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D M Granoff

Publications and source records attributed to D M Granoff.

At least 91 records · Page 5Linked to original sources

Subclass distribution of human antibodies to Haemophilus influenzae type b capsular polysaccharide.

The polysaccharide (PS) capsule of Haemophilus influenzae type b (Hib) is a "simple" antigen, polyribosylribitolphosphate. Although similar carbohydrate antigens have been reported to elicit IgG antibodies relatively restricted to the IgG2 subclass in man, we report here that Hib PS elicits substantial quantities of both IgG1 and IgG2 serum antibodies in most individuals. Because the determination of IgG subclass distribution can be technically difficult, we used four different approaches to establish our finding. First, we used an IgG subclass-specific, antigen-specific "sandwich assay." Second, we measured IgG subclasses of purified antibodies to Hib PS. Third, we showed that significant amounts of IgG anti-PS can be absorbed with a monoclonal anti-IgG1 affinity column. Fourth, we showed that IgG1 and IgG2 fractions of immune sera have clonally restricted anti-Hib PS antibodies that are easily distinguishable by their isoelectric points. The data indicate that both IgG1 and IgG2 contribute substantially to the IgG antibody response of most adults to immunization with Hib PS.

Adult↗

Concentrations of antibodies in paired maternal and infant sera: relationship to IgG subclass.

Previous studies comparing IgG subclass concentrations in cord and maternal sera have indicated that IgG1 is transported across the placenta to a greater extent than is IgG2. The purpose of our study was to examine the relationship between the transport of IgG1 and IgG2 and the transport of specific antibodies that are relatively restricted to a particular subclass, either IgG1 or IgG2. The concentrations of total serum IgG1 and IgG2 and those of IgG-anti-tetanus toxoid (TT) and anti-group A streptococcal carbohydrate (GAC) were measured in 30 paired maternal and cord sera. Previous studies have shown that anti-TT in adults is predominantly IgG1, whereas anti-GAC is predominantly IgG2. The mean cord/maternal concentration ratios of IgG1 and anti-TT were similar (1.77 +/- 0.56 and 1.93 +/- 0.67, respectively), but differed significantly (P = 0.0001) from those of IgG2 and anti-GAC (0.99 +/- 0.39 and 1.01 +/- 0.45, respectively). We confirmed the difference in cord/maternal concentration ratios of IgG1 and IgG2 antibodies by measuring IgG1 and IgG2 antibodies specific for Haemophilus influenzae type b capsular polysaccharide; the mean cord/maternal concentration ratio of IgG1-anti-Hib PS was significantly higher than that of IgG2-anti-Hib PS (2.23 +/- 0.83 compared with 0.94 +/- 0.49, P = 0.01). These results indicate that placental transport of IgG antibodies is related to their subclass composition.

Adult↗

Immunologic priming to capsular polysaccharide in infants immunized with Haemophilus influenzae type b polysaccharide-Neisseria meningitidis outer membrane protein conjugate vaccine.

Thirty children vaccinated at 2 to 17 months of age with Haemophilus influenzae type b polysaccharide linked to a partially purified 40,000 dalton outer membrane protein of Neisseria meningitidis were revaccinated 10 to 14 months later with conventional H. influenzae type b polysaccharide vaccine. The geometric mean anti-type b antibody concentration before reimmunization was 0.68 micrograms/mL, and rose to 31 micrograms/mL in sera obtained 1 month later. The mean level after immunization was not significantly different than that in sera from 12 adults immunized with type b polysaccharide vaccine (51 micrograms/mL, P = 0.3), and was 10-fold higher than that of 13 control children immunized with type b polysaccharide for the first time (2.7 micrograms/mL, P less than 0.001). The IgG responses of the children first given conjugate vaccine and then conventional type b polysaccharide vaccine were of a similar magnitude as those in the immunized adults. Further, the children maintained high levels of serum antibody 6 to 8 months later. Ten other children vaccinated in infancy with conjugate vaccine, and again with conjugate vaccine 10 to 15 months later, showed similar antibody responses to those of the group given conjugate vaccine in infancy, and booster with conventional polysaccharide vaccine. Thus vaccination with H. influenzae type b polysaccharide-outer membrane protein conjugate vaccine primes the immune system to an IgG memory antibody response to either type b polysaccharide or conjugate vaccine. Post-booster sera from all children tested showed high titers of functional activity in a complement-mediated bactericidal assay. These data suggest that protection of most infants from type b Haemophilus disease may be achieved by a combination of immunization at 2 to 4 months of age with this conjugate vaccine, and reimmunization 1 year later with conjugate or conventional type b polysaccharide vaccine.

Adult↗

Second episodes of Haemophilus influenzae type b disease following rifampin prophylaxis of the index patients.

Patients treated for Haemophilus influenzae type b disease frequently remain nasopharyngeal carriers of that organism and fail to develop protective concentrations of serum antibody. It has been suggested that rifampin prophylaxis of the index patient may prevent recurrence of disease by eliminating type b Haemophilus carriage. We report nine children who developed second episodes of disease 1 week or more after receiving rifampin prophylaxis. The median interval between the last dose of rifampin and admission to the hospital for the second episode was 70 days (range, 9 to 138). Analysis of biotypes and outer membrane protein polyacrylamide gel electrophoresis patterns of paired isolates from eight cases revealed that the second episodes in two of the children were caused by acquisition of new type b Haemophilus strains, whereas the second episodes in the remaining six children were caused by isolates which were indistinguishable from the respective isolates from the first episodes. Rifampin prophylaxis of the index patient may prevent some episodes of recurrent disease. However, in some patients who have received prophylaxis, second episodes can occur, probably as a result of reacquisition of the organism from contacts who did not receive rifampin or from acquisition of new type b strains.

Bacterial Outer Membrane Proteins↗

Immunogenicity of Haemophilus influenzae type b polysaccharide-Neisseria meningitidis outer membrane protein conjugate vaccine in 2- to 6-month-old infants.

Fifty infants, 2 to 6 months of age, were vaccinated with Haemophilus influenzae type b capsular polysaccharide covalently linked to an outer membrane protein from Neisseria meningitidis group B. Subjects were given two injections and were randomly assigned to receive the injections separated by 1 or 2 months. Each dose contained 15 micrograms of polysaccharide and 51 micrograms of protein, or approximately twice the amount of polysaccharide as used in our previous trial (Lancet 1986;2:299). Fevers of 38.0 degrees to 38.8 degrees C developed in three infants (6%) within 24 hours after vaccination, but there were no other notable reactions. Following one injection, the geometric mean antibody concentration increased from 0.13 micrograms/mL in preimmune serum to 1.50 micrograms/mL in serum obtained 1 to 2 months later (P less than .001). After a second injection, there was a further increase in serum antibody (geometric mean = 3.11 micrograms/mL, P less than .007). The geometric mean antibody concentration of the group reimmunized 2 months after the first injection was higher than that in the group reimmunized after 1 month (3.95 v 2.32 micrograms/mL, P = .05, by analysis of covariance with age as the covariant). These data confirm our previous preliminary observations on the safety and immunogenicity of this new conjugate vaccine in infants 2 to 6 months of age. The data suggest that a 2-month interval between the first and second injections results in higher levels of serum antibody than a 1-month interval.

Antibody Formation↗

Prospective surveillance of Haemophilus influenzae type b disease in Dallas County, Texas, and in Minnesota.

Among children less than 12 years of age residing in Dallas County, Texas, and in the state of Minnesota we conducted prospective, active surveillance of invasive Haemophilus influenzae disease. During 18 months, 616 cases were identified, of which 600 were caused by type b organisms. The annual incidence of disease was significantly greater in Dallas than in Minnesota (109 v 68/100,000 children younger than 5 years of age, P less than .001) and was greater in Dallas, even when rates for white children in the two regions were compared (P less than .001). Other regional differences were observed. In Dallas, a larger proportion of cases were in children attending day-care centers (27% compared with 12% in Minnesota, P less than .001) and more patients attended day care for greater than 40 h/wk (56% compared with 30% in Minnesota, P less than .001). Outer membrane protein subtyping of isolates revealed that in Dallas 6U isolates were associated significantly with cases in black children who attended day care. In Minnesota, but not in Dallas, isolates with subtype 1H were associated significantly with cases in children in day care. These data indicate that there are regional differences in the epidemiology of type b Haemophilus disease that may relate to differences in strains, day-care practices, or other unknown cultural or environmental factors. Finally, because only 15% of systemic Haemophilus disease in these regions occurred in children in the age groups recommended for vaccination (24 to 59 months), the new Haemophilus type b polysaccharide vaccine is expected to have a limited impact on the overall incidence of disease.

Bacterial Outer Membrane Proteins↗

Hemophilus influenzae type B disease in children vaccinated with type B polysaccharide vaccine.

We studied 55 cases of invasive Hemophilus influenzae type b disease occurring in children at least three weeks after vaccination with type b polysaccharide vaccine. Their mean age at the time of immunization was 27.8 months (range, 18 to 47). Meningitis developed in 39 patients, of whom 3 died and 6 had neurologic sequelae. We investigated certain host factors that may have contributed to the failure of the vaccine. The geometric mean concentration of antibody to type b polysaccharide in convalescent-phase serum from 31 of the vaccinated patients who had hemophilus disease was significantly lower than that in serum from 25 patients of similar age with the disease who had never been vaccinated (0.59 vs. 3.46 micrograms per milliliter, P less than 0.001). However, only 3 of 46 patients in whom the vaccine failed and who were tested for hypogammaglobulinemia had this finding, and none of 33 children tested for IgG2 had low serum concentrations of this immunoglobulin subclass, which is thought to be important in the immune response to polysaccharide antigens. In addition, all but 1 of the 46 patients in whom the vaccine failed and who were tested for IgG antibody to tetanus toxoid protein, a thymic-dependent antigen, had normal values, and 19 of 20 tested for hemolytic complement activity had normal levels. In white children, the presence of the Gm immunoglobulin phenotype (1,2,3, 17; ;5,13,21) was associated with a sevenfold increase in the relative risk of vaccine failure (P less than 0.003). We conclude that vaccine failure may be related in part to genetic factors, and that most vaccinated children in whom Hemophilus influenzae disease develops have deficient antibody responses to the type b polysaccharide despite normal serum concentrations of immunoglobulin and normal antibody responses to tetanus toxoid.

Antibodies, Bacterial↗

Immunogenicity in infants of Haemophilus influenzae type B polysaccharide in a conjugate vaccine with Neisseria meningitidis outer-membrane protein.

63 children, aged 2-17 months, were given a new conjugate vaccine composed of the capsular polysaccharide of Haemophilus influenzae type b linked to a Neisseria meningitidis outer-membrane protein. Subjects under 7 months received two injections separated by 1 month, and older subjects received either one or two injections. There were no systemic reactions to this vaccine when it was given with aluminium hydroxide. A single injection of vaccine was highly immunogenic; the geometric mean serum anticapsular antibody concentrations before immunisation and 1 month later were 0.35 microgram/ml and 0.98 microgram/ml for babies of 2-3 months, 0.12 microgram/ml and 1.85 micrograms/ml for those of 4-6 months, and 0.15 microgram/ml and 4.1 micrograms/ml for those of 8-17 months (p less than or equal to 0.003 for each age group). After a second injection of vaccine, 80% and 76% of infants of 2-3 and 4-6 months, respectively, had antibody concentrations greater than 1.0 micrograms/ml. Most subjects showed evidence of IgG responses as measured by enzyme-linked immunosorbent assay. 6-12 months after immunisation, serum antibody levels had fallen (p less than 0.05) but they remained higher than those of unimmunized controls (p less than 0.001).

Adult↗

Functional activity of different IgG subclass antibodies against type b capsular polysaccharide of Haemophilus influenzae.

The biologic activity of different human IgG subclass antibodies directed against the Haemophilus influenzae type b (Hib) capsular polysaccharide (PRP) was compared by using an in vitro complement-mediated bactericidal assay and an in vivo passive protection assay in infant rats. An IgG pool was made by Sephacryl S-300 chromatography of sera from adults immunized with PRP vaccine. An IgG2 subclass fraction was prepared by column immunoabsorption of the IgG pool with anti-IgG1 monoclonal antibody. An IgG1 subclass fraction was eluted from the affinity matrix. IgG1, IgG2, IgG3, and IgG4 concentrations in the fractions were measured by solid-phase competitive radioimmunoassays, and anti-PRP antibody was measured by a modified Farr assay. Each fraction was greater than 90% pure IgG2 or IgG1, respectively. There were no significant differences in the minimal anti-PRP antibody concentrations required to kill 50% of Hib cells in vitro (IgG, 0.22; IgG1, 0.21; and IgG2, 0.42 microgram/ml). Similarly, equivalent amounts of anti-PRP antibody of the IgG1 or IgG2 fractions protected against bacteremia (IgG1, 0.12; IgG2, 0.24 microgram per rat). IgG absorbed to remove anti-PRP antibody was neither bactericidal nor protective. Thus IgG1 and IgG2 anti-PRP antibody have equivalent functional activities against Hib as determined by these biologic assays.

Adult↗

Pathogenicity of a rifampin-resistant cerebrospinal fluid isolate of Haemophilus influenzae type b.

Two children in a day care facility developed Haemophilus influenzae type b meningitis. The second child was enrolled in the facility after rifampin had been administered to the other attendees. The isolate from the first child was susceptible to rifampin, but the isolate from the second was resistant. Both isolates had identical outer membrane protein PAGE profiles. To investigate the virulence of these isolates, we inoculated infant rats intranasally with either the rifampin-resistant or rifampin-susceptible CSF isolate. The rates of nasal colonization (14 of 20 and eight of eight animals inoculated with the rifampin-resistant and rifampin-susceptible isolates, respectively) did not differ significantly. However, bacteremia occurred less frequently in pups inoculated with the rifampin-resistant strain than in animals inoculated with the susceptible strain (four of 20 vs eight of eight, P less than 0.0001). Nasal washings, blood, and CSF obtained from animals inoculated with the rifampin-resistant isolate were divided and plated on media containing rifampin (1 microgram/ml) or without rifampin. Except for those from one animal, organisms isolated from blood and CSF grew only on medium lacking rifampin, whereas H. influenzae type b growing from nasal washings was frequently found on both media. We conclude that mutation of H. influenzae to rifampin resistance is a hazard of rifampin chemoprophylaxis. Rifampin-resistant isolates have the potential to cause disease in patients and experimental animals, although they may be relatively less pathogenic than the parent, susceptible organism.

Animals↗

Use of rapidly generated results in patient management.

The usefulness of a rapid diagnostic test in patient management depends on the sensitivity of the test, the clinical consequences of false-negative or false-positive results, the ease and cost of performance, and the timely availability of results. A test that is sensitive, specific, inexpensive, and rapid is presumed to be useful clinically. However, there has been surprisingly little effort to measure the actual impact of the results on patient care. Since antigen detection for Haemophilus influenzae type b disease has been available for more than a decade, it will be used as a model to illustrate several factors that help determine the benefits, limitations, and pitfalls of antigen detection in the management of patients with serious bacterial infections. Herein we will compare the use of antigen detection in meningitis with that in other Haemophilus influenzae type b diseases. We also will review our experience with the impact of rapid diagnosis on the treatment of bacterial meningitis. Finally, other factors that influence the usefulness of antigen detection on patient care will be explored by comparing the potential consequences of laboratory error on the management of patients with Haemophilus influenzae type b infections with that of management of other kinds of infections, such as streptococcal pharyngitis, sexually transmitted diseases, and viral respiratory infections.

Antigens, Bacterial↗

Prospects for prevention of Haemophilus influenzae type b disease by immunization.

A vaccine consisting of the polysaccharide (PS) capsule of Haemophilus influenzae type b (Hib) has recently been licensed in the United States. This vaccine is safe and effective in preventing invasive Hib disease in children two years of age and older, but it is ineffective in younger children, the group at greatest risk of disease. The PS vaccine also may be ineffective in preventing disease in certain subgroups of the population that are genetically at increased risk of disease and show impaired antibody responses to immunization. Thus, new strategies need to be considered. Currently, several new Hib PS-protein conjugate vaccines are being evaluated. These vaccines differ in their method of preparation, carrier protein, and PS size. In contrast to the plain Hib PS vaccine, conjugate vaccines are immunogenic in infants and elicit boostable increases in antibody to PS upon reinjection of vaccine. However, some infants less than six months of age do not respond. To confer protection on all infants, it may be necessary to modify further the conjugate vaccines. One approach is to use outer membrane proteins (OMPs) as vaccine components. Five major OMPs have been purified from Hib, and three, P1 (50 kilodalton [kDa]), P2 (37 kDa), and P6 (16 kDa), contain antigens capable of eliciting strain-specific protective antibodies in experimental animals. In summary, PS-protein conjugate vaccines hold enormous promise for the prevention of Hib disease in infants, but further work is needed to define the optimal carrier protein, PS size, and method of coupling. Information is also needed on whether genetic factors influence responses to these vaccines.

Adolescent↗

Antibody responses to Haemophilus influenzae type b polysaccharide vaccine in relation to Km(1) and G2m(23) immunoglobulin allotypes.

The antibody responses of children immunized with Haemophilus influenzae type b polysaccharide vaccine were examined in relation to the absence or presence of the Km(1) or G2m(23) immunoglobulin allotype. Ninety-seven children, 12-83 months of age, were immunized. Sera were obtained before immunization and two months later. Total serum antibody to the type b capsule was detected by a radioactive antigen-binding assay. IgG and IgM antibody responses were measured by enzyme-linked immunosorbent assay. Antibody responses to the type b capsule were more than threefold higher in blacks with the Km(1) immunoglobulin allotype compared with those in blacks lacking this allotype (P less than .02). The isotype affected was IgG (P less than .01) and not IgM. Serum concentrations of IgG2, but not of IgG1, also were higher in blacks with Km(1) (P less than .003). In whites there were no significant differences in the total or IgG-specific antibody responses to the type b capsule in relation to the Km(1) or G2m(23) allotype.

Antibodies, Bacterial↗

Genetic relationships of serologically nontypable and serotype b strains of Haemophilus influenzae.

A collection of 242 strains of Haemophilus influenzae, including 65 nontypable (unencapsulated) isolates and 177 encapsulated serotype b isolates recovered largely from children with invasive and noninvasive diseases in the United States, was characterized by the electrophoretic mobilities of 15 metabolic enzymes presumably encoded by chromosomal genes. All enzymes were polymorphic for three to seven electromorphs, and 94 distinctive multilocus genotypes (electrophoretic types [ETs]) were distinguished, among which mean genetic (allelic) diversity was 0.500. Isolates recovered from cases of invasive or noninvasive diseases did not differ significantly in level of genetic variation. The observation that 29 ETs were represented exclusively by serotype b isolates and that each of the 65 nontypable isolates was of a unique ET strongly confirmed the hypothesis that unencapsulated clinical isolates are not merely phenotypic variants of the common serotype b cell lines. Rather, the two types of isolates are distinctive subsets of the multilocus chromosomal genotypes of the species as a whole. Serotype b capsule occurred in three groups of isolates that are distantly related in multilocus enzyme genotype. Isolates of four closely related nontypable biotype IV ETs associated with obstetrical infections or neonatal bacteremia were highly divergent from all others examined and may be specifically distinct. A phylogenetic scenario was proposed in which the ancestor of H. influenzae was encapsulated and the nontypable clones arose by convergent evolutionary loss of the ability to synthesize or extracellularly express a polysaccharide capsule.

Alleles↗

Lipopolysaccharide gel profiles of Haemophilus influenzae type b are not stable epidemiologic markers.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lipopolysaccharide (LPS) was performed to assess the usefulness of this technique for the epidemiologic analysis of Haemophilus influenzae type b isolates. LPS samples were prepared from isolates which had been passaged either in vitro or in infant rats. Preparations from paired isolates from a number of epidemiologically related clinical situations also were examined. The gel patterns of LPS prepared on different occasions from an individual isolate were stable. However, the LPS gel patterns changed in 5 of 14 (36%) of the passaged isolates, and differences in gel patterns also were observed among epidemiologically related isolates. The variability in LPS electrophoretic patterns of individual isolates indicated that this technique is not useful for the epidemiologic analysis of H. influenzae type b disease.

Animals↗

Haemophilus influenzae type b disease in an Amish population: studies of the effects of genetic factors, immunization, and rifampin prophylaxis on the course of an outbreak.

In 1982, an outbreak of Haemophilus influenzae type b disease occurred in a 379-member Amish community. In an attempt to control the outbreak after the occurrence of the second case of disease, we investigated the combination of (1) rifampin chemoprophylaxis of all carriers of H influenzae type b and their household contacts from 1 month to 5 years of age and (2) H influenzae type b polysaccharide vaccine immunoprophylaxis of all community members 12 months of age and older. Despite our intervention, two additional cases of bacteremic H influenzae type b disease occurred in the ensuing 5 months, one in a 22-month-old infant who had been immunized at 19 months of age and the other in a child who had not been immunized because she was younger than 12 months of age. The outbreak ended following rifampin prophylaxis of all community members younger than 15 years of age. All of the children with disease were genetically related to one another, and three of the four were inbred. However, analysis of their coancestry revealed that neither the average level of kinship nor the average inbreeding level of the affected children differed significantly from those of the other children in the community. Furthermore, none of the four children with disease shared a human leukocyte antigen haplotype. Our observations suggest that inbreeding was not a risk factor in this community.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗