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Biomedical subjects

D M Granoff

Publications and source records attributed to D M Granoff.

At least 109 records · Page 6Linked to original sources
PubMed1985

Correlation of serum immunoglobulin subclass concentrations with antibody responses of children to immunization with Haemophilus influenzae type b polysaccharide-pertussis vaccine.

Children less than 24 months of age respond poorly to immunization with the capsular polysaccharide of Haemophilus influenzae type b. Because human antibodies to polysaccharide antigens are relatively restricted to IgG2, a late-maturing subclass, we examined the relationship between serum subclass concentrations and anticapsular antibody responses of 41 healthy children, 9 to 38 months of age, following immunization with type b polysaccharide mixed with pertussis vaccine. Both total and IgG anticapsular antibody responses correlated significantly with preimmune serum concentrations of IgG2 but not with those if IgG1. This correlation was age dependent, however, and after the effect of age was removed by partial correlation, the correlation between anticapsular antibody responses and serum IgG2 concentrations was no longer significant. These findings indicate that the ability to respond to this vaccine coincides with maturation of the ability to secrete immunoglobulin of the IgG2 subclass; however, individual variation in IgG2 that is independent of age does not correlate with antibody response to the type b polysaccharide.

Age Factors↗
PubMed1985

A population genetic framework for the study of invasive diseases caused by serotype b strains of Haemophilus influenzae.

One hundred seventy-seven isolates of serotype b Haemophilus influenzae recovered largely from children with invasive disease in the United States were characterized by the electrophoretic mobilities of 16 metabolic enzymes, the NaDodSO4/PAGE pattern of outer-membrane proteins (OMP), and biotype. Thirty-two distinctive multilocus genotypes (electrophoretic types, ETs) were distinguished on the basis of allele profiles at the enzyme loci. Twenty-eight OMP types and five biotypes were identified, but only 55 distinctive combinations of ET, OMP type, and biotype were represented. The strong nonrandom associations of characters and the recovery of isolates with identical properties in widely separated geographic regions and over a 40-year period suggest that the population structure of H. influenzae is basically clonal. Examination of nonserotype b isolates indicated that clones of serotype b are a restricted subset of the genotypes in the species as a whole. Currently, most of the invasive H. influenzae disease in the United States is caused by serotype b strains of two related ETs, and, more specifically, much of it is attributable to two subclones marked by OMP type. There is evidence that the frequency of the ET-1/OMP 1H/biotype I subclone has increased dramatically in the United States since the 1939-1954 period. The hypothesis that populations of H. influenzae are subject to marked temporal variation in clonal composition is supported by evidence of major differences in the genetic structure of populations in the United States and the Netherlands.

Bacterial Outer Membrane Proteins↗
PubMed1985

Serum opsonic activity after immunization of adults with Haemophilus influenzae type b-diphtheria toxoid conjugate vaccine.

We measured the uptake of radiolabeled Haemophilus influenzae type b by human polymorphonuclear leukocytes. Haemophilus influenzae type b strains were preopsonized in individual sera from six adults immunized with type b polysaccharide vaccine (PRP) or six adults immunized with PRP covalently coupled to diphtheria toxoid (PRP-D vaccine). Serum was heat inactivated before use, and exogenous human complement was added. Of the 12 subjects, 3 had high levels of opsonic activity (greater than 40% of immune control) in their preimmunization serum. This activity did not correlate with the concentrations of anti-PRP antibody and was unaffected by absorption of anti-PRP antibody. At 1 month after vaccination, the serum of PRP-D subjects had higher opsonic activity than that from subjects who received PRP (5% serum, mean PRP-D = 86%, mean PRP = 53%, P = 0.001). After 12 months, both groups had higher serum opsonic activity than before immunization (P less than 0.02), but there was no difference between the two groups (mean PRP-D = 48%, mean PRP = 51%). In postimmunization serum, opsonic activity induced by PRP-D or PRP vaccines correlated directly with anti-PRP antibody concentrations as measured by a radioantigen binding assay. We conclude that both vaccines induce opsonic activity, opsonic activity induced by immunization of adults correlates well with the concentration of anti-PRP antibody achieved, and in preimmune sera with low concentrations of anti-PRP antibody, factors other than anti-PRP antibody contribute to opsonic activity.

Adult↗
PubMed1985

Purification and partial characterization of outer membrane proteins P5 and P6 from Haemophilus influenzae type b.

The major outer membrane proteins of Haemophilus influenzae type b (Hib), designated P5 and P6 (R.S. Munson, Jr., J.L. Shenep, S.J. Barenkamp, and D.M. Granoff, J. Clin. Invest. 72:677-684, 1983), were purified to homogeneity and partially characterized. P5 was insoluble in octylglucoside-NaCl and could be extracted with 1% sodium dodecyl sulfate (SDS) in 20 mM phosphate (pH 7.5). Solubilized P5 was further purified on hydroxylapatite in 0.1% SDS. The purified protein had an apparent molecular weight of 27,000 as determined by SDS-polyacrylamide gel electrophoresis after sample preparation at room temperature. The protein migrated with an apparent molecular weight of 35,000 after heating for 30 min at 100 degrees C in the presence of 10% beta-mercaptoethanol (beta ME). Rabbit antisera prepared against the purified preparation immunoprecipitated solubilized protein P5 but had no protective activity in the infant rat bacteremic model. The SDS-insoluble residue was further extracted with 1% SDS-0.5 M NaCl-0.1% beta ME at 37 degrees C. A single outer membrane protein, designated P6, with an apparent molecular weight of 16,000, remained insoluble under these conditions. Antiserum prepared against this insoluble fraction contained antibodies which, after removal of anti-lipopolysaccharide antibody, immunoprecipitated P6 and protected infant rats challenged with Hib. Protein P6 could be released from the insoluble cell wall in the presence of SDS-NaCl-beta ME at 60 degrees C. Thus, proteins P5 and P6 could be purified from the cell envelope of Hib. Based on the results from infant rat passive protection experiments, antigens in the P6-cell wall fraction merit further investigation as possible vaccine components. In contrast, epitopes on protein P5 did not appear to elicit protective antibody.

Animals↗
PubMed1985

Relation of age, race, and allotype to immunoglobulin subclass concentrations.

Concentrations of IgG1, IgG2, and total IgG were measured by a solid phase radioimmunoassay in sera from 36 healthy adults and 114 healthy children. As expected, IgG2 and total IgG had a positive correlation with age in children. In addition to age, several other factors were associated with significant differences in serum subclass concentrations. Female children had higher concentrations of IgG1 than males, and black subjects had significantly higher concentrations of IgG1, IgG2, and total IgG than whites. Although Km(1) and Gm(23) immunoglobulin allotypes had no relation to subclass concentrations when tested as single factors, the Km(1) allotype interacted significantly with race so that Km(1)-positive black children had higher IgG2 concentrations than other subjects. Our findings may explain, in part, recent observations of an association of the Km(1) allotype with altered immune responses of blacks to certain vaccines containing bacterial polysaccharides. In addition, our data indicate the need to control factors such as sex, race, and allotype in studies of subclass concentrations or immune responses.

Adolescent↗
PubMed1985

Effect of rifampin chemoprophylaxis on carriage eradication and new acquisition of Haemophilus influenzae type b in contacts.

We conducted a multicenter trial designed to assess the efficacy of three different drug regimens on eradication of Haemophilus influenzae type b (HIB) from the nasopharynx of household contacts of patients with invasive type b Haemophilus disease. The drug regimens studied were rifampin, 20 mg/kg, once daily for four days; rifampin, 10 mg/kg, twice a day for four days; and placebo, once daily for four days. Shortly after admission of the index patient to the hospital, 26% of 492 household contacts were found to be colonized with HIB. Both rifampin regimens eradicated carriage significantly better than placebo at 10 and 30 days (P = .001). However, among contacts whose cultures were initially negative, new acquisition of the organism occurred infrequently in this 30-day follow-up period regardless of the drug or placebo regimen prescribed. We also measured the concentration of anticapsular antibody in sera obtained from contacts younger than 6 years of age. Samples were obtained soon after admission of the index patient to the hospital and 30 days later. Several carriers younger than 2 years of age had low concentrations of antibody in both specimens. In contrast, nearly all carriers 2 to 5 years of age had high concentrations of antibody even in the first sample. Children who were not carriers usually had low antibody concentrations which did not increase during the period of observation. Our results suggest that most intrafamilial spread of HIB occurs prior to hospitalization of the index patient and stimulates immunity in contacts older than 2 years of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1984

Immunogenicity of Haemophilus influenzae type b polysaccharide--diphtheria toxoid conjugate vaccine in adults.

The capsular polysaccharide of Haemophilus influenzae type b is a poor immunogen in human infants. In an attempt to enhance immunogenicity, this polysaccharide was covalently coupled to diphtheria toxoid and the conjugate tested as a vaccine in adult volunteers. Two injections of PRP-D vaccine were given, separated by one month. The anti-PRP antibody responses in this group were compared with those in a group receiving a comparable dose (20 micrograms) of conventional PRP vaccine. Both vaccines were well tolerated. A single injection of PRP-D was significantly more immunogenic than PRP, eliciting higher serum concentrations of total anti-PRP antibody 1 month later (geo means of 248 and 62 micrograms/ml, respectively; P less than 0.001). In addition, higher concentrations of IgG anti-PRP antibody were observed in the PRP-D group (P less than 0.001). One month after reinjection of vaccine, subjects receiving PRP-D showed a small but significant decline in total antibody (P = 0.03), whereas the serum antibody concentrations in the group that received PRP remained unchanged. At 12 months, the antibody concentrations of the two groups were not significantly different. Bactericidal activity and passive protection activity (infant rat model) were tested in pooled sera from the three highest and three lowest responders in each vaccine group; both PRP and PRP-D vaccines induced biologically active anti-PRP antibody. Thus PRP-D was found to elicit biologically active serum antibody and to be more immunogenic in adults than PRP vaccine; however, the duration of higher concentrations of antibody was transient.

Adult↗
PubMed1984

Do children with recurrent Haemophilus influenzae otitis media become infected with a new organism or reacquire the original strain?

Using the techniques of outer membrane protein gel analysis and biotyping, we characterized nontypable Haemophilus influenzae isolates from middle ear aspirates of 30 children with recurrent nontypable H, influenzae (NTHI) otitis media. Nine of the 13 children with early recurrence of NTHI otitis (less than 30-day intervals) had respective first and second isolates that were identical. In contrast, 14 of 18 children with late recurrences of NTHI otitis (greater than 30-day intervals) had respective first and second isolates that were different, whereas four children had late recurrences with organisms that appeared to be identical with their respective initial infecting strains. These results suggest that early recurrent NTHI otitis usually is a result of relapse with the initial infecting NTHI strain. In contrast, late recurrent disease is usually the result of infection with a new organism. However, the observation that four children had late recurrences with the original strains suggests that strain-specific protective immunity may not uniformly develop after recovery from NTHI otitis.

Child, Preschool↗
PubMed1984

Histocompatibility leukocyte antigen and erythrocyte MNSs specificities in patients with meningitis or epiglottitis due to Haemophilus influenzae type b.

The frequencies of erythrocyte MNSs antigens and certain histocompatibility leukocyte antigen (HLA) specificities (HLA-A, HLA-B, and HLA-DR) were determined in white patients with meningitis or epiglottitis due to Haemophilus influenzae type b and in controls. The frequency of the erythrocyte MNSs genotype was significantly lower among patients with meningitis than among those with epiglottitis (P = 0.03); this observation confirms a trend observed previously. However, the frequencies of the HLA specificities did not differ significantly in the three groups studied; this result fails to confirm previous reports of disease associations with several HLA-A and HLA-B specificities. Although susceptibility to different clinical manifestations of haemophilus disease may be influenced by genetic factors, our studies indicate that the major loci conferring susceptibility are not in linkage disequilibrium with specificities in the major histocompatibility complex.

Antigens↗
PubMed1984

Antibody-dependent alternative pathway killing of Haemophilus influenzae type b.

The bactericidal activities of human complement and human antibody directed against specific Haemophilus influenzae type b cell surface determinants were investigated. Strain Eagan, a laboratory isolate, and strain Kn, a clinical isolate, were used as the test organisms and gave qualitatively similar results. In the absence of antibody, both isolates were resistant to killing by 60% agammaglobulinemic serum (AGS) containing normal complement levels. The addition of affinity-purified immunoglobulin G anticapsular antibody was bactericidal with 15% AGS as the complement source. Bactericidal activity was also demonstrated with this antibody when the complement source was AGS-Mg-EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid], C2-deficient human serum (alternative complement pathway), or AGS in which factor D and properdin had been selectively inactivated (classical pathway). Immunoglobulin G fractions from a human serum pool or from serum from an adult who had recovered from H. influenzae type b (Kn) sepsis were absorbed to remove anticapsular antibody. The absorbed fractions containing noncapsular antibodies also activated complement-dependent bactericidal activity. But, in contrast to the results with anticapsular antibody, noncapsular antibodies did not elicit alternative pathway bactericidal activity. Incubation of cells of H. influenzae type b in C2-deficient serum or AGS-Mg-EGTA did not cause complement consumption (total hemolytic complement and C3). The addition of immunoglobulin G anticapsular antibody (but not noncapsular antibody) increased consumption of total complement and C3, paralleling the results of the bactericidal assays. These studies demonstrated an absolute requirement for anticapsular antibody in alternative pathway activation and killing of H. influenzae type b.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1984

Response to immunization with Haemophilus influenzae type b polysaccharide-pertussis vaccine and risk of Haemophilus meningitis in children with the Km(1) immunoglobulin allotype.

In experimental animals, immune responses to certain antigens are regulated by immunoglobulin allotype-linked genes. In an effort to detect such genes in humans, we examined the antibody responses of 74 healthy children with different Km(1) or Gm(23) allotypes to a Haemophilus influenzae type b vaccine (type b polysaccharide capsule-pertussis vaccine). The anticapsular antibody responses of black or white children with the Km(1) allotype were 4.6- to 9.5-fold higher than those of children who lacked this determinant (P less than 0.004). No significant differences were found in antibody response with respect to the Gm(23) allotype. The frequencies of Km(1) and Gm(23) also were examined in 170 patients with Haemophilus meningitis, 71 patients with epiglottitis, and 173 control children. Km(1) was detected less frequently in black patients with meningitis (38%) than in those with epiglottitis (81%, P less than 0.002) or in controls (66%, P less than 0.0007). The relative risk of meningitis thus was 3.2-fold lower among black children with the Km(1) allotype than in those who lacked this allotype (odds ratio = 0.3, 95% confidence interval 0.2 to 0.6). However, the risk of meningitis was not decreased in white children with the Km(1) allotype (odds ratio = 1.0). There were no significant differences in the frequency of Gm(23) among the patient groups and controls. The Km(1) allotype but not the Gm(23) thus defines a subpopulation of children of both races who are high responders to this vaccine, and black children but not white children with the Km(1) allotype are at decreased risk of developing Haemophilus meningitis. These data indicate that in blacks, genes associated with Km(1) may affect immune response to a prototype type b Haemophilus vaccine, and perhaps interact with another factor related to race to affect susceptibility to Haemophilus meningitis.

Antibodies, Bacterial↗
PubMed1984

Infectious diseases and child day care.

The number of day care centers and home care facilities has steadily increased in the United States. Recent interest has focused on the possible relationship between attendance at child day care facilities and the occurrence of certain infectious diseases. A variety of infectious agents have been reported as causes of illness among children and staff in day care programs. In general, however, concurrent risks for these infections among children attending and those not attending day care programs have not been established by prospective studies. A review is made of the pathogens that have been associated with infections in day care settings, patterns of occurrence of infectious diseases in day care facilities, aspects of control and prevention of these diseases, and controversies related to infectious diseases in child day care facilities. Aspects of this problem that warrant further research are outlined.

Child Day Care Centers↗
PubMed1983

Nosocomial spread of Haemophilus influenzae type b infection documented by outer membrane protein subtype analysis.

This report provides evidence of nosocomial transmission of Haemophilus influenzae type b infection between two children in an acute care hospital. In the past, transmission of infection between these children would have gone unrecognized, because the isolates from the respective cases differed in susceptibility to ampicillin. However, the outer membrane protein profile of the two isolates was identical and has previously been observed in less than 2% of type b Haemophilus isolates from patients with invasive disease in the St. Louis area. Thus the likelihood is very low that these two children were both infected with this strain by chance alone (P less than 0.0004). From previous experience, the risk of transmission of Haemophilus in an acute care setting would appear to be very low. However, nosocomial transmission may occasionally occur.

Ampicillin↗
PubMed1983

Siblings of patients with Haemophilus meningitis have impaired anticapsular antibody responses to Haemophilus vaccine.

Siblings of patients with type b Haemophilus influenzae meningitis are at increased risk of developing Haemophilus disease. We immunized 26 healthy siblings and 25 control subjects using a vaccine containing the type b polysaccharide capsule (10 micrograms PRP) and pertussis vaccine (4 opacity units) (Lederle Laboratories) to determine whether siblings of patients with Haemophilus meningitis had an impaired antibody response to PRP. Using two intramuscular injections one month apart, we found that the siblings had a lower response to PRP. One month after the second injection, 12 of 24 of the siblings had serum concentrations of anticapsular (PRP) antibody thought to be sufficient to confer protection against Haemophilus disease (greater than or equal to 300 ng/ml), compared with 19 of 24 of the control children tested (50% vs 79%, P = 0.035 by chi-square analysis). In comparison with the normal controls, the siblings produced significantly less IgG anti-PRP antibody but similar amounts of IgM. The impaired responsiveness to PRP was most evident among the 16 children born after their sibling had meningitis and who were not known to have been exposed to type b Haemophilus infection previously. These data indicate that siblings of some patients with type b Haemophilus meningitis have reduced ability to form IgG anti-PRP antibody, which may be associated with increased susceptibility to Haemophilus disease.

Antibodies, Bacterial↗
PubMed1983

Further studies of the role of noncapsular antibody in protection against experimental Haemophilus influenzae type b bacteremia.

Serum antibody against polyribosylribitol phosphate, the capsular antigen of Haemophilus influenzae type b, confers protection against experimental Haemophilus infection. Antibodies against noncapsular antigens are also protective, but the antigenic specificity of the protective antibodies remains unknown. Antilipopolysaccharide antibody was prepared by immunization of rabbits with boiled H. influenzae type b cells. Antilipopolysaccharide antibodies present in these sera did not protect against experimental Haemophilus bacteremia in infant rats. Antisera were also prepared by immunization of rabbits with live H. influenzae type b bacteria. After absorption of anticapsular and antilipopolysaccharide antibodies, these sera contained antibody to several outer membrane proteins which were accessible on the intact bacterial surface as detected by radioimmune precipitation. These absorbed sera prevented experimental Haemophilus bacteremia in infant rats. Thus, antibodies against noncapsular, non-lipopolysaccharide determinants, possibly against one or more outer membrane proteins, confer protection against experimental H. influenzae type b disease. In contrast, antibodies against lipopolysaccharide are ineffective.

Animals↗