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J B Robbins

Publications and source records attributed to J B Robbins.

At least 145 records · Page 8Linked to original sources
PubMed1983

Further studies on the immunogenicity of Haemophilus influenzae type b and pneumococcal type 6A polysaccharide-protein conjugates.

Conjugates were prepared by carbodiimide-mediated coupling of adipic acid hydrazide derivatives of Haemophilus influenzae type b (Hib), Escherichia coli K100, and pneumococcal 6A (Pn6A) polysaccharides with tetanus toxoid (TT), as an example of a "useful" carrier, and horseshoe crab hemocyanin (HCH), as an example of a "nonsense" carrier. These conjugates were injected into NIH mice, and their serum antibody responses to the polysaccharides and proteins were characterized. As originally reported, Hib conjugates increased the immunogenicity of the capsular polysaccharide and elicited greater than the estimated protective levels of anti-Hib antibodies in most recipients after one injection and in all after the third injection (Schneerson et al., J. Exp. Med. 152:361-376, 1980). Both Hib conjugates induced similar anti-Hib responses. The K100-HCH conjugate was more immunogenic than the K100-TT conjugate and elicited anti-Hib responses similar to the Hib conjugates after the third injection. Simultaneous injection of the K100 and the Hib conjugates did not enhance the anti-Hib response. The Pn6A-TT conjugate induced low levels of anti-Hib antibodies; when injected simultaneously with the Hib conjugates, the anti-Hib response was enhanced, as all mice responded after the first injection and with higher levels of anti-Hib than observed with the Hib conjugates alone (P < 0.05). The Pn6A conjugates were not as immunogenic as the Hib conjugates. Pn6A-TT was more effective than was Pn6A-HCH; it elicited anti-Pn6A (>100 ng of antibody nitrogen per ml) in 6 of 10 mice after the third injection. The addition of the Hib-HCH conjugate to the Pn6A-TT conjugate increased the anti-Pn6A response with a higher geometric mean antibody titer, and 9 of 10 mice responded after the third injection. A preparation of diphtheria toxoid, TT, and pertussis vaccine increased the anti-Hib antibody levels after the first injection only in mice receiving Hib-TT, but not in mice receiving Hib-HCH, suggesting that additional carrier protein (TT) enhanced the anti-polysaccharide response. Simultaneous injection of Hib and Pn6A conjugates with the same or different carriers resulted in an enhanced serum antibody response to each polysaccharide. The anti-tetanus toxin response reached protective levels (>0.01 U/ml) in most mice after the first injection and in all mice after the second and third injections of TT conjugates. A progressive increase in the anti-HCH response with each additional injection was noted in animals receiving HCH conjugates. Animals receiving the diphtheria toxoid-TT-pertussis vaccine preparation responded with a greater increase in anti-carrier antibody than those receiving the conjugates alone. This method of synthesis provided conjugates capable of inducing protective levels of antibodies to both the polysaccharides and carrier proteins.

Animals↗
PubMed1983

Bacterial meningitis in Egypt: analysis of CSF isolates from hospital patients in Cairo, 1977-78.

Bacterial meningitis remains a major cause of mortality and morbidity in many countries of the world despite effective antimicrobial therapy. Studies of the etiology and some laboratory characteristics of bacterial meningitis in Egypt were conducted during 1977-1978. All patients suspected of having bacterial meningitis were studied at the time of admission to the two fever hospitals of Cairo. Direct culture, serological identification of the capsular type, and countercurrent-immunoelectrophoresis of 1627 CSF specimens were done. Of these, 276 had bacteria identified either by culture or Gram stain. Pneumococci were the most common and the serotype distribution was similar to that reported from other parts of Africa; second were meningococci with groups C and B predominating; in third place was Haemophilus influenzae type b which caused the highest mortality and had an unusually young age distribution. There were 77 bacterial isolates (22%), including 11 species, designated as "other" because there was no predominant species. There were many "clear" CSF specimens that were found to contain pneumococci, meningococci or H. influenzae type b, confirming the need for more comprehensive laboratory facilities for accurate diagnosis of the etiology of bacterial meningitis.

Adolescent↗
PubMed1982

Differential complement resistance mediates virulence of Haemophilus influenzae type b.

Studies were undertaken to gain insight into the virulence of type b in contrast to the other Haemophilus influenzae capsular types. A relationship was found between the comparative virulence of H. influenzae types in humans and their resistance to the bactericidal effect of antibody-free complement. Type b was most resistant to the bactericidal effect of complement. The other types could be divided into three groups based upon their susceptibility to complement; this grouping was also related to their structural similarities. No association between virulence and either the biotype, source of isolate, in vitro association with peripheral polymorphonuclear leukocytes, or the total amount of capsular polysaccharide was found. However, among the type b strains, higher levels of cell-associated polysaccharide were associated with increased resistance to complement. The relative virulence of the six H. influenzae types in the infant rat model was generally similar to that in humans. After intraperitoneal challenge, type b and type a strains had the lowest 50% effective doses for bacteremia, removed by several logs from the values of the other types. By intranasal challenge, type b strains produced higher rates and levels of bacteremia than did type a strains. High levels of natural bactericidal antibodies to types c and e were found in adult female rats; this finding alone could not account for the differences in virulence among the H. influenzae types in the infant rat model. We propose that the virulence of type b strains is due to their greater resistance to the bactericidal activity of serum complement alone. Resistance to type b disease requires serum antibody to induce the complement-mediated reaction.

Animals↗
PubMed1982

Cross-immunogenicity of pneumococcal group 9 capsular polysaccharides in adult volunteers.

Group 9 organisms (types 9N, 9A, 9L, and 9V) account for about 3 to 4% of pneumococcal disease isolates throughout the world. Types 9N and 9V comprise about 90% of the group 9 disease isolates. Type 9N is more common than type 9V in adults, and type 9V predominates in infants and children. In the United States there have been eight reported cases due to group 9 pneumococci in individuals previously vaccinated; six were type 9V and two were type 9N. To ascertain the cross-immunogenicity of group 9 polysaccharides, volunteers were injected with vaccines of monovalent types 9N, 9A, 9V, or 9L, or bivalent (9N and 9A) or trivalent (9N, 9A, and 9V) polysaccharide vaccines. Monovalent types 9N, 9V, and 9L each stimulated a 5.8- to 7.5-fold geometric mean rise, and at least 80% of the volunteers responded with a twofold or greater homologous antibody rise. Type 9V induced a 5.8-fold geometric mean rise, but only 66% of the volunteers responded with a twofold or greater homologous antibody rise. Type 9N induced only a 2.1-fold geometric increase, and only 54% of the volunteers responded with a twofold or greater rise in anti 9V antibodies. Types 9L and 9A were the most cross-immunogenic. The trivalent preparation (9N, 9A, and 9V) gave the highest geometric mean titer and seroconversion rate to each of the group 9 polysaccharides. These results suggest that the polyvalent pneumococcal vaccine with its type 9N does not induce a satisfactory anti-type 9V response and should contain additional components in order to achieve greater protection against group 9 organisms.

Adult↗
PubMed1982

Antibody responses in adult volunteers to pneumococcal polysaccharide types 19F and 19A administered singly and in combination.

The immunogenicity of the two major group 19 pneumococcal polysaccharides, types 19F and 19A, was studied in human volunteers. Antibody responses after vaccination with either 50 micrograms of type 19F polysaccharide, 50 micrograms of type 19A polysaccharide, or 25 micrograms of both polysaccharides premixed in a single injection revealed that the combined vaccine was immunogenic for both types, but that the optimal immunogen for each was the homologous polysaccharide. We concluded that vaccination with type 19F polysaccharide antigen, included in the current pneumococcal vaccine, does not induce a satisfactory heteroimmunogenic response to type 19A.

Antibodies, Bacterial↗
PubMed1982

Hydrolytic stability of pneumococcal group 6 (type 6A and 6B) capsular polysaccharides.

The hydrolyses of the immunologically cross-reactive and constitutionally isomeric group 6 pneumococcal polysaccharides, types 6A and 6B, were investigated by 31P nuclear magnetic resonance spectroscopy, gel filtration through Sepharose 4B, reducing-sugar analysis, and rocket immunoelectrophoresis. Phosphorus nuclear magnetic resonance spectroscopy showed that cleavage of the repeating-unit phosphodiester linkages at pH 10, 60 degrees C was considerably faster (greater than 10(3) ) for the type 6A than the type 6B polysaccharide. Under these reaction conditions, 31P nuclear magnetic resonance kinetic measurements showed that the Na+ form of the type 6A polysaccharide underwent phosphodiester-linkage hydrolysis two times slower than the corresponding Ca+2 form; a stoichiometrically excess amount of Ca+2 caused a 30-fold enhancement of the latter hydrolysis rate. The spectroscopic characterization of phosphorus-containing end groups resulting from hydrolysis of the type 6A polymer provided additional mechanistic information. Heating the type 6A and 6B polysaccharides at 56 degrees C for various times led to gel filtration coefficients of distribution (Kd values) which indicated that the type 6A material underwent size reductions considerably faster than did the type 6B antigen; these increased Kd values qualitatively correlated with the loss of immunochemical reactivity measured by rocket immunoelectrophoresis. The application of a statistical theory to the depolymerization of the type 6A and 6B polysaccharides was consistent with random bond cleavage, as evidenced by the calculated versus measured gel filtration patterns. Although the molecular changes causing the size reductions were not fully elaborated, it was established that the acetal linkages of the type 6A and 6B polysaccharides were comparatively resistant to hydrolysis and that depolymerization by hydrolysis of the phosphodiester linkage was a major factor only in the type 6A structure. It was concluded that the hydrolytic stability of the type 6B antigen would favor its use in the polyvalent pneumococcal vaccine rather than the cross-reactive, but comparatively unstable, type 6A polysaccharide, if all other factors are equal.

Chemical Phenomena↗
PubMed1982

Identification of immunoglobulin heavy-chain isotypes of specific antibodies of horse 46 group B meningococcal antiserum.

Hyperimmune horse serum from a single animal (horse 46) immunized with group B (strain B-11) meningococcal vaccine provides a standardized, readily available diagnostic reagent used in primary isolation medium and for serogrouping of meningococci. Identification of the heavy-chain isotypes of specific anticapsular polysaccharide and anti-lipopolysaccharide isolated from horse 46 serum revealed a differential distribution in the occurrence of immunoglobulin classes. Meningococcal anticapsular antibodies of horse 46 serum were restricted predominately to the immunoglobulin M (IgM) class, with only trace amounts of IgGa present, whereas anti-lipopolysaccharide concomitantly produced showed a heterogeneity in its heavy-chain isotypes, consisting of IgM, IgGa, IgGb, moderate amounts of IgB, and a small amount of IgA.

Animals↗
PubMed1981

Lipid on capsular polysaccharides of gram-negative bacteria.

Hydrolysis of the meningococcal group A, B, and C, and Escherichia coli K92 polysaccharides by 60% aqueous hydrofluoric acid liberated various 1,2-diacylglycerols. These were extracted with chloroform, trimethylsilylated, and analyzed by gas chromatography-mass spectrometry. Two 1,2-diacylglycerols were the major components isolated. In each polysaccharide, 80 to 90% dipalmitoyl glycerol and 10 to 20% distearoyl glycerol were identified. No monoacylglycerols or mixed diacylglycerols were noted. The presence of the hydrophobic end causes the polysaccharides to aggregate in a micellar form and may be the entity by which the polysaccharide remains attached to the outer membrane of the bacterium giving rise to the structure recognized as a capsule.

Diglycerides↗
PubMed1981

Group C Neisseria meningitidis variant polysaccharide vaccines in children.

The currently United States-licensed group C Neisseria meningitidis vaccine, composed of the O-acetyl-positive capsular polysaccharide, is poorly immunogenic and does not afford protection from disease to infants and young children. Group C N. meningitidis O-acetyl-negative polysaccharide vaccine induces higher titers in adults than does the O-acetyl-positive vaccine. We compared the immunogenicity of these vaccines in 2-year-old children. Reactions were minimal and did not differ between the two vaccines. The postvaccination geometric mean titer was twofold greater in the O-acetyl-negative group (1.58 versus 0.73 micrograms of antibody per ml). The rates of decline in titer were similar in both groups. Further study regarding immunogenicity of and the anamnestic response to the O-acetyl-negative vaccine is warranted in the age group (less than 18 months) at highest risk for invasive meningococcal disease.

Antibodies, Bacterial↗
PubMed1980

Preparation, characterization, and immunogenicity of Haemophilus influenzae type b polysaccharide-protein conjugates.

A method is presented for covalently bonding Haemophilus influenzae type b capsular polysaccharide (HIB Ps) to several proteins. The method is efficient and relies upon the use of adipic dihydrazide as a spacer between the capsular polysaccharide and the carrier protein. In contrast to the poor immunogenicity of the purified HIB Ps in mice and rabbits, the HIB Ps-protein conjugates induced serum anti-type b antibodies having bactericidal activity at levels shown to be protective in humans when low doses were injected subcutaneously in a saline solution. The antibody response in mice was related to the dose of the conjugates, increased with the number of injections, and could be primed by the previous injection of the carrier protein. The HIB Ps-protein conjugates were immunogenic in three different mouse strains. The importance of the carrier molecule for the enhanced immunogenicity of the HIB Ps-protein conjugates was shown by the failure of HIB Ps hybrids prepared with either the homologous polysaccharide or pneumococcus type 3 polysaccharide to induce antibodie in mice. Rabbits injected with the HIB Ps-protein conjugates emulsified in Freund's adjuvant produced high levels of serum anti-type b antibodies which induced a bactericidal effect upon H. influenzae type b organisms. It is proposed that the HIB Ps component of the polysaccharide protein conjugates has been converted to a thymic-dependent immunogen. This method may be used to prepare protein-polysaccharide conjugates with HIB Ps and other polysaccharides to be considered for human use.

Animals↗
PubMed1980

Identification of Salmonella typhi in faecal specimens by an antiserum-agar method.

An antiserum-agar technique was evaluated as a method for detecting Salmonella typhi in faeces. Thirty-one laboratory strains of S. typhi produced immunoprecipitate haloes during overnight growth on SS agar and blood-agar-base infusion agar (BAB) containing donkey antiserum to a vaccine strain of S. typhi. Other salmonella species sharing O serogroup antigens with S. typhi also produced haloes when streaked in pure culture on SS-antiserum agar but not on BAB-antiserum agar. One hundred and forty-one consecutive faecal specimens were cultured on SS-antiserum agar. Results with this method were concordant with those of established isolation techniques on specimens from six of seven suspected carriers of S. typhi. Ten other salmonellas were isolated from the faecal specimens but only S. javiana, like S. typhi a serogroup-D organism, yielded false-positive haloes on antiserum agar. The antiserum-agar technique offers promise as a means of screening for S. typhi in faecal cultures.

Agar↗
PubMed1979

Form variation in Escherichia coli K1: determined by O-acetylation of the capsular polysaccharide.

The chemical basis for the alternating antigenic change called form variation noted for the Escherichia coli K1-capsular polysaccharide has been shown by 13C nuclear magnetic resonance to be a result of random O-acetylation of C7 and C9 carbons of the alpha-2-8-linked sialic acid homopolymer. A serologic method (antiserum agar) was developed to identify and isolate the form variants. The O-acetyl positive and O-acetyl negative K1 polysaccharides had unique biochemical and immunologic properties. The O-acetyl-positive variants resisted neuraminidase hydrolysis in contrast to the susceptibility of the O-acetyl negative variant to this enzyme. In addition, O-acetylation altered the antigenicity of the O-acetyl polysaccharides. When injected as whole organisms, O-acetyl positive organisms produced anti-K1 -antibodies in rabbits specific for this polysaccharide variant. O-acetyl negative organisms were comparatively less immunogenic; however, antibodies induced by these organisms reacted with both K1 polysaccharide variants. Burros, injected with either variant, produced antibodies reactive with both K1 polysaccharides.

Acetylation↗