Biomedical subjects
Publications and source records attributed to B Błaszczyk.
Allergic encephalomyelitis characterized by relapses and remissions was induced in chickens. The contents of M and G immunoglobulins in the serum was observed to increase at disease attacks and to decrease at its remissions. Similar correlation--though not so clear cut was noted with antitrypsin level. The usefulness of this model in pharmacotherapeutic investigations is discussed.
In the course of allergic encephalomyelitis (EAE) in chickens, azimexone suppressed the production of the specific IgM immunoglobulins. Moreover, it decreased the level of haptoglobin and sialic acid but stimulated the activity of antitrypsin. It was also observed to reduce the production of the plasmatic cells in the spleen. L-cysteine hydrazide hydrochloride decreased the level of sialic acid in EAE; intensified the activity of trypsin inhibitor and exerted no effect upon the level of antimyelin antibodies.
Specific anti-T and anti-B cell sera were used in treatment of allergic encephalomyelitis in chickens. The therapeutic effectivity of anti-T cell serum was very low. Unexpectedly, anti-B as well as normal serum amplified inflammatory reactions in the central nervous system. T and B cell count in peripheral blood was reduced according to the type of antiserum used. Alterations in the anti-myelin IgG, IgM and IgA antibodies level were typical for the course of allergic encephalomyelitis. Mechanisms of activity of anti-T and anti-B cell sera are discussed. We also show that the disease within the central nervous system is accompanied by the symptoms resulting from repeated administration of the xenogeneic sera.
The immune response measured by the level of specific anti-myelin antibodies and inflammatory process indicator-haptoglobin, were studied in chickens immunized with myelin basic protein (MBP). The histopathological changes in lymphoid organs were also investigated. The occurrence of specific anti-myelin antibodies was shown in sera of immunized animals. High level of antibodies was found in IgG class. The concentration of anti-myelin antibodies in the remaining immunoglobulin classes showed different levels. High concentration of haptoglobin (up to 2 g/l) was regularly noted in the course of allergic encephalomyelitis. All the above parameters were also evaluated in the course of experimental therapy with thymus factor X (TFX). This preparation distinctly reduced haptoglobin level in serum and had only slight effect on humoral immunity. In the course of inflammatory process plasmatic cells stimulation in the spleen was observed microscopically: this was even enhanced by the thymus hormone. The lymphoid system did not undergo any significant changes. The Harder's gland did not seem to have any influence upon the induction or therapy of MBP-induced inflammation.
Serological activity of swine IgM and IgG against Brucella abortus in RBPT was determined in relation to four other reactions used in Poland for diagnosing brucellosis standard agglutination test, complement fixation test, antiglobulin test, 2-mercaptoethanol test). Isolation of IgG was performed by the method of filtration on Sephadex gel G-200 of swine sera raised against Brucella abortus S19 by double immunization with suspension of killed bacteria. The presence of a certain Ig class in the fractions thus obtained was confirmed by immunoelectrophoresis and immunodiffusion tests. RBPT revealed the reaction of antibodies of IgM and IgG class which proves usability of this reaction diagnosis both early (IgM) and chronic (IgG) infection with brucellosis. Both classes of antibodies mentioned above were active also in SAT and CTT. Also the results obtained in AGT and MET were found interesting. In one of the sera, the absence of incomplete antibodies was observed, whereas positive reaction in antiglobulin test was found in its fractions containing IgG. This phenomenon was determined as concealment of incomplete agglutinins through higher level of complete antibodies in normal serum. In swine (the results were different from those obtained for cattle), apart from incomplete antibodies in IgG class, the presence of these agglutinins in IgM class was noted. On the other hand, the results obtained in MET proved that IgM antibodies of swine were not totally reduced when affected by 2-mercaptoethanol.
The influence of thymus hormone preparations on NK cell activity was studied. The following preparations were used: crude extract from calf thymus (TFX), its fraction V (TFX V) and synthetic pentapeptide (TP-5) fragment of native hemopoietin. It was shown that all preparations under study activate NK cells in vitro. The weakest effect was observed in the case of TFX, while both TFX V and TP-5 showed equally good stimulatory properties. The obtained results are discussed. The weak effect of the crude TFX is probably caused by the fact that it contains only 15% of fraction V which is responsible for affinity to the immune system.
Different times and temperatures of the 1st phase incubation of the complement fixation test (CFT) were used. It was found that a similar decrease in complement (C') activity occurred after 2 h at 37 degrees C, after 8 h at 20 degrees C and after 18 h at 4 degrees C. Simultaneously, in the same periods a double increase in standard serum titre at 4 and 20 degrees C was noted in comparison with titre at 37 degrees C. This increase of sensitivity of the CFT at 4 and 20 degrees C appeared to be a simulating one and resulted from the different pattern of reaction in each temperature in the block titration of antigen. The positive reactions in the range from the lowest to the highest dilutions of antigen were appearing and increasing and then disappearing in sequence at 4, 20 and 37 degrees C, creating the typical C' fixation area for each temperature. Similar results were obtained in four different C' fixation techniques, most frequently used in diagnosis of animal brucellosis. On the basis of our own examination results and literature data the warm and short C' fixation (37 degrees C, 30 min) was chosen in the standard technique of the CFT used in routine diagnostic survey for animal brucellosis in Poland.
The purpose of this paper was to prepare an indigenous standard of anti-Brucella abortus serum for the complement fixation test (CFT) as a homologue of the secon International Standard of anti-Brucella abortus Serum (ISABS-II), which contains 1000 international units of complement-fixing antibodies in 1 cm3. The indigenous Standard of anti-Brucella Abortus Serum (ISABAS) was prepared in 1975. The material used was the serum of a cow infected with Br.abortus, biotype 1, under natural conditions. The cow was beheaded on the 54th day after abortion. The serum obtained was filtered through a Seitz EK filter and appropriately diluted with normal bovine serum. The preparation was lyophilized in 8500 ampules, each of which contained 1 cm3 of serum. They were filled with nitrogen before closing them. It was shown that ISABAS activity in CFT was approximate to that of ISABS-II. The studies in CVL, Weybridge showed that one ampule contained 1115 international units of complement-fixing antibodies. ISABAS titres in the agglutination test (AT) and antiglobulin test (AGT) were 320+ + and 2560+ + + respectively. Reduction with 2-mercaptoethanol and separation on a column with Sephadex G-200 gel showed that anti-Brucella antibodies occurred in ISABAS mainly in IgG class. ISABAS lyophilizate is characterized by stability of CFT titre, good solubility and a low anticomplementary activity. The average weight of the lyophilizate in an ampule is 79.57 mg, standard deviation - 0.36%. ISABS may serve as the basis of antigen standardization and other elements of indigenous CFT technique in brucellosis.
The activity of cellular antigens and those as aqueous extracts prepared from 4 laboratory strains of Brucella abortus: 99, S-19, 119-3 and "Krotz" has been compared in this paper. The purpose of the studies was to select an antigen of the highest activity and to test it by the Polish Standard of anti-Brucella abortus serum. It was shown that cellular antigens are more active in CFT than aqueous extract antigens of the same brucella strain. Cellular suspension of Br. abortus strain S-19 was chosen for production of the domestic antigen. A chessboard system was used in standardization of this antigen. In the working dilution of the antigen in CF such an amount of it was used that gave the titre 250++ according to the Polish standard of anti-Brucella abortus serum. The multiplier 4 (1000:250) was obtained which serves to calculate CFT titres in terms of international units of the complement fixing antibodies (icftu) in 1 cm3 of the serum studied. In examinations of all species of farm animals by the standard CFT technique, 10-20 icftu were assumed as a doubtful result and greater than or equal to 20 icftu as a positive result. A definite excess of the antigen used in working solution limits the prozone range and permits to carry out CFT in mass diagnostic examinations initially using one lowest serum dilution.
A differentiated time and temperature of incubation were used in the first CFT phase. It was found that the activity of complement fixation (C') decreased in the reaction medium. This was observed in the control system in which at 1 C'H50 hemolysis inhibition increase from 50 to 90% was found after 2 hr at 37 degrees C, after 8 hr at 20 degrees C and after 18 hr at 4 degrees C. Simultaneously, a double increase of the standard serum titre was obtained at 4 and 20 degrees C as compared with that at 37 degrees C. At stable concentration of the antigen and complement the effect of C' fixation was the strongest at 4 degrees C, slightly weaker at 20 degrees C and the weakest at 37 degrees C. The above dependence resulted from a different reaction distribution in the chessboard system at each temperature. In the order from the lowest to the highest antigen dilutions positive reactions were increasing and then subsequently expired at 4, 20 and 37 degrees C, forming an optimal antigen dilution typical for each temperature. In our own standard CFT technique a heat and short C' fixation (37 degrees C for 30 min) were chosen which provided the required reaction sensitivity and permitted to make mass diagnostic examinations. It was also shown that the temperature and time of initial incubation of the antigen-antibody complex had no influence on the effect of C' fixation by this complex. This is supported by joint introduction of the antigen and complement into sera.
The purpose of this paper was to developed simple and accurate methods for standardization of the particular components of a hemolytic system and to choose optimal amounts of these components in CFT. On the basis of comparative studies and the data from literature the following procedure was chosen: 1. Sheep's blood was conserved with Alsever liquid and used after 5 days of stabilization. A 2% suspension of erythrocytes was standardized by centrifuging in calibrated test tubes at 1000 r for 10 min and spectrophotometrically, assuming the following criterion: the suspension of erythrocytes dissolved in distilled water at 1:10 ratio should show 0.500 extinction (E541), which corresponds to 5 X 10(8) erythrocytes in 1 cm3. 2. Lyophilized hemolysin was dissolved, conserved with glycerol and used after 5 days of stabilization. Hemolysin was titrated with 100% hemolysis in the presence of the excess of complement. The suspension of erythrocytes was sensitized with 5 units of hemolysin directly before using it. 3. Lyophilized complement (C) was dissolved, conserved with boric acid +K2SO4 containing liquid and used after 5 days of stabilization. Titration of C was done by 50% hemolysis; 3C'H50 was used in CFT. 4. A barbital buffer was used as diluent at pH 7.3-7.4, alternatively 0.85% NaCl with an addition of Ca and Mg at optimal concentration. The hemolytic system was incubated at 37 degrees C for 30 min.
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The studies concerned the effect of cyclophosphamide on immunological and biochemical indices of MBP-induced encephalomyelitis. Unlike isoprinosine, cyclophosphamide was observed to exert irregular effect. Contrary to the expected, the results obtained in some experimental systems show increased level of the parameters tested-IgG, antitrypsin and sialic acid. A suggestion has been put forward that the reason of the above phenomenon may be attributed not only to the type of drug applied but also to different course of treatment with regard to various affinity of the drug (different doses) to functionally different lymphocytes (suppressor, helper).
Concentration of haptoglobin (Hpc) in serum of chickens with inflammation and allergic encephalomyelitis (EAE) was determined. Chickens with aseptic inflammatory state revealed 2.5 times higher concentration of Hpc while in the course of EAE only small increase was observed. Administration of immunomodulator--isoprinosine, resulted in elevation of Hpc level as compared to nontreated chickens.
It was observed that in the course of allergic encephalomyelitis (EAE) in chickens. Isoprinosine reduced the production of the specific IgA and IgM antibodies against encephalic antigen. Its influence on IgG production was slight. It was also found that isoprinosine reduced the antitrypsine activity in EAE. Its effect upon the level of antibodies and biochemical indices of inflammation was particularly pronounced on days 28-35 after immunization, i.e. at the time of the most intensive microscopic inflammatory changes in the brain. Histological analysis revealed an inhibition of plasmatic cells reproduction in the spleens of chickens receiving isoprinosine. No effect of the drug on the GvH reaction in chickens was found.
The effect of thymectomy performed on the day of hatching has been studied in chickens rendered tolerant by 100 mg of HSA injected also on the day of hatching. After challenge at the age of 6 and 9 weeks, no significant difference was observed between the group of tolerant chickens, in which the thymectomy was complete or incomplete, and the non-operated tolerant one. The delayed recovery from tolerance observed in mammals was not seen even in chickens in which thymus was removed completely and no thymic residue remained. The anti-HSA antibodies were even slightly higher in thymectomized birds than in the non-operated ones but the difference was statistically non-significant.
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