WlADOMOSCI PAHAZYTOLOGlCZNE T. V I I, NR 4/5/6, 1961
SOME PHYSIOPATHOLOGICAL PROBLEMS IN THE PARASITIC INVASIONS
BY
HELIODOR SZWEJKOWSKI
Depattment of Pathological Anatomy, Veterin·ary Faculty, Warsaw
As we have hear<i it from Professor Stefański's Paper it is not so easy to become a parasite and that a „career" of an invasive form of the obligate parasite is full of difficulties. The paper by Kozar, published in that last number (3) of the „Parasitological Review" (Wiadomości Para-
zytologiczne) is introducing us to ~
the enzymatological problems of the parasite's mechanics, of infesta-
tion, which the parasite has to overcome when it is primarily breaking into a tissue, and in this further migration as well.
The infestation mechanisms, that could be defined often as action of the enzymes, that are contained in the excreta of the pa- rasite upon resisting them the tis- suler barriers of the host, have not been, as yet recognised exactly enough to allow for the explana- tion of all the phenomena that, from the stand point of pathology, could be defined as the action of noxa and pathogenic agents upon the organism being susceptibie to the action of the stimulus.
fot. J. Waluga Paper read at the 7th Congress of the Polish Parasitological Society, Olsztyn- Kortowo, 11-14 VI 1961.
t388 H. SZWEJKOWSKI
Michajłow (1960) emphasizes that: "numerous endoparasites, as it has been stated, are producing the buffer, preserving the antigenous sub- stances. Their evolucional origin is not always elear. According to some authors, the buffering substances, produced nowadays by the parasites, arose during their free life under saprobiosis or coprobiosis conditions, these substances rendered possible the transition to the parasitic way of living and in some measure they predestinated to it."
The same regularities in the evolutionary development have to evoke in the organism of the host the graduał formation of the protection mechanisms. The protection, as it is already known today, is a very complicated set of biochemical, physical and chemical phenomena, however it is continually described on the resistance and immunity. It is known that the protection symptoms were the best recognised in the infections diseases.
Michajłow (1960) states that: ,,the host-parasite system can be, as it emphasized Szichobalowa (1960), a good pattern for the immunological investigations. The antigens, that cause the production of antibodies, are the products of the parasites metabolism !.:nd the excreta from their glands. The antibodies originate under the influence -of both intestinal and tissuler rparasites, they are formating in the cells close to the place of living of the parasite".
In the main, the genesis of immunity, regardless of the kind of factor that is causing it, is a very complicated matter both morphologically and biochemically. These problems, from the immunological-functional point of view, were presented by Kozar in his paper entit;led: "Some Problems of łmmunity in Parasitic Diseases" delivered at the Third Congress of the Polish Society for Parasitology at Wrocław in 1952. The mentioned paper, as well as the subsequent elaborations dealing with the subject of immunity, were completely or almost completely apart of the cyto- logical or histological problems, which were strn waiting to be an object of interest of ours researchers. The problem is the more worth to be taken into cons'ideration as from the date of publication of an interesting work by Pflugfelder (1950) have already rpassed over ten years, likewise as from the appearing of the book "The Problems of Immunity in the Helminthozoa" by Szichobałow (1950).
"Becker draws attention that he development of knowledge in the sphere of parasites, immunology may be not only of theoretical im- portance, but of practical one as well. If we shall recognize exactly what are the means of protection which parasites of human being and those of farm animals have to their disposal, then we will be able to start the researches after the measures for "disarming" them, and more effective control methods used as yet (Michajłow, 1950) ."
PROBLEM PASOŻYT-ŻYWICIEL fl8}1
I do not intend to quote the well known facts concerning the cellular reactions of immunity. I am obEged, however, to mention that a support for it is Miecznikoff's theory of phagocytosis, which is, as Zabłocki (1959)
~ays: "a wide bio-logical generalisation synthetizing a huge materiał
obtained when examining the diversiform mechanisms of no:1rishing and inflammation and infection in animals in different stage of their development". As it is known, phagocytosis relies upon inhibition of foreign bodies by a cell. In lower animals it is one of fundamental vital signs, and in the h:gher ones it is preserved in the cells of meso- dermal origin, it plays also a great role in numerous physiological and pathological processes, among others in the inflammatory and immuno- logical ones. Evidently, the later investigations proved the inflammatory process to be transgressing considerably with its symptoms beyond the mechanism of phagccytosis .. The learning on inflarnmation was especially enriched by Menkin (1935).
,_;rwrtly, the inflammation process in contemporary seizure and fur ther consequences resulting from it, with regard to morphology could be presented as follows:
The pathogenic factor, viz. bacteria, protozoa, helminthes, that pene- trated into the organism of the host and were retained in the tissues, causcs the damage of them, nor necessarily of mechanical (traumatic) character, but the most often of biochemical (necrohiotic changes), or simultaneously of biochemical and morphological character. The presence of protein originating from the decomposed cells causes the appearance of an inflammatory exudation. In the ex:1dation Menkin has found the following substances:
1. leucotoxin being a polypeptide causing an increased vessels per- meability for leucocytes or microphages and their positive chemotaxis as well;
2. leucocytosis promoting factor which is a pseudoglobulin that causes leucocytosis by acting upon marrow;
3. leucopenin - whkh reduces the leucocytes quantity when they are in excess;
4. necrosin - an albuminous su'bstance causing the necrosis pheno-- mena in the inflammatory centre;
5. pyrexin - a pyrogenic s:.rbstance;
6. exudin - callcd the leucocytosis promoting factor II;
7. leucopenic factor II.
According to Menkin, the mentioned factors cause the generał re- actions for the part of organism of the host. They appear in form of irritation of smaller blood vessels adjacent for the given centre, in form of slackened blood run in them, and in taking of a ma,rginal position in
tl90 H. SZWEJKOWSKI
lumen of the vessel of the white space blood elements and of passing by them to the lesion. Certain number of the cellular elements of leuco- cytes appereauer, oontaining the oxydase-positive grnnulas, is originating from cells of the vessel's adventitia. Hence, as result from it, some part of microphages descent from the haemopoietic area, and the other part is of local descent from the transformed cells. They are relatively less numerous in number as compared with the first ones.
Products of the deoomposed cells at the site of injury cause an increased acidity within the environment, and this in turn exerts a chemo- taxically positive influence :ipon macrophages or autochthonous histio- cytes or descended from blood. These elements, together with the preced- ing ones, are capable of developing the phagocytic activity; the remotion is their aim. As the leucocytes are adjusted to the extracellular proteo- lytic activity, the macrophages with relation to the decomposed cells are adjusted to the intracellular phagocytosis, which is far wider than the proteolitic activities of the microphages. The macrophages in certain order surround the parasite involved in the centre, and, according to Pflugfelder, have to supply a nourishment to the parasite. Moreover, there appear, especially when the process is lasting longer, the giant cells of foreign bodies.
Behind the inflammatory process phase that could be called the in- filtrating-exudative one, as the process is pro}onging, there follows the infiltrating...:proliferative phase characteriz.ed by appearance of cellular infi1tration that is converting in gradually maturing non specific gra- nulaticn in which a lot of fibroblasts occure.
The picture presented here very shortly, corresponds wilh the opinions which arose on the basis of classical Miecznikow's theory enriched by investigations of numerous succeders-Ribbert, Klemensiewicz, Aschoff, Kiono, Menkin, and others.
It is beyond the question that phagocytosis, as one of the factors of pathological regulation is the most primitive phylogenetically early way of protection of the organism. On a higher evolutionary level begins t·o produce nor only a cellular protection but humoral as well which corresponds the opinion of cellular origin of antibodies.
For oertain it doesn't happen this way as one believed formerly that antibodies are produced directly by the cells ,of the reticulo-endothelial system. Basing on ,Miecznikoff's theory and upon later Aschoffs's opinions, it way plainly supposed that the cells that are phagocytising foreign bodies (viz. bacteria, parts of parasites) are producing simultaneously antibodies that are adjusted for neutralizing or combating them. ln- vestigations carried into the antibodies content of micro- and macro-
PROBLEM PASOŻYT-ŻYWICIEL (_j!) 1
phages proved that these cells did not contain anti'bodies, although they are phagocytising the antigens.
In connection with this, the participation of macrophages in anti- bodies production is damitted in such a meaning that they are able to do same introductory activities to be relied on the antigens „disintegra- tion", but they have no direct connection with the synthesis of anti- bodies. The investigations have eliminated the opinion, which has had numerous disciples, conoerning the p1°oduction of antigenes by the reticulo-endothelial system.
Further investigations of which it resulted that the organs containing great quantities of lymphoidal tissue, first of all lymphatic glands and spleen, contained a great quantity of antiibodies (and extirpation of the spleen reduced the quantity remarkably) contributed to the theory of lymphocytic origin of these substances.
When investigating deeper into the matter it has been proved that within the lymphoid tissue the lymphocytes and similar to them plasmatic cells were not differentiated and sometimes the latter ones were accepted as separate developmental stages of lymphocytes. But later on it has been stated that they are separate products, especially when it has been convin.ced that ,1fter collecting blood from an immunized animal and putting in vitro of lymphocytes and plasmatic cells together with the antigene. the abs,Jrbtion of the latter one has appeared almost ex- clusively on the surface of plasmatic oells. The cortical part of 'the thymus, in which abund;..,ntly appear small lymphocytes, did not prove to contain antibodies. It agues against the lymphocytic theory of antibo- dies origin.
When it became clfar that antibodies are connected with alpha-, beta-, or gamma- globvlins the investigations have been directed to the place ::ind mer~hanism of their origin. Globulins are produced by the cells cyto- pla'.;m when stimulated by ribonucleic acid (RNA). The pnuuction of rib0nu.cleic acid is unde1· the influence of heterochromatin.
lt has been provE'd also that cells of high nucleoproteidy eontent have protoplasma eharacterized by an inereased affinity to 1the basie dyes (haematoxylin), they possess also an increased capability of being stained in comparision with pyronin. This feature is characteristie first of all for plasmatic cells, and only to a low degree possessit limphocytes.
As it is known the plasmatic cells are characterized by polygonal shape and by relatively small nucleus sit:1ated excentrically and of distinct capsule, of chromatin nodules appearing on i:ts internal side, from towarids the central part of nucleus, round about which appears a brigh- ter area while cytoplasm is distinctly basophilic. The specific peculiarity of the plasmatic cells is the staining with methylic green
+
pyronin.lifl'.! H. SZWEJKOWSKI
They appear in an increased quantity under normal conditions in sub- mucous membrane of intestines, in mucosa of oviductus and of uterus as well as in some inflammatory processes (an old granulation). In marrow as well as in lymphatic nodes they are thought to be d:fferentiated forms of reticulum plasma cells. Since they do not attain the circulating blood and likewise the loose connecutive tissue, nevertheless they appear in it - it remained only to accept the appearance of them as the result of transportation (histogenie plasmatic cells).
As the immun:ty was growing stronger it could be proved that the number of plasmatic cells was increasing prominently as well. The multi- plication of them was observed in the regions rich in reticular cells having, as it was mentioned just before, the greatest inclinations and possibilities to transform in plasmatic cells (or in macrophagcs when the bacteriaemia occurs), within the spleen, limphatic nodes, thymus and marrow. The areas in various animals prove to a different degree developed achvation ability; e. g. in rabbit-marrow and in mouse-spleen and limphatic nodes. According to more recent opinions, the expression of increased cytoplasmic (exported) albumen prod:J.ction would be in- creased pyronophily.
It was succeeded to prove that the increase in plasmatic cells number (for revealing of which the staining with methylic green and pyronin is considered as specific) runs paraUelly to the increased antibodies quantity of serum. Decomposition of the plasmatic cells causes the increase of gamma-globulins or of globulins antibodies in man, monkeys, and rabbits.
The greatest quantities of plasma-cells appear in the course of immunisa- tion in the spleen. The place and time of origin of the cells are differ- entiated and depend on the kind and place of introducing the antigen, on the kind of animal etc.
The origin of juvenile forms of the plasmatic cells of plasmoblasts and of proplasmocytes was observed already after 1-2 days in the limphatic nodes, regional for the place of introduction of the antigen, as -well as in the spleen. In rabbits the summit of plasmatic cell forma- tion fell on 5-7th day from the introducing of antigen (there were Pneumococci used).
As it vesults trom the respective 1iterature, the fragmentary antigen, divided by mac:rophages, has been found in the lymph flowing away out of inflammatory lesion after ca 12 hours. According to a theory represented by Breinl and Haurowitz-Mudd-Pauling, the antigen as penetrating into the plasmatic cells, influence upon suitable formation of globulin molecules, granting them a ·character of specyfic antigens.
The genetic theory by Ehrich assumes that the antigens drawing near to the active areas of mesenchyma, which are reticulum in the
PROHLEM PASOZYT-ZYWICIEL
spleen and lymphatic nodes, cause the transformation of the retic:1lum cells in specyfic plasmatic cells adjusted to produce the specific anty- bodies.
In hyperimmunisation of animals it is to be observed an abundant multiplication of the plasmatic cells that in a micros:oopic preparation, e.g. out of the lymphatic node, occupy the whole field of vision under mean magnification.
It has been observed, as well, that the pyronin absorbing cells of an exterior of the plasmatic cells are apt to occur also in the environs of the blood vessels in such organs as lungs, liver, kidneys, it would point out to the transformation of the adventitia ce11s into the cells serving for the increased production of the plasmatic protein.
The picture, I presented in my paper, is principally based on the rcsult of experiments conducted on rabbits with pneumococci applied as an antigen. Similar rcsults in my Institute has obtained P:usiński using as an antigen a weakened strain of Erysipelothrix (Staub's strain).
Because under the influence of antigens from the parasitic objects immunological _bodies have been obtained. based on s:milar biochemical substances, that arise when producing antibodies in the bacterial proces- ses, it could be assumed per analogiam that the similar regularities are operating here. Unfortunatcly, with respcct to the parasitolog:cal immuno- Jogy \Ne know only two effecis, viz. the initial one, corresponding with the irruption of the parasite, eventually corresponding with the introduction of the antigen, and the finał el'fect -- viz. the occurrence of the antigens in the- organie fluids. Yet v./e dcm·t kncw hov.' the anti- gcns ar L' c1·iginat.ing j n the prccf:'ss causcd by p,.ll'a!:i.t2s (perhaps by the participation of similar elements?), we dori"t know also whether the processcs arc accompanie<l by the proliferous phenomena v,rithin the de fi ni te of mesenchyma areas joined with the transformation of definite cellular forms.
May be it would be possible by the way to clrnl.' up the ,,:nvolvement"
of the cnrrcsponding tissue reacticns appearing parallelly and stimulating the pr'oduction of antibodics. This way it -could be possible also to make an attcmpt to determine the local production of antigens as well as the production :n the areas distant from the place of infestation of the parasite.
I mentioned in the beginning of my paper that nowadays the in- flammatory process, which expresses the adaptation of the organism.
could be characterized at the first exudative-infiltrating phase as the- remotic one. The next phase-exudative-proliferative one - has been se- parnted and is atributed to have a great significance in the adaphibility process.
fH)4 H. SZWEJKOWSKI
Although already Marchand exactly enough had examined the morphology of the proliferous reactions, however, the understanding of the stage as an adaptability phase became possible only when the reticulor-endothelial theory of the antibodies origin has been shadowed by the new introduced lymphatic-cellular or plasmatic-cellular theory.
Today it is beyond ,the question that the exudative-proliferative phase in the inflammatory process is a sign of the antibodies production, and thus a sign of adaptability (Ehrich, 1956).
The processes of antibodies production in a great measure are in- fluenced by the hormones. For instance the adrenocorticotropical hor- mone mobilizes the cortisone and hydrocortisone. As regards the first of the hormones, it is known that it reduces the vital function of the cells that are producing the antibodies, and is handicap:ng their pro- duction.
The vitamins and especially ascorbic acid seem to stimulate the pro- duction of antibodies.
Author's address : Warszawa. Grochowska 272
