Type-specific Serum Research Articles

Synthetic peptide vaccine against mucosal colonization by group A streptococci. I. Protection against a heterologous M serotype with shared C repeat region epitopes.

M protein is an antigenically variable virulence determinant present on the surface of group A streptococci, and it provides the basis for the serologic typing scheme. Type-specific serum antibodies afford strong protection against infection by the homologous serotype. Non-type-specific antigenic epitopes also exist within the surface-exposed portion of M protein. Previous studies demonstrated that intranasal immunization with Ag corresponding to sequences within the non-type-specific pepsin-susceptible site and adjacent C repeat regions of M6 protein, evoke protective immunity against pharyngeal colonization by type 6 streptococci in a mouse model. Although the protective immunogens are not type-specific, the pepsin site region of M6 is shared among less than 20% of serotypes examined. Therefore it was necessary to determine whether more highly conserved M protein epitopes elicit mucosal protection against group A streptococci, and if protective immunity extends to heterologous serotypes. In this report, peptides were synthesized that correspond to sequences completely contained within the highly conserved C repeat region of M6 protein. Peptide Ag were covalently coupled to the mucosal adjuvant, cholera toxin B subunit (CTB), and mice immunized intranasally and orally with peptide-CTB conjugates were compared to control groups that received CTB only. Immunization with the peptide-CTB conjugates led to significant protection against pharyngeal colonization by group A streptococci. Furthermore, protection was observed against the heterologous M serotype, type 14. These findings suggest that protection against multiple serotypes of group A streptococci can be achieved with a vaccine consisting of the widely shared C repeat region of M6 protein.

Localized outbreak of enteroviral meningitis in adults.

During a thorough surveillance of viral infections of the central nervous system an outbreak of aseptic meningitis was discovered in the western part of Finland in late 1985. The 21 diseased young adults were carefully studied by different virological methods. A presumed viral etiology, in all cases of enteroviral origin, was found in 16 of 20 (80%) with adequate specimens. Four different enteroviruses were associated with this episode; in 9 cases the presumed etiological agent was echovirus 5, while coxsackie B5, echo 25 and echo 17 viruses appeared to be responsible for 4, 2 and 1 case, respectively. Sensitivity of different diagnostic methods as regards detection of the echovirus 5 infections was in order: increase of type-specific neutralizing serum antibodies, isolation of virus from faeces, isolation from throat and group diagnosis by demonstrating an increase in complement-fixing antibodies to coxsackie B5 virus antigen.

One-dose immunization against paralytic poliomyelitis using a noninfectious vaccine.

Recent advances in production and standardization of noninfectious poliovirus vaccine now make it feasible to induce durable immunity against paralytic poliomyelitis with one dose of a suitably standardized vaccine. A single dose of a vaccine containing 40, 8, and 32 D-antigen units of type 1, 2, and 3, respectively, administered to six-month-old infants, was observed to induce antibody levels of greater than or equal to 1:4 in greater than 90% and immunologic memory in all. Since protection against paralysis is associated with the presence of either type-specific serum antibody or type-specific immunologic memory, and since immunologic memory once induced is irreversible, then lifelong immunity to paralytic poliomyelitis can be induced with a single dose of a suitably standardized vaccine administered at five to seven months of age. In areas of the world where exposure to poliovirus can occur before this age, vaccine should be administered earlier. Until the influence of age and/or maternal antibody has been further studied, infants immunized before the age of six months should receive a second dose after six months of age.

Biotechnological approach to a new foot-and-mouth disease virus vaccine.

Major contributions towards the development of an absolutely safe FMDV vaccine are evident. With the identification of VP1 as the immunogenic protein, it is possible to manufacture a subunit vaccine via biotechnology. DNA sequences encoding the VP1 protein can be introduced into a bacterium with ease; under the appropriate conditions, large amounts of VP1 can be produced in a short time. The accumulation of amino acid sequences generated by recombinant DNA techniques allows identification of antigenic domains, which are the basis of variability among serotype and subtype viruses. As a result, vaccine production by chemical synthesis of short peptides corresponding to the antigenic determinants is greatly facilitated. At present, results from experimental vaccines employing genetically engineered or chemically synthesized VP1 antigens against homologous virus infection are encouraging. The current approach of preparing vaccine is to utilize the antigenic specificity of the virus. Since FMDV undergoes antigenic drift, variants not neutralized by type-specific serum will arise. An alternative approach is to prepare vaccines based on antigenic sites shared among all serotype and subtype viruses.

Enhanced susceptibility of mice with streptozotocin-induced diabetes to type II group B streptococcal infection.

Since diabetes mellitus predisposes adults to group B streptococcal (GBS) bacteremia, a murine model of streptozotocin-induced diabetes and type II GBS bacteremia was developed to assess certain immune factors which might influence susceptibility to infection. In diabetic mice, the 50% lethal dose for two strains of type II GBS was significantly lower (greater than 1 log10 decrease in CFU per milliliter) than in control animals. This enhanced virulence of GBS for diabetic animals was associated with prolonged bacteremia, persistent sequestration of organisms in the splanchnic reticuloendothelial system, and a shift from splenic to hepatic clearance. Although immunization of control and diabetic animals resulted in high concentrations of type-specific serum antibody, it had no effect on late reticuloendothelial system sequestration in diabetics. In contrast, depletion of complement by treatment of mice with cobra venom factor blocked reticuloendothelial system clearance and resulted in fatal infection in both diabetic and control mice. These results indicate that neither type-specific antibody nor an intact complement system is adequate for effective clearance of type II GBS bacteremia in mice with experimentally induced diabetes. This clearance deficit could be the result of a defect in hepatocyte membrane receptors necessary for removal of this encapsulated microorganism.

Type-specific immunity and pharyngeal acquisition of group A Streptococcus.

A prospective study of spread of M-type 1, 2, 13, 14, 25 and 60 group A Streptococcus in 64 families in Qalyub, Egypt, in 1972-1974 showed that type-specific serum bactericidal antibody does not protect against pharyngeal acquisition of homologous organisms. The presence of type-specific antibody also does not appear to affect duration of carriage of the organism. Type-specific immunity must be mediated in another way, such as by local antibody or trough prevention of infection (as evidenced by a host response) following acquisition. This study also confirms the observations of others that administration of penicillin lowers the probability that a person who acquires group A Streptococcus will develop type-specific antibody.

Type-specific serum antibodies against group B streptococci among pregnant women: relation to urogenital carriage and age.

AbstractSerum levels of type-specific antibodies against group B streptococci (GBS) were investigated in 53 pregnant urogenital carriers of GBS and in 20 pregnant non-carriers. The distribution of serotypes of GBS isolated from the urogenital tract of the 53 culture-positive patients was: type Ia, 20 patients (38%), type II, 10 (19%), and type III, 23 (43%). Antibodies to GBS types Ia, Ib, II and III were quantitated by the use of radiolabelled protein A. It was found that the pregnant GBS carriers had in general higher levels of type-specific antibodies to GBS than non-carriers, irrespective of the type of GBS harboured in the urogenital tract. Furthermore, it was found that younger GBS carriers (20 to 29 yr of age) had higher serum levels of type-specific antibodies than older carriers (30 to 39 yr), a difference that did not occur among the non-carriers.

1035 FACTORS INFLUENCING DEVELOPMENT OF GROUP A STREPTOCOCCAL TYPE-SPECIFIC ANTIBODY

Because factors influencing the immune response to streptococcal M protein following upper respiratory tract (URT) and skin infections are poorly understood, we evaluated epidemiologic data from 4 groups of patients (408 serially collected sera from 119 children; m age = 6.5 yrs.) followed carefully for 2 yrs. The date and site of first acquisition of 4 serotypes (6,31,49,52), and the type-specific serum bactericidal activity (SBA) from pre-and post-acquisition sera were determined. The effects of patient age, serotype, site of isolation and number of culture-positive visits for each M type on the development of SBA were studied. M-6 was isolated from only the URT; while M-31, 49, and 52 were isolated from both skin and URT. Significant SBA developed in 27 of 41 (66%) patients with M-6; 22 of 41 (54%) with M-31; 13 of 31 (42%) with M-49; and 4 of 6 (67%) with M-52. The magnitude of the SBA response was generally greater for those with M-6. Of those with M-31 or M-49, no difference in SBA was noted between those with recovery only from the URT and those with recovery from both the URT and skin lesions. The development of SBA was related to number of isolations, although different for those acquiring each type. The increase in SBA was age-related in those with M-6, but not in those with M-31 or M-49. These data define several complex variables which may influence development of SBA following group A streptococcal infections.

Bluetongue virus hemagglutination and its inhibition by specific sera.

Bluetongue-virus (BTV) was found to agglutinate a variety of erythrocytes including sheep-, chicken-, guinea pig- and mouse-erythrocytes. Hemagglutination was inhibited specifically with type specific serum. A temperature dependence was only found for chicken erythrocytes, which showed a hemagglutination optimum at 37 degrees C. The hemagglutination was lost upon treatment of the virus with 0.4% trypsin as well as after treatment with 0.01 M KJO4. Heating of the virus preparation to 56 degrees C resulted in the loss of the HA-activity. Gelchromatographic studies indicated that the hemagglutinating capacity is associated with the complete virion. Whereas virulent strains of BTV hemagglutinate a number of different erythrocytes the avirulent type tested produced only a slight hemagglutination with sheep red blood cells. However, specific antiserum produced with the avirulent strain yielded strong hemagglutination inhibition (HI) with the corresponding virulent strain. Treatment of sera prior to their use in the HI proved necessary to remove nonspecific inhibitors. The efficiency of KJO4 in removing nonspecific inhibitors. The efficiency of KJO4 in removing nonspecific inhibitors indicates that carbohydrates represent the major group of nonspecific inhibitors. The data represented recommended the hemagglutination inhibition tests as a new method to identify the various BTV serotypes.

Purification of adenovirus 4 type-specific hemagglutinins for use in diagnostic counterelectrophoresis tests.

Three type-specific hemagglutinins of adenovirus (AV) 4 were purified, tested for various activities, and used as immunogens in rabbits. The components were purified by anion exchange and exclusion chromatographies to purification factors of 33, 20, and 37, respectively, and were identified as fiber (polypeptide, mol wt 70,000; incomplete hemagglutinin), penton (mol wt greater than 400,000; incomplete hemagglutinin; possession of cytotoxic activity), and dodecon (mol wt greater than 400,000; complete hemagglutinin; appearance by electron microscopy). Rabbit antisera to these hemagglutinating components possessed type-specific hemagglutination-inhibition and serum neutralization antibodies and were thoroughly evaluated for potency and specificty in immunodiffusion (ID), immunoelectrophoresis, and counterelectrophoresis (CE) tests. The anti-fiber and anti-dodecon sera were highly type-specific in ID and CE tests with a battery of commonly isolated AV serotypes; the anti-penton serum was similarly specific after it was absorbed with AV 2 hexons. These sera were thus proved to be suitable reagents for use in both the routine diagnostic ID test and the more rapid CE test.

Spread of Streptococcus pneumoniae in Families. II. Relation of Transfer of S. pneumoniae to Incidence of Colds and Serum Antibody

Factors that affect the spread of Streptococcus pneumoniae and the antibody responses associated with colonization were studied in 64 families for periods of eight to 52 weeks. Surveillance included daily recording of respiratory symptoms and bimonthly pharyngeal cultures for identification of the pneumococcal carrier state. Rhinovirus cultures were included for a portion of the study period. Intrafamilial carriage of a single type of S. pneumoniae and simultaneous spread to more than one family member were commonmspread often occurred in association with an upper respiratory tract infection; simultaneous transmission of S. pneumoniae and a rhinovirus was documented. Preexisting, type-specific serum antibody did not prevent acquisition of homotypic S. pneumoniae but did appear to shorten the duration of pharyngeal carriage. Sera of all 11 adults had greater than 150 ng of antibody nitrogen/ml of homotypic serum antibody (measured by a radioimmunoassay) before colonization. In contrast, only one of 13 preschool children had homotypic antibody concentrations of this magnitude before colonization. A threefold or greater rise in the concentration of homotypic antibody occurred in 13 of 24 children (54%) after acquisition of S. pneumoniae; the increase in antibody concentration was associated with illness in six of the children. On the other hand, acquisition of S. pneumoniae in adults was not associated with an increase in concentration of homotypic serum antibody.

Identification of Infectious Bronchitis Virus by Interference with the B-1 Isolant of Newcastle Disease Virus. Waxing and Waning of Interference

The Massachusetts and the Connecticut types of infectious bronchitis virus (IBV) were identified by interference in embryonating chicken eggs (ECE) with the production of hemagglutinin by the B-1 isolant of Newcastle disease virus (NDV). This interference test appears to be specific because the above interference was eliminated by adding type-specific anti-IBV serum to the IBV-NDV system; however, interference was not detectable when fowlpox virus (FPV) and infectious laryngotracheitis virus (LTV) were substituted for IBV. Specificity of the interference test was dependent upon a system involving IBV, NDV, FPV, and LTV. The test can be done in 3 days and requires minimum laboratory facilities. Most of the experiments were done with the Massachusetts type of IBV. Only a few were with the Connecticut type. The interfering action of the above two types of IBV over the B-1 isolant of NDV waxed between the 24th and 54th hr after inoculation of NDV; it was waning at the 54th-60th hr postinoculation and was undetectable by the 66th-72nd hr.

Application of immunohistochemistry to study of avian leukosis virus.

Summary A modification of the peroxidase-labelled antibody technique was applied to study the distribution of virus antigens in chicken cells infected with two serotypes (A and B) of avian leukosis virus (ALV). Type-specific chicken antisera reacted only with cells infected with virus of the homologous envelope serotype. When unfixed cells were exposed to type-specific antivirus serum, only antigens located at the cell surface were stained, while cells exposed to type-specific antibodies after fixation revealed both surface and intracytoplasmic virus antigen. Cytoplasmic antigen was usually concentrated in discrete granules which often had a vesicular structure. Hamster antibodies against ALV group-specific (gs) antigen reacted with fixed infected cells regardless of envelope serotype, and the distribution of antigen was as shown with chicken type-specific antibodies. Intranuclear gs antigen may have been present in a few cells. Unfixed infected cells did not react with hamster gs antibody and confirmed the location of ALV gs antigen within the virus particle. Some chicken antisera with neutralizing antibodies against a single envelope serotype of ALV contained both type-specific and gs antibodies. These antisera reacted with fixed cells infected with virus of either the homologous or heterologous serotype and stained both surface and cytoplasmic antigens. With unfixed infected cells, these antisera combined only with the surface of cells infected with virus of the same serotype as the neutralizing antibodies in the serum. Thus, the reaction in heterologously infected fixed cells was with internal gs antigen. This confirms independently that chickens are not naturally tolerant to their homologous ‘C type’ gs antigens.

Isolation and Characterization of Virulent Strain of Infectious Bursal Disease Virus from Broiler Birds in Bangladesh

The bursa Fabricius of 50 dead broiler birds aged between 2 to 5 weeks were collected from three different private and Bangladesh Agricultural University (BAU) poultry farms for isolation of infectious bursal disease (IBD) virus during the period from August to September 2002. Each of the collected bursa was stored at -20°C until processed for the isolation of virus in chicken embryo fibroblast (CEF) cell culture. Of the 50 bursal samples cultured in the chicken embryo fibroblast (CEF) cells, of which 40 (80%) bursal samples were found positive for IBD virus. Each of the 40 isolated IBD virus was characterized as IBD virus M6 strain using type specific polyclonal serum raised in chickens by agar gel immunodiffusion test (AGIDT). These isolated IBD virus M6 strain caused 100% mortality in five weeks old layer chickens on experimental infection through intramuscular route of inoculation. The results of this study indicate that the local isolates of IBD virus are associated with high mortality in chickens under both the natural and experimental conditions in Bangladesh. Key words: Isolation, characterization; virulent strain; IBDV; broiler birds doi: 10.3329/bjvm.v2i1.1930 Bangl. J. Vet. Med. (2004). 2 (1) : 27-29

Induction of specific chromosomal aberrations by adenovirus type 12 in human embryonic kidney cells.

Nonrandom chromosomal breaks in chromosomes 1 and 17 were provoked in human embryonic kidney cells 24 hr after infection with adenovirus type 12. These chromosomal changes disappeared in persistently infected cultures. Neutralization of the virus with type-specific antiviral serum prior to infection prevented the occurrence of chromosomal aberrations. No viral deoxyribonucleic acid (DNA) synthesis, as determined by autoradiography, was seen in metaphases containing adenovirus type 12-induced chromosomal aberrations. Ultraviolet irradiation of the virus reduced chromosomal aberrations linearly. This reduction in aberrations was fourfold slower than the inactivation of viral infectivity. At 24 hr after infection of cells with purified (3)H-labeled adenovirus type 12, the isotope was found to be associated with the nuclei. The uptake of isotope was reduced ninefold when the labeled virus was neutralized with type-specific antiviral serum. This difference is considered to account for neutralization of labeled virions. In metaphases infected with labeled viruses, most of the clustered grains were seen only on one arm of the chromatid, even after 72 hr. Isochromatid labeling was found, however, in a small percentage of chromosomes, and increased with time after infection. This increase was threefold between 24 and 72 hr after infection, whereas the mean grain counts decreased twofold during the same period. This has been tentatively interpreted to mean that most of the viral DNA molecules or parts thereof are merely attached to cellular chromatin, but a small fraction of them becomes gradually integrated as time proceeds. Certain chromosomal sites appeared to be preferentially labeled when chromosome 2 was used as a model for evaluation.

Congenital infections with reovirus.

Congenital reovirus, type 2 infections were produced after intraperitoneal inoculations of brood mothers on the 1st, 3rd, 6th, 10th, and 15th day of gestation. The offspring presented with a varied syndrome. About a quarter of a total of over 200 mice showed symptoms within the first 14 days of life; namely, lassitude, retarded growth, and roughening of fur. Some died, apparently of respiratory or renal failure. Post mortem examination showed marked interstitial pneumonia and subcortical renal tubular necrosis. Reovirus was isolated in high titer from the kidney and lungs as well as from blood, hearts, hind limbs, and brains in lesser titer. At 3 wk of age over 50 apparently well mice were sacrificed, and virologic, serologic, and pathologic study was done. High titers of virus were again found in the kidney, lung, blood, brain, heart, and skeletal muscle, but all tissues appeared normal histologically. Type-specific serum antibody titers in these mice were approximately those of their mothers. Another half of these mice showed decreased spontaneous activity and growth retardation which appeared between the 15th and 36th days of life. Three of these mice with late illnesses had marked proptosis and conjunctivitis. A subepidermal conjunctival and extraocular muscle lymphocytic infiltrate was observed on section, and reovirus was isolated from these eyes in tissue culture. Again blood, brain, kidney, liver, spleen, myocardium, and skeletal muscle were studied, and were found to be normal histologically and not to contain reovirus. Finally, the rest of the mice remain well to date. At 3 months of age, 10 of them were sacrificed. All had lost their maternal antibody and contained no reovirus, type 2 hemagglutinating inhibiting antibodies. No developmental abnormalities were observed. These data suggest that prolonged reovirus infections may be established by means of congenital inoculation of the developing fetus. Tolerant infection with immune paralysis seems to have been established.

The isolation of deoxyribonucleic acid from virulent and avirulent strains of Haemophilus pertussis.

SUMMARY: Haemophilus pertussis cells contain a relatively large amount of deoxy-ribonucleic acid which may be extracted from the organisms with 2% sodium cholate. A method is described for the isolation of deoxyribonucleic acid from the mechanically disintegrated organisms, which yields an apparently undegraded preparation free from ribonuclcic acid and protein, but containing a polysaccharide component, probably in combination with the nucleic acid. This finding gives further evidence of the differences in biological properties of virulent and avirulent strains such as agglutinability by type-specific serum. precipitability by aluminium phosphate, and solubility in bile and sodium hydroxide.

Cytopathogenic effect of poliomyelitis viruses in vitro on, human embryonic tissues.

Summary and discussionThe experiments which have been described demonstrate the capacity of the Lansing and Brunhilde strains of poliomyelitis virus to cause cell injury and death. This cytopathogenic property is revealed (1) by degenerative changes produced in infected tissue fragments in flask cultures and apparent on histologic examination; (2) by failure of such tissue fragments to exhibit normal cell migration when explanted to plasma cultures; (3) by degeneration of newly emigrated cells in roller-tube cultures; (4) by decreased acid production by infected cells. The conclusion that certain of these manifestations of injury are induced as a result of infection by the virus is further supported by the fact that type specific immune serum prevents their development. These phenomena are of interest from two general points of view. First, they leave no doubt that poliomyelitis virus in vitro can multiply in cells other than those of the nervous system and cause profound injury of such cells. Secondly, t...

Attempts to produce lymphocytopenia in rabbits following the intravenous inoculation of certain viruses.

prevention of this response with typespecific immune serum has recently been reported by Harris and Henle.1 Similar lymphocytopenic responses were also demonstrated after the injection of mumps virus, rough pneumococci, and nucleoproteins derived from hemolytic streptococci, t The possibility existed that this phenomenon might be evoked by a wide number of infectious agents. Of particular interest to us was the consideration that it might be useful in the study of the causative agents of poliomyelitis, infectious hepatitis, and infectious mononucleosis. The purpose of this paper is to report the results of experiments with

Appraisal of treatment of Hemophilus influenzaetype B meningitis with specific rabbit serum and sulfonamides: Based on observation of sixty cases

Summary Sixty cases of H. influenzae type B meningitis in infants and children treated with Alexander's type-specific rabbit serum and sulfonamides were analyzed. These cases were divided into two groups: Group I is comprised of all cases up to Oct. 1, 1944, and Group II from Oct. 1, 1944 to May 31, 1945. The following data were obtained: 1.Bacteriemia was present in thirty-nine of fifty cases (78 per cent). 2.The case fatality rate was 33 1/3 per cent for the entire series, 38.7 percent for Group I, and 18.7 per cent for Group II; for children under 2 years of age it amounted to 45.7 per cent. 3.Of the sixty patients, thirty-two had completely recovered at the timeof discharge from the hospital. There was no appreciable difference in the rate of complete recoveries between the two groups. 4.The following complications and sequelae were encountered: purulent arthritis, facial paralysis, partial and complete deafness, foot drop, and hydrocephalus associated with mental retardation. 5.The total amount of intravenously injected serum was not the sole factorresponsible for the outcome of the disease. Of particular significance is the fact that eight of twenty-six patients succumbed in spite of the administration of more than 200 mg. of antibody nitrogen, including three of seven patients who received more than 300 mg. 6.All four patients who received serum intrathecally, at a time whenmany microorganisms and a correspondingly large amount of specific soluble substance were present in the spinal fluid, succumbed to the infection. The possible deleterious effects of this therapy have been discussed. Intrathecal injection of serum in patients whose spinal fluid contained but a small number of microorganisms or none at all was followed, in several instances, by clinical improvement and sterilization of the spinal fluid. The possibility that this form of therapy may have been responsible, in part at least, for the development of hydrocephalus in two patients has been considered. 7.Immediate reactions to the type-specific serum injected intravenously were encountered in seven instances; delayed reactions, in eight. Intrathecal injection of serum resulted in convulsions in two patients and fever in one. 8.Two cases, presenting, respectively, the initial stage and the classicalpicture of the Waterhouse-Friderichsen syndrome as seen on post-mortem examination, have been reported.