Reactogenicity and immunogenicity of a double-strength aceUular pertussis vaccine Dean A. Blumberg*, Patricia C. Chatfield*, James D. Cherry *§, Ricki G. Robinson t, Kathleen Smith t, Laura Mabie t, H. James Holroyd t, Leonard R. Baker t, the late Frank E. Dudenhoeffer t, Noelani Apau t, Jill G. Hackell * and Andrea Cawein * The reactogenieity and immunogenieity of a double-strength acellular pertussis vaccine were evaluated after administration to 16 4-6-year-old children. The vaccine contained toxoided lymphocytosis-promoting factor (6.0 #o/dose), filamentous haemaoglutinin (70 #9~dose), aoglutinooens (1.4 #9~dose) and the 69 kDa protein ( ,,~8.0 #9/dose). The vaccine was extremely well tolerated with few minor side effects followin9 immunization. Significant increases in antibodies to all pertussis vaccine components were noted. In summary, this double-strength acellular pertussis vaccine, containin9 a very hioh dose of filamentous haemagolutinin, had minimal reactogenicity and was immunooenic. These findings, as well as other studies with this vaccine, indicate that filamentous haema99lutinin is not a major determinant of vaccine reaetogenicity. Keywords: Pertussis; acellular vaccine; children; filamentous haemagglutinin; reactogenicity; immunogenicity INTRODUCTION Several acellular pertussis-component (APDT) vaccines have been developed in Japan 1. One Japanese acellular pertussis vaccine produced by Takeda Chemical Industries has been combined with diphtheria and tetanus toxoids produced by Lederle Laboratories and has undergone extensive testing in the United States 2-s. The Lederle/Takeda APDT vaccine is less reactogenic than Lederle whole-cell diphtheria-tetanus-pertussis (DTP) vaccine 2-4. When administered as a booster dose to 18-24-month-old and 4-6-year-old children previously immunized with DTP vaccine, there were similar antibody responses to lymphocytosis-promoting factor (LPF; also known as pertussis toxin) and agglutinogens, and better responses to the 69 kDa protein and filamentous haemagglutinin (FHA) when compared with DTP vaccination2's'5. However, when studied in a primary vaccination series in infants, APDT recipients had reduced responses to LPF and agglutinogens, a better response to FHA, and an equivalent response to the 69 kDa protein compared with DTP recipients4. In order to evaluate the effect of an increased dose of the acellular pertussis component on the serological responses to LPF and agglutinogens and the safety of the vaccine, the acellular pertussis component content *Department of Pediatrics, UCLA School of Medicine, Los Angeles, California, USA. tDescanso Pediatrics, La Canada, California, USA. tLederle Laboratories, Pearl River, New York, USA. §To whom correspondence should be addressed. (Received 31 October 1991; revised 27 January 1992; accepted 4 February 1992) 0264~, 10X/92/090614-03 © 1992 Butterworth-Heinemann Ltd
614 Vaccine, Vol. 10, Issue 9, 1992
was doubled and this double-strength APDT vaccine was studied in 4-6-year-old children.
METHODS
Subjects Sixteen healthy 4-6-year-old children received APDT vaccine in an open manner. All children had received four prior immunizing doses of DTP vaccine at the regularly scheduled intervals in the first 2 years of life. Vaccine was administered in a volume of 0.5 ml into the deltoid muscle with a 5/8 inch ( 1.56 cm) 25-gauge needle. The parents were requested to maintain a diary of local and systemic reactions for 10 days postimmunization. Oral temperature was recorded prior to immunization and 0.5, 3, 6, 24, 48 and 72 h after immunization. The presence or absence of the following reactions was recorded at 0.5, 3 and 6 h postvaccination and daily thereafter for 10 days: local tenderness, erythema, induration, swelling, site temperature and vesicles; irritability, drowsiness, anorexia and vomiting, Prophylactic antipyretic medication was not recommended, but was advised for temperature >/38.5°C. Reactogenicity data and information regarding antipyretic administration were obtained by telephone contact 1, 2, 3 and 14 days after immunization. This study was approved by the UCLA Human Subject Protection Committee. Informed consent was obtained from the parents after the design and purpose of the study were explained.
Acellular pertussis vaccine: D.A. Blumberg et al.
Laboratory techniques
RESULTS
Blood samples were obtained by venipuncture on day 0 (prior to immunization), day 1 and day 30. Antibody values were determined at days 0 and 30 at Lederle Laboratories. Enzyme-linked immunosorbent assays (ELISAs) were used to measure LPF, FHA and 69 kDa protein antibodies 3'6. A microagglutination assay was employed to measure pertussis agglutinating antibodies6; the antigens used were equal quantities Of Bordetella pertussis strains 130 and 138. StrainS 130 and 138 each present all serotypes (1.2.3.4.6). Complete blood counts with differentials and serum chemistry assays (sodium, potassium, chloride, bicarbonate, glucose, creatinine, urea n.i(rogen, phosphorus, total protein, albumin, total bilirubin, calcium, alkaline phosphatase, asparate aminotransferase, alanine aminotransferase and cholesterol) were performed by standard methods on days 0, 1 and 30 at the UCLA Clinical Laboratories.
Vaccine The adsorbed APDT vaccine was formulated by Lederle Laboratories such that each dose contained 7.5 Lf of diphtheria toxoid, 5 Lf of tetanus toxoid, and 600 haemagglutinating units (twice the previously tested dose) of Takeda acellular pertussis component. The •diphtheria and tetanus toxoids were produced by Lederle Laboratories. The acellular pertussis component was manufactured by Takeda Chemical Industries, Osaka, Japan from the Tohama strain of B. pertussis, predominantly serotype 1.21. Components of this double-strength vaccine include toxoided LPF (6.0 #g/dose), FHA (70#g/dose), agglutinogens (1.4 #g/dose) and 69 kDa protein (approximately 8.0#g/ dose).
Statistical methods
Sixteen 4-6-year-old Caucasian children received this double-strength APDT vaccine (seven girls and nine boys). Their mean (±s.d.) age was 4.5 (+__0.5) years. Very few side effects were noted. Three subjects experienced mild tenderness at the injection site which resolved within 48 h, and one subject had moderate local induration ( 3 - 4 c m ) which resolved after 72h. No • injection site erythema, swelling, warmth or vesicles were noted. Two subjects had vomiting (two or three episodes), and in both cases other family members had been ill with gastroenteritis. Mild anorexia and irritability postimmunization were each experienced by one subject. No subject had a temperature ~>38°C, drowsiness or used antipyretics. One subject had uncomplicated varicella (onset 13 days postimmunization). No serious adverse events occurred. Serological responses to vaccination _are shown in Table 1. Significant increases were noted ih all antibody values assayed. LPF antibody values increased 7.2 times, FHA values increased by a factor of 19.5, and 69 kDa protein values increased 18.7 times. The postimmunization geometric mean agglutinin titre was 7.1 times higher than the pre-immunization values, and 13 (81%) subjects had >~fourfold agglutinin titre rises. No changes in serum chemistry values were noted after vaccination other than an increase in total protein on day 1 ( m e a n _ s . d . : day 0 = 6 . 7 _ 0 . 2 g d 1 - 1 , day 1 = 6 . 9 _ 0 . 3 g d 1 - 1 , day 3 0 = 6 . 7 _ 0 . 3 g d 1 - 1 ; day 0 versus day 1, p = 0.04). The total white blood cell count increased on day 1 due to an increase in neutrophils and these values returned to baseline at day 30 (geometric mean + s.d. total white blood cell count" day 0 = 6.8 + 1.3 x 10 3 mm-3, day 1 = 8.5 ___1.3 x 103 mm-3, day 30 = 7.0 _ 1.3 x 103 mm -3, day 0 versus day 1 and day 1 versus day 30 both p < 0.02; neutrophil count: day 0 = 2.7 __+1.4 x 10 3 ram-3, day 1 = 3.8 _ 1.5 x 10 3 m m - 3 , day 30 = 2.4 + 1.8 x 10 3 mm -3, day 0 versus day 1 and day 1 versus day 30 both p < 0.03).
All antibody values, white blood cell counts and differentials were log-transformed prior to statistical analysis. Antibody comparisons of geometric means were made by paired t-test (similar results were obtained using Wilcoxon's signed rank test), and proportions of subjects reaching threshold values were compared using McNemar's test for paired proportions. Complete blood count and chemistry panel comparisons were made by paired t-test. No corrections were made for multiple comparisons.
Table 1
DISCUSSION Minimal reactions occurred despite administering twice the previously tested dose of the acellular pertussis component of this APDT vaccine. No severe or delayed reactions were noted. Serum chemistry and complete blood counts added to the assessment of the vaccine's safety profile. The statistically significant increase in total protein on day 1 was clinically insignificant, and most
Serological responses to double-strength APDT vaccine (n = 16)
Antibody
Pre-immunization (day 0)
Postimmunization (day 30)
P Value
LPF GMT'(EU ~ m1-1) n ( % ) /> 30 EU
22.1 (12.2, 40.0) 8 (50)
159.2 (109.9, 230.7) 16 (100)
30 EU
7.9 (4.1, 15.1) 2 (13)
152.9 (110.0, 212.6) 16 (100)
614 Vaccine, Vol. 10, Issue 9, 1992
was doubled and this double-strength APDT vaccine was studied in 4-6-year-old children.
METHODS
Subjects Sixteen healthy 4-6-year-old children received APDT vaccine in an open manner. All children had received four prior immunizing doses of DTP vaccine at the regularly scheduled intervals in the first 2 years of life. Vaccine was administered in a volume of 0.5 ml into the deltoid muscle with a 5/8 inch ( 1.56 cm) 25-gauge needle. The parents were requested to maintain a diary of local and systemic reactions for 10 days postimmunization. Oral temperature was recorded prior to immunization and 0.5, 3, 6, 24, 48 and 72 h after immunization. The presence or absence of the following reactions was recorded at 0.5, 3 and 6 h postvaccination and daily thereafter for 10 days: local tenderness, erythema, induration, swelling, site temperature and vesicles; irritability, drowsiness, anorexia and vomiting, Prophylactic antipyretic medication was not recommended, but was advised for temperature >/38.5°C. Reactogenicity data and information regarding antipyretic administration were obtained by telephone contact 1, 2, 3 and 14 days after immunization. This study was approved by the UCLA Human Subject Protection Committee. Informed consent was obtained from the parents after the design and purpose of the study were explained.
Acellular pertussis vaccine: D.A. Blumberg et al.
Laboratory techniques
RESULTS
Blood samples were obtained by venipuncture on day 0 (prior to immunization), day 1 and day 30. Antibody values were determined at days 0 and 30 at Lederle Laboratories. Enzyme-linked immunosorbent assays (ELISAs) were used to measure LPF, FHA and 69 kDa protein antibodies 3'6. A microagglutination assay was employed to measure pertussis agglutinating antibodies6; the antigens used were equal quantities Of Bordetella pertussis strains 130 and 138. StrainS 130 and 138 each present all serotypes (1.2.3.4.6). Complete blood counts with differentials and serum chemistry assays (sodium, potassium, chloride, bicarbonate, glucose, creatinine, urea n.i(rogen, phosphorus, total protein, albumin, total bilirubin, calcium, alkaline phosphatase, asparate aminotransferase, alanine aminotransferase and cholesterol) were performed by standard methods on days 0, 1 and 30 at the UCLA Clinical Laboratories.
Vaccine The adsorbed APDT vaccine was formulated by Lederle Laboratories such that each dose contained 7.5 Lf of diphtheria toxoid, 5 Lf of tetanus toxoid, and 600 haemagglutinating units (twice the previously tested dose) of Takeda acellular pertussis component. The •diphtheria and tetanus toxoids were produced by Lederle Laboratories. The acellular pertussis component was manufactured by Takeda Chemical Industries, Osaka, Japan from the Tohama strain of B. pertussis, predominantly serotype 1.21. Components of this double-strength vaccine include toxoided LPF (6.0 #g/dose), FHA (70#g/dose), agglutinogens (1.4 #g/dose) and 69 kDa protein (approximately 8.0#g/ dose).
Statistical methods
Sixteen 4-6-year-old Caucasian children received this double-strength APDT vaccine (seven girls and nine boys). Their mean (±s.d.) age was 4.5 (+__0.5) years. Very few side effects were noted. Three subjects experienced mild tenderness at the injection site which resolved within 48 h, and one subject had moderate local induration ( 3 - 4 c m ) which resolved after 72h. No • injection site erythema, swelling, warmth or vesicles were noted. Two subjects had vomiting (two or three episodes), and in both cases other family members had been ill with gastroenteritis. Mild anorexia and irritability postimmunization were each experienced by one subject. No subject had a temperature ~>38°C, drowsiness or used antipyretics. One subject had uncomplicated varicella (onset 13 days postimmunization). No serious adverse events occurred. Serological responses to vaccination _are shown in Table 1. Significant increases were noted ih all antibody values assayed. LPF antibody values increased 7.2 times, FHA values increased by a factor of 19.5, and 69 kDa protein values increased 18.7 times. The postimmunization geometric mean agglutinin titre was 7.1 times higher than the pre-immunization values, and 13 (81%) subjects had >~fourfold agglutinin titre rises. No changes in serum chemistry values were noted after vaccination other than an increase in total protein on day 1 ( m e a n _ s . d . : day 0 = 6 . 7 _ 0 . 2 g d 1 - 1 , day 1 = 6 . 9 _ 0 . 3 g d 1 - 1 , day 3 0 = 6 . 7 _ 0 . 3 g d 1 - 1 ; day 0 versus day 1, p = 0.04). The total white blood cell count increased on day 1 due to an increase in neutrophils and these values returned to baseline at day 30 (geometric mean + s.d. total white blood cell count" day 0 = 6.8 + 1.3 x 10 3 mm-3, day 1 = 8.5 ___1.3 x 103 mm-3, day 30 = 7.0 _ 1.3 x 103 mm -3, day 0 versus day 1 and day 1 versus day 30 both p < 0.02; neutrophil count: day 0 = 2.7 __+1.4 x 10 3 ram-3, day 1 = 3.8 _ 1.5 x 10 3 m m - 3 , day 30 = 2.4 + 1.8 x 10 3 mm -3, day 0 versus day 1 and day 1 versus day 30 both p < 0.03).
All antibody values, white blood cell counts and differentials were log-transformed prior to statistical analysis. Antibody comparisons of geometric means were made by paired t-test (similar results were obtained using Wilcoxon's signed rank test), and proportions of subjects reaching threshold values were compared using McNemar's test for paired proportions. Complete blood count and chemistry panel comparisons were made by paired t-test. No corrections were made for multiple comparisons.
Table 1
DISCUSSION Minimal reactions occurred despite administering twice the previously tested dose of the acellular pertussis component of this APDT vaccine. No severe or delayed reactions were noted. Serum chemistry and complete blood counts added to the assessment of the vaccine's safety profile. The statistically significant increase in total protein on day 1 was clinically insignificant, and most
Serological responses to double-strength APDT vaccine (n = 16)
Antibody
Pre-immunization (day 0)
Postimmunization (day 30)
P Value
LPF GMT'(EU ~ m1-1) n ( % ) /> 30 EU
22.1 (12.2, 40.0) 8 (50)
159.2 (109.9, 230.7) 16 (100)
30 EU
7.9 (4.1, 15.1) 2 (13)
152.9 (110.0, 212.6) 16 (100)
