Prevention of reaginic antibody production and anaphylactic gastric ulcer by pesticides and by a polychlorinated biphenyl

EkVIRONMENTAL

RESEARCH

Prevention Anaphylactic

25, 381-385 (1981)

of Reaginic Antibody Production and Gastric Ulcer by Pesticides and by a Polychlorinated Biphenyll

F. ANDRE,* J. GILLON,? *Unit& Frclncc,

de Physioputhologie crnd +Depurtment

Digestirse

of Medicine.

C. ANDRE,* INSERM Western

AND G. JOURDAN*

U 45, Hhpital Edward Herriot. Lyon. General Hospitcd. Edinburgh. Scotland

Received November 4. 1980 The effects of different pesticides. lindane, phosalone, and carbaryl. and of one polychlorinated biphenyl, pyralene, were studied on the development of reaginic antibody production and the formation of allergic gastric ulceration in Mustomys natnlensis following prior sensitization and subsequent challenge with ovalbumin. The results indicate that lindane, phosalone, and pyralene have a pronounced suppressive effect on type I hypersensitivity reaction when ingested orally. These results give further cause for concern about the hazards to health which might result from continued indiscriminate use of such substances.

INTRODUCTION

There is increasing concern about the biological effects of various pesticides, which have been used extensively in agriculture and which may persist in the environment for long periods of time. Human exposure to these agents may occur as a result of ingestion of foodstuffs, and as a consequence of the use of such products in pharmaceutical preparations and in public-health pest control. Research into this problem has so far tended to be focused on the potential carcinogenic properties of these agents, but more recently their effects on immune responses have received particular attention, and it is becoming clear that important changes in host immunity may occur after pesticide ingestion. We have chosen to study the effects of different families of pesticides (organochlorine, organophosphate, carbamate), namely, lindane, phosalone, and carbaryl, and of one polychlorinated biphenyl, pyralene, on the development of reaginic antibody production and the formation of allergic gastric ulcers in the African rodent Mastomys natalensis following prior sensitization and subsequent challenge with ovalbumin. The results suggest that these substances have important effects on type 1 hypersensitivity reactions, and we discuss the implications of these findings in relation to their other known biological effects, and in particular to their carcinogenic properties. MATERIALS

AND METHODS

The experiments were carried out using our previously described model of anaphylactic gastric ulceration in the small African rodent, M. ‘natalensis (An&-C et ’ This work was supported by a grant from the DelCgation G&&rale Technique (77.7.0444).

B la Recherche Scientifique et

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5: 1981 hy Academic Press. Inc. of reproductive in any form rcvzrved

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al., 1979a). These animals are particularly suitable for the study of type 1 hypersensitivity reactions in view of their well-developed mast cell system. Outbred animals were reared in our laboratory and were used when over 3 months old. They received water ad libitum and were maintained on ovalbumin-free UAR mouse food (Villemoisson/Orge, France) until the time of the experiments. Sensitization and induction ofgastric ulcer. Ovalbumin recrystallized five times was obtained from Calbiochem. Animals were sensitized by intraperitoneal injection of 3 pg ovalbumin in a volume of 0.2 ml with 1 mg aluminum hydroxide. They were subsequently challenged by intramucosal injection of 1 mg ovalbumin in 0.01 ml saline in the corpus region of the stomach under general anesthesia. The animals were sacrificed 3 days later to determine whether gastric ulceration had occurred. Detection of reuginic antibodies. Reaginic antibodies were sought by measuring the degranulation of peritoneal mast cells in the presence of the antigen, as described by Prouvost-Danon et al. (1972). Mast cells were collected from the peritoneal cavity of sensitized and unsensitized Mustomys. Passive sensitization of normal mast cells in vitro was performed by incubation with 25 ~1 of heated (60 min at 56°C) or nonheated sera collected from animals sensitized 7 days previously. These passively sensitized cells and those from sensitized animals were incubated with 100 pg/ml ovalbumin at 37°C for 15 min. Degranulation of mast cells was determined after staining with toluidine blue. Experiment I. Effect of pesticides on the induction of unuphyluctic gastric ulcer. Four groups of 12 animals received a diet containing lindane (150 ppm), phosalone (50 ppm), carbaryl(l50 ppm), or pyralene (150 ppm) for 1 month prior to sensitization and induction of gastric ulceration as described above. A group of 12 control animals was also studied. Experiment 2. Effect of pesticides on reuginic antibody production. In a second experiment to test the effect of pesticides and biphenyls on reaginic antibody production, four different groups of 12 animals received the above pesticidecontaining diets for 1 month before sensitization. The diets were continued during the 7-day period after sensitization. The animals were then sacrificed and peritoneal mast cells and serum were collected. Reaginic antibodies were sought using the test of degranulation of mast cells as described above. A control group of 12 animals was again included. RESULTS

Effect of pesticides on unuphyluctic gastric ulcer. The results of the first experiment are shown in Table 1. All 12 control animals had developed a welldefined gastric ulcer at the challenge site on Day 3. Ten of the 12 animals on carbaryl had developed ulcers, but only 1 of those on lindane, none on phosalone, and 4 in the pyralene group had ulcers. These results are statistically highly significant for lindane (P < O.OOOl), phosalone (P < O.OOOOl), and pyralene (P < 0.005) using the x2 test. Effect of pesticides on reuginic antibody production. The results of the second experiment are shown in Table 2. The mast cell degranulation test was positive in all 12 control animals. In contrast no evidence of reaginic antibody production was

PESTICIDES

AND

TABLE THE

EFFECTS

Dietary

OF PRETREATMENT WITH DIETS ON THE DEVELOPMENT

group

I

VARIOUS PESTICIDEOF ANAPH~LACTIC

12 12 I2 I2 I2

Note. The organochlorine phenyl pyralene significantly hypersensitivity.

lindane, reduced

OR PYRALENE-CONTAINING GASTRIC

No. developing gastric ulcer

No.

Control Lindane Phosalone Pyralene Carbaryl

383

IMMUNOSUPPRESSION

ULCERS

Significance (x2)

I2 I 0 4 IO

P < 0.0001 P < 0.00001 P < 0.005 N.S.

the organophosphate phosalone. and the polychlorinated bithe incidence of gastric ulcer. indicating an inhibition of type I

found in the lindane and phosalone groups, while 9 of the carbaryl group and 4 of those on pyralene showed positive reactions. The results are statistically significant for lindane (P < O.OOOOl), phosalone (P < 0.00001). and pyralene (P < 0.005). Our results indicate that these groups of pesticides and biphenyls, in particular the organophosphate phosalone and the organochlorine lindane, have a pronounced suppressive effect on type 1 hypersensitivity reactions when ingested orally. The in vitro test of mast cell degranulation was rendered negative in sensitized animals fed on one of these substances, and the extent of this negative effect was such that the development of anaphylactic gastric ulcers was also prevented. DISCUSSION

The increasing amounts of various insecticides in the environment have already stimulated a large volume of research, with the emphasis being directed toward potential carcinogenic effects and direct toxicity. In recent years it has also been realized that these compounds have important immunosuppressive properties affecting both humoral and cellular immune mechanisms. Large oral doses of various pesticides including DDT have been shown to impair the humoral immune TABLE THE

EFFECTS

OF PRETREATMENT

WITH

DIETS ON REAGINIC

THE

ANTIBODY

No. Dietary Control Lindane Phosalone Pyralene Carbaryl

group

No. I? I2 I2 I2 I2

2 PESTICIDE-

-\ND

PYRALENE-CONTAINING

PRODUCTION

in which MCDT” positive I2 0 0 4 9

Significance (XL) P -c 0.00001 P ‘z 0.0ooo1 P < 0.005 N.S.

” MCDT = mast cell degranulation test. Ni>te. The reaginic antibody production was measured by the mast cell degranulation complete inhibition of the normal reaginic antibody response is seen in the lindane. pyralene groups.

test. Almost phosalone. and

384

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ET

AL.

response in mice (Wiltrout el al., 1978), while oral carbaryl causes a reduction in y-globulin production after antigenic challenge in New Zealand white rabbits (Street and Sharma, 1975). The latter study also demonstrated impairment of cellular immunity after 4 weeks of dietary treatment with clinically subtoxic amounts of four pesticides. Roux et al. (1979) showed that lindane had a suppressor effect on the responses to phytohemagglutinin of human lymphocytes in vitro. Phagocytic functions in the reticuloendothelial system have been shown to be affected by intravenous carbaryl (Pipy et al., 1978). The effect of dietary DDT on the anaphylactic response to intraperitoneal challenge with diphtheria toxoid in albino rats was studied by Gabliks et al. (1973, and they showed that DDT had a suppressive action on type 1 hypersensitivity responses. The same group showed that mice infected with the influenza virus while on a DDT-containing diet had more severe infections and a higher mortality than did controls (Gabliks and Utz, 1979). With the exception of this study, however, attempts to relate the immunosuppressive effects of pesticides to disease states have been few. Our results confirm the ability of certain pesticides and biphenyls to suppress type 1 hypersensitivity reactions as measured by reaginic antibody production. A blocking effect of exactly similar magnitude on the production of experimental gastric ulcers has also been demonstrated. This model of gastric ulceration undoubtedly depends mainly on type 1 hypersensitivity and the release of histamine and the other vasoactive amines from sensitized mast cells, and we have shown in previous experiments that these effects can be prevented by treatment with the mast cell degranulation inhibitor disodium cromoglycate (Andre et al., 1978) and the histamine HZreceptor antagonist cimetidine (Andre et al., 1979a). It seems likely that anaphylaxis may play an important role in the etiology of gastritis and gastric ulceration in humans (Andre et al., 1979b), and an increased incidence of these disorders has been reported in workers chronically exposed to organochlorine compounds (Paramonchik and Platonova, 1968). The discrepancy between our results showing a protective effect of these substances on experimental gastric ulceration and this report of an increased incidence of such disorders in humans probably indicates that chronic low-grade exposure can affect the immune response differently from short-term exposure to high doses. Differences between species may also be important. While at first sight a possible protective action against anaphylaxis might therefore seem beneficial, any immunosuppressive property in a substance present in the environment should be regarded as disquieting. Relative suppression of one or more limbs of the immune response is thought to be the mechanism underlying various disease processes, and the dangers are highlighted by the tendency of patients on long-term immunosuppressive agents, for example, following renal transplants, to develop neoplasms of the reticuloendothelial system. It has been suggested that anaphylactic reactions may be important in the body’s resistance to the development of solid tumors (Lyncher al., 1978), and this may indeed be their most important function. Taken with the fact that lindane in high doses has been shown to be highly carcinogenic in mice (Reuber, 1979) and that large quantities of this substance have been demonstrated in the extracted lipids of gastric carcinomas (Wassermann et al., 1978), the results presented here give further cause

PESTICIDES

AND

IMMUNOSUPPRESSION

for concern about the hazards to health which might result from continued criminate use of such substances.

385

indis-

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