Agglutinability of cattle red cells. 3. Conglutination.

Addition of both complement and conglutinin was necessary to conglutinate the cattle red cells (CRC) when they were sensitized by different blood typing reagents. Although the degree of conglutinability of the CRC was influenced by the particular blood factor-reagent combination the average conglutinability (i.e. average titre scores) of CRC from different MZ pairs varied from 1.9 to 16.2. The titres of complement varied from zero to 1:32, while the titres of conglutinin ranged from 1:8 to 1:1024 in the different sera from MZ cattle twins. The variance due to differences in the titre scores between MZ pairs was 82.2% for conglutinin and 68.3% for complement. There was no evident association between the titres of conglutinin and complement.     

Agglutinability of cattle red cells. 4. The effect of antiglobulin in comparison with treatment by pronase.

The average titre scores varied from zero to 24.7 for the cattle red cells (CRC) from different MZ pairs. These CRC were sensitized with different blood-typing reagents and were titre-tested against the double dilution series of an anti-bovine gamma-globulin serum. A significant negative correlation (r = - 0.82; P less than 0.001) was found between the degree of agglutinability and the amount of neuraminic acid of the surface component of CRC cleaved by pronase. After pronase treatment of the CRC it could be demonstrated that (1) the activity of the V and E'3 blood factors became destroyed; (2) three new specific receptors became evolved; (3) the degree of 'direct' agglutinability due to the A2, O3, W, S2 and Z anti-sera did not parallel with the titre scores obtained in the anti-globulin tests.     

Agglutinability of cattle red cells. 4. The effect of antiglobulin in comparison with treatment by pronase

The average titre scores varied from zero to 24.7 for the cattle red cells (CRC) from different MZ pairs. These CRC were sensitized with different blood-typing reagents and were titre-tested against the double dilution series of an anti-bovine y-globulin serum. A significant negative correlation (r = - 0.82; P <0.001) was found between the degree of agglutinability and the amount of neuraminic acid of the surface component of CRC cleaved by pronase.
After pronase treatment of the CRC it could be demonstrated that (1) the activity of the V and E'₃ blood factors became destroyed; (2) three new specific receptors became evolved; (3) the degree of 'direct' agglutinability due to the A₂, O₃ W, S₂ and Z anti-sera did not parallel with the titre scores obtained in the anti-globulin tests.     

Agglutinability of cattle red cells. 1. Agglutination by lectins of enzyme-treated red cells

Pronase-treated cattle red cells (CRC) from different monozygous (MZ) twin pairs could be classified as weakly (titre 1: 2, 1: 4) or strongly (titre 1: 1024, 1: 4096) agglutinable when Phytohemagglutinin-M (Phy-M) was used as agglutinin. In the presence of concavalin-A (Con-A), the CRC from different MZ pairs treated with pronase, A-chymotrypsin or trypsin showed a gradation from low (titre 1: 32) to high (titre 1: 256 000) agglutinability. The trypsin-treated CRC which had A₁, A₂ blood factors usually had a titre of 1: 8000 or higher with Con-A. Both the intact CRC and the CRC treated with proteolytic enzymes were capable of absorbing the Phy-M or Con-A lectins.     

Agglutinability of cattle red cells. 2. Agglutination by isologous and heterologous sera of enzyme-treated red cells

Neuraminidase treatment made cattle red cells (CRC) agglutinable by the agglutinins of the different heterologous but not of the homologous sera. These agglutinins were, however, absorbed by both the nauraminidase-treated and the intact CRC.
Proteolytic treatment made CRC agglutinable also by the normal cattle, isoimmune and autologous sera. Agglutination titres of the CRC ranged from 1: 2 to 1: 256, but the variation between CRC from members of monozygous (MZ) pairs was not greater than ± 2 agglutination score units the range of experimental error.
Treatment with trypsin made the A₁, A₂ factors more emergent on the surface of CRC for agglutination by anti-A₂, while pronase treatment had a similar effect upon agglutination of Z-positive cells by anti-Z.     

Evidence that bovine conglutinin reacts with an early product of C3b degradation, and an improved conglutination assay.

When EAC43b were treated with heated serum in EDTA, reactivity with bovine conglutinin appeared rapidly, even at 0 degrees C, and almost simultaneously with the loss of C3b rosetting capacity. At the time conglutinability first appeared, there was no detectable decrease in I-A or hemolytic C3 activity, and no detectable C3 antigen release from the cells. With prolonged exposure to heated serum in EDTA, I-A (immune adherence) and hemolytic C3 activity were lost. If this exposure was at 37 degrees C, C3 antigen became strongly detectable in the supernatant fluid, and eventually conglutinability was markedly reduced or lost, whereas C3d rosettes were unaffected. We suggest that bovine conglutinin reacts with some early product of C3b degradation, rather than with C3d, and propose that this intermediate be designated C3k. We have developed a semi-quantitative assay for bovine conglutinin, utilizing a Coulter Counter to register the decrease in total particles due to red cell aggregation. By using this method, we have detected conglutination with mouse complement (C) as well as with that from man and the guinea pig.     

Conglutinin, immunoconglutinins and heterophile antibodies in the sera of apparently healthy moose (Alces alces) and caribou (Rangifer tarandus) in Quebec

Serum samples from apparently healthy wild populations of moose and caribou in the province of Quebec, Canada were screened for the presence of conglutinin (K), immunoconglutinins (IKS) and heterophile antibodies. The conglutinating factor in moose and caribou sera was characterized utilizing the necessity of calcium ions for its reaction with sensitized sheep erythrocytes which had been alexinated with horse complement. The conglutinating substance in these animals did not require calcium ions for its activity. The conglutinating activity in both moose and caribou sera was characterized due to IKS as those present in sheep, dog and rabbit sera. Both moose and caribou had non-agglutinating type of heterophile antibodies. Their titres varied from 0 to 80. None of the animals tested had K in their blood. The titre of IKS varied from 0 to 640 with a mean value of 41 in moose, whereas it varied from 0 to 80 with a mean value of 18 in caribou. About 75% of all the animals in both the groups were positive for IKS. The specificity of IKS was demonstrated by the total removal of its activity on absorption with alexinated cells. Presence of IKS in these animals is suggestive of latent infection(s) possibly of bacterial, viral or parasitic origin.     

Some properties of piglet,calf, pig and bovine conglutinin

In sera of newborn piglets which were prevented from sucking maternal colostrum a low titre of conglutinin activity was demonstrated. For comparison of properties of this piglet conglutinin sera derived from adult pigs, cows and calves were used. Conglutinin from precolostral piglet serum behaves in different way as compared with immunoconglutinin from adult pig following reduction with 2-mercaptoethanol and in reaction with EDTA and thus resembled bovine natural conglutinin. In density gradient ultracentrifugation and inhibition reaction with zymosan andn-acetyl-d-glucosamine reacts piglet conglutinin as pig immunoconglutinin. Conglutinin present in precolostral calf serum behaves like a typical natural conglutinin of adult cattle.     

Significance of an increase in the titer of agglutinins in the microagglutination test in leptospirosis

The authors considered different views of investigators on the diagnostic antibody titre level in the microagglutination test (MAT) in leptospirosis of man and animals. Some of them took into consideration MAT in low titres (1:10-1:20), and others - in high only (1:400, 1:1000), which gave no possibility to assess the state of leptospirosis morbidity. The authors suggest that the assessment of the level of the titres in single and repeated studies should be approached differentially. In single examination 1:100 and over should be considered as a positive MAT titre for man, 1:200 and over - for cattle, 1:20 and over - for swine, and 1:20 and over for murine rodents. In repeated investigations any level of the titre in case of its dynamics should be taken into consideration.     

The serodiagnosis of human hydatid disease: 1. The routine use of latex-agglutination and complement-fixation in diagnosis

The combined use of complement fixation (CF) and latex agglutination (LA) tests is reported on sera from 6328 patients with suspected hydatid disease; 191 were confirmed positive at operation ('known positives'). Results by LA are related to CF titres. Both tests were negative in 90% of specimens. Nine patients were subsequently found infected of whom 3 became positive in tests after operation. Of sera positive in both tests, 75% were from 'known positives'. The remainder were almost certainly from infected patients. Half the patients whose sera were LA positive/CF less than or equal to 1/4 were follow-up 'known positives' in whom CF titres had waned; 2 were early infections. Only 3% of the cases with an LA negative/CF titre of greater than or equal to 1/16 were 'known positives' and 6% where the CF titre was 1/8. The remaining CF results in the group were false positives and accounted for 1.2% of all sera tested. Findings show that a CF titre greater than or equal to 1/8 with positive LA indicates past or present infection; a negative CF test with positive LA usually indicates past infection; rarely, infection is present when a CF titre is greater than or equal to 1/8 and LA is negative. A rising CF titre and positive LA indicates current infection; reliable prognosis following treatment is given by CF.     

Variation in the haemolytic rate of red cells in cattle

A colorimetric method was used to study the variations in the haemolytic rates between red cells from different individuals and with different blood factors. Marked differences were observed between the heterozygous and homozygous genotypes of 18 out of 20 different blood factors and between the B₁ or E₃factors, when they occurred in heterozygous genotypes but in different phenogroups. Between MZ twin pairs a continuous variation was found in the haemolytic rates for 17 out of 18 different blood factors, which indicated a quantitative genetic variation depending on individuality, in addition to the dosage- and pheotype-dependent variation.
The similar rankings of the haemolytic rates of the blood factors of the B₁O₃Y₂A'E₃phenotype in red cells from 11 MZ twin pairs suggested their simultaneous regulation. When the haemolytic rates of different blood factors at four or more loci were compared for 15 MZ twin pairs, the ranking results of a given locus were independent of those of the other loci in all pairs, except one.     

Immunization of Cattle Against Rabies Using Inactivated Cell Culture Vaccines

Twenty-one heads of cattle were vaccinated with Madibovin, 31 with Rabdomun and 127 with Rabisin on 4 different farms. Rabies neutralizing antibody titre (≥0.5 IU/ml) was detected in 80% of 163 animals tested about 1 month and in 42% of 133 animals tested about 1 year after primary vaccination. On 3 of the farms 86 animals received booster vaccination about 1 year after primary vaccination. All these animals had antibody titre (≥0.5 IU/ml) about 1 month after booster and antibody levels were higher than after the primary vaccination. Rabies antibody titres (≥0.5 IU/ml) were detected in 96% of 50 animals tested 1 year after the booster. No significant differences (p>0.05) in antibody levels were detected between animals vaccinated with Madibovin or Rabisin (farm C) respectively with Rabisin or Rabdomun (farm D) at any collection time. Responses to rabies vaccines varied considerably between the farms. After primary vaccination of the animals on 2 farms with the same batch of Rabisin, the antibody levels clearly differed (p<0.0001) between the farms. Our results indicate that booster is always necessary after primary vaccination to ensure that all animals are protected.     

X chromosome inactivation patterns correlate with fetal-placental anatomy in monozygotic twin pairs: implications for immune relatedness and concordance for autoimmunity.

BACKGROUND: Monozygotic (MZ) twinning is a poorly understood phenomenon that may result in subtle biologic differences between twins, despite their identical inheritance. These differences may in part account for discordant expression of disease in MZ twin pairs. Due to their stochastic nature, differences in X chromosome inactivation patterns are one source of such variation in female MZ twins. MATERIALS AND METHODS: We investigated X chromosome inactivation patterns in the blood of 41 MZ twin pairs based on methylation of the androgen receptor gene using a Hpa II-PCR assay. Twenty-six female MZ twin pairs with autoimmune disease (rheumatoid arthritis or multiple sclerosis) were studied. In addition, we studied 15 newborn female MZ twin pairs who were characterized at birth with respect to the anatomy of chorionic membranes (dichorionic versus monochorionic). RESULTS: We found a strong correlation between dichorionic fetal anatomy and differences in X chromosome inactivation patterns between members of an MZ twin pair. In contrast, all monochorionic twin pairs had closely correlated patterns of X chromosome inactivation. X chromosome inactivation patterns did not distinguish between MZ twin pairs who were concordant or discordant for autoimmune disease. CONCLUSIONS: The highly similar patterns of X chromosome inactivation among monochorionic twin pairs may result from their shared placental blood supply during intrauterine life. Alternatively, these patterns may indicate that X chromosome inactivation occurs before the twinning event in this anatomic subgroup of MZ twins. The data further suggest that these factors do not make a major contribution to the high discordance rates for autoimmune disease in MZ twin pairs.     

Persistence of antibodies of the IgG class against cytomegaloviruses in blood donors

High titre plasmas gained from blood donors are the initial material used for producing human immunoglobulin containing cytomegalovirus antibodies (CMV-HIG). For this purpose the sera gained from 467 permanent donors at the District Institute for Blood and Transfusion Service in Berlin were investigated on their CMV antibody content of IgG class by means of an indirect immunofluorescence test (IFT). The infection rate of blood donors amounted to 62% (291/467). CMV-IgG titre greater than or equal to 1:40 was determined in 88 sera (18.8%) and greater than or equal to 1:160 in 14 sera (3%). Two CMV-HIG laboratory samples (charges 3113 and 3117) were produced from these plasmas. Particularly immunoglobulin fraction of charge 3117 (No. 3117 N II) revealed excellent antibody titres (IFT 1:640 [CMV-IgG] or 1:40 [CMV-IgM] respectively, neutralisation test 1:32). Checks made with the donors' CMV-IgG titres after repeated application of plasmapheresis resulted in maximal titres changes of two dilution stages in a period of 15 months. Thus, in producing CMV-HIG a sure, tested pool of donors can be resorted to.     

Interaction between the ABO and Rh-Hr blood group systems in pairs of monozygotic (MZ) and dizygotic (DZ) twins

Previous studies have revealed significant deviations of twin pairs' blood group distributions. A comparison between monozygotic (MZ) and dizygotic (DZ) pairs in a sample of 688 twin pairs as to interaction between ABO and Rh indicates that the main contribution to total deviation comes from the MZ pairs, thus confirming a different behaviour of the two twin types in this respect.     

Immunity to equine herpesvirus 1 infection in foals during the first year of life.

A band of 23 pregnant mares on a Thoroughbred breeding farm all had serum virus-neutralizing antibody titres to equine herpesvirus 1 (EHV-1). Antibody was not transferred to their foals in utero. All foals received antibody from colostrum and developed antibody titres similar to their dams. The serum virus-neutralizing antibody titres were observed in 10 of these foals for 1 year. Decay of passive immunity occurred at the rate of 3.25 two-fold dilutions in 100 days and reached zero at the mean time of 180 days. The foals were exposed to EHV-1 twice. Foals with a geometric mean titre of 1 : 25 experienced infection and a rise of titre, while those with a geometric mean titre of 1 : 76 resisted infection.     

Use of the indirect hemagglutination reaction for detecting antibodies to Pseudomonas pyocyanea in acute suppurative destructive pneumonia]

No less than 4-fold increase in the antibody titres to Pseudomonas aeruginosa during the infectious process served as laboratory confirmation of its participation in the infectious process. In 49 of 91 patients hemagglutining titre exceeded the diagnostic one (1:640). In 9 patients (chiefly in infants) hemagglutinin titre remained low, but there was a rise of antibody level to Pseudomonas aeruginosa during the disease. High hemagglutinin titres were noted in 12 patients on admission to the clinic, with reduction of the antibody titres at the late periods of the disease. Antibody titres remained unchanged during the disease in 15 patients. In 3 cases the indirect hemagglutination test was assesed as negative. In the rest of the patients hemagglutinin titres varied within the range of the diagnostic titre. Thus, the indirect hemagglutination test with erythrocytic diagnostic agent permitting to determine antibodies to Pseudomonas aeruginosa could be used at the clinic in combination with bacteriological and other investigations for establishing etiology of the destructive process.     

The choice of adjuvants in Mycoplasma vaccines

The use of adjuvants in vaccine production is an important aspect of potent vaccines. This investigation was concerned with finding the most efficient adjuvants for use in Mycoplasma vaccines produced in Nigeria. Four different vaccines were produced from the Gladysdale strain of Mycoplasma mycoides subspecies mycoides. They differed depending on the type of adjuvants used. Each vaccine was used to vaccinate eight cattle using a dose of 1 ml. Two other groups of eight cattle were used as controls. One of the two groups received 1 ml dose of inactivated Gladysdale vaccine without adjuvant while the second group received 1 ml dose of saline. The number of cattle that had the peak complement fixing (CF) antibody titres of 1/80 in each group of cattle was four for vaccine containing aluminium hydroxide gel, eight for vaccine containing liquid paraffin, one for vaccine containing sodium alginate and one for vaccine without adjuvant. Seven cattle from the group vaccinated with vaccine containing Freund's incomplete adjuvant had peak CF antibody titres of 1/80 or higher. The two groups vaccinated with vaccine containing liquid paraffin and Freund's incomplete adjuvant survived challenge at 6 months post vaccination. Freund's incomplete adjuvant and liquid paraffin containing 10% Arlacel A are the most efficient adjuvants.     

Haemagglutination-inhibiting and neutralizing antibodies to type 3 and 5 rhinoviruses in human sera and nasal washings.

Presence of antibodies to RV 3 and RV 5 was tested by HIT and NT in 60 human sera. Antibodies to RV 3 were detected in 23 sera by HIT in a titre range of 1:4--1:64 and in 19 sera by NT in a titre range of 1:4--1:256. Antibodies to RV 5 were detected in 31 sera by HIT in titres of 1:4--1:268 and 27 sera by NT in the same titre range. In a group of 22 persons with unequivocal serum antibodies nasal secretory antibodies were found in 11 subjects in titres of 1:4--1:32. In a group of 16 persons without detectable serum antibodies, presence of secretory antibodies (titre 1:4) was only found in four cases.     

Persistence of influenza serum antibodies in humans following immunization with a bivalent A/Victoria and A/New Jersey vaccine.

The persistence of serum antibodies 1 year after immunization with a bivalent vaccine containing recombinant viruses that were antigenically identical with A/Victoria/3/75 (H3N2) and A/New Jersey/8/76 (Hsw1N1) viruses was measured in 128 persons aged 18 to 65 years. Serum samples were tested with the hemagglutination inhibition assay against the two vaccine antigens and against A/Texas/1/77 (H3N2) and A/USSR/90/77 (H1N1) viruses. Prior to vaccination 56% and 79% of the participants had been found to be seronegative to A/Victoria and A/New Jersey antigens respectively; the geometric mean antibody titres were low (1:5 to 1:11) except in persons aged 51 to 65 years, whose mean titre of antibody to the A/New Jersey antigen was 1:23, and persons aged 26 to 35 years, whose mean titre of antibody to the A/USSR antigen was 1:25. By 3 weeks after vaccination 85% of the seronegative persons had a fourfold or greater rise in titres of antibodies to the viruses in the vaccine, and 70% had a fourfold increase in titre of antibody to the A/Texas antigen. Of the persons aged 26 to 35 years (seronegative and seropositive) 68% had a fourfold or greater increase in titre of antibody to the A/USSR antigen. There was no change in the mean titres of 19 unvaccinated control subjects during the observation period. At 6 and 12 months after vaccination the titres of antibodies to the A/Victoria and A/New Jersey antigens had declined moderately in all age groups from those observed 3 weeks after vaccination. The rate of decline was similar for the various antibodies except that to the A/USSR antigen in persons 26 to 35 years of age, in whom the decline was much slower.