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Biomedical subjects

L A Bergmeier

Publications and source records attributed to L A Bergmeier.

At least 37 records · Page 2Linked to original sources
PubMed1995

T cell responses in macaques after vaginal immunization with particulate SIV p27 antigen.

Rhesus monkeys were immunized by the vaginal and oral routes using a recombinant particulate simian immunodeficiency virus (SIV) antigen. Augmenting vaginal by oral immunization in macaques elicits proliferative CD4+ T cells in the circulation which are specific to the immunizing p27 antigen. Reconstitution of enriched CD4+ T cells, B cells and macrophages from circulating mononuclear cells help B cells in specific IgA anti-p27 antibody synthesis. The results suggest that augmented vaginal immunization induces systemic CD4+ T and B cell responses which may play a part in the protective immunity against SIV (HIV) infection.

Administration, Intravaginal↗
PubMed1994

Targeted lymph node immunization with simian immunodeficiency virus p27 antigen to elicit genital, rectal, and urinary immune responses in nonhuman primates.

A s.c. route of immunization was developed in non-human primates, which targets the genitourinary-rectal associated lymphoid tissue. A vaccine consisting of rSIV gag p27, expressed as hybrid Ty virus-like particles (p27: Ty-VLP) was administered in the proximity of the internal iliac lymph nodes. Secretory IgA and IgG Abs to the p27 Ag were elicited in the vaginal, male urethral, rectal and seminal fluids, urine and serum. Two or more immunodominant B cell epitopes were identified within peptides 51-90 and 121-170 of the sequence of p27, using serum or biliary IgA and IgG Abs. CD4+ T cell proliferative responses to p27 were elicited predominantly in the targeted internal iliac, as well as the inferior mesenteric lymph nodes and the spleen, but not in the unrelated lymph nodes. These cells were then studied for helper function in p27 specific B cell Ab synthesis. Specific IgA and IgG Abs were detected in the same lymphoid tissues as those that displayed proliferative responses. However, cross-over reconstitution experiments between splenic and iliac lymph node B and CD4+ T cells suggest that the iliac B cells are essential for specific IgA Ab synthesis, whereas splenic B cells preferentially synthesize IgG Ab. The targeted lymph node (TLN) route of immunization gave comparable B cell, proliferative T cell, and Th cell responses to the vaginal, male genitourinary, and rectal mucosal routes, which were augmented by oral immunization. However, the TLN route induced urinary and seminal fluid sIgA and IgG Abs in addition to genital and rectal Abs. Generating secretory IgA and IgG Abs at the mucosal surfaces, and T and B cell immunity in the regional draining lymph nodes, spleen and circulation by TLN immunization may prevent transmission of virus through the mucosa, dissemination of the virus, and the formation of a latent reservoir of infection.

Animals↗
PubMed1994

Mucosal model of genital immunization in male rhesus macaques with a recombinant simian immunodeficiency virus p27 antigen.

Human immunodeficiency virus (HIV) can be transmitted through infected seminal fluid or vaginal or rectal secretions during heterosexual or homosexual intercourse. To prevent mucosal transmission and spread to the regional lymph nodes, an effective vaccine may need to stimulate immune responses at the genitourinary mucosa. In this study, we have developed a mucosal model of genital immunization in male rhesus macaques, by topical urethral immunization with recombinant simian immunodeficiency virus p27gag, expressed as a hybrid Ty virus-like particle (Ty-VLP) and covalently linked to cholera toxin B subunit. This treatment was augmented by oral immunization with the same vaccine but with added killed cholera vibrios. Polymeric secretory immunoglobulin A (sIgA) and IgG antibodies to p27 were induced in urethral secretions, urine, and seminal fluid. This raises the possibility that the antibodies may function as a primary mucosal defense barrier against SIV (HIV) infection. The regional lymph nodes which constitute the genital-associated lymphoid tissue contained p27-specific CD4+ proliferative and helper T cells for antibody synthesis by B cells, which may function as a secondary immune barrier to infection. Blood and splenic lymphocytes also showed p27-sensitized CD4+ T cells and B cells in addition to serum IgG and IgA p27-specific antibodies; this constitutes a third level of immunity against dissemination of the virus. A comparison of genito-oral with recto-oral and intramuscular routes of immunization suggests that only genito-oral immunization elicits specific sIgA and IgG antibodies in the urine, urethra, and seminal fluid. Both genito-oral and recto-oral immunizations induced T-cell and B-cell immune responses in regional lymph nodes, with preferential IgA antibody synthesis. The mucosal route of immunization may prevent not only virus transmission through the genital mucosa but also dissemination and latency of the virus in the draining lymph nodes.

Animals↗
PubMed1994

Antibodies to human and non-human primate cellular and culture medium components in macaques vaccinated with the simian immunodeficiency virus.

Inactivated simian immunodeficiency virus (SIV) grown in a human T-cell line induces protection from infection by the virus in macaques. However, observations that immunization with uninfected human T cells or with SIV-1 prepared in human T cells can also induce protection, has raised the possibility that protective antigens could be of human cellular origin. Sera from animals immunized with fixed infected and uninfected human T cells, as well as from animals immunized with partially purified cell-free SIV have been examined for their ability to bind to human and macaque peripheral blood mononuclear cells (PBMC) and to-components present in fetal calf serum (FCS) in which the cells were grown. Analysis by flow cytometry suggests that antibodies to human cell surface antigens can be elicited with both inactivated SIV grown in human T cells and by uninfected T cells. There was a significant association between the presence of anti-cell antibodies and protection from infection. However, anti-cell surface antibodies were not detected with macaque mononuclear cells by flow cytometry or by immunoprecipitation, unless these cells were first treated with FCS or activated by a mitogen. Immunoprecipitation of resting human PBMC with sera from immunized animals suggests the presence of antibodies to class I heavy and light chains [beta 2-microglobulin (beta 2 m)] and to bovine beta 2m, which may originate in FCS used to grow the cell line. Antibodies to CD4 were also found in sera from animals immunized with SIV grown in human T cells. We suggest that human cellular components augmented by FCS elicit anti-class I heavy chain, beta 2m, CD4 and FCS antibodies which may be responsible for protection against SIV infection in macaques.

Animals↗
PubMed1992

Induction of mucosal and systemic immunity to a recombinant simian immunodeficiency viral protein.

Heterosexual transmission through the cervico-vaginal mucosa is the principal route of human immunodeficiency virus (HIV) infection in Africa and is increasing in the United States and Europe. Vaginal immunization with simian immunodeficiency virus (SIV) had not yet been studied in nonhuman primates. Immune responses in macaques were investigated by stimulation of the genital and gut-associated lymphoid tissue with a recombinant, particulate SIV antigen. Vaginal, followed by oral, administration of the vaccine elicited three types of immunity: (i) gag protein p27-specific, secretory immunoglobulin A (IgA) and immunoglobulin G (IgG) in the vaginal fluid, (ii) specific CD4+ T cell proliferation and helper function in B cell p27-specific IgA synthesis in the genital lymph nodes, and (iii) specific serum IgA and IgG, with CD4+ T cell proliferative and helper functions in the circulating blood.

Administration, Oral↗
PubMed1991

Isolation of a human T cell line specific for a streptococcal cell surface antigen.

A streptococcal cell surface antigen of Mr 185,000 (SAI/II) expressed by Streptococcus mutans has previously been well characterised. A T cell line specific for native SAI/II has been isolated from peripheral blood mononuclear cells (PBMC) of a naturally sensitised normal individual. This line has been maintained in culture for several months and was shown to be highly specific, not only for different preparations of native antigen but also for recombinant SAI/II protein. It did not respond to a homologous antigen SpaA (Mr 210,000), extracted from Strep. sobrinus. The phenotype of the line was CD3+ CD4+ CD8- TcR alpha beta +. HLA typing and inhibition studies showed that the response was restricted by both DR and DP encoded class II.

Antibodies, Monoclonal↗
PubMed1990

Sequencing and characterization of the 185 kDa cell surface antigen of Streptococcus mutans.

The gene spa P (formerly designated as spa P1) encoding the Mr 185,000 surface antigen I/II of Streptococcus mutans, serotype c (strain NG5) has been sequenced. The deduced amino acid sequence of antigen I/II (1561 residues) includes a putative signal peptide (residues 1-38), as well as a transmembrane region (residues 1537-1556). The N-terminal part of the protein (residues 39-550) is particularly rich in alanine and includes three tandem repeats of a sequence of 82 residues. This region is predicted to be alpha-helical, adopting a coiled-coil formation, and may account for the cell surface hydrophobicity associated with expression of antigen I/II. In contrast the C-terminal region (residues 800-1549) is proline-rich, favouring an extended conformation. Comparison with the sequence determined from Strep. mutans strain MT8148 showed that antigen I/II is highly conserved with the exception of a short central region (residues 750-805). N-terminal sequencing of purified antigens I and II components indicated that antigen I extends from the amino-terminus of the intact Mr 185,000 surface antigen while antigen II extends from residue 996.

Amino Acid Sequence↗
PubMed1990

Identification of T- and B-cell epitopes in synthetic peptides derived from a Streptococcus mutans protein and characterization of their antigenicity and immunogenicity.

Natural immunity to synthetic peptides (SP) derived from the sequences of a 3800 Mr Streptococcus mutans antigen was found in human subjects. Significant serum IgG antibodies were detected both to the native streptococcal antigen and to the SP17, containing essentially residues 1-15. A series of short peptides with deletions at the amino- and carboxy-termini were then tested to identify the B-cell epitopes. Residues 8-13 and 1-6 bound significant serum IgG antibodies but only the former consistently inhibited human antibodies, suggesting that residues 8-13 constitute a major B-cell epitope. The human CD4 subset of T-cells was then examined and this showed a significant uptake of [3H]-thymidine when stimulated with both the native streptococcal antigen and the SP17. The series of short peptides was then used to stimulate CD4 cells, in order to determine the T-cell epitope. The synthetic peptide with residues 6-15 was the shortest peptide that stimulated significant [3H]-thymidine uptake and this peptide was designated as a T-cell epitope. The immunogenicity and antigenicity of SP17 was also investigated in macaques. Immunization of monkeys with the free SP17 failed to elicit serum antibodies or T-cell responses. However, immunization with SP17 linked to tetanus toxoid as a carrier elicited serum antibodies and proliferative responses of lymphocytes, not only to the synthetic peptide but also to the native streptococcal antigen. As in the human studies a B-cell epitope was found in residues 8-13, whereas an overlapping T-cell epitope was located in residues 7-15.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗
PubMed1989

Immunogenicity of synthetic peptides derived from the sequences of a Streptococcus mutans cell surface antigen in nonhuman primates.

The immunogenicity and antigenicity of synthetic peptides (SP) derived from the sequences of a cell surface Ag of Streptococcus mutans were investigated in macaque monkeys. Immunization with the free peptides of 11, 17, and 21 residues failed to elicit serum antibodies or T cell responses. However, immunization with the SP17 and SP21 linked to tetanus toxoid (TT) as a carrier elicited serum antibodies and proliferative responses of lymphocytes, not only to the SP but also to the native streptococcal Ag. In vivo recall of SP-TT immunized monkeys with suboptimal doses of the native streptococcal Ag resulted in a significant increase in antibodies, both to the SP and the streptococcal Ag, confirming that the SP shares antigenic epitopes with the native Ag. B and T cell epitopes were then determined and a B cell epitope was found in residues 8-13, whereas an overlapping T cell epitope was located in residues 7-15. The T cell epitope has an amino-terminal leucine and carboxy-terminal glycine and alanine added to residues 8-13 of the B cell epitope. In spite of the B and T cell epitopes being expressed in SP17 (residues 1-15), the monomer failed to induce serum antibodies without a carrier. However, immunization with a dimer of SP17 elicited both serum antibodies and proliferative responses of lymphocytes without a carrier. The results suggest that the monomeric SP17 is not immunogenic and needs to be dimerised in order to elicit antibodies and T cell responses, both to the SP and to the streptococcal Ag.

Amino Acid Sequence↗
PubMed1989

Induction of experimental autoimmune thyroiditis in B cell-depleted mice.

The effect of B cell depletion on the induction and severity of murine experimental autoimmune thyroiditis was investigated. Thirteen CBA mice were given repeated intraperitoneal doses of 700 micrograms purified rabbit anti-mouse Ig antibody from 24 hours to 8 weeks after birth. Controls were given normal rabbit IgG (14 mice) or were left uninjected (10 mice). At six weeks all mice received two doses of 70 micrograms murine thyroid extract in complete Freund's adjuvant. Only 2/13 of the anti-Ig treated mice were fully B cell-deficient as determined by serum IgM, spleen cell immunofluorescence and responsiveness to LPS; however, the levels of anti-thyroglobulin autoantibodies were very low in 7/13 mice. The results demonstrate that thyroiditis can be actively induced in the absence of B cells and autoantibodies but that B cells may play a role in increasing disease severity.

Animals↗
PubMed1989

Local oral immunization with synthetic peptides induces a dual mucosal IgG and salivary IgA antibody response and prevents colonization of Streptococcus mutans.

A small cell surface antigen of Streptococcus mutans was partially sequenced and the amino terminal peptides of 11, 15 and 20 amino acid residues and a dimer of the 15 and 20 residues peptides were synthesized. The synthetic peptides (SP) were used in topical oral immunization of the gingivomucosal epithelium of macaque monkeys. Sequential examination for antibodies over a period of up to 30 weeks revealed that six applications of the linear or cyclized SP11 and a random SP11 induced negligible or very low antibody levels. In contrast, the SP17 (SP15 with added cysteine at each terminus), SP21 (SP20 with one cysteine) and the dimer (SP35) induced significant anti-SP as well as anti-native streptococcal antibodies in the gingival fluid and in saliva. The functional significance of this immune response was examined by studying its effect on oral colonization of S. mutans following feeding of a carbohydrate-rich diet. Whereas control animals, sham-immunized with a random SP of 11 residues, showed increased colonization of the teeth by S. mutans, there was no colonization or a significant reduction in colonization of animals immunized with the cyclized SP17, linear SP21 or dimerized SP35. These experiments suggest that local immunization with SP derived from the sequences of a streptococcal cell surface antigen induce a dual local immune response of gingival IgG and salivary IgA antibodies against the SP and native SA. These antibodies may be involved in preventing colonization of S. mutans, which is the principal agent in the development of dental caries.

Animals↗
PubMed1988

The effect of immunization with a 14-kDa streptococcal antigen on primate T cell and B cell responses.

A streptococcal antigen (SA) of 185 kDa was isolated from Streptococcus mutans and this antigen induced in vitro helper, suppressor and contrasuppressor activities with primate peripheral blood lymphocytes. The 185-kDa SA was then treated by sodium dodecyl sulfate and yielded a 4-kDa SA which was capable of eliciting only helper activity. We have now cleaved the 185-kDa SA with cyanogen bromide, in an attempt to identify suppressor and contrasuppressor determinants. A 14-kDa SA was separated from the cyanogen bromide digest and its ability to elicit T cell and B cell functional activities was tested in rhesus monkeys. Whereas the 185-kDa SA (and 4-kDa SA) elicited high serum anti-SA antibodies and the CD4 cells showed an increase in DNA synthesis, this was not demonstrable with the 14-kDa SA. However, the 14-kDa SA, unlike the 185-kDa SA, activated a significant proportion of CD4 and CD8 cells to bind the Vicia villosa lectin (VV) and this is a characteristic feature of contrasuppressor cells. We then studied the effect of sequential immunization of monkeys with the 14-kDa SA, followed by the 185-kDa SA. The results of this showed suppression of the CD4 proliferative response, in the presence of a normal antibody production. We suggest that the split tolerance between the T cell proliferative and B cell differentiating functions might be interpreted on the basis of suppressor CD8 cells inhibiting the CD4 proliferative phase and the VV-adherent CD8 cells contrasuppressing B cell antibody formation.

Animals↗
PubMed1988

Separation and characterization of a 14,000-dalton cyanogen bromide-generated peptide from a 185,000-dalton streptococcal antigen.

The cell surface streptococcal antigen (SA) (185,000 molecular weight [185K SA]) was isolated from Streptococcus mutans and digested with cyanogen bromide. Three major products with molecular weights of 100,000, 50,000, and 14,000 appeared within 1 h of digestion. Time course studies of digestion by polyacrylamide gel electrophoresis showed maximal intensity of the 14K band after 8 h. However, other bands appeared as well, notably 70K and 20K bands. Several bands were eluted from the gels, and their antigenicity was studied. They reacted with antisera to the native 185K SA I/II, as well as with those to the SA I and SA II antigens, though antibody binding by radioimmunoassay was significantly lower than that with the native SA. The 14K SA was identified on Western blots (immunoblots) with anti-SA I/II, I, and II antisera. The digested SAs were then tested for their immunogenicity by injecting CBA mice with the separated SA mixed in complete Freund adjuvant. Whereas the unseparated cyanogen bromide-treated SA and separated 70K and 20K SAs were immunogenic, the 14K SA failed to elicit serum antibodies. Further investigation of the 14K SA revealed that although it is apparently not immunogenic, it can induce a primary antibody response in mice when followed by the native 185K SA and a secondary response when mice are immunized first with the 185K SA.

Animals↗
PubMed1984

Natural antibodies in man to a protein antigen from the bacterium Streptococcus mutans related to dental caries experience.

Serum antibodies to a cell-wall protein antigen derived from Streptococcus mutans (SA I/II) were examined in 108 subjects by a solid-phase radioimmunoassay. As a control, antibodies to lipoteichoic acid and to Strep. mutans whole cells were examined and the relationship to dental caries of isotype-specific antibodies to both antigens determined. Serum-IgG antibodies to SA I/II and to Strep. mutans were significantly greater in subjects of low caries experience than in subjects of high caries experience, whereas serum-IgG antibodies to LTA showed no relationship. Serum-IgM antibodies to SA I/II were depressed in subjects with carious lesions, whereas IgM antibodies to Strep. mutans in the same subjects were raised. IgA antibodies against any antigen were not related to caries. Serum antibodies of each isotype to SA I/II were generally positively-correlated with antibodies to LTA but inhibition experiments demonstrated specificity of antibodies for each antigen. The relationship between antibodies to SA I/II and dental caries are consistent with those reported with antibodies to whole cells of Strep. mutans and suggest that high-serum IgG antibodies to SA I/II may be associated with protection against dental caries in man.

Adolescent↗
PubMed1984

Natural antibodies in man to Streptococcus mutans: specificity and quantification.

Antibodies to whole cells of Streptococcus mutans were examined in 108 subjects by a solid-phase radioimmunoassay and quantified by reference to isotype-specific affinity-purified antibodies. Serum antibodies of each isotype were present in all subjects examined. The mean concentration of serum antibodies to S. mutans was calculated as about 84 micrograms/ml of IgG (range 33-140 micrograms/ml), 26 micrograms/ml of IgA (range 12-43 micrograms/ml) and 9 micrograms/ml of IgM (range 4-15 micrograms/ml). The mean antibody values accounted for about 0.7, 1 and 0.8% of the total IgG, IgA, and IgM, respectively. Overall the antibody binding to whole cells of S. mutans accounted for about 0.8% of the total immunoglobulin. Inhibition experiments using a variety of purified cell wall antigens revealed that the binding of antibodies to whole cells could be inhibited by about 30% with a purified protein antigen (SA I/II) and with glucosyltransferase (GTF), by 25% with c polysaccharide and by 16% with lipoteichoic acid. The protein antigens GTF and SA I/II appear to be major immunogenic cell wall antigens, but natural antibodies in man that bind to S. mutans whole cells have been induced by several antigens, some of which are specific to S. mutans and some of which are shared with other Gram-positive bacteria.

Adolescent↗
PubMed1983

Lack of antibodies to human heart tissue in sera of rhesus monkeys immunized with Streptococcus mutans antigens and comparative study with rabbit antisera.

Immunization of rabbits and rhesus monkeys with Streptococcus mutans whole cells, cell walls, and defined streptococcal antigens (SAs) SA I/II, SA I, and SA II resulted in high antibody titers to SA I/II (10(-4) to 10(-6)) when tested by the solid-phase radioimmunoassay. Cross-reactive antibodies to heart homogenates (HH) were not elicited in rhesus monkeys. A few rabbits immunized with cell wall or SA II preparation in Freund complete adjuvant followed by the incomplete adjuvant yielded low antibody titers (up to 10(-2)) to the HH. The specificity of the putative heart cross-reactive antibodies was tested by immunoadsorption with related and unrelated antigens. Whereas antibody to SA I/II showed specific adsorption with SA I/II but not with HH, immunoglobulin G, or the unrelated antigens, antibody to HH seemed to have been adsorbed with all of the related and unrelated antigens. There was no evidence for the development of heart cross-reactive antibodies on immunization of rhesus monkeys or rabbits with SA I/II and aluminium hydroxide or Freund incomplete adjuvant, administered by the subcutaneous or intramuscular route in doses of up to 13 mg of the immunogen.

Animals↗
PubMed1983

The detection and specificity of class specific antibodies to whole bacterial cells using a solid phase radioimmunoassay.

A solid phase radioimmunoassay has been developed which can be used for the detection of isotype specific antibodies to whole bacteria and other particulate antigens, and is applicable to a variety of species. Bacteria are bound to the solid phase by the use either of antibodies, or of methyl glyoxal. Both methods result in a sensitive and reproducible assay, and bacteria do not appear to desorb from the solid phase. The specificity of antibodies to whole bacteria was examined by absorption of antisera with various species of bacteria and retesting, or by determining the binding of antisera to various bacteria bound to the solid phase. Both methods revealed specificity for the bacteria examined. Inhibition studies showed that antibodies to Streptococcus mutans whole cells could be inhibited by purified cell surface antigens glucosyltransferase and antigen I/II, but only minimally by lipoteichoic acid, c polysaccharide or dextran. In murine antisera antibodies of the IgG, IgM, and IgA classes could be detected at amounts of less than 1 ng/ml.

Actinomyces↗