• Clinical and Experimental Pediatrics
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• v.49 no.3
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• pp.242-250
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• 2006

Haemophilus influenzae type b(Hib) conjugate vaccines prevent Hib disease in individuals and reduce the carriage and transmission of the organism in the community. The incidence of Hib disease has been decreased dramatically in a diverse range of countries through the use of a variety of conjugate vaccines and vaccine schedules. In some countries, the vaccine has caused a near-disappearance of invasive Hib disease through a combination of direct protection and herd immunity. The effectiveness of the vaccine was not modified by the type of conjugate vaccine, the number of doses given(two, three or four), age at first vaccination(two months, 42 to 90 days, three months) and whether the vaccine was tested in an industrialized or developing country. Over 15 years of international experience with vaccines has also demonstrated that they are safe. In 2004, Hib vaccines were adapted in routine immunization in 92 countries in the world. Decisions regarding the use of the Hib vaccine in routine immunization schedules depend not only on the effectiveness and efficacy of the vaccine but also on factors such as burden of disease, vaccine cost, and competing priorities. In Korea, Hib disease burden seemed to be lower than other developed countries(~10/100,000). Moreover Hib vaccines showed excellent immunogenicity in Korean children in many studies. Therefore, a potential approach to economize the cost of Hib vaccines could be to explore the possibilities of using reduced vaccine doses for immunization as some other countries.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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• pp.215-222
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• 2006

The Lowry method was used in this study to measure protein in Haemophilus influenzae type b (Hib) conjugate vaccines (polyribosylibitol phosphate-tetanus toxoid; PRP-TT) using deoxycholic acid (DOC) to induce protein precipitation. Trichloroacetic acid (TCA) did not induce precipitation adequately from the Hib conjugate bulk and the freeze-dried Hib conjugate product. Its yield was approximately 50%. The matrix structure of Hib conjugate inhibits precipitation by TCA. Although the Lowry method can be carried out without precipitation in Hib conjugate bulk when no residual impurities (adipic acid dihydrazide [ADH], 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide-HCI [EDAC], phenol and cyanogens bromide [CNBr], etc.) are present, it cannot be used for Hib conjugate products that contain sucrose 8.5%, because 8.5% concentration of sucrose enhanced the protein concentration. DOC- and HCl-induced precipitation is an alternative method for evaluating the protein content of the Hib conjugate bulk and the Hib conjugate product. The precipitation was optimal with a final concentrate of 0.1% for DOC at $4^{\circ}C$ and pH 2. This Lowry method, using DOC/HCI precipitation to induce protein precipitation, was confirmed a consistent, reproducible, and valid test for proteins in Hib conjugate bulk and its freeze-dried product.

• Pediatric Infection and Vaccine
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• v.24 no.3
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• pp.125-133
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• 2017

Purpose: After the introduction of Haemophilus influenzae type b (Hib) vaccine in 1995 in Korea, it was included in the national immunization program in 2013. In the post-Hib vaccine era, some studies in other countries reported that invasive Hib disease affects adults, especially the elderly and immunocompromised persons, more often than it affects children. To evaluate disease susceptibility, quantitative and qualitative analysis of anti-polyribosylribitol phosphate (PRP) antibodies were carried out in Korean adults aged 20 to 85 years. Methods: Sera were collected from 39 healthy adults (20 to 50 years of age) and from 30 elderly adults (75 to 85 years of age) who did not have immune-compromising conditions. The concentration of anti-PRP immunoglobulin G (IgG) and serum bactericidal indices (SBIs) were measured by enzyme-linked immunosorbent assay and serum bactericidal assay. Results: Geometric mean concentrations of anti-PRP IgG and geometric mean SBIs were $0.88{\mu}g/mL$ (95% confidence interval [CI], 0.17 to 3.85) and 354 (95% CI, 50 to 2,499) in young adults and $1.67{\mu}g/mL$ (95% CI, 0.53 to 5.24) and 449 (95% CI, 146 to 1,376) in elderly adults, respectively. When the threshold of seropositivity for anti-PRP IgG was applied as 0.15 or $1.0{\mu}g/mL$, which is the protective antibody level in children, seropositive rates were 87.2% or 53.8% in young adults and 100% or 60% in elderly adults. The seropositivity rates of the SBI ($SBI{\geq}4$) were 82.1% and 100% in the groups, respectively. Conclusions: Most subjects in the adult and elderly adult groups display immunity to Hib based on quantitative and qualitative antibody levels, but not all. Because high immunization and low Hib circulation rates may reduce the natural Hib immunity in the population, monitoring Hib immunity as well as disease are needed continuously.

• BMB Reports
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• v.33 no.3
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• pp.249-255
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• 2000

The Haemophilus influenzae type b (Hib) has been a major cause of bacterial meningitis in children who are less than two years old. The variable (V) region repertoire of adult Caucasian antibodies (Abs) to Hib polysaccharide (PS) has been characterized well. The majority of adult antibodies against Hib uses VL that is derived from the Vk gene A2 and have arginine at the N region. In order to explore the possibility those antibody responses to Hib-PS is variable in various age groups, we examined the VL regions of the antibodies to Hib-PS in Korean adults and children. We immunized Korean adults (n = 8) and children (n = 39) with Hib tetanus conjugated vaccines, isolated RNAs from the peripheral lymphocytes, and amplified the A2-derived VL regions by RT-PCR. The PCR products were subcloned and sequenced. Forty-seven out of 54 independent clones from children used the $J{\kappa}2$, or $J{\kappa}3$ gene in preference. The adults, however, used all of the $J{\kappa}$ genes evenly. With respect to the amino acid sequences of variable regions, adult $A2-J{\kappa}$ recombinants have a germline sequence. But, the 76th codon (AGC) of child $A2-J{\kappa}2$ recombinants was substituted with CGC (arginine) in most cases (88 %) and 77 percent of child clones using the $A2-J{\kappa}3$ genes have isoleucine-109 at the junction of $J{\kappa}-C{\kappa}$ instead of threonine that is found in a germline sequence. These results suggest that the mechanism of antibody production in young children is different from that of adults.

• Clinical and Experimental Pediatrics
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• v.50 no.5
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• pp.449-456
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• 2007

Purpose : Antibody persistence after primary series of Haemophilus influenzae type b (Hib) vaccine and responses to a boosters are little known in Korean children. We performed this study to evaluate the antibody titer in relation with a booster immunization of Hib vaccine in Korean children. Methods : One hundred forty-four children aged 12-23 months old were enrolled in three university hospitals. The immunogenicity of a boosters with Hib vaccine was assessed in children previously primed with Hib vaccine. Antibody persistence was also assessed in children who had received 3 doses of Hib vaccine without a booster. Anti-polyribosylribitol phosphate (PRP) IgG antibody levels and bactericidal titers were determined by enzyme immunoassay and bactericidal assay at the Center for Vaccine Evaluation and Study, Medical Research Institute, Ewha Womans University. Results : Prior to a booster in the second year of life, geometric mean antibody concentrations were $2.39{\mu}g/mL$ and the percent of subjects who had a anti-PRP antibody level ${\geq}1{\mu}g/mL$ was 68.6%. After boosting, antibody concentration was $19.09{\mu}g/mL$ and the percent of subjects who had a anti-PRP antibody level ${\geq}1{\mu}g/mL$ was 96.5%, which reflects previous immune priming. In subjects who had finished primary immunization only, the bactericidal titer was 3,946 and in subjects who had a booster, it was 11,205. Anti-PRP antibody level was correlated with serum bactericidal titer. Conclusion : Many children aged 12-23 month old still had protective antibodies after recommended primary immunization only. A booster dose seemed to induce good anamnestic antibody responses in Korean children.

• Clinical and Experimental Pediatrics
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• v.46 no.11
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• pp.1085-1088
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• 2003

Purpose : This study was performed to assess the difference of organisms causing bacterial meningitis according to time. Methods : We analyzed retrospectively 40 medical records of bacteriologically proven meningitis from 1992 to 2002. We divided them into two groups; neonate's group(14 cases), and children's group(26 cases). The results of the neonate's group were compared with those of previously reported articles in Korea, in 1970s-1980s. The causative agents of the children's group were analyzed according to the stage before and after the introduction of H. influenza type b(Hib) vaccine. Results : In neonates, Group B streptococci(GBS) was the most common cause of bacterial meningitis. There was a trend in Korea that major causative agents of neonatal bacterial meningitis have changed from gram negative bacteria including E. coli to gram positive bacteria including GBS. In children, H. influenzae was isolated in six out of 11 cases(55%) in 1992-95, before the introduction of Hib vaccine, while two out of seven(29%) were isolated in 1999-2002, after the introduction of the Hib vaccine. Conclusion : Our study showed that the most common agent of neonatal bacterial meningitis was GBS. There was a trend that after the introduction of Hib vaccine, the incidence of H. influenza meningitis decreased in children.

• Pediatric Infection and Vaccine
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• v.10 no.2
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• pp.208-214
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• 2003

Purpose : We performed a retrospective study on bacterial meningitis in children, pertaining to the causative organisms, bacterial resistance to antibiotics and the effect of recent introduction of Haemophilus influenzae type b(Hib) vaccine. Methods : We analysed the forty-three cases of bacterial meningitis which had been treated at the Inha University Hospital from June 1996 to June 2003. Results : Nineteen cases(44.2%) of them were infants younger than 2 months of age, and 29 cases(67.5%) younger than 1 year of age. The common causative organisms under 2 months of age were group B streptococcus(GBS)(47.4%), E. coli(21.1%), and Klebsiella pneumoniae(21.1%). In the age group beyond 2 months of age, S. pneumoniae were seen in 50 %, H. influenzae in 16.7% and N. meningitidis in 16.7%. All of the five cases of Hib meningitis had not been vaccinated for Hib. There has been no Hib meningitis cases since 2001. Overall fatality rate was 4.5%, and complication occurred in 39%. Complications was significantly less frequent in patients resumed to be treated within 48 hours after onset compared to after 48 hours after onset. Penicillin resistance of S. pneumoniae and GBS isolated among bacterial meningitis cases was high. Conclusion : Timing of treatment after the onset of the disease appeared the most important factor for prognosis of bacterial meningitis. The cases of H. influenzae meningitis have decreased probably due to Hib vaccination.

• Pediatric Infection and Vaccine
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• v.21 no.2
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• pp.81-95
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• 2014

Purpose: Hematopoietic cell transplantation (HCT) recipients are vulnerable to invasive infection by Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (Sp). This study was performed to evaluate immune responses after Hib and Sp vaccination in Korean pediatric HCT recipients. Methods: Patients were prospectively enrolled at Samsung Medical Center during 2009-2011. ELISA tests to detect anti-PRP IgG antibody and antibodies to Sp serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were performed at the Center for Vaccine Evaluation and Study, Ewha Medical Research Institute. Results: Ten patients (two allogeneic, eight autologous recipients) with median age 5.4 years (range 2.7-12.2 years) were enrolled. Before Hib vaccination, 60% of patients' anti-PRP IgG titers were below $0.15{\mu}g/mL$. After vaccination, 100% of patients' anti-PRP IgG titers increased above $0.15{\mu}g/mL$ (cut-off value for detection) and $1.0{\mu}g/mL$ (cut-off value for seroprotection). For pneumococcus, in 2-5 year-old patients, pre-vaccination geometric mean concentrations (GMCs) of IgG for six serotypes (4, 6B, 9V, 14, 18C, and 23F) were below $0.35{\mu}g/mL$ and at 5 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. In patients older than 5 years, pre-vaccination GMCs of IgG for four serotypes (4, 9V, 14, and 23F) were below $0.35{\mu}g/mL$ and at 3 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. Conclusion: Most HCT recipients had low or no protective antibodies to Hib and Sp before vaccination, but showed good immune responses to protective levels after vaccination.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.490-494
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• 2004

An efficacy test of PRP (polyribosylribitol phosphate)-TT (Tetanus toxoid) conjugate vaccines was carried out using BALB/c mice as an animal model by inoculating Haemophilus in­fluenzae type b (Hib) with a virulence enhancement factor (VEF). Three administrations of the conjugate vaccines at 2-week intervals elicited a significantly high level of PRP antibodies (P>0.0001). The protective activity of the PRP immunization was challenged with either Hib with iron dextran (Hib/) or with a combination of mucin and hemoglobin (Hibmh) as a VEF. The me­dium lethal dose $(LD_{50})$ for Hibmh and Hibiwas measured as 10 CFU (Colony Forming Unit) and $2.5{\times}10^{8}$ CFU respectively. Each immunized animal was challenged with five or ten times the $LD_{50}$ level of bacteria with a VEF. A significant difference in mortality between the immunized and control mice (P> 0.01) was observed with the Hibmh challenge inoculation but not with the Hibi challenge inoculation. These results show that a combination of mucin and hemoglobin was able to enhance the virulence of Hib in BALB/c mice to cause a lethal infection, thus suggesting that BALB/c mice introduced to this method can be an effective model animal for testing the protective efficacy of H. influenzae conjugate vaccines.

• Pediatric Infection and Vaccine
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• v.13 no.2
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• pp.130-136
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• 2006

Purpose : The purpose of this study is to confirm the epidemiologic change of the causative organisms of neonatal and pediatric bacterial meningitis in Korea. And we tried to evaluate the risk factors correlated with prognosis which was available on the day of admission. Methods : Retrospectively, we reviewed the medical records of 57 patients admitted for bacterial meningitis at six hospitals affiliated with Catholic Medical Center for 6 years(Jan. 2000~Dec. 2005). Results : 22 cases(38.6%) of them were neonates under 28 days and 35 cases were infants and children ; 16 cases(28.1%), under 1 year ; 6 cases(10.5%), under 5 years ; 13 cases (22.8%), under 15 years. In neonates, 16 cases(72.7%) were caused by group B streptococcus (GBS). In infants and children, S. pneumoniae(25.7%), H. influenzae type b(Hib)(22.8%) and N. meningitidis(22.8%) were common cause of bacterial meningitis in order. In the informations available on the day of admission, weight deficit for age under 3 percentile, increased CRP level and decreased glucose level of CSF were related to poor prognosis(P<0.05). Conclusion : GBS became a leading cause of neonatal bacterial meningitis. Though, pneumoccocal, Hib and meningococcal meningitis were confirmed as major causes of bacterial meningitis. The routine immunization of pneumococcal and Hib vaccines will be considered, and it is necessary to introduce meningococcal vaccines to our country in the future.