Immunoregulatory activity of the lungs in relation to corpuscular and soluble antigens

The immunoregulatory activity of the lungs in normal Wistar rats has been evaluated by difference between primary and secondary immune response to the same dose of the antigen introduced into the respiratory tract or intravenously. As shown in this investigation, intratracheal immunization with corpuscular antigens is accompanied by faintly pronounced antibody formation and a high degree of delayed hypersensitivity, while the introduction of soluble antigens into the respiratory tract leads to the active production of antibodies. The immunoregulatory activity of the lungs is T-dependent. The preliminary introduction of corpuscular or soluble antigen into the respiratory tract is accompanied by the formation of the local mechanism in the lungs for suppressing immune response to the subsequent intratracheal immunization.     

Bovine babesiosis: Partial purification and characterization of blood culture-derived Babesia bovis

Morphologic, biologic and immunologic properties of corpuscular and soluble fractions of Babesia bovis purified from in vitro blood cultures were studied. Supernatant fluids obtained during routine medium exchange were studied. Supernatant fluids obtained during routine medium exchange were submitted to differential centrifugation to separate soluble and corpuscular babesial antigens from erythrocyte stroma. Extracellular babesiae were sedimented with infected and uninfected erythrocyte ghosts. The majority of babesiae were found in erythrocyte ghosts. Clumps of extracellular parasites were sometimes formed in vitro and generally could not be separated from uninfected erythrocytes. Centrifugation over a discontinuous Ficoll density gradient did not improve separations. Parasites remained viable throughout the purification procedure but were killed by freezing and rapid thawing. Both corpuscular and soluble antigen fractions elicited the production of specific anti-babesial antibodies when injected into calves. Electron microscopy of corpuscular antigen revealed the presence of intra- and extraerythrocytic babesial merozoites. A surface coat was visible loosely adhering to the plasma membrane of the parasites. Parasite suspensions and cell-free supernatant fluids obtained from in vitro cultures of B. bovis should provide a variety of unique antigens for further in vitro and in vivo studies.     

Physico-chemical properties of microbial antigens and the immuno-allergic tests]

Experiments were conducted on guinea pigs sensitized by the delayed and immediate types of allergy. Different antigens obtained from the strain of Brucella abortus BA-19 were used for sensitization and the resolving action. Comparison of the resolving properties of the corpuscular, soluble (ultrasound treated) antigens and purified protein fractions, polysaccharide and RNA was carried out in the skin reactions of the immediate and delayed type, passive skin anaphylaxis, acute anaphylactic shock, and the Schults-Dale test. Immediate reactions to the purified protein fraction were weaker than those to the whole soluble antigen, by which the animals were sensitized. Polysaccharide and the RNA-fractions proved to be inactive in the allergic reactions.     

An immunoenzyme method for demonstrating Coxiella burnetii antigens

Materials on the development of an enzyme immunoassay (EIA) system for the detection of the antigens of C. burnetii, the causative agent of Q rickettsiosis, are presented. The system is highly specific and effective with respect to both corpuscular antigens of phases 1 and 2 and soluble antigen (lipopolysaccharide). The sensitivity of this method varies within the range 5-100 ng/ml. The effectiveness of EIA as a quantitative (semiquantitative) control test used in the process of the production of Coxiella preparations has been demonstrated.     

Monoclonal antibodies to the species-specific and group antigens of Rickettsia prowazekii]

A number of hybridomas to different R. prowazekii determinants were obtained by the hybridization of spleen cells of BALB/c mice immunized with R. prowazekii corpuscular and soluble antigens. Some of the monoclonal antibodies (McAb) reacted with R. prowazekii thermolabile species-specific protein and did not react with R. typhi antigens (McAb of batches B4/4 and A-D3). McAb C5/2 and A3/2 reacted with the group thermostable antigen, common for R. prowazekii and R. typhi. McAb to the species-specific thermolabile antigen belonged to IgG2a. The McAb thus obtained permit the identification of R. prowazekii and R. typhi and the solution of the problem of the intragroup differentiation of rickettsiae belonging to the typhus group.     

Luminescent-serologic analysis of the antigenic interrelationship between R. canada and classic representatives of rickettsiae of the typhus group

The results of studying the antigenic relationships of R. canada, a new Rickettsia species, and classical Rickettsia species of the typhus group are presented. The study was carried out by luminescent serological analysis with the use of corpuscular antigens and the live infectious agent cultures. R. canada and Rickettsiae of the typhus group were similar in their antigenic structures; this, however, could be revealed only in the study of the live cultures of the infectious agents. The study of corpuscular antigens revealed unilateral relationship: R. prowazeki antigen could be detected with homologous and heterologous sera, R. canada antigen with homologous serum only. In the CFT and the agglutination test corpuscular R. canada antigen reacted with homologous and heterologous sera. The study of the live cultures of the infectious agents revealed that different R. prowazeki and R. typhi strains vary in the degree of their similarity to R. canada.     

Expression of multivalency in the affinity chromatography of antibodies. Appendix: Derivation and evaluation of equations for independent bivalent interacting systems in quantitative affinity chromatography.

The expression of multivalency in the interaction of antibody with immobilized antigen was evaluated by quantitative affinity chromatography. Zones of radioisotopically labeled bivalent immunoglobulin A monomer derived from the myeloma protein TEPC 15 were eluted from columns of phosphorylcholine-Sepharose both in the absence and presence of competing soluble phosphorylcholine. At sufficient immobilized phosphorylcholine concentration, the variation of elution volume of bivalent monomer with soluble ligand was found to deviate from that observed for the univalent binding of the corresponding Fab fragment. In addition, the apparent binding affinity of the bivalent monomer increased with immobilized antigen density. Use of equations relating the variation of elution volume with free ligand concentration for a bivalent binding protein allowed calculation of microscopic single-site binding parameters for the bivalent monomeric antibody to both immobilized and soluble phosphorylcholine. The chromatographic data not only demonstrate the effect of multivalency on apparent binding affinity but also offer a relatively simple means to measure microscopic dissociation constants for proteins participating in bivalent interactions with their ligands.     

Immunologic memory for Staphylococcus studied by adoptive transfer

The basic regularities of the formation and realization of immunological memory to staphylococcal corpuscular antigen were studied in adoptive transfer experiments on CBA mice. The capacity of spleen cells for generating anamnestic response to staphylococci in the body of irradiated syngeneic recipients appeared on day 3 after the immunization of donors. The formation of immunological memory to staphylococci in mice was shown to be directly related to the dose of the antigen. The study also revealed that intact splenocytes did not suppress the realization of immunological memory to staphylococci in the system of adoptive transfer. The conclusion of the absence of the "isogeneic barrier" for memory cells specific to staphylococcal corpuscular antigen was made.     

Use of the immunofluorescent microagglutination reaction for the purpose of studying ornithosis infection. 1. Production and approbation of a corpuscular luminescent diagnosticum]

A method of obtaining the fluorescent ornithosis corpuscular diagnostic agent providing for the extraction of the corpuscular antigen with its subsequent conjugation with fluorochrome-fluoresceine isothiocyanate. The use of this preparation permits to stage the agglutination reaction on the basis of immunofluorescent analysis, this facilitating the recording of the reaction results and considerably decreasing consumption of the diagnostic agent. As shown, the suggested immunofluorescent microagglutination reaction was characterised by a sufficiently high sensitivity, specificity, and can be used for detection of antibodies to the causative agent of ornithosis.     

Mitogenic activity of the antigens isolated from different Staphylococcus aureus strains in relation to mouse and guinea pig splenocytes

The mitogenic effect of corpuscular antigens with respect to the splenocytes of animals was found to depend on the strain of Staphylococcus aureus. The maximum synthesis of DNA in the cells was induced by corpuscular antigen Smith and the minimum synthesis, by Wood-46. The synthesis of DNA was activated in both B- and T-splenocytes in response to corpuscular antigens Wood-46, Cowan-1 and Smith, as well as to the cell wall and protein A. Peptidoglycan produced a mitogenic effect only in B-lymphocytes, and teichoic acid showed no mitogenic activity in mouse splenocytes. The mitogenic effect of staphylococcal antigens on splenocytes depended on the dose of the antigen and the time of cultivation. After 48-hour cultivation the incorporation of 3H-thymidine into the DNA of mouse cells was 5 times higher than into the DNA of guinea-pig cells. The optimum mitogenic dose in thymectomized BALB/c mice with respect to splenocytes was higher than in normal BALB/c mice practically by one order.     

Age-related changes in hematological and serum biochemical values in cats]

Nineteen hematological and serum biochemical values were analyzed for 91 healthy cats of both sexes (aged 1 to 48 months) that were bred and reared in our laboratory. Age-related changes were found for many parameters. Red blood cell counts (RBC), hemoglobin (Hb), hematocrit (Ht), Mean corpuscular constants, GPT, total protein (TP) and albumin (ALB) initially were low but increased then stabilized. White blood cell counts (WBC), alkaline phosphatase (ALP), inorganic phosphorus (Pi), total bilirubin (TBil), total cholesterol (TC), glucose (GLU), and triglyceride (TG) initially were high, but decreased then stabilized. No age-related changes were found for GOT, blood urea nitrogen, or calcium. Of the parameters that changed with age, the mean corpuscular constants, GPT, GLU, and TG became stabilized during the first 3 to 4 months of life, but others (RBC, Hb, Ht, TP, ALB) became stabilized after 9 to 11 months, during which period body weight reached a plateau. Some parameters (WBC, ALP, TG, Pi) showed change up to 18 months of age. These results suggest that cats 9 to 11 months old can be regarded as adults; but for some parameters, cats aged 18 months, or older, are better regarded as adults. Sex-related differences in the values for mean corpuscular volume, mean corpuscular hemoglobin, and WBC that were found after 11 months of age were higher in females. ALB was higher in males.     

Toxicity of Bordetella pertussis suspensions associated with the preparation of pertussis vaccine

The process of the detoxication of B. pertussis suspensions during their storage has a wave-like character and is determined by changes in the levels of the toxicity of the soluble and corpuscular fractions. Conditions facilitating the transition of toxic cellular products into the soluble state may lead to the increase of the toxic activity of pertussis vaccines.     

Relationship of the sensitizing and protective properties of pertussis antigens

When comparing delayed hypersensitivity (DN) to B. pertussis corpuscular antigens with the agglutinogenic composition of B. pertussis, as well as to its histamine-sensitizing, leukocyte-sensitizing, adjuvant and hemagglutinating activity, no correlation was detected between the specific sensitizing properties of these antigens and the serotype and studied nonspecific properties of B. pertussis. The DH level correlated with the protective activity of B. pertussis corpuscular antigens and the ultrasonic fractions of B. pertussis. The close correlation of these two phenomena suggest that DH-inducing and protective B. pertussis antigens are identical, though their action has different manifestations, depending on the method of administration of the antigen preparation. On these grounds a new method for evaluating the immunogenicity of B. pertussis antigens is proposed. This method consists in comparing the sensitizing activity of the antigen under test and that of the national standard.     

Specificity of suppressor cells of the delayed hypersensitivity type for Staphylococcus studied by immunoadsorption

The immunological specificity of T-suppressors obtained from mice after intravenous immunization with corpuscular antigen was shown. The splenocytes of such a mice suppressed DH to staphylococcal antigens, but not to sheep red blood cells. The suppressor cells under study were specifically adhesive to staphylococci.     

Detection of brucella antigens with colloid metal particles as markers for specific antigens

Gold and silver sols were comparatively approved as markers of specific IgG isolated from hyperimmune Brucella antisera for the detection of brucellar antigens. The sensitivity of the test system using gold immunosol proved to be some higher (3.1-9.8 ng/ml of soluble and 2.0 x 10(4)-5.3 x 10(6) CFU/ml of corpuscular brucellar antigens) than that achieved with the use of silver immunosol (5.7-18.4 ng/ml of soluble and 6.1 x 10(4)-8.0 x 10(6) CFU/ml of corpuscular brucellar antigens). At the same time silver sol was a cheaper and more available marker. Both test systems were found to be highly specific. False positive results were observed only with Yersinia enterocolitica O:9 at high concentrations due to the fact that they had common polysaccharide haptens incorporated into lipopolysaccharides of these microorganisms. The proposed test systems with colloid metal particles used as markers of specific antibodies for the detection of Brucella antigens are technologically simple, economic, rapid, highly sensitive and specific. Their use in combination with other serological methods will make the results of analyses more informative, thus improving the quality of laboratory diagnostics of brucellosis.     

Reactivity of Two Selected Antigens of Neisseria gonorrhoeae

Two antigen preparations, the soluble antigen and a fraction 1 thereof, isolated in the course of a systematic study of the various antigens of the virulent gonococcus, have been investigated for their ability to serve as antigens for the detection of antibody in patients infected with the gonococcus. The soluble antigen was reactive with 88.2% of the sera from infected females, and fraction 1 was reactive with 71.6% of the sera. Of sera from infected males, only 27.6% reacted with the soluble antigen and only 20.4% with fraction 1. Of sera from individuals presumed free of gonococcal infection, approximately 4% reacted with the soluble antigen; none reacted with fraction 1. This study suggests that these antigens might be adaptable to the detection of human gonococcal antibody, especially in the female.     

Immunological Response of the Rabbit to Vi Antigen

Vi antibody response of rabbits varied depending on whether Vi antigen was administered in particulate or soluble state. Vi antigen in particulate form induced hemagglutinins, bacterial agglutinins, and passive cutaneous anaphylaxis (PCA) antibodies, whereas soluble Vi antigen induced only hemagglutinins. Guinea pigs passively sensitized with antisera against particulate Vi antigen gave PCA reactions when challenged with either soluble or cellular Vi antigen; antisera against soluble Vi antigen were negative for PCA. The specificity of PCA was demonstrated by its dependence on the Vi concentration and by absorption of PCA activity from antisera with V-form cells of Salmonella typhosa.     

Immune response to ram erythrocytes and the cellular state of the monocytic phagocytosing system in the early period of a thermal injury

The degree of immune response to corpuscular antigen and macrophage reaction within 24 hours after thermal injury were examined in a comparative study. The number of antibody-producing cells, antigen engulfment and elimination by the cells of peritoneal exudate, their adhesiveness to glass, and cathepsin activity in the cells of peritoneal exudate and the spleen were determined. Mice subjected to thermal burns showed suppressed immune response and decreased functional activity of the cells of the monocytic phagocyte system within the first 24 hours, which was manifested by a decrease in the adhesiveness of these cells to glass, and their ability to engulf and destroy xenogenic erythrocytes.     

Immunogenic complexes of the soluble surface antigens of Sh. sonnei]

The comparative study of the immunogenic properties of Sh. sonnei (phases I and II) soluble surface antigens obtained by the modified method of aqueous-saline extraction and Sh. sonnei (phase I) antigen obtained by Boivin's method was made with the use of the keratoconjunctival test in guinea pigs. The protective activity of a high molecular fraction obtained by the fractionation of phase I soluble surface antigens in Sepharose 4B was studied. Boivin's antigen, when used for immunization in optimum doses, was found to have pronounced protective properties, whereas phase II soluble surface antigens showed no protective activity. A high molecular fraction obtained from phase I soluble surface antigen was found to be the most immunogenic. Protective activity was largely connected with protein antigen. The question whether protein antigen was an independent protective antigen or whether it constituted a part of a complex which determined the protective activity of a high molecular fraction remained unsolved.     

Identification of the soluble HVP-associated antigen of the lymphoblastoid cell line established from lymphomatous baboon (Papio hamadryas).

A new technique (indirect double immunodiffusion) for detection of EBV-associated soluble antigen and corresponding antibodies has been developed. This technique includes three steps: 1) simple double immunodiffusion with extracts of Raji cells (or other EBV-genome positive cells) and human sera containing antibodies against EBV-associated soluble antigen; 2) extensive washing and treatment with anti-human globulin; 3) extensive washing and treatment with tannic acid. Using this test it was shown that the soluble antigen indistinguishable from EBV-associated soluble antigen was present in KMPG-1 cells producing HVP.