HS (1986) Immune response during coccidiosis in SC and FP chickens. I. In vitro assessment of T cell proliferation response to stage-specific parasite antigens. Veterinary Immunology and Immunopathology 13, 321330.Google Scholar
Martin, A, Lillehoj, HS, Kaspers, B and Bacon, LD (1993) Antigen-specific T cell proliferation following coccidia infection. Poultry Science 72, 20842094.Google Scholar
Muul, LM, Heine, G, Silvin, C, James, SP, Candotti, F, Radbruch, A and Worm, M (2011) Measurement of proliferative responses of cultured lymphocytes. Current Protocols in Immunology, Chapter 7, Unit7 10, pp. 126. doi: 10./0471142735.im0710s94.Google Scholar
Norup, LR, Dalgaard, TS, Pedersen, AR and Juul-Madsen, HR (2011) Assessment of Newcastle disease-specific T cell proliferation in different inbred MHC chicken lines. Scandinavian Journal of Immunology 74, 2330.Google Scholar
Pieper, J, Methner, U and Berndt, A (2008) Heterogeneity of avian gammadelta T cells. Veterinary Immunology and Immunopathology 124, 241252.Google Scholar
Pierce, AE, Long, PL and Horton-Smith, C (1962) Immunity to Eimeria tenella in young fowls (Gallus domesticus). Immunology 5, 129152.Google Scholar
Renaux, S, Quere, P, Buzoni-Gatel, D, Sewald, B, Le Vern, Y, Coudert, P and Drouet-Viard, F (2003) Dynamics and responsiveness of T-lymphocytes in secondary lymphoid organs of rabbits developing immunity to Eimeria intestinalis. Veterinary Parasitology 110, 181195.Google Scholar
Rose, ME (1974) The early development of immunity to Eimeria maxima in comparison with that to Eimeria tenella. Parasitology 68, 3545.Google Scholar
Rose, ME and Hesketh, P (1982) Immunity to coccidia in chickens: adoptive transfer with peripheral blood lymphocytes and spleen cells. Parasite Immunology 4, 171185.Google Scholar
Rose, ME and Hesketh, P (1987) Eimeria tenella: effects of immunity on sporozoites within the lumen of the small intestine. Experimental Parasitology 63, 337344.Google Scholar
Rose, ME and Long, PL (1962) Immunity to four species of Eimeria in fowls. Immunology 5, 7992.Google Scholar
Rose, ME, Lawn, AM and Millard, BJ (1984) The effect of immunity on the early events in the life-cycle of Eimeria tenella in the caecal mucosa of the chicken. Parasitology 88(Pt 2), 199210.Google Scholar
Schmatz, DM, Crane, MS and Murray, PK (1984) Purification of Eimeria sporozoites by DE-52 anion exchange chromatography. Journal of Protozoology 31, 181183.Google Scholar
Suhwold, A, Hermosilla, C, Seeger, T, Zahner, H and Taubert, A (2010) T cell reactions of Eimeria bovis primary and challenge-infected calves. Parasitology Research 106, 595605.Google Scholar
Svahn, A, Linde, A, Thorstensson, R, Karlen, K, Andersson, L and Gaines, H (2003) Development and evaluation of a flow-cytometric assay of specific cell-mediated immune response in activated whole blood for the detection of cell-mediated immunity against varicella-zoster virus. Journal of Immunological Methods 277, 1725.Google Scholar
Talebi, A and Mulcahy, G (1995) Correlation between immune responses and oocyst production in chickens monospecifically infected with Eimeria maxima. Avian Pathology 24, 485495.Google Scholar
Trout, JM and Lillehoj, HS (1996) T lymphocyte roles during Eimeria acervulina and Eimeria tenella infections. Veterinary Immunology and Immunopathology 53, 163172.Google Scholar
Waldenstedt, L, Elwinger, K, Lunden, A, Thebo, P, Bedford, MR and Uggla, A (2000) Intestinal digesta viscosity decreases during coccidial infection in broilers. British Poultry Science 41, 459464.Google Scholar
Wattrang, E, Dalgaard, TS, Norup, LR, Kjaerup, RB, Lunden, A and Juul-Madsen, HR (2015) CD107a as a marker of activation in chicken cytotoxic T cells. Journal of Immunological Methods 419, 3547.Google Scholar
Wattrang, E, Magnusson, SE, Näslund, K, Thebo, P, Hagström, Å, Smith, AL and Lundén, A (2016a) Expression of perforin, granzyme A and Fas ligand mRNA in caecal tissues upon Eimeria tenella infection of naïve and immune chickens. Parasite Immunology 38, 419430.Google Scholar
Wattrang, E, Thebo, P, Lunden, A and Dalgaard, TS (2016b) Monitoring of local CD8b-expressing cell populations during Eimeria tenella infection of naive and immune chickens. Parasite Immunology 38, 453467.Google Scholar
Williams, RB (2001) Quantification of the crowding effect during infections with the seven Eimeria species of the domesticated fowl: its importance for experimental designs and the production of oocyst stocks. International Journal for Parasitology 31, 10561069.Google Scholar
Witcombe, DM and Smith, NC (2014) Strategies for anti-coccidial prophylaxis. Parasitology 141, 13791389.Google Scholar