Activation and migration of CD4 + lymphocytes induced by different foreign bodies

Transcription

1 Activation and migration of CD4 + lymphocytes induced by different foreign bodies Бобрышев Д. В., F. Entschladen 2, В.Д. Гримм, W.-D. Grimm 1 Ставрополь State Medical University, Россия, University of Witten/Herdecke, Germany Institute of Immunology 1 Department of Periodontology 2

2 Introduction Methoden Ergebnisse Diskussion Immune System Extracellular pathogens such as bacteria (stars) and intracellular pathogens such as viruses (hexagons) enter the body T helper cells are essential for the activation of B cells (6), and provide cytotoxic T cells with interleukin-2

3 Monocytes and Periodontitis The secretion of PGE 2, IL-1β, IL-6, and TNFα by adherent mononuclear cells from early onset periodontitis patients. Shapira L, Soskolne WA, Sela MN, Offenbacher S, Barak V. J Periodontol Feb;65(2):

4 T Lymphocytes and Periodontitis Destructive periodontitis lesions are determined by the nature of the lymphocytic response. Gemmell E, Yamazaki K, Seymour GJ. Crit Rev Oral Biol Med. 2002;13(1):17-34.

5 Monocytes and T Lymphocytes and Periodontitis Antigen-presenting cells in human periodontal disease tissues. Gemmell E, Carter CL, Hart DN, Drysdale KE, Seymour GJ. Oral Microbiol Immunol Dec;17(6): Functional studies are required to determine the roles of different APC subpopulations in periodontal disease.

6 3-D Lymphocyte Migration is a sensible indicator for assessing biocompatibility and immunological reactions as to periodontal regenerative procedures. Influence of Two Dental Alloys on Primary Human CD8+-Lymphocyte- Migration Grimm WD, Entschladen F, Gassmann G, Zaenker KS J Dent Res Spec Iss: 156 In vitro 3D-Lymphocyte Migration as a Periodontal Healing Model Gassmann G, Entschladen F, Zaenker KS, Grimm WD J Dent Res 2001 Mar 80 Spec Iss: 769 Activation and Migration of CD4+ Lymphocytes Induced by Different Foreign Bodies v Möller S, Entschladen F, Gassmann G, Zaenker KS, Grimm WD Parodontologie 2003 Nov 14, 4: 432 CD4+ Migration and CD25 Activation Due to EMD Gassmann G, Schwenk B, Entschladen F, Zaenker KS, Grimm WD J Dent Res 2003 June 82 Spec Iss B: 293

7 Goal of the study Can our sequentional monocyte T lymphocyte co-culture migration model serve to provide further insight into the cellular mechanisms that may play a role in the link between periodontitis and foreign bodies?

8 LPS LPS 24 h Inkubation RPMI Supernatant Supernatant Control Supernatant Supernatant CD4+ CD4+ 1 h Migration 10 esults

9 Migration System Video-Camera 37 C 37 C time-lapse

10 Migration Chamber Wax Wall PBS Cover Slip Object Slide Collagen with T-lymphocytes in Supernatant

11 Migration Chamber 10µm

12 Migration Chamber 10µm

13 Cell Tracking 30 cells selected randomly Digitalization One minute steps Mean calculated for whole population

14 Einleitung Methoden Results Diskussion Activation and Migration of T-Lymphocytes induced by dental alloys CD25 Flow-cytometric analysis of the expression of CD25 of T-Lymphocytes after contact with Heralloy or Albabond compared to the expression of CD 25 of the control group. (Gm= 3,13; M1=99%) Gm = 3.17 M1= 99.0% Gm = 3.09 M1= 98.9% Gm = 3.20 M1= 98.8% Gm: 3.24 M1= 99.0% Gm = 3.18 M1= 98.7% Gm = 3.40 M1= 98.8%

15 Migratory activity of T- lymphocytes

16

17 Einleitung Methoden Results Diskussion Activation and Migration of T-Lymphocytes induced by dental alloys CD45RO The antibody bound FITC-Fluorescence of specific antibodies (filled grey) is compared to the expression of CD 45R0 of the control group. (Gm= 6,18; M1=99%) (black line) Gm= 4.60 M1= 99.1% Gm= 5.47 M1= 98.0% Gm= 5.58 M1= 99.7% Gm= 6.11 M1= 99.4% Gm= 6.24 M1= 99.7% Gm= 6.38 M1= 99.7%

18

19

20 Conclusions CD4+ lymphocyte migration is enhanced by monocyte supernatant. Enhancement is more pronounced in the supernatant of the autologous monocytes. LPS in the supernatant leads to a reduction of the enhancement caused by the monocyte supernatant. CD4+ migration is more enhanced in the diseased group than in the healthy control

21 Conclusions II Cytokine/Chemokine profiles of the monocyte supernatants have to be assessed in order to elucidate the pathway of migration activation. Receptor expession (CD25, CXCR, CCR) has to be analyzed on the surface of the lymphocytes.

22 Acknowledgements StGMU Dr. Бобрышев Д. В., Prof. Dr. Щетинин Е.В., Prof. Dr. Сирак С.В., Prof. Dr. В.Д. Гримм Institute of Immunology: Prof. Dr. Kurt S Zaenker PD Dr. Frank Entschladen Internal Medicine Private Pratice Dr. Koch, Witten, Germany Department of Periodontology: Prof. Dr. Wolf-Dieter Grimm, Dr. J. Engl-Schmücker, Dr. A. Krajewski, Dr. B. Schwenk, Dr. Fi. Shihabi, Dr. S. von Möller, Dr. E. Wlachmbei