- Email: [email protected]
Cytokine transcription in T cell subsets from healthy, ReA, and Chlamydia-infected individuals
Cytokines in injlammation
25 June 1997 - Poster presentations
measured by change in UV absorption of diethyldithiocatbamate at the wave length of 282 nm. Reauftaz It has been shown that IgG subfraction obtained at pH-7.2 actively bound the copper ions from CuSQ saline while IgG subfraction obtained at pH-8.6 was found to bind the copper ions with two fold lower efficiency. The study of proliferation of human peripheral blood lymphocytes (PBL) in presence of the fractions compared demonstrated a stimulatory action of IgG subfraction actively bound the copper ions on the in v&o PBL proliferation whereas IgG pH-8.6 was not found to serve as PBL proliferation stimulator. Conclusion: Because of correlation of IgG stimulatory action on human PBL proliferation with a quantity of IgG aggregates, the data obtained may indicate that transition metal ions are involved in IgG aggregation, and thereby in activation of the in vitro human PBL proliferation.
1P.3.06.31 1 The immunological status-surface evaluation through natural and immune antibodies in the ABO blood group system - Preliminary results V. Hafner ‘, C. Turcas I, A. Z&grean ‘, L. Cazacu *. ’ BloodTransfusion Cenfre of Bucharest Romania,*National Institute of Blood Tmnsfusion, Romania The immune response, a genetically determined process, may suffer external influences, such as psychological, social, economkzalaggressions. The present study, was conducted on 600 blood donors (healthy individuals). It tries to evaluate the possible variations of natural and immune antibodies in the ABO system, linked to age and sex, in the today local context. Based on the principles of classical blood serology, the methods we used (titration with treated and untreated cells and Bombail-Girard technique with extemporaneous addition of bromelin), have been adapted to micro plates (“U” bottom), for an easier way to handle. The results confirm, once more, that the capacity of antibody production (secondary response) diminishes with age, even in healthy individuals. To be noticed: an apparent change of the immunological status related to sex, taking into account that women present statistically, in normal conditions, a higher capacity of the immune response (reversed pattern in the present study).
P.3.10
Cytokines in inflammation
P.3.10.01
Cytokine transcription In T cell subsets from healthy, ReA, and CMafnyd/&MHecMJ lndlviduals
H. Beacock-Sharp, J.S.H. Gaston. Department ofMedicine, CamM&?e University Clinical S&w/, Cambridge, UK Introduction: Reactive arthritis (ReA), which develops after infection with intracellular bacteria such as chlamydia, is associated with HlA-B27. However, whether organism-specific HLA-B27-restricted CDB+ T cells are involved in the pathogenesis of ReA is unresolved. The expression of the activation malkers CD69 and HlA-DR on CD8+ Tcells isolated from synovial fluid (SF) suggests an important role. To elucidate futther the role of the CD8+ T cells in the pathogenesis of ReA we have examined the cytokine profile of these cells immediately ax viw, and compared this wtth CD4+ T cells, which can readily be shown to respond to the biggetfng bacteria. Peripheral blood (PB) T cell subsets stimulated with PHA or Chlamydia Wacbomafi~infected macrophages from either healthy controls or indlviduals with an uncomplicated C. trachomatfs infection (GU) were also investigated. Methods: Mononuclear cells were separated from PB and SF. CD4+ and CD8+ T cell subsets were isolated, using a fluorescence activated cell sorter, either immediately ax viw or after stimulation with PHA or C. trachomafisinfected macrophages. mRNA was extracted from these cells and RT-PCR performed to detect mRNA of cytoklnes of interest. Results: Stimulation
IL-2 IL-4
10
30
20
-.--Average tltre level of immune antibodies-ABC
system.
The level of natural anti-A and anti-B antibodies (involved in cellular recognition patterns - onto genetic level) is maintained within limits in every sequence of age, demonstrating that a major impact of today stress factors could not be detected at the present time.
P 3 06 I...
32
1Immune-receptowndocrine
interactions in the pathophysiology of major depression
SergeiMiner&o. Khadrov /n&We of Clinical and Experimental Neurology and PsychiatryLwv Psychiatric Hospital, Ukraine The role of the Adrenergic and Benzodiazepine receptors in development of immunosuppmssion on leukocyte-pyruvatdehydrogenase activity was studied in 60 patients with Primary Major Depressive Episode. Investigation included administration of u-adrenergic “recepto? drug L-DOPA and benzodiazopine “recepto? dtugDlASEPAM before dexamethazone immunosuppression test (DIST). In the cases of Ptfmary Major Depressive Episode in separation with an Anxiety Disorder (30 patients - comparison subjects) after L-DOPA and DIASEPAM administration actfvity of leukocyte-pyruvatde-gydrogenasis increased more than 25%. DIASEPAM of a 10 mg dose had a higher immunosuppressive effect than L-DOPA (0.5 g) on alteration of leukocyle-pymvatdegydmgenase actfvity (more than 5 mmol/lhour, p < 0.05). DIASEPAM immunosuppressive action did not correlate with positive dynamics of T-and-B-cells lymphocytes. From other side, mechanism of L-DOPA actfon on adrenetgic receptors stimulated T-cell receptors and made them hyperactive (produced increasing tangs of T-cell lymphocytes). It means that adrenergic and benzodiazepine receptors are interacting with each other and influencing T-and-B-cells receptors in different ways during immunosuppression. Finally, it is important to study immune-receptor-Endocrine interactions and major depression.
Forum lounges
09:80-18:30/12:8&14:88
IFN-y
c
405
ControlPS
Control PB
GU PB
FleA SF
none
PHA
C. trachomatis
norm
CD4
CD8
CD4
CDS
CD4
CD8
CD4
CDS
+ +
+
+ +
+ +
+ +
+ +
+ +
+ +
+ +
+ +
” ”
+ +
” ”
” +
+
+
”
+
”
+
IL-10
” +
” ” ”
IL-17
+
”
+ = PCR product visualised on agarcse gel by ethidium bromide staining: ” = undetectable
Condusions: PHA stimulation of PB induces transcription of all cytokine genes investigated in both CD4+ and CD8+ T cells. CD4+ T cells from unstimulated ReA SF, and from GU PB stimulated wm the infecting organism, were found to transcribe IFN-y and IL-2. Interestingly, CD8+ T cells isolated from both these cell populations were also found to produce IL-10 and 11-17.Thus, both C. trachomalisstimulated CD8+ T cells and ax viva CDE+Tcells from ReA SF are likely to secrete both pm-inflammatory (e.g. 11-17)and regulatory (e.g. IL-lo) cytokjnes. These results point to a complex role for CD8+ T cells in the pathogenesis of ReA.
1P.3.10.02 1 Blood cells and bacterial inflammation T. Matsumoto. Matsumoto Living Cell Research Laboratory Tokyo,Japan Introduction and Alm: My research on erythrocytes (red blood cell = Rbc) and leucocyte, aims to disclose the essential function of them in bacterial inflammation. Method and Mateflals: The tissue culture method based on MatsumotoHagiya method has been conducted using 7% Dextran-salin solution, 5% CQ incubator, pseudomonas sp., blood cells, and others. Resultst (1) Rbc are classified into at least three groups (top, middle and bottom layers) through the Matsumoto-Hagiya method. (2) When top Rbc are exposed to pseudomonas sp.. Rbc and Rbcderived cells demonstrate rotating or reversive movement on or around pseudomonas sp. (video) which should be interpreted BStheir bacterfo-static influence. (3) Rbcs in the middle layer remain but the multiplication of pseudomonas sp. is suppressed. (photo) (4) Rbcs in the bottom layer are static and pseudomonas sp. very rapidly multiplies. (photo) (5) Leucocytes show amoeboid movement but do not pmve phagocytcsis. (video) Conclusion: The inflammation occurs when immunological balance is bmken by many factors. Such as: immuno-imbalance induced by influenza, hepatitis, excessive fatigue, malnuttftion and many other diseases: General factors: bleeding, ischaemia, trauma etc.: Local factors.
25 June 1997 - Poster presentations
measured by change in UV absorption of diethyldithiocatbamate at the wave length of 282 nm. Reauftaz It has been shown that IgG subfraction obtained at pH-7.2 actively bound the copper ions from CuSQ saline while IgG subfraction obtained at pH-8.6 was found to bind the copper ions with two fold lower efficiency. The study of proliferation of human peripheral blood lymphocytes (PBL) in presence of the fractions compared demonstrated a stimulatory action of IgG subfraction actively bound the copper ions on the in v&o PBL proliferation whereas IgG pH-8.6 was not found to serve as PBL proliferation stimulator. Conclusion: Because of correlation of IgG stimulatory action on human PBL proliferation with a quantity of IgG aggregates, the data obtained may indicate that transition metal ions are involved in IgG aggregation, and thereby in activation of the in vitro human PBL proliferation.
1P.3.06.31 1 The immunological status-surface evaluation through natural and immune antibodies in the ABO blood group system - Preliminary results V. Hafner ‘, C. Turcas I, A. Z&grean ‘, L. Cazacu *. ’ BloodTransfusion Cenfre of Bucharest Romania,*National Institute of Blood Tmnsfusion, Romania The immune response, a genetically determined process, may suffer external influences, such as psychological, social, economkzalaggressions. The present study, was conducted on 600 blood donors (healthy individuals). It tries to evaluate the possible variations of natural and immune antibodies in the ABO system, linked to age and sex, in the today local context. Based on the principles of classical blood serology, the methods we used (titration with treated and untreated cells and Bombail-Girard technique with extemporaneous addition of bromelin), have been adapted to micro plates (“U” bottom), for an easier way to handle. The results confirm, once more, that the capacity of antibody production (secondary response) diminishes with age, even in healthy individuals. To be noticed: an apparent change of the immunological status related to sex, taking into account that women present statistically, in normal conditions, a higher capacity of the immune response (reversed pattern in the present study).
P.3.10
Cytokines in inflammation
P.3.10.01
Cytokine transcription In T cell subsets from healthy, ReA, and CMafnyd/&MHecMJ lndlviduals
H. Beacock-Sharp, J.S.H. Gaston. Department ofMedicine, CamM&?e University Clinical S&w/, Cambridge, UK Introduction: Reactive arthritis (ReA), which develops after infection with intracellular bacteria such as chlamydia, is associated with HlA-B27. However, whether organism-specific HLA-B27-restricted CDB+ T cells are involved in the pathogenesis of ReA is unresolved. The expression of the activation malkers CD69 and HlA-DR on CD8+ Tcells isolated from synovial fluid (SF) suggests an important role. To elucidate futther the role of the CD8+ T cells in the pathogenesis of ReA we have examined the cytokine profile of these cells immediately ax viw, and compared this wtth CD4+ T cells, which can readily be shown to respond to the biggetfng bacteria. Peripheral blood (PB) T cell subsets stimulated with PHA or Chlamydia Wacbomafi~infected macrophages from either healthy controls or indlviduals with an uncomplicated C. trachomatfs infection (GU) were also investigated. Methods: Mononuclear cells were separated from PB and SF. CD4+ and CD8+ T cell subsets were isolated, using a fluorescence activated cell sorter, either immediately ax viw or after stimulation with PHA or C. trachomafisinfected macrophages. mRNA was extracted from these cells and RT-PCR performed to detect mRNA of cytoklnes of interest. Results: Stimulation
IL-2 IL-4
10
30
20
-.--Average tltre level of immune antibodies-ABC
system.
The level of natural anti-A and anti-B antibodies (involved in cellular recognition patterns - onto genetic level) is maintained within limits in every sequence of age, demonstrating that a major impact of today stress factors could not be detected at the present time.
P 3 06 I...
32
1Immune-receptowndocrine
interactions in the pathophysiology of major depression
SergeiMiner&o. Khadrov /n&We of Clinical and Experimental Neurology and PsychiatryLwv Psychiatric Hospital, Ukraine The role of the Adrenergic and Benzodiazepine receptors in development of immunosuppmssion on leukocyte-pyruvatdehydrogenase activity was studied in 60 patients with Primary Major Depressive Episode. Investigation included administration of u-adrenergic “recepto? drug L-DOPA and benzodiazopine “recepto? dtugDlASEPAM before dexamethazone immunosuppression test (DIST). In the cases of Ptfmary Major Depressive Episode in separation with an Anxiety Disorder (30 patients - comparison subjects) after L-DOPA and DIASEPAM administration actfvity of leukocyte-pyruvatde-gydrogenasis increased more than 25%. DIASEPAM of a 10 mg dose had a higher immunosuppressive effect than L-DOPA (0.5 g) on alteration of leukocyle-pymvatdegydmgenase actfvity (more than 5 mmol/lhour, p < 0.05). DIASEPAM immunosuppressive action did not correlate with positive dynamics of T-and-B-cells lymphocytes. From other side, mechanism of L-DOPA actfon on adrenetgic receptors stimulated T-cell receptors and made them hyperactive (produced increasing tangs of T-cell lymphocytes). It means that adrenergic and benzodiazepine receptors are interacting with each other and influencing T-and-B-cells receptors in different ways during immunosuppression. Finally, it is important to study immune-receptor-Endocrine interactions and major depression.
Forum lounges
09:80-18:30/12:8&14:88
IFN-y
c
405
ControlPS
Control PB
GU PB
FleA SF
none
PHA
C. trachomatis
norm
CD4
CD8
CD4
CDS
CD4
CD8
CD4
CDS
+ +
+
+ +
+ +
+ +
+ +
+ +
+ +
+ +
+ +
” ”
+ +
” ”
” +
+
+
”
+
”
+
IL-10
” +
” ” ”
IL-17
+
”
+ = PCR product visualised on agarcse gel by ethidium bromide staining: ” = undetectable
Condusions: PHA stimulation of PB induces transcription of all cytokine genes investigated in both CD4+ and CD8+ T cells. CD4+ T cells from unstimulated ReA SF, and from GU PB stimulated wm the infecting organism, were found to transcribe IFN-y and IL-2. Interestingly, CD8+ T cells isolated from both these cell populations were also found to produce IL-10 and 11-17.Thus, both C. trachomalisstimulated CD8+ T cells and ax viva CDE+Tcells from ReA SF are likely to secrete both pm-inflammatory (e.g. 11-17)and regulatory (e.g. IL-lo) cytokjnes. These results point to a complex role for CD8+ T cells in the pathogenesis of ReA.
1P.3.10.02 1 Blood cells and bacterial inflammation T. Matsumoto. Matsumoto Living Cell Research Laboratory Tokyo,Japan Introduction and Alm: My research on erythrocytes (red blood cell = Rbc) and leucocyte, aims to disclose the essential function of them in bacterial inflammation. Method and Mateflals: The tissue culture method based on MatsumotoHagiya method has been conducted using 7% Dextran-salin solution, 5% CQ incubator, pseudomonas sp., blood cells, and others. Resultst (1) Rbc are classified into at least three groups (top, middle and bottom layers) through the Matsumoto-Hagiya method. (2) When top Rbc are exposed to pseudomonas sp.. Rbc and Rbcderived cells demonstrate rotating or reversive movement on or around pseudomonas sp. (video) which should be interpreted BStheir bacterfo-static influence. (3) Rbcs in the middle layer remain but the multiplication of pseudomonas sp. is suppressed. (photo) (4) Rbcs in the bottom layer are static and pseudomonas sp. very rapidly multiplies. (photo) (5) Leucocytes show amoeboid movement but do not pmve phagocytcsis. (video) Conclusion: The inflammation occurs when immunological balance is bmken by many factors. Such as: immuno-imbalance induced by influenza, hepatitis, excessive fatigue, malnuttftion and many other diseases: General factors: bleeding, ischaemia, trauma etc.: Local factors.