Background: Serologic data on diseases that are preventable by vaccine are useful to evaluate the success of immunization programs and to identify susceptible subgroups.Objective: To provide national estimates of immunity to diphtheria and tetanus by measurement of serum antibody levels.Design: Examination of data from the Third National Health and Nutrition Examination Survey, a representative cross-sectional sample of the U.S. population.Setting: 89 randomly selected locations throughout the United States.Participants: 18 045 persons 6 years of age or older who were examined from 1988 to 1994.Measurements: Serum samples obtained at a single time point were tested for diphtheria and tetanus antitoxin.Results: Overall, 60.5% of Americans 6 years of age or older had fully protective levels of diphtheria antibody (greater than or equal to0.10 IU/mL) and 72.3% had protective levels of tetanus antibody (>0.15 IU/mL). Ninety-one percent of Americans 6 to 11 years of age had protective levels of both diphtheria and tetanus antibody; this proportion decreased to approximately 30% among persons 70 years of age (29.5% for diphtheria and 31.0% for tetanus). Adult Mexican-Americans were slightly less likely to have protective levels of antibody to both toxins. Only 47% of persons 20 years of age or older had levels that were protective against both diseases, and only 63% of adults who were protected against tetanus were also protected against diphtheria.Conclusions: A substantial proportion of adults in the United States do not have antibody levels that are protective against diphtheria and tetanus. In addition, although the recommended vaccine is a combination of tetanus and diphtheria, only 63% of adults with protective antibody to tetanus also had protective antibody to diphtheria.

After 30 years of control, epidemic diphtheria returned to the Soviet Union in 1990. To develop control strategies, the immunogenicity of the tetanus and diphtheria toxoids (Td) vaccine was assessed. Workers who were 18-67 years old received two Td immunizations separated by 30 days. A neutralization assay determined diphtheria antitoxin (DAT) on enrollment and on days 7, 30, 60, and 425. On enrollment, 43.0% of 488 workers had DAT <0.1 IU/mL. After one dose, 88.5% had DAT >/=0.1 IU/mL, after two doses, 92.2% had >/=0.1 IU/mL and >90% of participants <30 or >/=50 years of age attained >/=1.0 IU/mL; however, only 78.4% of those who were 30-39 years old and 51.8% of those who were 40-49 years old achieved >/=1.0 IU/mL after two doses. To control the epidemic in Ukraine, one Td dose should be administered to virtually the entire population (persons 30-49 years old require three doses of Td for optimal individual protection and to maximize population immunity).

To determine the immunogenicity and safety of a single dose of diphtheria toxoid among adults, blood samples for detecting serum antitoxin levels were obtained from 18- to 59-year-old subjects (n=248) before and 30 days after immunization with Td (tetanus-diphtheria toxoids; manufactured by Serum Institute of India). By day 30, the seroprevalence of antitoxin levels >/=0.1 IU/mL increased from 22.6% to 81.5%; median antitoxin levels increased from 0.01 to 4.0 IU/mL. These parameters were lowest among subjects who were 40-59 years old, especially among those 40-49 years old. Adverse reactions (local redness, swelling, induration, fever>39 degrees C) were reported by 5.3% of participants. Our findings suggest that, in general, one dose of the Indian-produced Td vaccine is efficacious and safe in inducing an adequate immune response against diphtheria in adults; however, in Georgia, persons 40-59 years old, especially those 40-49 years old, will require additional doses of toxoid to achieve protective levels of antitoxin.

Epidemic diphtheria spread to Ukraine in 1991, where it peaked in 1995 with >5000 reported cases, To refine epidemic control strategies, immunogenicity of a tetanus-diphtheria toxoids vaccine (Td) containing 2 limits of flocculation (Lf) diphtheria toroid was evaluated. During a mass vaccination campaign, adults at a clinic in Odessa received one dose of Td, At enrollment, 57.2% of 341 study participants had levels of diphtheria antitoxin (DAT) greater than or equal to 0.1 IU/mL. Thirty and 180 days after receiving one dose of Td, 91.5% and 84.5% of the participants, respectively, had DAT levels greater than or equal to 0.1 IU/mL, However, among 40- to 49-year-old participants, only 78.8% and 73.8% had DAT levels greater than or equal to 0.1 IU/mL at 30 and 180 days, respectively. This study suggests that one dose of 2 Lf diphtheria toroid is highly effective in raising DAT to protective levels in most adults; however, the study also shows that certain age groups, particularly persons 40-49 and, to a lesser degree, 30-39 years old may require additional doses or a complete three-dose primary vaccination series for optimal protection against diphtheria.

363 A life-threatening complication of transplantation is B cell post-transplant lymphoproliferative disorder (BPLD). Pediatric transplants are exquisitely sensitive to this spectrum of EBV-associated (pre)neoplastic disorders because of the higher susceptibility of the EBV-naive child to develop primary EBV infection in the early posttransplant period, a clinical setting associated with the highest risk of developing fulminant disease. The most likely source of EBV is the donor organ. Recently, the EBV load was shown to be clinically significant in the management of BPLD. Purpose-To determine the sensitivity of semi-quantitative dilution (SD-PCR) and competitive (SC-PCR) methods in determining EBV load in organ donor PBMCs. Methods-Sequential plasma specimens obtained from UNOS donors (n=88) were measured for EBV serology by ELISA (Gull Laboratories, USA) resulting in 97% positive for EBV IgG and EBNA 1, while all were negative for EBV IgM. WBCs from parallel specimens (n=51) were used for DNA isolation or further purified by CD19 coated magnetic beads (Miltenyi Biotech); the efficiency of this method was determined to be >95% by FACS analysis. DNAs were then analyzed by SD-PCR and/or SC-PCR using EBV type-specific sequences cloned into pCRII to determine EBV subtype and load. Results/Conclusion-EBV detection in donor PBMCs was enhanced by enriching the B cell population and by increasing the DNA amounts. Using T cell with Namalwa DNA (1-2 copies per genome) or cloned target DNA, we found that a 25-45 fold increase in the EBV target allowed detection. In the donor population, this was accomplished by B cell enrichment (range 6-9 fold) and by increasing the input amount of cells to approximately 6 × 105 B cells. This allowed detection in donor specimens that were ordinarily infected below PCR detection levels. Other viral analyses having a subset of cells as infective targets will benefit from these methods. While detecting EBV in the donor, the question remains whether either the threshold or the competitive method will discriminate between organs at risk for transmitting active infection.

In an effort to determine the optimal dose of pertussis toxoid (PT) and filamentous haemagglutinin (FHA) for use in acellular pertussis vaccines we compared the immunogenicity and safety of acellular pertussis vaccine combined with diphtheria and tetanus toxoids containing 12.5 μg (DTaP-12.5) or 25 μg (DTaP-25) each of PT and FHA with a whole cell pertussis vaccine in infants immunized at 2, 4 and 6 months of age. Recipients of accellular vaccines developed higher anti-FHA concentrations and more rapid anti-PT serological responses that infants who received whole cell pertussis vaccine combined with diphtheria and tetanus toxoids (DPT). A dose response was noted; infants immunized with DTaP-25 developed significantly (P<0.03) higher anti-FHA and anti-PT levels than infants who received DTaP-12.5. No rise in the agglutinin titres was noted for recipients of the acellular vaccines although this vaccine stimulated increases in agglutinins when given as the fourth or fifth dose to children who had received three doses of DTP. The rates of erythema, induration, pain, irritability, crying, increased sleepiness, and decreased appetite were significantly (P≤0.05) lower in infants who received acellualr vaccines than in infants who received DTP. When the data from the injections at 2, 4 and 6 months of age were combined, no significant differences in the rates of any adverse event were noted for recipients of DTaP-12.5 or DTaP-25. The rates of most adverse reactions following DTP decreased from the first to the third immunization except fever, which increased. For acellular vaccine recipients, the rates of fever and erythema increased somewhat from the first to the third injection but remained far below the rates following DTP. The acellular vaccine was safe and immunogenic, and a dose-response effect was demonstrated.

OBJECTIVE:To assess immunity to diphtheria in a sample of Canadian adults.DESIGN:A seroprevalence study of a group of plasmapheresis donors was performed over a four-month period in 1996. A convenience sample of 1619 sera was collected to obtain a good distribution by age groups and centres. The determination of diphtheria antitoxin concentrations was performed by neutralization of diphtheria toxin in cell culture.SUBJECTS:A total of 1619 plasmapheresis donors from Halifax, Quebec City, London, Calgary and Edmonton were studied.RESULTS:Of the 1619 sera, 20.3% tested showed susceptibility to diphtheria (antitoxin concentration less than 0.01 IU/mL). The proportion of susceptibles increased from 9.5% in subjects 30 to 39 years of age to 36.3% in those 60 years of age or more. The age group 20 to 29 years demonstrated a higher proportion of susceptibles (18.3%) than the next age group (30 to 39 years) in four of the five centres. Significant differences in antibody levels were also observed among the centres. There was no statistically significant difference between sexes.CONCLUSIONS:Overall, detectable antibody and presumably immunity to diphtheria in the present sample of Canadian adults is relatively good. However, reason(s) for the relatively high proportion of susceptibles in those aged 20 to 29 years of age in certain centres, as well as why Canada has not experienced any diphtheria outbreaks in the past 20 years given these susceptibility levels, should be investigated further.

Varicella vaccine was administered to seven children with corticosteroid-sensitive nephrotic syndrome. Immunization was not associated with any significant reactions or with increased frequency of relapse. The antibody response was, however, variable and a second dose was necessary before seroconversion was achieved in four patients. The findings indicate that immunization with varicella vaccine is safe in children with nephrotic syndrome in remission, but that a two-dose vaccine schedule should be considered. (J Pediatr 1997;131:688-90)

OBJECTIVE:To compare the safety and immunogenicity of 12 different acellular pertussis vaccines combined with diphtheria and tetanus toxoids (DTaP) with one licensed diphtheria, tetanus, and whole-cell pertussis vaccine (DTwP) as a fourth-dose booster in children who had previously received DTaP or DTwP primary vaccinations.METHODS:Healthy 15- to 20-month-old children were enrolled at six National Institutes of Health Vaccine Treatment and Evaluation Units. All had been randomly assigned to receive three primary doses of DTaP or DTwP at 2, 4, and 6 months of age as part of an earlier National Institutes of Health multicenter trial of DTaP vaccines in the same Vaccine Treatment and Evaluation Units. Parents recorded the occurrence and magnitude of fever; irritability; and injection site redness, swelling, and pain for 3 days after vaccination. Sera obtained before and 1 month after the booster vaccination were analyzed for antibody to pertussis toxin (PT), filamentous hemagglutinin (FHA), fimbriae (FIM), and pertactin (PRN). Diphtheria and tetanus toxoid as well as PT neutralizing (Chinese hamster ovary cell) and whole-cell agglutinating antibodies were measured on a subset of sera.RESULTS:A total of 1293 children contributed fourth-dose reaction data. Reactions were less frequent after DTaP than after DTwP. For children vaccinated with a fourth dose of DTaP, which was the same DTaP as received in the primary series, fever and injection site redness, swelling, and pain increased in prevalence compared with the third dose in the primary series. For children receiving DTaP as a fourth dose, injection site redness and swelling occurred more frequently in DTaP-primed than in DTwP-primed children. Variation in the occurrence of reactions among DTaP vaccines was observed. A total of 1160 paired pre- and postvaccination sera were available for analysis. Serum antibody concentrations before boosting were lower than those obtained 1 month after the primary immunization. After the fourth dose, significant increases in antibodies directed against the included antigens were observed for all vaccines; postbooster vaccination antibody titers differed significantly among the DTaP vaccines. For children primed and boosted with the same DTaP, antibody levels were not directly related to the quantity of antigen included for PT, FHA, and FIM; for PRN, there was a closer relationship. Some DTaP vaccines given as fourth-dose boosters elicited antibody to PRN or FIM in some vaccinees, although the DTaP vaccines were not reported to contain these antigens; these responses were observed more frequently in DTwP-primed children. Agglutinin antibody rises were observed in all groups immunized with four doses of a DTaP vaccine containing FHA or PRN, regardless of whether the vaccine included FIM. Diphtheria and tetanus antibody levels exceeded the presumed protective concentration (0.1 IU/mL for diphtheria and 0.01 IU/mL for tetanus) after the fourth dose for all vaccinees.CONCLUSION:Although differences were observed in reaction rates among the DTaP vaccines given as a fourth dose, the DTaP vaccines were, in general, associated with fewer adverse events than a US-licensed DTwP. For DTaP vaccines, fever; irritability; and injection site pain, redness, and swelling occurred more frequently after the fourth dose than after the third dose of the same vaccine in the primary series. No DTaP was consistently most or least reactogenic or immunogenic. Although serologic correlates of pertussis immunity are not defined, it is clear that most DTaP vaccines can stimulate comparable or higher serum antibody responses than DTwP for those antigens contained in the vaccine.

Cytomegalovirus (CMV), the most significant infectious cause of morbidity following renal transplantation, may be a greater problem for children than for adults due to their relative lack of experience with this virus. Therefore, we prospectively gave Gammagard as prophylaxis to CMV-negative children who received CMV-positive allografts and compared the results to our experience with similar high-risk recipients transplanted prior to our use of intravenous immunoglobulin G (IvIgG). Symptomatic CMV disease developed in 17% of the IvIgG recipients as compared with 71% of the untreated patients (p = 0.01). The CMV infections that did occur in IvIgG recipients developed significantly later than in untreated children (median time of onset after transplantation 2.60 vs. 1.35 months; p < 0.05) and generally were less severe, although 1 IvIgG recipient died despite prophylaxis. IvIgG administration did not affect the frequency of rejection or graft or patient survival. We conclude that IvIgG administration to high-risk pediatric renal transplant recipients may protect against posttransplantation CMV disease and may lessen the severity of infections that do develop in patients who receive it.

Six NIH Vaccine Treatment and Evaluation Units (VTEUs) enrolled 1340 15- to 20-month-old children to receive one of 12 DTaP or a DTwP vaccine as a booster after they had received 3 primary doses of the same DTaP or a DTwP at 2, 4, and 6 months of age. Fever, irritability, and injection site redness, swelling, and pain was assessed for 3 days after vaccination. Pre- and post- sera were analyzed for antibody (Ab) to pertussis toxin (PT), filamentous hemagglutinin (FHA), fimbriae (FIM), pertactin (PRN), and diphtheria (D) and tetanus (T) toxoids. Pertussis toxin neutralization (CHO) and agglutinating antibodies (AGG) were also measured for a subset of children. Common reactions increased after the booster dose compared with the primary series for both DTaP and DTwP; however, DTaP vaccines produced fewer reactions than DTwP. Significant variation in the occurrence of reactions among DTaP groups was observed. Priming with DTaP or DTwP did not influence the rate of reactions to DTaP boosting. Significant increases in Abs directed against the included antigens were observed for all vaccines; post booster Ab titers differed among the DTaP and DTwP vaccines. For children primed and boosted with the same DTaP, Ab levels correlated poorly with the quantity of antigen included for PT, D and T toxoid; for FHA, PRN and FIM there was a closer correlation. Children primed with DTwP and boosted with DTaP had higher Ab responses to PT than children who were primed with DTaP, indicating enhanced DTwP priming. Some DTaP vaccines given as a fourth dose in DTwP primed children elicited Ab to pertussis components not reported to be in the vaccines; suggesting the probable presence of trace quantities of these components. Although the highest AGG titers were observed in children immunized with DTaP vaccines containing FIM, AGG Ab responses occurred for all vaccines containing FHA. Use of DTaP for boosters in 15- to 20-month-old children has revealed reaction rates and immunogenicity results generally consistant with those following the primary series; however moderate but measurable increases in reactogenicity and some unexpected immunogenicity results were observed. Spon: NIH/NIAID& the vaccine companies.

SAFETY AND IMMUNOGENECITY OF THE VARICELLA VACCINE(VARIVAX®) IN CHILDREN WITH NEPHROTIC SYNDROME (NS). 2190

Viral infections such as influenza are an important cause of morbidity following organ transplantation. We evaluated the immunogenicity of a commercially available influenza vaccine in pediatric renal transplant recipients in a two-phase, prospective study. In phase one, 47 transplant patients and seven control subjects with bronchopulmonary dysplasia received influenza vaccine. Sera were collected at the time of vaccination and 6 wk later. In phase two, sera from 18 transplant recipients and 47 healthy adults who had received the same vaccine were collected 6-12 months after vaccination. Antibody titers to the A/Taiwan/1/86 antigen were measured with hemagglutination inhibition assay in both phases of the study. Vaccine was well tolerated in all subjects. No vaccinated patient required hospitalization for complications of influenza infection. Vaccination did not increase the frequency of acute allograft rejection. In phase one, 43 patients (91%) and 5 controls (71%) either seroconverted (developed a fourfold or greater rise in titer), or developed post-vaccination titers > or = 1:160 (p = NS). Among the transplant recipients, non-seroconverters had a higher pre-vaccination geometric mean antibody titer (GMT) than those who seroconverted. Seroconversion developed independently of whether patients received double or triple immunosuppression. In phase two, post-vaccination GMT were similar for patients and control subjects at 11.5 and 8 months post-vaccination, respectively. In our study, influenza vaccination produced equivalent humoral immunity in transplant recipients and normal subjects. Routine influenza vaccination should be performed annually in this high-risk population.

OBJECTIVE:To compare the immunogenicity of a licensed conventional whole-cell (WCL) and 13 diphtheria-tetanus-acellular pertussis (DTaP) vaccines that differed in source, method of manufacture, and included antigens; all vaccines included diphtheria and tetanus toxoids.METHODS:Healthy infants were enrolled through six university-based vaccine and treatment evaluation units and were randomized to receive one of the study vaccines at 2, 4, and 6 months of age. Sera were obtained before the first immunization and 1 month after the third immunization and were analyzed for antibody to pertussis toxin (PT), filamentous hemagglutinin, fimbriae, pertactin, and diphtheria and tetanus toxins. Chinese hamster ovary cell toxin neutralization assays were performed, and levels of agglutinating antibodies were determined.RESULTS:Of 2342 infants enrolled, 1942 contributed usable preimmunization and postimmunization serum specimens. Each vaccine produced significant increases in antibodies directed against the included antigens; postimmunization antibody titers differed significantly among the DTaP vaccines. For each evaluated antigen, the majority of DTaP vaccines produced antibody responses that equaled or exceeded those produced by WCL. For some antigens (eg, PT), mean antibody levels by vaccine correlated poorly with the quantity of antigen included in each vaccine; for others (eg., fimbriae), there was a close correlation.CONCLUSION:Although serologic correlates of pertussis immunity are not defined, it is clear that DTaP vaccines can stimulate immune responses that exceed those of licensed whole-cell vaccine with respect to the measured antibodies. Particularly for PT, immunogenicity seems to depend on factors in addition to antigen concentration, possibly including antigen derivation and formulation. No DTaP was most or least immunogenic with respect to all included antigens.

OBJECTIVE:To evaluate the effect of maternally derived antibody on the immunogenicity and reactogenicity of acellular (DTaP) or whole-cell (DTP) pertussis vaccine with diphtheria and tetanus toxoids combined.METHODS:A total of 2342 infants were randomized to receive one of 13 DTaP or 2 DTP vaccines at 2, 4, and 6 months of age. The correlation between preimmunization and postimmunization antibody after three doses of vaccine and the relation between preimmunization antibody and adverse reactions after the first immunization were modeled by linear regression.RESULTS:After DTP but not DTaP, higher levels of preexisting antibody were associated with substantial (28% to 56%) reductions in the subsequent antibody response to pertussis toxin (PT). For other pertussis antibodies, modest inverse correlations were seen between preexisting antibody concentrations and most postimmunization antibody responses (resulting in 8% to 18% reductions in postimmunization antibody) for both DTP and DTaP. There was no consistent association in any DTP or DTaP group between adverse reactions and preimmunization antibody levels.CONCLUSION:The PT antibody response to DTaP, unlike DTP, is not adversely affected by preexisting antibody to PT. Inhibitory effects with respect to other antibodies, seen with both DTP and DTaP, were relatively modest. Our data suggest that the use of acellular pertussis vaccines in adults, which could confer higher levels of antibody in women before pregnancy, would be unlikely to adversely affect pertussis antibody responses after DTaP among infants born to mothers with high antibody levels.

OBJECTIVE:To evaluate the effect of simultaneous Haemophilus influenzae type b conjugate (Hib) vaccination on the safety and immunogenicity of selected acellular (DTaP) and whole-cell (DTP) pertussis vaccines with diphtheria and tetanus toxoids combined.METHODS:Enrollment of infants into a large multicenter study of the safety and immunogenicity of 13 DTaP and 2 DTP vaccines was partially completed when the first Hib vaccine, HbOC (Haemophilus b oligosaccharide conjugate vaccine), was licensed for use in infants. Thereafter, at each immunization most infants received HbOC simultaneously with DTaP (or DTP), administered in opposite thighs. Postvaccination geometric mean titers or concentrations (GMTs) of pertussis antibodies as measured by six different assays were compared pairwise among groups of infants receiving 0, 1, 2, or 3 simultaneous HbOC immunizations. The incidence of reactions was compared between infants who received only DTaP or DTP and those who received HbOC simultaneously.RESULTS:Comparison of postvaccination GMTs was possible among groups of infants receiving different numbers of simultaneous immunizations for 10 of the 13 DTaP and both DTP vaccines. Increased HbOC exposure had no consistent dose-response effect on antibody titers for DTaP or DTP vaccines in any assay. Significant differences between groups in postvaccination GMTs were observed with 4 DTaP vaccines in 1 to 2 assays each; the GMTs were higher with increasing HbOC exposure for 2 DTaP vaccines and lower for 2 others. There was no significant increase in reactions with simultaneous HbOC and DTaP immunization.CONCLUSIONS:Based on these retrospective analyses, there did not seem to be an interference in pertussis immunogenicity or alteration in reactogenicity associated with the simultaneous administration of HbOC and DTaP. These findings are encouraging with respect to the development of DTaP-Hib combination vaccines.