Flow Cytometric analysis of Th1 and Th2 cytokines in PBMCs as a parameter of immunological dysfunction in patients of Superficial Transitional cell carcinoma of bladder

Springer Science and Business Media LLC - Tập 55 - Trang 734-743 - 2005
A. Agarwal1, S. Verma1, U. Burra1, N. S. Murthy1, N. K. Mohanty2, S. Saxena1
1Institute of Pathology, ICMR, Safdarjung Hospital Campus, New Delhi, India
2Department of Urology, Safdarjung Hospital Campus, New Delhi, India

Tóm tắt

Transitional cell carcinoma (TCC) is the commonest cancer of the bladder. Although majority of TCC can be diagnosed at an early stage and removed easily by transurethral resection of tumor (TURT), the management of this carcinoma is complicated due to frequent recurrences usually within 6 months to one-year period. An imbalance between the Th1 and Th2 immune responses has been attributed to immune dysregulation in various malignancies. The present study aims to evaluate the Th1 and Th2 balance in Peripheral Blood Mononuclear Cells of 41 TCC patients (20 recurrent and 21 non-recurrent) using flow cytometry. It also further assesses immunological and cellular factors influencing the anti-neoplastic activity of the TCC patients and in 21 normal healthy subjects in terms of their cytokine expression and various cell surface markers. The findings of the study revealed that the cell surface markers CD3+, CD4+ and CD8+ along with NK cells were found to be significantly lower in patients than healthy controls (p<0.01). The mean percent expression of CD4+ was significantly lower in patients showing recurrence (23.9±9.84) as compared to patients with non-recurrence (31.1±12.27). The percentage of CD4+T-cells (mean±SD) producing IFN-γ, IL-2 and TNF-α were statistically significantly reduced in patients (19.1±4.94, 52.3±20.86 and 12.8±4.49) as compared to healthy controls (23.3±3.67, 67.5±12.0 and 17.6±5.96 respectively), (p<0.01, 0.018, 0.001). On the contrary, the mean levels of IL-4, IL-6 and IL-10 in patients (63.8±17.01, 60.4±14.79 and 65.7±14.84 respectively) were significantly higher as compared to healthy controls (24.4±8.77, 26.5±5.28 and 20.6±3.81 respectively), (p<0.001). No statistically significant difference was observed in the cytokine expression between patients showing recurrence and non-recurrence. Patients with bladder cancer seem to develop a Th2 dominant status with a deficient type1 immune response. The lymphocyte evaluation along with cytokine measurement can provide a sensitive and valuable tool for evaluating the function of cell-mediated immunity in these patients and can also find application in therapeutic monitoring of bladder cancer patients as new targets for immunotherapy.

Tài liệu tham khảo

Agarwal A, Rani M, Saha GK, Valarmathi TM, Bahadur S, Mohanti BK, Das SN (2003) Disregulated expression of the Th2 cytokine gene in patients with intraoral squamous cell carcinoma. Immunol Invest 32:17–30

Assenmacher M, Schmitz J, Radbruch A (1994) Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-gamma and in interleukin-4-expressing cells. Eur J Immunol 24:1097–101

Banat GA, Christ O, Cochlovius B, Pralle HB, Zoller M (2001) Tumour-induced suppression of immune response and its correction. Cancer Immunol Immunother 49:573–86

Boyum A (1968) Isolation of leucocytes from human blood. Further observations. Methylcellulose, dextran, and ficoll as erythrocyteaggregating agents. Scand J Clin Lab Invest 97:31–50

Chen LK, Tourvieille B, Burns GF, Bach FH, Mathieu-Mahul D, Sasportes M, Bensussan A (1986) Interferon: a cytotoxic T lymphocyte differentiation signal. Eur J Immunol 16:767

Cher DJ, Mosmann TR (1987) Two types of murine helper T cell clone. II. Delayed-type hypersensitivity is mediated by TH1 clones. J Immunol 138:3688–94

Elson LH, Nutman TB, Metcalfe DD, Prussin C (1995) Flow cytometric analysis for cytokine production identifies T helper 1, T helper 2, and T helper 0 cells within the human CD4+CD27- lymphocyte subpopulation. J Immunol 154:4294–301

Erard F, Wild MT, Garcia-Sanz JA, Le Gros G (1993) Switch of CD8 T cells to noncytolytic CD8-CD4-cells that make TH2 cytokines and help B cells. Science 260:1802–5

Gajewski TF, Fitch FW (1988) Anti-proliferative effect of IFN-gamma in immune regulation. I. IFN-gamma inhibits the proliferation of Th2 but not Th1 murine helper T lymphocyte clones. J Immunol 140:4245–52

Goto S, Sato M, Kaneko R, Itoh M, Sato S, Takeuchi S (1999) Analysis of Th1 and Th2 cytokine production by peripheral blood mononuclear cells as a parameter of immunological dysfunction in advanced cancer patients. Cancer Immunol Immunother 48:435–42

Halak BK, Maguire HC Jr, Lattime EC (1999) Tumor-induced interleukin-10 inhibits type 1 immune responses directed at a tumor antigen as well as a non-tumor antigen present at the tumor site. Cancer Res 59:911–7

Hernandez-Pando R, Rook GA (1994) The role of TNF-alpha in T-cell-mediated inflammation depends on the Th1/Th2 cytokine balance. Immunology 82:591–5

Jackson AM, Ivshina AV, Senko O, Kuznetsova A, Sundan A, O’Donnell MA, Clinton S, Alexandroff AB, Selby PJ, James K, Kuznetsov VA (1998) Prognosis of intravesical bacillus Calmette-Guerin therapy for superficial bladder cancer by immunological urinary measurements:statistically weighted syndrome analysis. J Urol 159:1054–63

Jung T, Lack G, Schauer U, Uberuck W, Renz H, Gelfand EW, Rieger CH (1995) Decreased frequency of interferon-gamma- and interleukin-2-producing cells in patients with atopic diseases measured at the single cell level. J Allergy Clin Immunol 96:515–27

Jung T, Schauer U, Heusser C, Neumann C, Rieger C (1993) Detection of intracellular cytokines by flow cytometry. J Immunol Methods 159:197–207

Kaempfer R, Gerez L, Farbstein H, Madar L, Hirschman O, Nussinovich R, Shapiro A (1996) Prediction of response to treatment in superficial bladder carcinoma through pattern of interleukin-2 gene expression. J Clin Oncol 14:1778–86

Kavanaugh DY, Carbone DP (1996) Immunologic dysfunction in cancer. Hematol Oncol Clin North Am 10:927–51

Klausner RD, Donaldson JG, Lippincott-Schwartz J (1992) Brefeldin A: insights into the control of membrane traffic and organelle structure. J Cell Biol 116:1071–80

Klein LT, Miller MI, Ikeguchi E, Buttyan R, Connor JP, Katz A, Raffo AJ and Olsson C (1996) Anti-fas antibody mediated apoptosis in bladder tumor cells: a potential intravesical therapeutic agent. Proc Annu Meet Amer Assoc cancer Res 37:A103

Kurisu H, Matsuyama H, Ohmoto Y, Shimabukuro T, Naito K (1994) Cytokine-mediated antitumor effect of bacillus Calmette-Guerin on tumor cells in vitro. Cancer Immunol Immunother 39:249–53

Lattime EC, Gomella LG, McCue PA (1992) Murine bladder carcinoma cells present antigen to BCG-specific CD4+ T-cells. Cancer Res 52:4286–90

Lee KY, Goedegebuure PS, Linehan DC, Eberlein TJ (1995) Immunoregulatory effects of CD4+ T helper subsets in human melanoma. Surgery 117:365–72

Lewis, C.E (1991) Detecting cytokine production at the single-cell level. Cytokine 3:184–8

Lord EM, Frelinger JG (1998) Tumor immunotherapy: cytokines and antigen presentation. Cancer Immunol Immunother 46:75–81

Luo Y, Szilvasi A, Chen X, DeWolf WC, O’Donnell MA (1996) A novel method for monitoring Mycobacterium bovis BCG trafficking with recombinant BCG expressing green fluorescent protein. Clin Diagn Lab Immunol 3:761–8

Maeda H, Kuwahara H, Ichimura Y, Ohtsuki M, Kurakata S, Shiraishi A (1995) TGF-beta enhances macrophage ability to produce IL-10 in normal and tumor-bearing mice. J Immunol 155:4926–32

Manson LA (1991) Does antibody-dependent epitope masking permit progressive tumor growth in the face of cell-mediated cytotoxicity? Immunol Today 12:352–5

Meyaard L, Hovenkamp E, Keet IP, Hooibrink B, de Jong IH, Otto SA, Miedema F (1996) Single cell analysis of IL-4 and IFN-gamma production by T cells from HIV-infected individuals: decreased IFN-gamma in the presence of preserved IL-4 production. J Immunol 157:2712–18

Mosmann TR, Coffman RL (1989) TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties. Annu Rev Immunol 7:145–73

O’Donnell MA, Luo Y, Chen X, Szilvasi A, Hunter SE, Clinton SK (1999) Role of IL-12 in the induction and potentiation of IFN-gamma in response to bacillus Calmette-Guerin. J Immunol 163:4246–52

Old LJ (1987) Tumour necrosis factor. Polypeptide mediator network. Nature 326:330–1

Olsson CA, Rao CN, Menzoian JO, Byrd WE (1972) Immunologic unreactivity in bladder cancerpatients. J Urol107:607–9

Openshaw P, Murphy EE, Hosken NA, Maino V, Davis K, Murphy K, O’Garra (1995) A Heterogeneity of intracellular cytokine synthesis at the single-cell level in polarized T helper 1 and T helper 2 populations. J Exp Med 182:1357–67

Paul WE, Seder RA (1994) Lymphocyte responses and cytokines. Cell 2876:241–51

Pellegrini P, Berghella AM, Del Beato T, Cicia S, Adorno D, Casciani CU (1996) Disregulation in TH1 and TH2 subsets of CD4+ T cells in peripheral blood of colorectal cancer patients and involvement in cancer establishment and progression. Cancer Immunol Immunother 42:1–8

Picker LJ, Singh MK, Zdraveski Z, Treer JR, Waldrop SL, Bergstresser PR, Maino VC (1995) Direct demonstration of cytokine synthesis heterogeneity among human memory/effector T cells by flow cytometry. Blood 86:1408–19

Powrie F, Menon S, Coffman RL (1993) Interleukin-4 and interleukin-10 synergize to inhibit cell-mediated immunity in vivo. Eur J Immunol 23:3043–9

Prescott S, James K, Hargreave TB, Chisholm GD, Smyth JF (1992) Intravesical Evans strain BCG therapy: quantitative immunohistochemical analysis of the immune response within the bladder wall. J Urol 147:1636–42

Pryor K, Stricker P, Russell P, Golovsky D, Penny R (1995) Antiproliferative effects of bacillus Calmette-Guerin and interferon alpha 2b on human bladder cancer cells in vitro. Cancer Immunol Immunother 41:309–16

Pyrah LN, Raper FP, Thomas, GM (1964) Report of a follow-up of papillary tumours of the bladder. Br J Urol 36:14–25

Scott, P (1991) IFN-gamma modulates the early development of Th1 and Th2 responses in a murine model of cutaneous leishmaniasis. J Immunol 147:3149–55

Seder RA, Paul WE (1994) Acquisition of lymphokine-producing phenotype by CD4+ T cells. Annu Rev Immunol 12:635–73

Silverberg E, Lubere JA (1988) Cancer statstics 1988. CA Cancer J clin 38:5–22

Stevens TL, Bossie A, Sanders VM, Fernandez-Botran R, Coffman RL, Mosmann TR, Vitetta ES (1988) Regulation of antibody isotype secretion by subsets of antigen-specific helper T cells. Nature 334:255–8

Swain SL, Weinberg AD, English M, Huston G (1990) IL-4 directs the development of Th2-like helper effectors. J Immunol 145:3796–806

Torti FM, Lum BL (1986) Superficial carcinoma of bladder: natural history and the role of interferons. Semin oncol 13:57–60

Tripp CS, Wolf SF, Unanue ER (1993) Interleukin 12 and tumor necrosis factor alpha are costimulators of interferon gamma production by natural killer cells in severe combined immunodeficiency mice with listeriosis, and interleukin 10 is a physiologic antagonist. Proc Natl Acad Sci U S A 90:3725–9

Zaks TZ, Chappell DB, Rosenberg SA, Restifo NP (1999) Fas-mediated suicide of tumor-reactive T cells following activation by specific tumor: selective rescue by caspase inhibition. J Immunol 162:3273–9

Zielinski CC, Mueller C, Tyl E, Tichatschek E, Kubista E, Spona J (1990) Impaired production of tumor necrosis factor in breast cancer. Cancer 66:1944–8

Zlotta AR, Drowart A, Huygen K, De Bruyn J, Shekarsarai H, Decock M, Pirson M, Jurion F, Palfliet K, Denis O, Mascart F, Simon J, Schulman CC, Van Vooren JP (1997) Humoral response against heat shock proteins and other mycobacterial antigens after intravesical treatment with bacille Calmette-Guerin (BCG) in patients with superficial bladder cancer. Clin Exp Immunol 109:157–65

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Springer Science and Business Media LLC

Tập 55

734-743

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