- Email: [email protected]
Apoptosis as possible mechanism of cytotoxicity induced by toxics agents implicated in toxic oil syndrome (TOS)
Poster Session 2H. Metals I
P1 G-1 95 [ t
T CELL REACTION AGAINST ANILINE METABOLITES AND THEIR ENDOGENOUS FORMATION IN WHITE BONE MARROW CELLS
Marty B.F. Wulferink *, Jos~ B. Gonz~ilez, Carsten Goebel, Ernst Gleichmann. Medical Institute of Environmental Hygiene at
the Heinrich Heine University, Division of lmmunology, Duesseldorf Germany Aniline derivatives, also termed para-compounds or arylamines, are used for different purposes, e.g., as drugs, dyes, and for tanning, and they are notorious for inducing allergy and autoimmunity. Previously we showed that, in the case of the aniline derivative procainamide (PA), phagocytes can generate reactive intermediate metabolites which are immunogenie for mouse T cells. Here we show that the proteinreactive aniline metabolite nitrosobenzene (NB), other than the non-reactive aniline itself, can induce a popliteal lymphnode (PLN) response. A secondary response in the same animal was obtained with suboptimal dose of NB. Evidence for T-cell dependence of the reaction was obtained with T cell-deficient, nu/nu mice, which failed to react to NB. While bone marrow cells (WBMC) incubated with aniline in vitro for 2 days induced a PLN response, suggesting the formation of immunogenic aniline metabolites in WBMC. This is further supported by the finding that T cells, primed to NB, could specifically be restimulated in vitro by offering WBMC preincubated with aniline as antigen. We therefore propose that WBMC can generate reactive aniline metabolites, such as NB, which become immunogenic by altering self proteins. Generation of such metabolites by the ubiquitous WBMC-derived cells could be responsible for the great variety of adverse immune reactions, such as SLE and agranulocytosis, seen after exposure to aniline derivatives.
K+eywords: aniline; nitrosobenzene; T cells; immunogenicity; phagocytes; metabolism !
P1G-196 1 IMMUNOGENICITY OF SYNTHETIC OIL ANILIDES, I
THE REPUTED ETIOLOGIC AGENTS OF SPANISH TOXIC OIL SYNDROME (TOS)
55
Preliminary results show that mice primed with linolenylanilide can mount a specific secondary PLN response to the aniline metabolite NB. Taken together, these results are consistent with the hypothesis outlined above.
Keywords: oil anilides; nitrosobenzene; immunogenicity 1
I P1G-1 971 J
APOPTOSIS AS POSSIBLE MECHANISM OF CYTOTOXlClTY INDUCED BY TOXlCS AGENTS IMPLICATED IN TOXIC OIL SYNDROME (TOS)
Soledad Gallardo, Victoria del Pozo, Blanca Cardaba, Aurora Jurado, Isabel Cortegano, Ana del Amo, Ignacio Arrieta, Pilar Palomino, Carlos Lahoz *. Immunology Department,
Fundacidn Jim~nez Dfaz, Av. Reyes Cattlicos 2, 28040 Madrid, Spain Toxic Oil Syndrome caused by ingestion of rapeseed oil denaturated with aniline presented clinical features which are compatible with an activation of the immune system [1-3]. We wanted to know the effect of some of the implicated toxics (aniline, 3-(N-phenylamino)l,2-propanediol and its mono and dioleyl esters on human lymphocytes. For this purpose, we used several standardized methods to analyze their possible triggering effect on the of the programmed cell death: Morphological evaluation by fluorescence microscopy staining the cells with Acridine Orange and Propidium Iodide, study of DNA degradation by flow cytometry, labeling with Merocianine 540 and DNA fragmentation assay by gel electrophoresis. Our results indicated that only the di-oleyl ester of 3-(Nphenylamino)-l,2-propanediol induced apoptosis in human lymphocytes, in concentration and time dependent way. [1] Gallardo S., del Pozo V., C~trdaba B. et al Toxicology 93: 289-299, 1994 [2] Lahoz C., Rose N. R., Goter Robinson C. J. Immunology of TOS WHO Regional Publications, No. 42, pages 143-152, 1992. [3] LahozC.,TricasL.,VelaC. et aI. Eur. J. Resp. Diseases 62:415-418, 1983
Keywords: apoptosis; toxic oil syndrome; cytotoxicity; TOS and immune activation
Jos6 B. Gonzfdez *, Marty Wulferink, Mark Goebel, Carsten Goebel, Ernst Gleichmann. Medical Institute for
Enviromental Hygiene at Heinrich Heine University, Division of Immunology, Diisseldorf, Germany It has been proposed that TOS is a graft-versus-host-like disease in which T lymphocytes react to unknown self-proteins, that were altered by oil anilides contained in the toxic oil. More specifically, T cells should be sensitized to self-proteins altered by protein-reactive metabolites of the aniline component of oil anilides: nitrosobenzene (NB) and/or N-hydroxylaniline (HA). The fatty acid components of oil anilides, in contrast would target the compounds to phagocytes (possessing lipid receptors) and activate them like adjuvants. Phagocytes, in turn, might generate the reactive metabolites mentioned, as we have shown for the aniline derivative procainamide. Using the politeal lymph node assay (PLNA) in mice we have tested the immunogenicity of linolenylanilide, linoleylanilide, and a mixture of three fatty acid esters of 3-(N-phenylamino)-1, 2- propanediol (PAP): 1-oleyl, 2- linoleyl-PAP; 1-oleyl, 2-1inolenyl-PAP and. 1-1inolenyl, 2-1inoleyl-PAP. These substances were previously blind coded and obtained via the WHO/FIS scientific committe for TOS. The following results were obtained: (i) linolenylanilide, linoleylanilide, and the mentioned PAP ester mixture induced in that order significant primary PLN responses when compared with linolenic acid, linoleic acid, and triolein, respectively, used as aniline-free controls. (ii) Compared with oil anilides, the reactive aniline metabolites NB and HA induced significant primary PLN responses even at 100-fold lower molar doses. (iii) PLN reactions to linolenylanilide and NB proved to be T cell-dependent since athymic mice failed to respond. (iiii)
Poster Session 2 P2H. Metals I
MODEL OF INORGANIC MERCURY IP2H-1 98 I PERFUSION KINETICS [
I.B. Tokin *, P.A. Smirnova. Faculty of Applied Mathematics, State
University, St. Petersburg 198904, Russia The proposed model considers the kinetics of mercury in man at the vapour inhalation of elementary mercury. In contrast with the previous models [1] the present one constructed on the different basis on consideration of the inorganic mercury balance and it metabolites in each organs. The mathematical model is submitted by the system of the differential equations, describing mercury incoming and distribution, its biotransformation and outputting from the body. The solution of the equations system allows to determine the change of concentration of inorganic mercury and its metabolites in organ in the different periods of time and the level of the metabolites outputting with urine and excrements. The necessary condition for the model decision is the preliminary evaluation of the parameters meaning included in the equation. The equation for the estimation of the inorganic mercury concentration change in organ is as follows: VdC/dt = QC - QC/R.
P1 G-1 95 [ t
T CELL REACTION AGAINST ANILINE METABOLITES AND THEIR ENDOGENOUS FORMATION IN WHITE BONE MARROW CELLS
Marty B.F. Wulferink *, Jos~ B. Gonz~ilez, Carsten Goebel, Ernst Gleichmann. Medical Institute of Environmental Hygiene at
the Heinrich Heine University, Division of lmmunology, Duesseldorf Germany Aniline derivatives, also termed para-compounds or arylamines, are used for different purposes, e.g., as drugs, dyes, and for tanning, and they are notorious for inducing allergy and autoimmunity. Previously we showed that, in the case of the aniline derivative procainamide (PA), phagocytes can generate reactive intermediate metabolites which are immunogenie for mouse T cells. Here we show that the proteinreactive aniline metabolite nitrosobenzene (NB), other than the non-reactive aniline itself, can induce a popliteal lymphnode (PLN) response. A secondary response in the same animal was obtained with suboptimal dose of NB. Evidence for T-cell dependence of the reaction was obtained with T cell-deficient, nu/nu mice, which failed to react to NB. While bone marrow cells (WBMC) incubated with aniline in vitro for 2 days induced a PLN response, suggesting the formation of immunogenic aniline metabolites in WBMC. This is further supported by the finding that T cells, primed to NB, could specifically be restimulated in vitro by offering WBMC preincubated with aniline as antigen. We therefore propose that WBMC can generate reactive aniline metabolites, such as NB, which become immunogenic by altering self proteins. Generation of such metabolites by the ubiquitous WBMC-derived cells could be responsible for the great variety of adverse immune reactions, such as SLE and agranulocytosis, seen after exposure to aniline derivatives.
K+eywords: aniline; nitrosobenzene; T cells; immunogenicity; phagocytes; metabolism !
P1G-196 1 IMMUNOGENICITY OF SYNTHETIC OIL ANILIDES, I
THE REPUTED ETIOLOGIC AGENTS OF SPANISH TOXIC OIL SYNDROME (TOS)
55
Preliminary results show that mice primed with linolenylanilide can mount a specific secondary PLN response to the aniline metabolite NB. Taken together, these results are consistent with the hypothesis outlined above.
Keywords: oil anilides; nitrosobenzene; immunogenicity 1
I P1G-1 971 J
APOPTOSIS AS POSSIBLE MECHANISM OF CYTOTOXlClTY INDUCED BY TOXlCS AGENTS IMPLICATED IN TOXIC OIL SYNDROME (TOS)
Soledad Gallardo, Victoria del Pozo, Blanca Cardaba, Aurora Jurado, Isabel Cortegano, Ana del Amo, Ignacio Arrieta, Pilar Palomino, Carlos Lahoz *. Immunology Department,
Fundacidn Jim~nez Dfaz, Av. Reyes Cattlicos 2, 28040 Madrid, Spain Toxic Oil Syndrome caused by ingestion of rapeseed oil denaturated with aniline presented clinical features which are compatible with an activation of the immune system [1-3]. We wanted to know the effect of some of the implicated toxics (aniline, 3-(N-phenylamino)l,2-propanediol and its mono and dioleyl esters on human lymphocytes. For this purpose, we used several standardized methods to analyze their possible triggering effect on the of the programmed cell death: Morphological evaluation by fluorescence microscopy staining the cells with Acridine Orange and Propidium Iodide, study of DNA degradation by flow cytometry, labeling with Merocianine 540 and DNA fragmentation assay by gel electrophoresis. Our results indicated that only the di-oleyl ester of 3-(Nphenylamino)-l,2-propanediol induced apoptosis in human lymphocytes, in concentration and time dependent way. [1] Gallardo S., del Pozo V., C~trdaba B. et al Toxicology 93: 289-299, 1994 [2] Lahoz C., Rose N. R., Goter Robinson C. J. Immunology of TOS WHO Regional Publications, No. 42, pages 143-152, 1992. [3] LahozC.,TricasL.,VelaC. et aI. Eur. J. Resp. Diseases 62:415-418, 1983
Keywords: apoptosis; toxic oil syndrome; cytotoxicity; TOS and immune activation
Jos6 B. Gonzfdez *, Marty Wulferink, Mark Goebel, Carsten Goebel, Ernst Gleichmann. Medical Institute for
Enviromental Hygiene at Heinrich Heine University, Division of Immunology, Diisseldorf, Germany It has been proposed that TOS is a graft-versus-host-like disease in which T lymphocytes react to unknown self-proteins, that were altered by oil anilides contained in the toxic oil. More specifically, T cells should be sensitized to self-proteins altered by protein-reactive metabolites of the aniline component of oil anilides: nitrosobenzene (NB) and/or N-hydroxylaniline (HA). The fatty acid components of oil anilides, in contrast would target the compounds to phagocytes (possessing lipid receptors) and activate them like adjuvants. Phagocytes, in turn, might generate the reactive metabolites mentioned, as we have shown for the aniline derivative procainamide. Using the politeal lymph node assay (PLNA) in mice we have tested the immunogenicity of linolenylanilide, linoleylanilide, and a mixture of three fatty acid esters of 3-(N-phenylamino)-1, 2- propanediol (PAP): 1-oleyl, 2- linoleyl-PAP; 1-oleyl, 2-1inolenyl-PAP and. 1-1inolenyl, 2-1inoleyl-PAP. These substances were previously blind coded and obtained via the WHO/FIS scientific committe for TOS. The following results were obtained: (i) linolenylanilide, linoleylanilide, and the mentioned PAP ester mixture induced in that order significant primary PLN responses when compared with linolenic acid, linoleic acid, and triolein, respectively, used as aniline-free controls. (ii) Compared with oil anilides, the reactive aniline metabolites NB and HA induced significant primary PLN responses even at 100-fold lower molar doses. (iii) PLN reactions to linolenylanilide and NB proved to be T cell-dependent since athymic mice failed to respond. (iiii)
Poster Session 2 P2H. Metals I
MODEL OF INORGANIC MERCURY IP2H-1 98 I PERFUSION KINETICS [
I.B. Tokin *, P.A. Smirnova. Faculty of Applied Mathematics, State
University, St. Petersburg 198904, Russia The proposed model considers the kinetics of mercury in man at the vapour inhalation of elementary mercury. In contrast with the previous models [1] the present one constructed on the different basis on consideration of the inorganic mercury balance and it metabolites in each organs. The mathematical model is submitted by the system of the differential equations, describing mercury incoming and distribution, its biotransformation and outputting from the body. The solution of the equations system allows to determine the change of concentration of inorganic mercury and its metabolites in organ in the different periods of time and the level of the metabolites outputting with urine and excrements. The necessary condition for the model decision is the preliminary evaluation of the parameters meaning included in the equation. The equation for the estimation of the inorganic mercury concentration change in organ is as follows: VdC/dt = QC - QC/R.