新生幼犬的被动免疫研究

新生幼犬通过初乳获得的被动免疫,对它的生存非常重要,本研究用犬的多价免疫血清代替初乳作为幼犬保护性免疫球蛋白,对生后2 d内不能获得初乳的幼犬,以及虽然获得初乳但初乳中特异性免疫球蛋白含量较低的幼犬进行了被动免疫试验。对5窝35只幼犬在生后,随机分为吃初乳的对照组、口服血清组、皮下注射血清组、以及口服和皮下注射血清又吃初乳5个组,在幼犬出生时、及出生后48 h和第5天采血,测定犬细小病毒、犬传染性肝炎、和犬副流感的血凝抑制（HI）抗体效价。结果表明：对没有吃到初乳的仔犬,通过早期口服或皮下注射途径给予成年犬的血清,均能代替初乳供给幼犬免疫球蛋白。对吃到初乳的仔犬,能进一步提高幼犬体内特异性抗体水平,但以皮下注射途径最佳。     

Antigenic studies of a VH fragment: demonstration of three sets of antigens, idiotypic, VH subgroup, and VH framework-specific antigens.

The antigenic properties of the VH region of immunoglobulin heavy chains were studied by means of a fragment corresponding to the variable part of the heavy chain of an IgG3 myeloma protein (KUP) and an antiserum made against this fragment. By hemagglutination, hemagglutination inhibition, and immunofluorescence techniques, it was shown that the anti-VH antiserum detected three sets of antigens in the VH region, namely idiotypic antigens, VH subgroup-specific antigens, and VH domain-(framework) specific antigens. The VH fragment inhibited in a VHII subgroup-specific hemagglutination inhibition test system. The VH fragment was thus antigenically similar to the tvh region found in the intact molecules and the light chains were not needed to express the VH subgroup antigens or the VH framework antigens.     

Pili (fimbriae) of the causative agents of plague and pseudotuberculosis: their detection in the hemagglutination reaction, the passive hemagglutination reaction and in an immunoenzyme analysis system

Newly developed serological methods for the detection of pili in the passive hemagglutination (PHA) test with the use of immunoglobulin erythrocytic diagnosticum and in the enzyme immunoassay (EIA) with the use of specific immunoglobulins labeled with horse radish peroxidase have been found to exceed the method of detecting pili, based on the determination of their hemagglutinating activity, in sensitivity and specificity. Besides, the PHA test and EIA have proved to be capable of detecting low molecular fragments of pili, obtained by sonication and having lost their hemagglutinating activity.     

Erythrocytic diagnostic agents for determining antibodies to Proteus O antigens

Highly sensitive and specific erythrocyte diagnostic agents (ED) for the determination of antibodies to Proteus O-antigens have been obtained by the sensitization of formolated sheep red blood cells (SPBC) with activated lipopolysaccharides (LPS) without the use of mediators. The tannin treatment of formolated SRBC and/or the increase of temperature from 45 degrees C to 100 degrees C in the process of the preparation of ED have been found to produce no increase in effectiveness. Antibody ED permitting the detection of Proteus O- and H-antigens has been obtained by the sensitization of formolated chick red blood cells with immunoglobulin preparations to Proteus hydroxylamine antigens, carried out with the use of amidol. The experiments have shown the possibility of using this antibody ED for the determination of O-antibodies in the antigen neutralization test with nonactivated LPS used as an agglutinating agent. The passive hemagglutination test with antibody ED has proved to be a more sensitive method for the detection of O-antibodies than the antigen neutralization test with antigenic ED. The determination of Proteus etiology in the passive hemagglutination test with the use of antigenic ED has been shown to be highly effective in the examination of patients with chronic osteomyelitis at the stage of exacerbation.     

The results of a controlled field trial of a dried erythrocytic immunoglobulin diagnostic agent for detecting the hepatitis A virus antigen]

The results of the controlled field trial of lyophilized erythrocytic immunoglobulin diagnosticum for the detection of hepatitis A virus antigen in the urine and feces of patients are presented. This diagnosticum was used for the study of urine and fecal samples from 225 patients (of these, 176 had hepatitis A) and 54 healthy persons in the passive hemagglutination (PHA) test. Their blood sera were studied in the PHA test (to detect HBsAg) and the radioimmunoassay (to detect anti-HAV IgM). The immunoglobulin diagnosticum under study was found to be nonspecific and faintly sensitive and, therefore, unsuitable for use in medical practice.     

Use of the specific rosette formation test in tick-borne encephalitis for the purpose of laboratory diagnosis

In the study of blood samples collected from patients with tick-borne encephalitis, the modified antigen-specific rosette formation test with erythrocytes, loaded with tick-borne encephalitis virus antigen via the specific immunoglobulin, has been used. The number of rosette-forming cells has been the highest during the acute period of the disease. The use of this test has confirmed the clinical diagnosis of the disease, together with hemagglutination inhibition serving as the main diagnostic test, in 35% of cases. The results of this study make it possible to recommend the antigen-specific rosette formation test for the early diagnosis of tick-borne encephalitis.     

Detection of HBs antigen with latex particles coated with human antibody prepared by affinity chromatography on concanavalin A-sepharose.

Human HBs antibody was isolated by affinity chromatography on HBs antigen absorbed to concanavalin A linked to Sepharose 4B. When a human anti-HBs immunoglobulin preparation obtained by Cohn's cold ethanol fractionation method was used as a starting material, the antibody was concentrated about 10 times in terms of the passive hemagglutination titer with a recovery rate higher than 50%. Latex particles coated with human anti-HBs antibody thus prepared were proved to be useful in detecting HBs antigen in human blood samples. In its sensitivity and in rapidity of its performance, the antibody-coated latex agglutination test seems to be superior to conventional immunodiffusion techniques.     

Rubella Hemagglutination Inhibition: Removal of Nonspecific Agglutination Due to Manganous Chloride

Nonspecific inhibitors of rubella hemagglutination can be removed by treatment of sera with heparin-manganous chloride for use in the hemagglutination-inhibition test. After removal of nonspecific inhibitors by this procedure, an excess of manganous chloride may remain. This may cause the cells to agglutinate, thus obscuring the reading at low serum dilutions. This disadvantage can be overcome by the addition of sodium carbonate, which forms an insoluble compound with manganous chloride and does not interfere with antibody determination. The procedure presents a further refinement of the hemagglutination inhibition test for rubella by increasing specificity and sensitivity; it permits detection of antibody levels as low as 1:4 in sera.     

Measles antibodies in exocrine secretions

The results of the analysis of saliva samples taken from 157 persons aged 1-48 years for the presence of antimeasles antibodies in the neutralization test, the hemagglutination inhibition test and the passive hemagglutination test are presented. The data obtained in this study suggest that antimeasles antibodies can be detected in saliva for many years after the formation of immunity, but quickly disintegrate after a saliva sample is taken.     

Detection of postvaccination mumps virus antibody by neutralization test, enzyme-linked immunosorbent assay and sensitive hemagglutination inhibition test

A group of 251 children aged 2-3 years given live attenuated mumps virus vaccine PAVIVAC of Czechoslovak production were tested for antiparotitis antibody levels in pre- and postvaccination sera by neutralization test (NT), enzyme-linked immunosorbent assay (ELISA) and sensitive hemagglutination inhibition test, enhanced by heterologous antibody to human immunoglobulin G (E-HIT). The prevaccination findings were as follows: positive ELISA IgG titres, neutralization antibodies and hemagglutination inhibition antibodies were present in, respectively, 35%, 25.9% and 27.9% of the sera. Postvaccination seroconversions were evaluated in 159 susceptible vaccinees whose prevaccination sera had been negative by all three tests. The lowest seroconversion was detected by NT (74.2%), seroconversions by ELISA and E-HIT were appreciably higher (82.4% and 86.8%, respectively). The seven children showing a seroconversion by E-HIT but not by ELISA had a 4 fold increase of anti-mumps ELISA IgG antibodies as well, but the rise of antibody titres was at a level falling in the range below the positivity criterion for ELISA. The statistically evaluated detection rate for antibodies was significantly higher (significance test "t") by ELISA as compared with neutralization test. However, antibody levels (geometric mean titres) were 8-10 times lower in postvaccination sera than in convalescent sera of 30 children with mumps in all three tests.     

Release of bound immunoglobulin from SSPE brain by acid elution.

SSPE brain homogenate extracted at pH 7.4 yields immunoglobulin with a 4- to 5-fold greater hemagglutination inhibition activity per microgram of IgG than serum from the same patient. Serial washing of the homogenate results in a low level steady-state release of IgG. Elution of the washed sediment with pH 2.5, 0.1 M glycine buffer results in a 2- to 3-fold increase in recovery of hemagglutination inhibition activity with a greater hemagglutination inhibition activity per milligram of IgG than the IgG recovered by phosphate-saline extraction at pH 7.4.     

Immunochemical properties of immunoglobulin G conjugated with dextran

Antigen-binding activity and effector functions of immunoglobulin G from horse and rabbit sera have been investigated, using hemagglutination, kinetic immune lysis, immune lysis in microplates and rosette-forming test with peritoneal mononuclear cells of mice, after their conjugation with dextran, MW 35-50 kD. The formation of conjugates of two types has been demonstrated: protein-dextran and protein-dextran-protein. It has been revealed that protein-dextran-protein conjugates have high specific antigen-binding activity, as compared to native IgG from rabbit sera specific for SRBC, while interactions with the complement system and Fc receptors is depressed.     

Cervical secretory immunoglobulin A to human papillomavirus type 16 (HPV16) from HPV16-infected women inhibit HPV16 virus-like particles-induced hemagglutination of mouse red blood cells

Secretory immunoglobulin A (sIgA) antibodies are the first line of defence at the genital mucosa, and are thought to hinder viral infections by binding to conformational epitopes on the viral capsid. To investigate if cervical sIgA binds to conformational epitopes of the Human papillomavirus type 16 (HPV16), cervical mucus samples from 109 HPV16-infected patients were examined in a HPV16 virus-like particles-induced hemagglutination inhibition assay. 48 (44.1%) patients were able to inhibit hemagglutination. Inhibition of hemagglutination was associated with the presence of sIgA (P=0.001). In conclusion, naturally occurring cervical anti-HPV16 sIgA binds to and hinders conformational epitopes on the viral capsid, suggesting that these antibodies might have a neutralizing capacity.     

The possibility of using equine serum albumin in place of bovine serum albumin and ovalbumin in radioimmunological and immunoenzyme analyses and in virological practice

Horse serum albumin has been shown to meet the requirements to protein preparations for microanalysis and thus to be suitable for use in kits of reagents for the radioimmunological determination of insulin and myoglobin, for the determination of tick-borne encephalitis virus antigen by the method of the enzyme immunoassay and for the stabilization of proteins in the hemagglutination test and the hemagglutination inhibition test.     

A trial application of the latex test for evaluation of antibody level in rabbit immune sera]

A latex test was elaborated which served for evaluation of quality of rabbit immune sera for antigen 0 of selected Gram-negative bacteria. Sensitivity and specificity of this test in comparison with passive hemagglutination and immunoenzymatic DOT-ELISA reactions was evaluated. These studies were performed on immune sera for antigen O of Salmonella groups B, C1, C2, D and E, Yersinia pseudotuberculosis and in antigen preparations for above listed microorganisms both in homologous and heterologous systems. It was found that sensitivity of the latex test is 9 to 160 times lower than that of passive hemagglutination and 7 to 307 lower than for DOT-ELISA. Sensitivity of the latex test and passive hemagglutination reaction was evaluated on the basis of results of cross reaction between studied antigens and unabsorbed rabbit sera, establishing so called sensitivity indexes, which were informing how many times heterologous titer is lower than homologous titer. So evaluated sensitivity of the latex test was close to sensitivity of the passive hemagglutination reaction. It was found that slide latex test is characterized by satisfactory sensitivity and good sensitiveness and may be used for evaluation of antibody level 0 antigens of Salmonella and Yersinia. The value of this test is characterized by high repeatability of results, as well as low work and time-consuming.     

Detection of Foot-and-Mouth Disease Virus Antibodies: II. Use of Fractionated Bovine Antisera for Improving the Specificity of a “Passive” Hemagglutination Test1

Because 7S immunoglobulin (Ig) G antibodies of low type specificity were present in mixtures with highly specific 19S IgM antibodies, many bovine antisera to foot-and-mouth disease virus (FMDV) type A(12), strain 119 cross-reacted with type O of FMDV and to some degree with type C in the passive hemagglutination (HA) test. After 19S IgM antibodies were separated by density gradient centrifugation or precipitated with 4% (w/v) polyethylene glycol, the antigen could be determined with "block" HA tests. Such tests used several antigen concentrations in the titration of each antiserum. Adding 4% (w/v) polyethylene glycol to the serum was especially convenient for rapid precipitation of 19S IgM antibodies for the test. Similar results were obtained with bovine 19S IgM antibodies to FMDV type O, subtype 1, strain Caseros and type C strain Rezende.     

Use of the indirect hemagglutination inhibition reaction for evaluating the specific activity of allergens

To evaluate the specific activity of house-dust allergen, the passive hemagglutination inhibition test was used. Nine commercial batches of house-dust allergen were studied by means of this test. The results of the determination of the activity of house-dust allergen in the passive hemagglutination inhibition test, the skin tests and the indirect mast-cell degranulation test were in good correlation.     

实时荧光核酸恒温检测技术（SAT）在甲型流感病毒动物模型检测中的应用

评价实时荧光核酸恒温检测技术（SAT）对H1N1和H3N2两种甲型流感病毒小鼠模型肺组织匀浆液的检测效能。小鼠滴鼻感染不同浓度H1N1、H3N2病毒液30μL,感染96h后取肺组织匀浆液分别进行SAT检测及抗原血凝效价测定。结果表明,SAT检测的病毒拷贝数基本与抗原血凝效价测定结果一致,但灵敏度提高10^4倍。与血凝实验比较,SAT检测具有灵敏度高、特异性强、结果准确、高通量自动化的优势,有利于甲型流感动物模型的建立,为实验室筛选抗甲型流感病毒药物奠定基础。     

A copper-binding immunoglobulin from a myeloma patient. Purification, identification, and physical characterization.

A copper.protein complex present in the serum of a hypercupremic myeloma patient has been purified to homogeneity using gel filtration, DEAE-cellulose chromatography, and concanavalin A/Sepharose affinity chromatography. Immunoelectrophoresis and hemagglutination inhibition tests showed the copper-bound protein to be an IgG1-type immunoglobulin with lambda light chains. The immunoglobulin is of normal molecular weight (150,000) with normal size light and heavy chains (28,000 and 56,000, respectively). The carbohydrate portion of the molecule appears to be abnormal in that it interacts with concanavalin A, whereas most immunoglobulins of the gammaG-type do not. The copper in the native copper.IgG complex is in an EPR-indeterminable valence state. Copper was efficiently removed from the copper.IgG complex by dialysis against 0.1 M potassium cyanide. The apo-IgG was separated from the copper.cyanide complex by gel filtration. The copper complex was reconstituted by equilibrating the apo-IgG with 7.7 muM cupric ions.     

Estimation and Identification of Streptococcus dysgalactiae Protective Antibody in Sera of Rabbits and Cattle

Rabbit and cow anti-Streptococcus dysgalactiae sera were tested by bacterial agglutination, complement fixation, hemagglutination, and immunodiffusion for the presence of antibody. The results of these tests were compared with mouse-protection studies on the same serum to estimate which in vitro test would best reflect the in vivo protective capacity of serum. Identification of the antibody constituents responsible for the mouse protection, hemagglutination, and complement fixation titers were established by reacting whole and diluted antisera with mercaptoethanol before and after testing. Results indicate that the complement fixation test may be a more accurate indicator of IgG protective bovine and rabbit antibody, whereas the hemagglutination test may more readily reflect a wider range of protective antibody levels and IgM. The complement fixation test showed some shared responses to IgG and IgM in both the rabbit and cow, whereas the IgM components seemed to be the predominant factor influencing hemagglutination titers in the rabbit and more so in the bovine. Mouse protection tests with mercaptoethanol-treated cow and rabbit sera indicate that the protective capacity of these antisera is shared between IgM and IgG components.