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Erschienen in: Immunologic Research 2/2018

14.02.2018 | Original Article

Chronic graft-specific cell-mediated immune response toward candidate xenogeneic biomaterial

verfasst von: Katherine V. Gates, Leigh G. Griffiths

Erschienen in: Immunologic Research | Ausgabe 2/2018

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Abstract

Despite rabbits becoming an increasingly popular animal model, a flow cytometry panel that combines T cell markers (CD3, CD4, CD8, CD25, FOXP3) with a method for monitoring proliferation is lacking in this species. It has been shown that the rabbit model can be used to identify xenoantigens within bovine pericardium (BP), a common biological heart valve replacement material; however, these methods rely on monitoring the humoral immune response. The development of a rabbit T cell proliferation assay has utility in monitoring graft-specific cell-mediated immune responses toward bovine pericardium. Isolation and culture conditions were optimized to avoid cell death, red blood cell contamination, and non-specific proliferation. Effect of cell culture and stimulation on distribution and intensity of T cell markers was analyzed and compared between cells isolated from naïve and BP-immunized rabbits. Submaximal levels (0.25 μg/mL) of concavalin A were used to stimulate proliferation toward BP extract, with resultant proliferation compared between naïve and BP-immunized rabbits. Density stratification followed by ammonium potassium chloride (ACK) lysis yielded the greatest number of viable peripheral blood mononuclear cells with the least amount of erythrocyte contamination. Flat-bottomed plates were necessary to reduce non-specific proliferation in culture. T cells responded appropriately to maximal mitogenic stimulation (5 μg/mL concavalin A). Interestingly, immunization increased the intensity of FOXP3 in T regulatory cells compared to cells from naïve animals. With addition of submaximal levels of concavalin A, T cells from immunized rabbits proliferated in response to BP protein extract, while cells from naïve rabbits did not. In immunized rabbits, not only did more CD4+ T cells proliferate in response to BP re-stimulation, but the intensity of CD25 was increased indicating cell activation. This research provides a functional cell-mediated screening assay for assessment of BP-based biomaterials in rabbits, overcoming the limitations of previous humoral immune system-based assessments of biomaterial antigenicity in this important experimental animal species.
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Literatur
14.
19.
Zurück zum Zitat Zaidi AH, et al. Preliminary experience with porcine intestinal submucosa (CorMatrix) for valve reconstruction in congenital heart disease: histologic evaluation of explanted valves. J Thorac Cardiovasc Surg. 2014;148(5):2216–4. 2225 e1CrossRefPubMed Zaidi AH, et al. Preliminary experience with porcine intestinal submucosa (CorMatrix) for valve reconstruction in congenital heart disease: histologic evaluation of explanted valves. J Thorac Cardiovasc Surg. 2014;148(5):2216–4. 2225 e1CrossRefPubMed
21.
Zurück zum Zitat Dahm M, Lyman WD, Schwell AB, Factor SM, Frater RW. Immunogenicity of glutaraldehyde-tanned bovine pericardium. J Thorac Cardiovasc Surg. 1990;99(6):1082–90.PubMed Dahm M, Lyman WD, Schwell AB, Factor SM, Frater RW. Immunogenicity of glutaraldehyde-tanned bovine pericardium. J Thorac Cardiovasc Surg. 1990;99(6):1082–90.PubMed
24.
Zurück zum Zitat National Research Council. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press; 2011. p. 1–246. National Research Council. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press; 2011. p. 1–246.
27.
Zurück zum Zitat Lin YZ, Deng XL, Shen N, Lü XL, Zhao LP, Kong XG, et al. Development of a CFSE-based flow cytometry for evaluating EIAV-stimulated proliferation of T lymphocytes. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008;24(11):1044–7.PubMed Lin YZ, Deng XL, Shen N, Lü XL, Zhao LP, Kong XG, et al. Development of a CFSE-based flow cytometry for evaluating EIAV-stimulated proliferation of T lymphocytes. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008;24(11):1044–7.PubMed
28.
Zurück zum Zitat Andersen MN, et al. Elimination of Erroneous Results in Flow Cytometry Caused by Antibody Binding to Fc Receptors on Human Monocytes and Macrophages. Cytometry Part A. 2016;89a(11):1001–9.CrossRef Andersen MN, et al. Elimination of Erroneous Results in Flow Cytometry Caused by Antibody Binding to Fc Receptors on Human Monocytes and Macrophages. Cytometry Part A. 2016;89a(11):1001–9.CrossRef
30.
Zurück zum Zitat de Baetselier P, Vaeck M, de Smet W, Ron Y. Antigen-induced proliferation assay for rabbit T lymphocytes. I Characteristics of the response. Immunology. 1980;41(4):997–1003.PubMedPubMedCentral de Baetselier P, Vaeck M, de Smet W, Ron Y. Antigen-induced proliferation assay for rabbit T lymphocytes. I Characteristics of the response. Immunology. 1980;41(4):997–1003.PubMedPubMedCentral
32.
Zurück zum Zitat Mendez-David I, et al. A method for biomarker measurements in peripheral blood mononuclear cells isolated from anxious and depressed mice: beta-arrestin 1 protein levels in depression and treatment. Front Pharmacol. 2013;4:124.CrossRefPubMedPubMedCentral Mendez-David I, et al. A method for biomarker measurements in peripheral blood mononuclear cells isolated from anxious and depressed mice: beta-arrestin 1 protein levels in depression and treatment. Front Pharmacol. 2013;4:124.CrossRefPubMedPubMedCentral
35.
Zurück zum Zitat Sloane ED, Muscoplat CC, Kaneene JM, Klausner DJ, Thoen CO, Johnson DW. In vitro stimulation of bovine peripheral blood lymphocytes: comparison of round- and flat-bottom microtiter plates for detection of tuberculin hypersensitivity. J Clin Microbiol. 1978;7(2):172–5.PubMedPubMedCentral Sloane ED, Muscoplat CC, Kaneene JM, Klausner DJ, Thoen CO, Johnson DW. In vitro stimulation of bovine peripheral blood lymphocytes: comparison of round- and flat-bottom microtiter plates for detection of tuberculin hypersensitivity. J Clin Microbiol. 1978;7(2):172–5.PubMedPubMedCentral
36.
37.
Zurück zum Zitat Uppal SS, Verma S, Dhot PS. Normal values of CD4 and CD8 lymphocyte subsets in healthy indian adults and the effects of sex, age, ethnicity, and smoking. Cytometry B Clin Cytom. 2003;52(1):32–6.CrossRefPubMed Uppal SS, Verma S, Dhot PS. Normal values of CD4 and CD8 lymphocyte subsets in healthy indian adults and the effects of sex, age, ethnicity, and smoking. Cytometry B Clin Cytom. 2003;52(1):32–6.CrossRefPubMed
39.
Zurück zum Zitat Rabinovitch PS, et al. Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry. J Immunol. 1986;137(3):952–61.PubMed Rabinovitch PS, et al. Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry. J Immunol. 1986;137(3):952–61.PubMed
43.
Zurück zum Zitat Sultani M, et al. Anti-inflammatory cytokines: important immunoregulatory factors contributing to chemotherapy-induced gastrointestinal mucositis. Chemother Res Pract. 2012;2012:490804.PubMedPubMedCentral Sultani M, et al. Anti-inflammatory cytokines: important immunoregulatory factors contributing to chemotherapy-induced gastrointestinal mucositis. Chemother Res Pract. 2012;2012:490804.PubMedPubMedCentral
46.
Zurück zum Zitat Croft M, Bradley LM, Swain SL. Naive versus memory CD4 T cell response to antigen. Memory cells are less dependent on accessory cell costimulation and can respond to many antigen-presenting cell types including resting B cells. J Immunol. 1994;152(6):2675–85.PubMed Croft M, Bradley LM, Swain SL. Naive versus memory CD4 T cell response to antigen. Memory cells are less dependent on accessory cell costimulation and can respond to many antigen-presenting cell types including resting B cells. J Immunol. 1994;152(6):2675–85.PubMed
50.
Zurück zum Zitat Ciabattini A, Pettini E, Medaglini D. CD4(+) T cell priming as biomarker to study immune response to preventive vaccines. Front Immunol. 2013;4:421.CrossRefPubMedPubMedCentral Ciabattini A, Pettini E, Medaglini D. CD4(+) T cell priming as biomarker to study immune response to preventive vaccines. Front Immunol. 2013;4:421.CrossRefPubMedPubMedCentral
Metadaten
Titel
Chronic graft-specific cell-mediated immune response toward candidate xenogeneic biomaterial
verfasst von
Katherine V. Gates
Leigh G. Griffiths
Publikationsdatum
14.02.2018
Verlag
Springer US
Erschienen in
Immunologic Research / Ausgabe 2/2018
Print ISSN: 0257-277X
Elektronische ISSN: 1559-0755
DOI
https://doi.org/10.1007/s12026-018-8985-8

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