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[Isolation of Ancylostoma duodenale antigens and production of immune sera].

Rabbits were immunized using intact larvae or homogenates from Ancylostoma duodenale. Antisera were tested by immunodiffusion. The homogenates promote the formation of antibodies but the intact worms were not able to induce them. The antisera were partially purified by precipitation with amonium sulphate 40% saturation and filtration through Sephadex G-200. The purified material was attached to Sepharose 6B and used as immunoabsorbent for the isolation of the antigens from the soluble extracts of parasites. The isolated antigens were used in order to obtain new antisera. These antisera were used for the preparation of more efficient immunoabsorbent which allow to isolate new antigens that gave three precipitation lines by immunodiffusion. The polyacrylamide gel electrophoresis of crude homogenate discriminate 12 components, and the electrophoresis of the isolated antigens gave only 3 bands.

Adjuvants, Immunologic↗
PubMed1975

The application of polyvalent horse immune sera for electroimmunodiffusion methods.

Horse immune sera do not give satisfactory results in immunochemical techniques based on electrophoresis of antigens through antibody-containing agarose gel. As the majority of precipitating horse antibodies belongs to the beta globulins, they migrate in the gel during electrophoresis. After enzymatic treatment the pepsin fragments work well in all electroimmunodiffusion methods.

Animals↗
PubMed1977

[Immunofluorescence with PI-3 virus--preparation and evaluation of conjugates from immune sera of various animal species].

Immune sera with a high content of antibodies to para-influenza-3 (PI-3) virus suitable for production of specific conjugated immunoglobulins were prepared on conventionally reared calves, rabbits, and guinea-pigs, applying antigen in different ways. When processing the sera of the above mentioned animal species to conjugates, the sera differed in pure IgG yield, in completed conjugate, and molar F/P ratios, but their ability to demonstrate antigens of the PI-3 virus in the dilution was approximately the same. The serum prepared on a colostrum deprived calf showed the lowest antibody content and the lowest pure IgG yield, the colouring ability of the conjugate prepared being as well somewhat lower.

Animals↗
PubMed1978

Experimental immune pancreatitis in the mouse by rabbit immune sera directed against purified enzymes of the exocrine pancreas.

An experimental xenogeneic immune pancreatitis was induced in AB-mice by repeated intraperitoneal injections of rabbit immune sera directed against purified pancreatic enzymes (alpha-amylase, lipase, trypsin) for 3 hours up to 8 days. Histologically, the immune pancreatitis is characterized by three different findings: 1. Multiple acinar cell necroses on the 2nd, 3rd and 5th day of immune serum application. 2. A dedifferentiating acinar cell atrophy with development of pseudocanalicular acini on the 5th and 9th day. 3. An increasing interstitial histiolymphoplasmocytic pancreatitis on the 5th and 9th experimental day. Ultrastructurally, the acinar cell necroses proved as the final stage of a step-by-step developing acute lethal cell damage. The dedifferentiating acinar cell atrophy corresponds to a chronic sublethal cell injury with alteration of different cytoplasmic components. The interstitial pancreatitis in immune serum treatment is characterized by differently activated histiocytes and lymphocytes as well as by mature plasma cells. Because of immune histological findings (peri- and intraacinar deposition of rabbit globulin, specific fixation of guinea-pig complement, and appearance of mouse globulin in the mouse exocrine pancreas) and control experiments with rabbit and mouse normal serum as well as with physiological saline, the pathogenesis of the induced xenogeneic immune pancreatitis is regarded as a twophase process: 1. The acinar cell necroses are mainly due to a cytotoxic immune reaction (possibly in combination with an immune complex reaction) caused by specific anti-pancreatic enzyme antibodies of the applied immune sera. The dedifferentiating acinar cell atrophy may be the result of a specific action of the anti-enzyme antibodies against the corresponding pancreatic enzymes in the apical secretion granules of the pancreatic acinar cells. 2. The interstitial histiolymphoplasmocytic pancreatitis is mainly the morphologic substrate of an extravascularly (intraperitoneally) induced serum sickness reaction (immune complex reaction) due to the foreign proteins applied with the xenogeneic immune sera.

Amylases↗
PubMed1978

Preparation of immune sera against carcinoembryonic antigen (CEA).

CEA antigen obtained by the Wrocław team and CEA standard antigen from Chester Beatty Cancer Research Institute, London were used for immunization of goats. Two immunization methods were adopted: one with decreasing antigen doses beginning from 500 microgram and the other with equal, small doses of 10 microgram. Anti-goat IgG1 + IgG2 immune serum was obtained by immunizing a horse with 10 mg of the antigen with complete Freund's adjuvant. Anti-CEA immune sera obtained after immunization with small doses were highly active; in the RIA test they showed half maximum CEA binding at 1:40,000 and 1:80,000 dilutions. Likewise, the anti IgG1 and IgG2 serum from the horse proved to be highly active.

Animals↗
PubMed1976

Studies on the Treponema pallidum immobilizing activity in normal human serum. 3. The kinetics of immobilization reaction of normal and immune sera.

The influence of immobilizing antibody, complement and lysozyme, on the T. pallidum immobilization reactions of normal and immune sera was studied. Lysozyme shortened the lag periods and increased the reaction rates of the reactions of normal and immune sera. At high concentrations of added lysozyme, variations in the concentrations of immobilizing antibody and complement, within a wide range, did not further influence the kinetics of the two reactions. Preincubation with lysozyme did not influence the treponemes in the following immune serum immobiliation reaction provided the lysozyme was removed before the addition of antibody and complement. Normal serum was found to immobilize T. pallidum more rapidly than immune serum. This was seen also if the reaction mixtures were almost identical, the only differences being the immobilizing IgM antibody involved in the normal and the IgG antibody involved in the immune serum reaction.

Blood↗
PubMed1975

Protection against scrub typhus infection engendered by the passive transfer of immune sera.

The passive transfer of convalescent sera did not protect the majority of mice against challenge with the homologous strain and was completely ineffective against challenge with strains unrelated by fluorescent antibody techniques. When the immune sera was incubated with the rickettsia in vitro and then inoculated into the mice a dramatic increase occurred in the number of surviving mice. The importance of these data in relation to published results with other species of rickettsia is discussed.

Animals↗
PubMed1979

Precipitation of visna viral proteins by immune sera of rabbits and sheep.

Eighteen polypeptides equivalent to 1.2 x 10(6) daltons of visna virus were specifically precipitated by immune sera from rabbits and sheep. The hyperimmunized rabbit antisera contained high concentrations of antibodies against p25 and p14, whereas the sera from sheep actively infected with visna virus showed a large quantity of anti-gp115 antibody. The results indicate that almost all the polypeptides reported previously (F. H. Lin, J. Virol. 25:207--214, 1978) are virus-specific components of visna. The presence of anti-gp115 antibody in sera of infected sheep may offer a simple and sensitive diagnostic procedure for visna.

Animals↗
PubMed1979

Discrimination of Antibodies to Hepatitis B surface antigen from antibodies to inherited serum protein variants in immune sera of human and animal origin.

Antisera to hepatitis B surface (HBs) antigen, both of human and rabbit origin, have been examined. The anti-HBs serum derived from a multiply transfused patient did, in addition, contain antibodies directed against the inherited beta-lipoprotein antigen Ag(x), whereas one of the rabbit immune sera also contained antibody to the inherited Lp(a) antigen. Thus, in human, as well as in animal anti-HBs sera, antibodies to inherited normal serum antigens may cause false positive reactions. This problem may be overcome by absorption procedures if appropriate control systems are available. False positive reactions caused by the Ag(x) antigen may also be avoided by use of agarose as supporting medium for the test, as Ag(x) precipitin lines do not appear in this medium. An undialysable high molecular weight component may be obtained from Oxoid "Ionagar" by washing. When this is added to agarose the Ag(x) antigen reaction appears also in this medium.

Absorption↗
PubMed1977

[Migration capacity of tumor cells treated with immune sera].

The action of antitumour sera on the adhesive properties of the L-cells and the cells of Ehrlich's ascitic carcinoma. It was shown that after the treatment with immune sera the tumour cells lost their capacity to form rosettes with sheep red blood cells, to adhere to plastics and glass; when administered into the mouse organism these cells were worse retained in the lungs, spleen and the liver. The data obtained indicated that the antibodies could play a significant role in the tumour cell metastasis.

Animals↗
PubMed1976

Purification and serological characterization of the major envelope glycoprotein from AKR murine leukemia virus and its reactivity with autogenous immune sera from mice.

The major envelope glycoprotein (gp71) from AKR murine leukemia virus (MuLV) was purified and its serological reactivity with heterologous and autogenous immune mouse sera was examined. Homologous and interspecies competition radioimmunoassays using antisera to Rauscher-MulV gp69/71 or Friend-MuLV gp71 or antisera to feline leukemia virus to precipitate 125I-labeled gp71 from various MuLV showed that distinct differences exist between Rauscher- or Friend-MuLV and AKR-MuLV glycoproteins. Characteristically the AKR-MuLV gp71, in contrast to FLV or RLV gp71, does not compete fully in homologous or interspecies radioimmunoassays with iodinated Friend of Rauscher glycoproteins. Purified 125I-labeled AKR-MuLV gp71, in contrast to the Rauscher- or Friend-MuLV glycoproteins, reacts with normal (autogenous immune) mouse sera in direct radioimmune precipitation assays. Competition experiments further demonstrate that this is a predominant immunological reactivity of normal mouse sera which had previously been detected by radioimmune precipitation assay against intact virions.

AKR murine leukemia virus↗
PubMed1979

Effect of immune sera upon enhanced in vitro antibody responses.

Immunization of rabbits with S marcescens bacteria in Freund's incomplete adjuvant resulted in development of anti-Serratia sera with bacterial agglutinating properties as well as LPS modulating properties. The antisera reacted with the LPS in counterimmunoelectrophoretic assays. Such antisera also stimulated the LPS induced enhancement of specific anti-SRBC responses from normal spleen cell cultures but did not further enhance the stimulated responses of similar cultures incubated with the smaller molecular weight PS-rich derivative. The antisera had no effect on LPS or PS enhanced nonspecific background anti-SRBC response as compared to normal rabbit serum-treated controls. These results point to a possible role for immune complexes as an enhancer of the immunostimulating activities of LPS antibody-treated cultures and also indicate a difference between LPS and PS as immunomodulators.

Animals↗
PubMed1977

Preparation of specific immune sera against rabbit skeletal, pig and human cardiac heavy meromyosins.

Antibodies were produced in guinea-pigs against heavy-meromyosin (HMM) and tested by immunodiffusion and microcomplement fixation (MCF). A similar response was obtained for HMM extracted either from human and pig left ventricles, or from rabbit white skeletal muscles. For each muscle, the antisera cross-react quantitatively with the native intact myosin and are highly specific. Thus, this procedure permits the preparation of muscle type-specific anti myosin antibodies, by a simpler means than those previously published.

Animals↗
PubMed1979

Effect of 2-deoxy-D-glucose on the binding of preimmune and immune sera to herpes simplex virus-infected cells as measured by quantitative immunofluorescence.

The binding of immune and preimmune rabbit sera to fixed VERO cells infected with herpes simplex virus type 1 (HSV 1) was measured by quantitative immunofluorescence. False positive binding of the preimmune serum (PS) occurred from 8 hr post infection (p.i.), while the immune serum (IS) revealed positive fluorescence starting with 4 hr p.i. The false binding of PS decreased significantly (P = 0.01) if the cells were cultured in the presence of 2-deoxy-D-glucose (DOG). Statistical analysis of data on the binding of IS to infected cells kept in the presence or absence of DOG showed that the mean values of immunofluorescence were not significantly different (P less than 0.1). Cells cultured for 8 hr p.i. in the presence of 6 mM DOG seem to be a suitable antigen for detection of the actual level of specific antibodies by indirect immunofluorescence.

Antibodies, Viral↗
PubMed1978

The characterization of antibacterial antibodies in bovine immune sera to Staphylococcus aureus.

Humoral antibody responses to the encapsulated Smith diffuse strain of Staphylococcus aureus were examined in cows immunized with the killed vaccine via different systemic routes. The sequential appearance of the antibody within different immunoglobulin classes in the sera during the course of immunization was followed by passive hemagglutination (PHA) and precipitation (PC) reactions and the mouse passive protection test. Repeated intravenous injections with the killed vaccine suspended in buffered saline stimulated production of IgM antibody exclusively during the whole period of immunization. On the contrary, following intramuscular administration with the vaccine incorporated in Freund's incomplete adjuvant, the antibodies appeared predominantly in IgG fractions of the sera. Specific antibody to the homologous strain used for vaccination was prepared from bovine immune sera by an absorption and elution process. The mouse passive protective activity of the antibody preparation was removed by absorption with the capsular polysaccharide antigen as well as by the whole cell adsorbent of the Smith diffuse strain, but not by the Smith compact and Cowan I strains of S. aureus. IgM, IgG1 and IgG2 proteins were isolated from the purified antibody and were compared, on a weight basis, with respect to their biological activities. Slightly higher activity of the IgG over the IgM antibody was demonstrated both in the mouse passive protection test and PC reaction, whereas in the PHA reaction, IgM antibody was shown to possess a significantly higher activity than IgG antibody. These studies suggest that IgG as well as IgM antibody might play an important role in protection against infection with encapsulated strains of S. aureus in cows.

Animals↗
PubMed1979

Specific precipitation of mouse and rat alpha 2-macroglobulin by immune sera directed against murine leukemia viruses.

Leukemia viruses preparations obtained from plasma of leukemic mice or rats were used to immunize rabbits. By immunoelectrophoresis, these non-absorbed antisera reacted specifically with one single mouse serum alpha 2 globulin which was further identified as alpha 2-macroglobulin by several criteria, including gel chromatography and nitracentrifugation. When mouse plasma derived virions were used as antigen, they gave rise to antibodies cross-reacting completely with the corresponding mouse and guinea pig protein and partially with rat and calf globulin. Immunization of rabbits with rat plasma derived virions induced antibodies directed preferentially against the rat alpha 2-macroglobulin.

Animals↗