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R J Genco

Publications and source records attributed to R J Genco.

At least 271 records · Page 15Linked to original sources
PubMed1976

Blastogenic response of human lymphocytes to oral bacterial antigens: characterization of bacterial sonicates.

Soluble sonicate supernatant preparations were made from Actinomyces viscosus (ATCC 19246), A. naeslundii (ATCC 12104), two strains of Veillonella alcalescens (strain HV-1 and a human oral isolate), Streptococcus sanguis (ATCC 10556), S. mutans (strain 6715-T2), Bacteroides melaninogenicus (strain K110), and Leptotrichia buccalis (isolated from human dental plaque). These supernatants were characterized with reference to their chemical and antigenic components and their biological activity determined by using in vitro lymphocyte blastogenesis as a measure of the host's cellular immune response. The sonicate supernatant of each bacterium contained protein, neutral sugars, methylpentose, and nucleic acids. Protein was the major component in all except L. buccalis, in which neutral sugars predominated. The antigenic components in each supernatant were detected by using rabbit antisera prepared against the whole bacteria and the sonicate supernatant. The supernatants showed a complex antigenic distribution on immunoelectrophoretic analysis. The supernatants were shown to be antigenic and not mitogenic in nature, since neither cord blood lymphocytes nor all adult lymphocytes were stimulated. The supernatant antigen preparations showed a reproducible, dose-dependent, and kinetic response in vitro, which was similar to that seen with the antigen preparation streptokinase-streptodornase.

Actinomyces↗
PubMed1976

Blastogenic response of human lymphocytes to oral bacterial antigens: comparison of individuals with periodontal disease to normal and edentulous subjects.

Cell-mediated immunity in humans to antigens derived from oral plaque bacteria was investigated by using the lymphocyte blastogenesis assay. Subjects with varying severities of periodontal disease including normal, gingivitis, periodontitis, and edentulous were compared. Mononuclear leukocytes were separated from peripheral blood and cultured with antigens prepared by sonication of Actinomyces viscosus (AV), Actinomyces naeslundii (AN), Veillonella alcalescens (VA), Leptotrichia buccalis (LB), Bacteroides melaninogenicus (BM), and homologous dental plaque (DP). The lymphocyte response of subjects with gingivitis or periodontitis was significantly greater than that of normal subjects to antigens of AV, AN, and DP, but did not differ from the response of edentulous subjects. Periodontitis subjects were significantly more reactive than edentulous and normal subjects in response to VA, LB, and BM. These findings suggest that the tested gram-negative bacteria and the host response they evoke are associated with advanced periodontal destruction.

Actinomyces↗
PubMed1976

Purification, characterization, and immunogenicity of cell-associated glucan from Streptococcus mutans.

Appropriate immunization with whole cell vaccines of S mutans appear to induce antibodies that inhibit implantation of S mutans on tooth surfaces and associated dental caries. To better understand the mechanisms by which vaccination prevents S mutans implantation and dental caries, and to prepare antigens whose effectiveness and safety can be tested in animal models of caries, we set out to purify and chemically characterize the CAG of S mutans. The CAG of S mutans strain 6715 was prepared by extracting cells with potasssium hydroxide at 100 C. After neutralization and extensive washing, the water-insoluble product was characterized by a battery of chemical analyses and found to be a relatively pure glucan. The CAG was weakly immunogenic in rabbits when administered in Freund's complete adjuvant. In monkeys (M irus) immunized via the parotid duct with an aqueous solution of CAG, a definite but weak serum IgG, IgM, IgA, and salivary IgA antibody response was observed. Absorption experiments showed that the CAG induced antibodies that cross reacted with Sephadex G-25 and others that reacted with unique determinants on CAG. Retention of native antigenic determinants through the purification procedures was verified by the observations that antiserums to CAG reacted with whole cells of S mutans and by the fact that antiserums to S mutans cells reacted with CAG.

Animals↗
PubMed1976

Effects of antibodies on adherence and cell-associated glucan production by Streptococcus mutans cells.

Studies reported here show that hyperimmune rabbit serums can have profound inhibitory effects on adherence of S mutans to smooth surfaces and that this reduction in adherence is correlated with the reduction of CAG. The latter was measured by an assay developed in our laboratory that estimates CAG production by measuring the uptake of 14C glucose-labeled sucrose into components which can be extracted by dilute alkali. This is a direct demonstration of the effect of antibody on a metabolic function of S mutans that is important in virulence. The direct correlation bewteen inhibition of CAG production, and inhibition of adherence by an antibody, suggests that the inhibition of adherence is brought about by reduction of CAG synthesis. Further studies reported here show that the inhibition of adherence and of CAG synthesis is much more effective in homologous antiserums as compared to heterologous antiserums. These results point to the need for in vivo experiments to test the effects of vaccines on colonization by heterologous as well as homologous S mutans strains.

Animals↗
PubMed1976

Immunization of Macaca fascicularis (Macaca irus) monkeys with Streptococcus mutans: specificity of antibody responses in saliva.

M fascicularis monkeys were immunized subcutaneously in the vicinity of the major salivary glands and by retrograde infusion into the parotid duct, with a vaccine containing Formalin-killed S mutans strain 6715 cells and culture-fluid antigens. Indirect immunofluorescent staining was used to titrate and classify antibodies. Subcutaneous immunization induced only a serum response, whereas intraductal infusion stimulated both an IgA antibody response in the parotid fluid and a serum response. Immunized and nonimmunized control groups were orally infected with S mutans strain 6715. The establishment in dental plaque was quantitated by recovery of the infecting organism on selective media and by immunofluorescent staining of plaque smears taken from individual tooth surfaces. The establishment of S mutans strain 6715 was noticeably inhibited in immune monkeys. Immunofluorescent assays for antibody also showed that serum and parotid fluid containing serum IgA antibodies cross reacted with other d serotype and a serotype strains but not representative b and c strains. Immune and control groups were then orally infected with S mutans strain GS-5, a c serotype strain, and no inhibition in establishment was detected of the non-cross-reacting type c organism in the immune group. A latter series of booster immunizations via the intraductal route resulted in a significant decrease in parotid fluid flow. Histological investigations showed inflammatory cell infiltration and replacement of epithelium by connective tissue in the glands from immunized monkeys. A separate group of monkeys, younger than the first, was immunized with the same vaccine via the duct only. In this group, immunizations were given at shorter intervals, but the immunization response was similar to that observed in the first group. The investigations reviewed here and new experiments reported show that immunization of monkeys with S mutan strain 6715 via the parotid duct elicited a reproducible IgA antibody response in the parotid fluid as well as a serum antibody response. Inhibition of colonization on tooth surfaces in immune monkeys showed specificity for the immunizing strain suggesting that inhibition was antibody mediated.

Age Factors↗
PubMed1975

Evaluation of human oral organisms and pathogenic Streptococcus for production of IgA protease.

IgA protease is a proteolytic enzyme found in whole human saliva and in dental plaque that cleaves both secretory and myeloma IgA of human origin to yield intact Fabalpha and Fcalpha fragments. To determine which bacteria are capable of producing this enzyme, we have examined a variety of strains normally found in the human oral cavity and a number of streptococci of known Lancefield group serotype. Streptococci of groups A, B, C, D, F, G, H, M, and N, Streptococcus mutans, Streptococcus sanguis, Streptococcus mitior, Streptococcus salivarius, Streptococcus faecalis, Veillonella, Lactobacillus, Actinomyces, Propionibacterium, Bacteroides, and Fusobacterium were grown in liquid medium, and fluids were examined for IgA protease activity. Only S. sanguis and clinically isolated group H streptococci elaborated IgA protease under the culture conditions used. Negative strains could not be stimulated to produce the enzyme when cultured in the presence of secretory IgA. Among the natural oral bacteria, capacity to produce IgA protease is restricted to certain species of Streptococcus, notably those of the group H serotype. Since secretory immunity is mediated by the IgA class of antibody, the presence of this enzyme at mucosal surfaces could modify the secretory immune function.

Actinomyces↗
PubMed1975

Antibody response in the parotid fluid and serum of Irus monkeys (Macaca fascicularis) after local immunization with Streptococcus mutans.

The antibody response of Macaca fascicularis in parotid saliva and serum to local immunization by two routes with Streptococcus mutans was studied and compared over 1 year. Antibodies were titrated and classified by indirect immunofluorescent staining using specific antiglobulin conjugates. Antiglucosyltransferase activity was assayed by an enzyme inhibition test. Animals were immunized first by injecting formalin-killed bacterial cells and cell products subcutaneously into the vicinity of the four major salivary glands. The monkeys were next immunized by retrograde instillation of antigen into the parotid duct. Extensive subcutaneous local immunization gave a serum response only. After parotid duct immunization, high titers of immunoglobulin A (IgA) antibody, along with traces of immunoglobulin G (IgG) immunoglobulin M (IgM) antibody, appeared in the parotid saliva, and in the serum high titers of IgG antibody were present along with lower titers of IgA and IgM. IgA antibodies in parotid fluid were shown by double immunofluorescent staining to be associated with antigenic determinants which cross-reacted with an antiserum directed to human secretory component. Titers in parotid fluids and sera fell sharply when immunization was stopped. This response pattern was reproducible. High concentrations of antibody capable of inhibiting glucosyltransferase prepared from S. mutans were found in the sera, but relatively little was detected in the parotid fluids. Extensive immunization via the parotid duct resulted in transient functional impairment of the gland, as evidenced by diminished salivary flow rates. We conclude that parotid ductal immunization can be an effective method for stimulating a salivary secretory IgA antibacterial antibody response.

Administration, Oral↗
PubMed1975

Prevention of Streptococcus mutans infection of tooth surfaces by salivary antibody in Irus monkeys (Macaca fascicularis).

Four irus monkeys (Macaca fascicularis) were immunized with Streptococcus mutans 6715 killed cells and cell products by injection in the vicinity of the major salivary glands and by instillation into the parotid glands via the ducts. After immune group and a sham-immunized control group of monkeys were infected orally with viable strain 6715 organisms. Dental plaque samples were taken at intervals from the buccal and lingual grooves of the first permanent molars. These samples were evaluated for recovery of strain 6715 by cultural methods. In addition, individual samples were taken from 10 representative tooth surfaces and were evaluated by indirect immunofluorescent staining for strain 6715. Results showed that immune monkeys had fewer infected surfaces and fewer organisms on the infected surfaces than the control animals. These studies indicate that salivary antibody to cariogenic streptococci inhibits implantation of these organisms in dental plaque and may be protective against dental caries.

Animals↗
PubMed1973

Inhibition of glucosyltransferase activity by antisera to known serotypes of Streptococcus mutans.

Using a recently developed assay for glucosyltransferase activity based on (14)C-glucose incorporation into an alcohol-insoluble polysaccharide, we were able to study inhibition by antibody of this enzyme activity. Rabbit antibody was relatively specific for the strain of Streptoccus mutans from which the enzyme was obtained. Absorption studies showed that neither removal of antibodies directed to dextran nor absorption with intact bacteria offset the enzyme-inhibitory capacity of these sera, whereas absorption with partially purified enzyme did result in removal of the inhibitory capacity.

Absorption↗
PubMed1973

Immunocyte response to experimental mumps virus infection in Rhesus monkeys.

Nineteen rhesus monkeys were inoculated with mumps virus by retrograde ductal instillation into the parotid gland. Evidence of infection was obtained in all instances. Virus was isolated from buccal swabbings and parotid biopsies for 1 week after inoculation. A vigorous serum-neutralizing antibody response occurred within 3 weeks, and there was marked monocytic infiltration of the parotid stroma. The monocytic infiltrate comprised as much as 60% of the total gland volume 1 week after infection. The predominant inflammatory cells were non-immunoglobulin-containing mononuclear cells resembling lymphocytes. Plasma cells containing immunoglobulin G (IgG), IgA, IgM, and IgE increased in numbers in the gland after infection, the greatest increase occurring in IgG-containing cells. Neutralizing antibodies and interferon were found in extracts prepared from the infected glands. Neutralizing activity was highest in samples taken 3 weeks after infection but was detectable in samples taken as soon as 36 to 48 h after infection. Interferon activity was detected in significant amounts 36 to 48 h after infection. Challenge of previously infected animals resulted in an increase in the monocytic infiltrate as well as an increase in numbers of immunoglobulin-containing plasma cells. However, reinfection did not occur as evidenced by the inability to culture shed virus after challenge. This model should be useful for in vivo study of biochemical mediators which evoke inflammatory cell infiltration and which may be significant both in protection and in tissue damage.

Absorption↗