The effect of paraquat lung on mononuclear cells

Exp. Pathol. 1988;34: 115-118 VEB Gustav Fischer Verlag Jena

l)Department of Hygiene and Epidemiology, University Medical School, Szeged (Head: Prof. ILLES DEsI) 2)Department of Pathology, Country Hospital, Kecskemet (Head: Prof. G. LUSZTIG) 3)Bioiogicai Isotope Laboratory, "A. J." University Szeged, Szeged, Hungary

The effect of paraquat lung on mononuclear cells

With 2 figures Address for correspondence: K. BARABAS, M.D., Dept. of Hygiene and Epidemiology, Univ. Med. School Szeged, Dom ter 10, Szeged, H - 6720, Hungary Key words: paraquat toxicity; macrophage activation; mouse footpad swelling test; mononuclear cells; lung

Summary The mouse footpad swelling test was used to clarify the possibility of the induction oflocal cellular reaction with cell suspension of mouse lungs treated with paraquat in a syngeneous animal. Among the inbred strains used, the highest, statistically significant cellular reactivity was observed in C3HI He strain mice. These results suggest indirect evidence of macrophage activation in the lung toxicity of paraquat.

Introduction We have performed experiments with Gramoxone®, containing paraquat (PQ) (I, l-dimethyl-4,4 dipyridilium dichloride) as active ingredient, for some years (5,6). Its toxic effect on the lungs of mammals was divided into two well-separated stages by SMITH and HEATH (8). The first is the initial, destructive stage, which is characterized by alveolar edema and hemorrhage; it results in early death, which occurs 2-3 days after intoxication. The survivors of the acute symptoms or mammals, poisoned with smaller doses reach the proliferative stage after 4-7 days. Mononuclear cells and spindle-shaped fibroblast -like cells then appear. The intraalveolar septum widens and finally the infiltration of fibrocytes and fibroblasts results in the restriction of the respiratory functions. Activation of the alveolar macrophages and fibroblasts is induced by the destruction of pneumocytes of type 2. In order to clarify the pathomechanism, the following question must be answered: Is it possible to induce cellular reaction in a syngeneous animal with the cell suspension of a mouse lung treated with PQ? To answer this the mouse footpad swelling test described by COOPER in 1972 (2) was applied. Our hypothesis is that if it is possible to bring about a reaction, then indirect evidence is given about different factors produced in the lung containing PQ, which can influence lymphocytes among others. One of these may be the migration inhibiting factor (MIF), produced by activated macrophages.

Materials and Methods Animals: C3H/He; BALB/C; and AKR inbred mouse strains were used. To perform semiallogenic transplantation, the (C3H XAKR)F4 hybrid was used. Male mice with a mean weight of30.0

±

1.4 g were inoculated.

The lung cell suspension in physiological saline was made from the lungs of animals 3 days after oral administration of paraquat. Cell counting was performed by Tiirk staining in a Biirker chamber. 2.4 x 105 cells in 0.05 ml were injected into the hind footpad of an animal belonging to a similar strain in the case of syngeneous transplantation. On Exp. Pathol. 34 (1988) 2

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Fig. 1. Histological picture of mouse footpad after inoculation of untreated syngeneous lung suspension. HE, x 130.

Fig. 2. Histological picture of mouse footpad after inoculation of syngenous cell suspension 01 iung treated with PQ. HE, x 130.

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semiallogenic transfer C3HIHe lung cells were injected into the footpad of a (C3HXAKR)F4 hybrid. The thickness of the footpad was measured with a micrometer before and 24 h after administration. Statistical evaluation was performed with Student's 2-tailed t-test. Histological examination was made with hematoxylin-eosin staining.

Results and Discussion Among the inbred strains the highest reactivity was observed in the mice from the C3H/He strain. Although the degree of footpad swelling was also significant in the AKR and BALB/C animals, the significance level in the group of C3H1He mice was equal to that in seffiiallogenic transplantation (table I). Table 1. Summary of measurements Mouse strain

Mouse lung cell susp. 2.4x 105 cells in 0.05 ml

BALB/C

C untreated lung PQ/60 mg/kg

Thickness of footpad

Extent of swelling

before inoculation

24 h after inoculation

2.14 ± 0.073 mm

2.12 ± 0.06 mm

2.14±0.12 mm

2.26 ± 0.09 mm

1.2xlO- l mm

p

<

0.05

p p

< <

0.001 0.001

AKR

C PQ

1.63 ± O.OS mm 1.62 ± 0.11 mm

1.95 ± O.OS mm 2.19 ± O.IS mm

3.2x 10- 1 mm 5.7xlO- 1 mm

C3H/He

C PQ

I.S2 ± 0.079 mm I.S2 ± 0.11 mm

I.S0 ± 0.06 mm 2.36 ± 0.16 mm

5.4x 10- 1 mm

p

<

0.001

PQ

I.S9 ± 0.04 mm

2.66 ± 0.16 mm

7.7xlO-1 mm

p

<

0.001

(C3HxAKR) F4

The histological picture shows that the untreated lung cell suspension resulted in the appearance of several round cells in the corium layer of the mouse footpad (fig. I). Round-cell infiltration developed in response to the lung cell suspension containing PQ in the AKR mouse strain, and it invaded the muscular coat as well. It was characteristic of the histological reaction of the animals from the BALB/C strain that there were many round cells both in the dermis and in the muscles, including many plasma cells. Many round cells, macrophages and granulocytes appeared both in the dermis and in the muscular layer in the histological picture of the mice from the C3H/He strain (fig. 2). The PQ2+ cation accumulating in the lungs participates in a redox cycle and generates a superoxide anion from molecular oxygen (1). The multiple connection of the oxygen radical and the immune system is well known (3, 4). Various immune reactants (immunoglobulins, immunecomplexes, complement components, lymphokines) are able to activate controlled migration and phagocytosis, but they also play important roles in modulating the functions of the lymphocytes. Our histological results mainly suggest that different tissue factors are produced in the lungs due to PQ, and these cause the migration of mononuclear cells (granulocytes, neutrophils and lymphocytes) to the lung tissue. References I. Bus, J. S., AUST, S. D., GILSON, J. E.: Superoxide and singlet oxygen catalysed lipid peroxidation as a possible mechanism for paraquat toxicity. Biochem. Biophys. Res. Commun. 1974; 58: 749-755. 2. COOPER, M. G.: Delayed-type hypersensitivity in the mouse. Scand. J. Immunol. 1972; 1: 167 - 172. 3. GOLDSTEIN, J. M., Roos, D. M., KAPLAN, H. B., WEISSMAN, G. B.: Complement and immunglobulins stimulate superoxide production by human leukocytes independently of phagocytosis. J. Clin. Invest. 1974; 56:

1155-1164. 4. KRANT, E. H., SAGONE, A. L.: The effect of oxidant injury on the lymphocyte membrane and functions. J. Lab. Clin. Med. 1981; 98: 697-703. Exp. Pathol. 34 (19SS) 2

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5. MATKOVICS, B., SZAB6, L., VARGA, SZ. I., NovAK, R., BARABAs, K., BERENCSI, G.: In vivo effects of paraquat on some oxidative enzymes of mice. Gen. Pharmac. 1980; 11: 267-270. 6. - BARABAs, K., SZAB6, L., BERENCSI, G.: In vivo study of the mechanism of protective effects of ascorbic acid and reduced glutathione in paraquat poisoning. Gen. Pharmac. 1980; 11: 455 -46l. 7. NAKAGAWARA, A., DE SANTES, N. M., NOGREIRA, N., NATHAN, C. F.: Lymphokines enhance the capacity of human monocytes to secrete reactive oxygen intermediates. J. Clin. Invest. 1982; 40: 1047-1048. 8. SMITH, P., HEATH, D.: Paraquat. Clin. Rev. Toxicol. 1976; 4: 411-445. (Received June 26, 1987)

Exp. Pathol. 1988; 34: 118 VEB Gustav Fischer Verlag Jena

Book Review VICC - International Union Against Cancer.

TNM Classification of Malignant Tumours edited by D. HERMANEK and L. H. SOBIN. 4th, fully revised edition, 197 pages, DM 22, -, ISBN: 3-540-17366-8. Springer-Verlag, Berlin-Heidelberg -New York-London-Paris-Tokyo 1987. (Origin ally published by VICC) This fourth edition of TNM system for the classification of malignant tumours is a fully revised one. The variations which were included in the former editions have been eliminated, previously unclassified tumours have been added, and the site classifications have been updated. The new edition is the result of the collaboration of the national American, British, Canadian, French, German, Italian and Japanese TNM committees. As an introduction the history and the principles of the TNM system are briefly dealt with, followed by its general rules including the anatomical regions and sites, the clinical and pathological classification, the histopathological grading, additional descriptors, e.g., multimodal therapy, recurrent tumours, residual tumour classification (new in the fourth edition), etc. - It is recommended that the International Histological Classification of Tumours is to be used for classification and definition of tumour types, and the WHO International Classification of Diseases for Oncology (ICD-O) is recommended to be used for storage and data retrieval. Under these aspects the tumours of the various regions are described: head and neck, digestive system, lung, bone and soft tissues, skin, breast, gynecological tumours, urological tumours, ophthalmic tumours, brain tumours. Hodgkin's disease and non-Hodgkin's lymphoma, pediatric tumours. - Each chapter is introduced by notes on the headings under which the regional subject is described, e.g., rules for classification with the procedures for assessing the T, Nand M categories, anatomical subsites, definition of the regional lymph nodes, TNM clinical classification, pTNM pathological classification, G histopathological grading, residual tumour after treatment (R) classification and stage grouping. Each region is finished by a summary. The TNM system deals primarily with the classification by anatomical extent of disease as determined clinically and histopathologically. The anatomical extent is based on the assessment of T = extent of primary tumour; N = absence or presence and extent of regional lymph node metastasis; M = presence or absence of distant metastasis. It is quite obvious that only a single internationally accepted classification as a basis of a "common language" in comparing tumour diseases, their diagnosis, course and results of treatment, can guarantee the cooperation in this field. The new edition of the TNM classification is the best solution of this problem and an irreplacable material for all physicians faced with tumour diseases. F. BOLCK, Jena

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