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Biomedical subjects

B B Diena

Publications and source records attributed to B B Diena.

At least 37 records · Page 2Linked to original sources
PubMed1978

The lipopolysaccharide (R type) as a common antigen of Neisseria gonorrhoeae. I. Immunizing properties.

The ability of R-type lipopolysaccharide (LPS), isolated from Neisseria gonorrhoeae colony type 4, to protect against infection with N. gonorrhoea colony type 1 (T1 isolates) in the mouse and chicken embryo was investigated. C57 black mice were immunized intraperitoneally with 50 microgram of LPS, and challenged intracerebrally with 10-20 LD50's of N. gonorrhoeae colony type 1. Immunized mice were significantly protected (P less than 0.01 to less than 0.05) against challenge with different T1 isolates of N. gonorrhoeae when compared with non-immunized mice. Mice, injected with succinylated or alkali-treated LPS were not protected against gonococcal challenges. In a second animal model, leghorn hens were immunized intravenously with three injections of 500 microgram of LPS followed by a booster of 2.5 mg 2 weeks later. Embryonated eggs obtained from immunized hens were protected against challenge with 5 x 10(3) - 1 x 10(4) LD50's of three different T1 isolates. When hens were injected with the chemically modified LPS, the embryos were not resistant to gonococcal challenge. The results of this study demonstrate the ability of R-type gonococcal LPS to provide protection against different T1 isolates of N. gonorrhoeae.

Animals↗
PubMed1978

The lipopolysaccharide (R type) as a common antigen of Neisseria gonorrhoeae. II. Use of hen antiserum to gonococcal lipopolysaccharide in a rapid slide test for the identification of N. gonorrhoeae from primary isolates and secondary cultures.

An antiserum has been prepared in hens to R-type gonococcal lipopolysaccharide (LPS) and used in a simple slide-agglutination test for the identification of Neisseria gonorrhoeae. Anti-LPS serum agglutinated gonococcal cells representative of the four colony types of N. gonorrhoeae. Absorption of the antiserum with LPS removed the agglutinating activity. Secondary cultures (1120) were tested without observation of the colony type and all were agglutinated. No agglutination occurred with strains of Neisseria meningitidis, Neisseria lactamica, non-pathogenic Neisseria. Pseudomonas aeruginosa, Branhamella catarrhalis, or with species of lactobacilli and Acinetobacter. Cross-reactivity of the antiserum occurred with some streptococci. The anti-LPS serum was used to identify N. gonorrhoeae in primary isolates from the cervix, urethra, and pharynx. Of 251 gonococcal isolates tested, 249 were agglutinated by the antiserum, while all of the corresponding second cultures were agglutinated. The antiserum did not agglutinate N. meningitidis found in primary isolates from pharyngeal specimens. Anti-LPS hen serum should be useful for the rapid identification of N. gonorrhoeae in primary isolates or secondary cultures.

Agglutination Tests↗
PubMed1978

Enzyme-linked immunosorbent assays for the detection of Neisseria gonorrhoeae specific antibodies.

An indirect enzyme-linked immunosorbent assay (ELISA) using rigid polystyrene microtiter plates was adapted to detect specific gonococcal antibodies against outer membrane-complex antigens extracted from Neisseria gonorrhoeae. The concentration of antigen to obtain maximum coating of the well was 10 micrograms protein per millilitre. The optimal binding of the primary antibody and enzyme-conjugated antimmunoglobulin was achieved after 1 h at 37 degrees C. Under these conditions using gonococcal antisera, no cross-reactivity was observed with outer membrane antigens extracted from Neisseria meningitidis serogroups B, C, X, Y, and W135. Neisseria meningitidis serogroup A demonstrated low levels of cross-reactivity. All the non-pathogenic Neisseria spp. tested were negative (absorbance value at 400 nm/30 min less than 0.15). The reaction of immune serum against outer membrane complex absorbed to the microwells was completely inhibited with soluble-specific antigen but not with purified N. gonorrhoeae lipopolysaccharide. Quantitative inhibition permitted the measurement of low levels of antigen (0.5 microgram/ml). The detection of N. gonorrhoeae antibody with ELISA is specific and highly sensitive.

Agglutination Tests↗
PubMed1977

In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells.

As a basis for studies of gonococcal pathogenicity, tissue culture cells were infected with type 1 or type 4 Neisseria gonorrhoeae to determine intracellular viability. A simple and objective means of measurement was devised, based on the uptake of tritiated protein and deoxyribonucleic acid precursors by cycloheximide-inhibited cells infected with gonococci. Cycloheximide was found to inhibit protein synthesis by over 97% tissue culture cells at a concentration of 100 microng/ml. In contrast, N, gonorrhoeae was found to be highly resistant to this antibiotic, and protein synthesis was unaffected by concentrations up to 1,000 microng/ml. Extracellular gonococci were eliminated by treatment with high concentrations of penicillin during cycloheximide inhibition and prior to the addition of radioisotope. Levels of protein and deoxyribonucleic acid synthesis by N. gonorrhoeae in the cycloheximide-treated cells were significantly higher in T1-infected cells (RE2, HeLa, or HEp-2) than in the corresponding T4-infected cells. No differences were observed in tissue cell susceptibility to gonococcal infection. Intracytoplasmic localization of N. gonorrhoeae was confirmed by electron microscopy.

Bacterial Proteins↗
PubMed1977

Homocytotropic antibodies (IgE) to Neisseria gonorrhoeae in the rat and a cross-reactivity of heterologous gonococcal strains.

Outbred Wistar rats were immunised with a single intraperitoneal injection of a mixture of 30 mg of A1(OH)3 and 100microng of gonococcal zeolite antigen (ZA). Ten days after immunisation, ZA prepared from T1 and T4 colonies of Neisseria gonorrhoeae strain GC6 (GC6-T1 ZA and GC6-T4 ZA) were able to elicit a reaginic (IgE) response which declined to low levels by 35 days. There was a significant amount of histamine release from the mast cells of actively-sensitised rats on challenge with a specific gonococcal antigen. The antisera heated at a temperature of 56 degrees C for four hours failed to elicit a passive cutaneous anaphylaxis (PCA) reaction, indicating a lack of IgGa antibody. In addition antisera to GC6-T1 ZA gave a positive PCA reaction with GC6-T4 ZA and ZA prepared from T1 colonies of six heterologous gonococcal strains suggesting that these strains of N. gonorrhoeae share common antigenic determinants.

Animals↗
PubMed1977

The effect of hyperimmunization with Neisseria gonorrhoeae on the presence of gonococcal antibody in serum, tissues, and secretions of the rabbit.

Antibody responses in sera, tissues, and secretions of the urogenital tract and lower respiratory tract of rabbits hyperimmunized with Neisseria gonorrhoeae were examined. Antibody was detected by passive hemagglutination, whole-cell agglutination, bentonite flocculation, and in some cases immunodiffusion-in-gel. Immunization of rabbits either intravenously or intramuscularly resulted in the presence of gonococcal antibodies in the sera, spleens, and tissue of the urogenital tract (vagina, cervix, uterus, and fallopian tubes). Gonococcal antibody was also found in secretions bathing the mucosa of the urogenital tract and lower respiratory tract. Antibodies were not detected in sera, tissues, and secretions of non-immunized rabbits. The spleen was shown to synthesize gonococcal antibody in vitro in response to hyperimmunization. Tissues of the urogenital tract did not appear to synthesize gonococcal antibody thus suggesting and antibodies present in secretions of the urogenital tract were derived mainly from serum.

Agglutination Tests↗
PubMed1977

Inhibition of attachment of Neisseria gonorrhoeae to tissue cells by goat milk antigonococcal immunoglobulin G.

An immunoglobulin G (IgG)-containing fraction was isolated from milk, obtained from a goat before and after instillation of the mammary gland with Neisseria gonorrhoeae colony type 1 (T1). The presence of IgG and the absence of immunoglobulins-A and -M in this fraction was confirmed by immunodiffusion in gel and immunoelectrophoresis. Postinstillation IgG inhibited the attachment of the homologous strain of N. gonorrhoeae (T1) to tissue cells. The percentage of Rhesus monkey kidney cells with gonococci attached was 38.9% +/- 6.4. Inhibition was significant (P less than 0.01) when compared to the inhibitory effect of preinstillation IgG (73.5% +/- 3.1) or buffer control )76.8% +/- 2.8). Absorption of postinstillation IgG with rabbit anti-goat IgG or whole cell gonococci removed the inhibitory effect. Postinstillation IgG gave little or no inhibition of attachment (maximun 13%) of seven heterologous (T1) strains of N. gonorrhoeae to tissue cells. These results may provide a reason for recurrent gonococcal infection in humans despite the presence of circulating antibody (IgG) to N. gonorrhoeae. Such an antibody, if present in secretions bathing the mucosa, may fail to prevent infection and colonization of the urogenital tract by a different immunotype of N. Gonorrhoeae.

Animals↗
PubMed1977

Mitogenic activity of Neisseria gonorrhoeae surface antigens in mouse splenic lymphocyte culture.

The mitogenic effects of Neisseria gonorrhoeae endotoxin, fractionated envelope componenents, and intact cells were examined on unsensitized mouse splenic lymphocytes in vitro. The stimulatory effect of these substances was measured by increased [3H]thymidine incorporation in spleen cell cultures. Intact cells, purified lipopolysaccharide (LPS), and cell envelope preparations were highly stimulatory and the stimulation index was dose dependent. Fractionated components of the envelope demonstrated variable stimulation when tested at identical LPS concentrations, reflecting the mitogenic activity of the protein moieties. The stimulatory dose responses for purified N. gonorrhoeae and Escherichia coli LPS were compared and mitogenicity was higher with gonococcal LPS at all concentrations tested. Alkaline detoxification or succinylation of N. gonorrhoeae LPS results in loss of ability to induce blast transformation. The mitogenicity of cell-surface components of N. gonorrhoeae is discussed in terms of LPS and protein content.

Animals↗
PubMed1977

The guinea pig as a model for the asymptomatic human typhoid carrier.

Studies were designed to develop an animal model which mimics the asymptomatic carrier state of typhoid fever, Salmonella typhi infection, in man. The gallbladder of the guinea pig [Can (HA)] was surgically cannulated to provide a route for infection and bile sampling. Organisms (2 X 10(8) S typh:) were introduced by way of the cannula and samples of bile removed and analyzed periodically. Salmonella typhi was recovered from the bile and feces of infected, yet apparently normal, guinea pigs for periods of up to 5 months thus mimicking the human symptomeles carrier. Some guinea pigs were treated with chloramphenicol after infection. Antibiotic therapy with chloramphenicol did not eliminate the carrier state. Microabscesses of the liver were induced by S typhi both in untreated and treated animals (cultures were positive from these abscesses). When blood antibiotic levels dropped, the organism was found in the bile fluid and was again excreted. Vaccinated guinea pigs developed elevated levels of 0 and H serum antibody, but were not protected against infection of the gallbladder with S typhi (2 X 10(8) organisms). Similarly, vaccination failed to eliminate the carrier state.

Administration, Oral↗
PubMed1976

Type and strain variation in induction of L forms of Neisseria gonorrhoeae.

Strains of Neisseria gonorrhoeae were inoculated onto brain heart infusion (Difco) agar supplemented with penicillin and polyvinylpyrrolidone (PVP) as a stabilizer and cultivated in a candle extinction jar. L-form colonies became visible after a few days. They continued to grow and were viable for up to 38 days. The extent of inducibility of L forms of N. gonorrhoeae was examined semiquantitatively. It was found to be constant for each type and strain and to depend only slightly on the amount of penicillin added to the medium. None of the types of one strain produced L-form colonies. In another strain, avirulent types (T3, T4) showed more ability to produce L forms than virulent types (T1, T2) and no L forms were produced by the subtypes of T1 and T2-T1a and T2a. In a third strain, only T4 produced L forms. Electron microscopic studies showed that the L forms consisted of a number of membranous vesicles and a variety of cell types such as those completely lacking cell walls and those with only remnants of cell walls. The results indicate the existence of subtypes of T1 and T2 of gonococci and the intrinsic inducibility of gonococcal types and strains to produce L forms.

Culture Media↗
PubMed1976

The goat mammary gland as a model infection site for Neisseria gonorrhoeae.

Live and formalin-killed gonococci were instilled into the mammary glands of lactating and nonlactating goats. In lactating goats viable gonococci elicited a limited inflammatory process whereas in non-lactating goats, severe inflammation and swelling appeared and peaked on the 3rd day after instillation and persisted for about 10 days. No viable gonococci were recovered after the first day, but fluorescent antibody staining showed gonococci in the exudate from non-lactating goats up to 7 days after instillation.

Animals↗
PubMed1976

Antibody production in milk serum of goats experimentally infected with Neisseria gonorrhoeae.

Instillation of the goat's mammary gland with Neisseria gonorrhoeae colony type T1 has elicited and antibody response in the goat milk serum (GMS). Purification, and characterization of the GMS by gel filtration, electrophoresis, immunodiffusion, analytical ultracentrifugation, and serological analyses demonstrated thtat the active immune component was mainly in the IgA and IgG fractions (F2 and F3) of GMS.

Animals↗