Abstract
BACKGROUND: The goal of our study was to examine spontaneous and stimulated apoptosis of peripheral blood MNC from allergic patients, sensitized to Der p I antigen as compared to cells from non-atopic subjects. Furthermore we aimed to investigate which populations of mononuclear cells (lymphocytes, monocytes) undergo the apoptosis and to determine relations between apoptosis and serum levels of sFas/APO-1, ICE/caspase-1 or TNF-alpha. METHODS: The study included 17 patients with perennial, allergic asthma and/or allergic rhinitis [6 male and 11 female; mean age 29,5 years; (range 15-49)]. Apoptosis was assessed by fluorescence technique and confirmed by flow-cytometric method and DNA ladder. Serum levels of sFas, ICE/caspase-1 or TNF-alpha were determined by immunoassays (ELISA). RESULTS: Apoptotic index of unfractionated mononuclear cells (MNC) and lymphocytes (but not monocytes) were significantly higher in allergic patients as compared to non-allergic subjects after 48 and 72 hours of culture (p<0.05). Incubation of cells with ConA (10 microg/ml) resulted in a significant increase in the proportion of apoptotic cells in all populations once the apoptotic index for MNC and lymphocytes (but not monocytes) was again significantly higher in allergic as compared to non-allergic subjects after 24, 48 and 72 hour of culture. In allergic patients, mean serum sFas level, was significantly lower then in non-allergic group (mean value 624.8 pg/ml +/- 25.67 versus 802.0 pg/ml +/- 31.91; p = 0.003) and in both groups sFas level correlated inversely with apoptosis of MNC. The mean ICE/caspase-1 concentration was significantly higher in sera of allergic patients as compared to non-allergic group (mean value 27.71 pg/ml +/- 3.79 vs 23.54 pg/ml respectively; p<0.01). ICE/caspase-1 levels in allergic patients correlated with apoptotic index of mononuclear cells (r = 0.57; p<0.001). CONCLUSIONS: An increased spontaneous and mitogen-induced apoptosis of MNC from peripheral blood of atopic patients as well as different serum levels of sFas and ICE/caspase-1 correlating with apoptosis, suggest different regulation of apoptotic process in peripheral blood mononuclear cells of patients with allergic asthma and/or rhinitis.
Full Text
The Full Text of this article is available as a PDF (317.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alam R. Chemokines in allergic inflammation. J Allergy Clin Immunol. 1997 Mar;99(3):273–277. doi: 10.1016/s0091-6749(97)70042-3. [DOI] [PubMed] [Google Scholar]
- Bøyum A. Separation of lymphocytes, granulocytes, and monocytes from human blood using iodinated density gradient media. Methods Enzymol. 1984;108:88–102. doi: 10.1016/s0076-6879(84)08076-9. [DOI] [PubMed] [Google Scholar]
- Cheng J., Zhou T., Liu C., Shapiro J. P., Brauer M. J., Kiefer M. C., Barr P. J., Mountz J. D. Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule. Science. 1994 Mar 25;263(5154):1759–1762. doi: 10.1126/science.7510905. [DOI] [PubMed] [Google Scholar]
- Chicheportiche Y., Bourdon P. R., Xu H., Hsu Y. M., Scott H., Hession C., Garcia I., Browning J. L. TWEAK, a new secreted ligand in the tumor necrosis factor family that weakly induces apoptosis. J Biol Chem. 1997 Dec 19;272(51):32401–32410. doi: 10.1074/jbc.272.51.32401. [DOI] [PubMed] [Google Scholar]
- Cohen G. M. Caspases: the executioners of apoptosis. Biochem J. 1997 Aug 15;326(Pt 1):1–16. doi: 10.1042/bj3260001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cormican L., O'Sullivan S., Burke C. M., Poulter L. W. IFN-gamma but not IL-4 T cells of the asthmatic bronchial wall show increased incidence of apoptosis. Clin Exp Allergy. 2001 May;31(5):731–739. doi: 10.1046/j.1365-2222.2001.01099.x. [DOI] [PubMed] [Google Scholar]
- Grzegorczyk J., Majkowska-Wojciechowska B., Kowalski M. L. The release of eosinophil chemotactic activity and eosinophil chemokinesis inhibitory activity by mononuclear cells from atopic asthmatic and non-atopic subjects. Mediators Inflamm. 2000;9(1):7–13. doi: 10.1080/09629350050024320. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Göke R., Göke A., Göke B., Chen Y. Regulation of TRAIL-induced apoptosis by transcription factors. Cell Immunol. 2000 May 1;201(2):77–82. doi: 10.1006/cimm.2000.1650. [DOI] [PubMed] [Google Scholar]
- Hansen G., Berry G., DeKruyff R. H., Umetsu D. T. Allergen-specific Th1 cells fail to counterbalance Th2 cell-induced airway hyperreactivity but cause severe airway inflammation. J Clin Invest. 1999 Jan;103(2):175–183. doi: 10.1172/JCI5155. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herrmann M., Lorenz H. M., Voll R., Grünke M., Woith W., Kalden J. R. A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res. 1994 Dec 11;22(24):5506–5507. doi: 10.1093/nar/22.24.5506. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jayaraman S., Castro M., O'Sullivan M., Bragdon M. J., Holtzman M. J. Resistance to Fas-mediated T cell apoptosis in asthma. J Immunol. 1999 Feb 1;162(3):1717–1722. [PubMed] [Google Scholar]
- Kato M., Nozaki Y., Yoshimoto T., Tamada Y., Kageyama M., Yamashita T., Kurimoto F., Nakashima I. Different serum soluble Fas levels in patients with allergic rhinitis and bronchial asthma. Allergy. 1999 Dec;54(12):1299–1302. doi: 10.1034/j.1398-9995.1999.00303.x. [DOI] [PubMed] [Google Scholar]
- Krug N., Tschernig T., Holgate S., Pabst R. How do lymphocytes get into the asthmatic airways? Lymphocyte traffic into and within the lung in asthma. Clin Exp Allergy. 1998 Jan;28(1):10–18. doi: 10.1046/j.1365-2222.1998.00197.x. [DOI] [PubMed] [Google Scholar]
- Mahmudi-Azer S., Velazquez J. R., Lacy P., Denburg J. A., Moqbel R. Immunofluorescence analysis of cytokine and granule protein expression during eosinophil maturation from cord blood-derived CD34 progenitors. J Allergy Clin Immunol. 2000 Jun;105(6 Pt 1):1178–1184. doi: 10.1067/mai.2000.106930. [DOI] [PubMed] [Google Scholar]
- Majno G., Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol. 1995 Jan;146(1):3–15. [PMC free article] [PubMed] [Google Scholar]
- Marsters S. A., Sheridan J. P., Pitti R. M., Brush J., Goddard A., Ashkenazi A. Identification of a ligand for the death-domain-containing receptor Apo3. Curr Biol. 1998 Apr 23;8(9):525–528. doi: 10.1016/s0960-9822(98)70204-0. [DOI] [PubMed] [Google Scholar]
- Marsters S. A., Sheridan J. P., Pitti R. M., Brush J., Goddard A., Ashkenazi A. Identification of a ligand for the death-domain-containing receptor Apo3. Curr Biol. 1998 Apr 23;8(9):525–528. doi: 10.1016/s0960-9822(98)70204-0. [DOI] [PubMed] [Google Scholar]
- McGahon A. J., Martin S. J., Bissonnette R. P., Mahboubi A., Shi Y., Mogil R. J., Nishioka W. K., Green D. R. The end of the (cell) line: methods for the study of apoptosis in vitro. Methods Cell Biol. 1995;46:153–185. doi: 10.1016/s0091-679x(08)61929-9. [DOI] [PubMed] [Google Scholar]
- Monteseirín J., Guardia P., Delgado J., Llamas E., Palma J., Conde A., Conde J. Peripheral blood T lymphocytes in seasonal bronchial asthma. Allergy. 1995 Feb;50(2):152–156. doi: 10.1111/j.1398-9995.1995.tb05072.x. [DOI] [PubMed] [Google Scholar]
- Movérare R., Elfman L., Stålenheim G., Björnsson E. Study of the Th1/Th2 balance, including IL-10 production, in cultures of peripheral blood mononuclear cells from birch-pollen-allergic patients. Allergy. 2000 Feb;55(2):171–175. doi: 10.1034/j.1398-9995.2000.00244.x. [DOI] [PubMed] [Google Scholar]
- Nuñez G., Benedict M. A., Hu Y., Inohara N. Caspases: the proteases of the apoptotic pathway. Oncogene. 1998 Dec 24;17(25):3237–3245. doi: 10.1038/sj.onc.1202581. [DOI] [PubMed] [Google Scholar]
- Owen-Schaub L. B., Angelo L. S., Radinsky R., Ware C. F., Gesner T. G., Bartos D. P. Soluble Fas/APO-1 in tumor cells: a potential regulator of apoptosis? Cancer Lett. 1995 Jul 20;94(1):1–8. doi: 10.1016/0304-3835(95)03834-j. [DOI] [PubMed] [Google Scholar]
- Spinozzi F., Fizzotti M., Agea E., Piattoni S., Droetto S., Russano A., Forenza N., Bassotti G., Grignani F., Bertotto A. Defective expression of Fas messenger RNA and Fas receptor on pulmonary T cells from patients with asthma. Ann Intern Med. 1998 Mar 1;128(5):363–369. doi: 10.7326/0003-4819-128-5-199803010-00004. [DOI] [PubMed] [Google Scholar]
- Thueson D. O., Speck L. S., Lett-Brown M. A., Grant J. A. Histamine-releasing activity (HRA). I. Production by mitogen- or antigen-stimulated human mononuclear cells. J Immunol. 1979 Aug;123(2):626–632. [PubMed] [Google Scholar]
- Treves A. J., Yagoda D., Haimovitz A., Ramu N., Rachmilewitz D., Fuks Z. The isolation and purification of human peripheral blood monocytes in cell suspension. J Immunol Methods. 1980;39(1-2):71–80. doi: 10.1016/0022-1759(80)90295-1. [DOI] [PubMed] [Google Scholar]
- Vermes I., Haanen C., Steffens-Nakken H., Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995 Jul 17;184(1):39–51. doi: 10.1016/0022-1759(95)00072-i. [DOI] [PubMed] [Google Scholar]
- Vignola A. M., Chanez P., Chiappara G., Siena L., Merendino A., Reina C., Gagliardo R., Profita M., Bousquet J., Bonsignore G. Evaluation of apoptosis of eosinophils, macrophages, and T lymphocytes in mucosal biopsy specimens of patients with asthma and chronic bronchitis. J Allergy Clin Immunol. 1999 Apr;103(4):563–573. doi: 10.1016/s0091-6749(99)70225-3. [DOI] [PubMed] [Google Scholar]
- Vignola A. M., Chiappara G., Gagliardo R., Gjomarkaj M., Merendino A., Siena L., Bousquet J., Bonsignore G. Apoptosis and airway inflammation in asthma. Apoptosis. 2000 Nov;5(5):473–485. doi: 10.1023/a:1009661406440. [DOI] [PubMed] [Google Scholar]
- Walsh G. M. Mechanisms of human eosinophil survival and apoptosis. Clin Exp Allergy. 1997 May;27(5):482–487. [PubMed] [Google Scholar]
- Wang J., Palmer K., Lotvall J., Milan S., Lei X. F., Matthaei K. I., Gauldie J., Inman M. D., Jordana M., Xing Z. Circulating, but not local lung, IL-5 is required for the development of antigen-induced airways eosinophilia. J Clin Invest. 1998 Sep 15;102(6):1132–1141. doi: 10.1172/JCI2686. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Neerven R. J. The role of allergen-specific T cells in the allergic immune response: relevance to allergy vaccination. Allergy. 1999 Jun;54(6):552–561. doi: 10.1034/j.1398-9995.1999.t01-1-00092.x. [DOI] [PubMed] [Google Scholar]
