Trial Outcomes & Findings for A Phase 1 Dose Escalation Study to Examine the Safety of the P2-VP8 Rotavirus Vaccine (NCT NCT01764256)

Last Updated: 2019-03-12

Results Overview

Adverse events were collected through 28 days following the final study injection and were graded for severity. Unsolicited adverse events were also assessed for relationship to vaccine. A final follow-up contact was attempted 6 months following the final study injection to inquire about new chronic health conditions, serious health events, and hospitalizations.

Recruitment status

COMPLETED

Study phase

PHASE1

Target enrollment

48 participants

Primary outcome timeframe

6 months after final vaccination (224 days)

Results posted on

2019-03-12

Participant Flow

Participant milestones

Participant milestones
Measure	Placebo 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.	10 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.
Received All Vaccinations STARTED	12	12	12	12
Received All Vaccinations COMPLETED	10	11	11	12
Received All Vaccinations NOT COMPLETED	2	1	1	0
Completed Study STARTED	10	11	11	12
Completed Study COMPLETED	10	11	11	11
Completed Study NOT COMPLETED	0	0	0	1

Reasons for withdrawal

Reasons for withdrawal
Measure	Placebo 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.	10 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.
Received All Vaccinations Adverse Event	1	1	0	0
Received All Vaccinations Did not meet inclusion criteria	0	0	1	0
Received All Vaccinations Lost to Follow-up	1	0	0	0
Completed Study Lost to Follow-up	0	0	0	1

Baseline Characteristics

A Phase 1 Dose Escalation Study to Examine the Safety of the P2-VP8 Rotavirus Vaccine

Baseline characteristics by cohort

Baseline characteristics by cohort
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.	Total n=48 Participants Total of all reporting groups
Age, Continuous	29.6 years n=5 Participants	34.2 years n=7 Participants	33.8 years n=5 Participants	32.5 years n=4 Participants	32.5 years n=21 Participants
Sex: Female, Male Female	7 Participants n=5 Participants	5 Participants n=7 Participants	6 Participants n=5 Participants	7 Participants n=4 Participants	25 Participants n=21 Participants
Sex: Female, Male Male	5 Participants n=5 Participants	7 Participants n=7 Participants	6 Participants n=5 Participants	5 Participants n=4 Participants	23 Participants n=21 Participants
Region of Enrollment United States	12 participants n=5 Participants	12 participants n=7 Participants	12 participants n=5 Participants	12 participants n=4 Participants	48 participants n=21 Participants
Height	172.7 centimeters n=5 Participants	173.1 centimeters n=7 Participants	168.8 centimeters n=5 Participants	171.9 centimeters n=4 Participants	171.6 centimeters n=21 Participants
Weight	85.5 kilograms n=5 Participants	90.0 kilograms n=7 Participants	81.5 kilograms n=5 Participants	85.6 kilograms n=4 Participants	85.6 kilograms n=21 Participants

PRIMARY outcome

Timeframe: 6 months after final vaccination (224 days)

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Maximum Severity of Adverse Events After Any Vaccination Possibly related to vaccine · Mild	1 Participants	2 Participants	0 Participants	2 Participants
Maximum Severity of Adverse Events After Any Vaccination Possibly related to vaccine · Moderate	0 Participants	1 Participants	0 Participants	0 Participants
Maximum Severity of Adverse Events After Any Vaccination Possibly related to vaccine · Severe	0 Participants	0 Participants	0 Participants	0 Participants
Maximum Severity of Adverse Events After Any Vaccination Possibly related to vaccine · None	11 Participants	9 Participants	12 Participants	10 Participants
Maximum Severity of Adverse Events After Any Vaccination All adverse events · Mild	6 Participants	2 Participants	6 Participants	5 Participants
Maximum Severity of Adverse Events After Any Vaccination All adverse events · Moderate	3 Participants	3 Participants	0 Participants	1 Participants
Maximum Severity of Adverse Events After Any Vaccination All adverse events · Severe	0 Participants	0 Participants	1 Participants	0 Participants
Maximum Severity of Adverse Events After Any Vaccination All adverse events · None	3 Participants	7 Participants	5 Participants	6 Participants

PRIMARY outcome

Timeframe: 7 days after each vaccination (Day 7, 35, 63)

For all cohorts, local and systemic reactogenicity data for all vaccinations were collected by subjects via diary card up to 7 days post each vaccination. Solicited systemic reactogenicity events included headache, muscle pain, fever, nausea, vomiting, fatigue, joint aches, and chills. Solicited local systemic reactogenicity events included injection site pain, tenderness, redness, swelling, itching, and local lymphadenopathy.

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Maximum Local or Systemic Reactogenicity After Any Vaccination Systemic reactogenicity · None	10 Participants	4 Participants	9 Participants	6 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination Systemic reactogenicity · Mild	1 Participants	7 Participants	3 Participants	5 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination Systemic reactogenicity · Moderate	1 Participants	1 Participants	0 Participants	1 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination All reactogenicity · None	7 Participants	3 Participants	6 Participants	4 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination All reactogenicity · Mild	3 Participants	7 Participants	6 Participants	6 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination All reactogenicity · Moderate	2 Participants	2 Participants	0 Participants	2 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination Local reactogenicity · None	7 Participants	4 Participants	7 Participants	4 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination Local reactogenicity · Mild	4 Participants	7 Participants	5 Participants	7 Participants
Maximum Local or Systemic Reactogenicity After Any Vaccination Local reactogenicity · Moderate	1 Participants	1 Participants	0 Participants	1 Participants

PRIMARY outcome

Timeframe: 7 days post Vaccination #1 on Day 0

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Maximum Local or Systemic Reactogenicity After the First Vaccination All reactogenicity · None	7 Participants	4 Participants	8 Participants	5 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination All reactogenicity · Mild	3 Participants	8 Participants	4 Participants	6 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination All reactogenicity · Moderate	2 Participants	0 Participants	0 Participants	1 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Local reactogenicity · None	7 Participants	4 Participants	8 Participants	5 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Local reactogenicity · Mild	4 Participants	8 Participants	4 Participants	6 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Local reactogenicity · Moderate	1 Participants	0 Participants	0 Participants	1 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Systemic reactogenicity · None	11 Participants	8 Participants	11 Participants	7 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Systemic reactogenicity · Mild	0 Participants	4 Participants	1 Participants	5 Participants
Maximum Local or Systemic Reactogenicity After the First Vaccination Systemic reactogenicity · Moderate	1 Participants	0 Participants	0 Participants	0 Participants

PRIMARY outcome

Timeframe: 7 days post Vaccination #2 on Day 35

Population: 47 subjects received the second vaccination (one less than Analysis Population Description). Subject in the placebo group withdrew Completed vaccination # 1 but withdrew due to moderate pain, tenderness and swelling, before Vaccination 2.

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=11 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Maximum Local or Systemic Reactogenicity After the Second Vaccination All reactogenicity · None	10 Participants	6 Participants	9 Participants	8 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination All reactogenicity · Mild	1 Participants	5 Participants	3 Participants	4 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination All reactogenicity · Moderate	0 Participants	1 Participants	0 Participants	0 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Local reactogenicity · None	11 Participants	7 Participants	9 Participants	8 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Local reactogenicity · Mild	0 Participants	4 Participants	3 Participants	4 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Local reactogenicity · Moderate	0 Participants	1 Participants	0 Participants	0 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Systemic reactogenicity · None	10 Participants	9 Participants	12 Participants	11 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Systemic reactogenicity · Mild	1 Participants	3 Participants	0 Participants	1 Participants
Maximum Local or Systemic Reactogenicity After the Second Vaccination Systemic reactogenicity · Moderate	0 Participants	0 Participants	0 Participants	0 Participants

PRIMARY outcome

Timeframe: 7 days post Vaccination #3 on Day 56

Population: 44 subjects received third vaccination (4 less than Analysis Population Description). 2 subjects lost in Placebo group, one due to pain, swelling (after vaccination #2) and 1 due to incarceration. 1 subject lost from 10 μg cohort due to illness, 1 subject lost in 30 μg cohort because did not continue to meet inclusion/exclusion criteria.

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=10 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=11 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=11 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Maximum Local or Systemic Reactogenicity After the Third Vaccination All reactogenicity · None	9 Participants	7 Participants	9 Participants	8 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination All reactogenicity · Mild	1 Participants	3 Participants	2 Participants	3 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination All reactogenicity · Moderate	0 Participants	1 Participants	0 Participants	1 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Local reactogenicity · None	10 Participants	8 Participants	10 Participants	8 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Local reactogenicity · Mild	0 Participants	3 Participants	1 Participants	4 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Local reactogenicity · Moderate	0 Participants	0 Participants	0 Participants	0 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Systemic reactogenicity · None	9 Participants	8 Participants	9 Participants	9 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Systemic reactogenicity · Mild	1 Participants	2 Participants	2 Participants	2 Participants
Maximum Local or Systemic Reactogenicity After the Third Vaccination Systemic reactogenicity · Moderate	0 Participants	1 Participants	0 Participants	1 Participants

SECONDARY outcome

Timeframe: 4 weeks post 3rd immunization (84 days)

Seroresponse was defined as as a four-fold increase in antibody titers between baseline and 4-weeks post-third injection.

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=11 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Number and Percentage of Subjects With Anti-P2-VP8 Immunoglobulin G (IgG) and Immunoglobulin A (IgA) Seroresponses IgG · Seroresponse	0 Participants	12 Participants	11 Participants	12 Participants
Number and Percentage of Subjects With Anti-P2-VP8 Immunoglobulin G (IgG) and Immunoglobulin A (IgA) Seroresponses IgG · No seroresponse	11 Participants	0 Participants	1 Participants	0 Participants
Number and Percentage of Subjects With Anti-P2-VP8 Immunoglobulin G (IgG) and Immunoglobulin A (IgA) Seroresponses IgA · Seroresponse	0 Participants	12 Participants	12 Participants	12 Participants
Number and Percentage of Subjects With Anti-P2-VP8 Immunoglobulin G (IgG) and Immunoglobulin A (IgA) Seroresponses IgA · No seroresponse	11 Participants	0 Participants	0 Participants	0 Participants

SECONDARY outcome

Timeframe: Days 0, 28, 56 and 84 (before the first injection and 4 weeks after each injection)

Population: Included subjects that received vaccinations (see participant flow for reasons why some subjects did not receive all vaccinations).

Measured from sera taken on Days 0, 28, 56 and 84 (before the first injection and 4 weeks after each injection).

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin G (IgG) Baseline	519 titer Interval 155.0 to 1740.0	362 titer Interval 164.0 to 800.0	457 titer Interval 270.0 to 773.0	285 titer Interval 135.0 to 604.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin G (IgG) Pre-vaccination 2	551 titer Interval 172.0 to 1761.0	2607 titer Interval 1168.0 to 5821.0	4314 titer Interval 2273.0 to 8190.0	2867 titer Interval 1070.0 to 7686.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin G (IgG) Pre-vaccination 3	513 titer Interval 140.0 to 1883.0	7373 titer Interval 3470.0 to 15666.0	8489 titer Interval 4670.0 to 15431.0	12438 titer Interval 6365.0 to 24307.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin G (IgG) 4 weeks post-vaccination 3	520 titer Interval 145.0 to 1870.0	13090 titer Interval 5881.0 to 29135.0	13932 titer Interval 7163.0 to 27097.0	19546 titer Interval 10231.0 to 37343.0

SECONDARY outcome

Timeframe: Days 0, 28, 56 and 84 (before the first injection and 4 weeks after each injection)

Population: Included subjects that received vaccinations (see participant flow for reasons why some subjects did not receive all vaccinations).

Measured from sera taken on Days 0, 28, 56 and 84 (before the first injection and 4 weeks after each injection).

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 Participants 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin A (IgA) Baseline	519 titer Interval 115.0 to 1740.0	362 titer Interval 164.0 to 800.0	457 titer Interval 270.0 to 773.0	285 titer Interval 135.0 to 604.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin A (IgA) Pre-vaccination 2	551 titer Interval 172.0 to 1761.0	2607 titer Interval 1168.0 to 5821.0	4314 titer Interval 2273.0 to 8190.0	2867 titer Interval 1070.0 to 7686.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin A (IgA) Pre-vaccination 3	513 titer Interval 140.0 to 1883.0	7373 titer Interval 3470.0 to 15666.0	8489 titer Interval 4670.0 to 15431.0	12438 titer Interval 6365.0 to 24307.0
Geometric Mean Titer (GMT) of Anti-P2-VP8 Immunoglobulin A (IgA) 4 weeks post-vaccination 3	520 titer Interval 145.0 to 1870.0	13090 titer Interval 5881.0 to 29135.0	13932 titer Interval 7163.0 to 27097.0	19546 titer Interval 10231.0 to 37343.0

SECONDARY outcome

Timeframe: 4 weeks post 3rd immunization (84 days)

Seroresponse was defined as as a four-fold increase in antibody titers between baseline and 4-weeks post-third injection.

Outcome measures

Outcome measures
Measure	10 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 Participants 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain WA · Seroresponse	8 Participants	5 Participants	7 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain WA · No seroresponse	4 Participants	7 Participants	5 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain 89-12 · Seroresponse	10 Participants	8 Participants	10 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain 89-12 · No seroresponse	2 Participants	4 Participants	2 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain DS1 · Seroresponse	0 Participants	6 Participants	7 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain DS1 · No seroresponse	12 Participants	6 Participants	5 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain P · Seroresponse	7 Participants	8 Participants	10 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain P · No seroresponse	5 Participants	4 Participants	2 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain ST3 · Seroresponse	0 Participants	1 Participants	2 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain ST3 · No seroresponse	12 Participants	11 Participants	10 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain BRB · Seroresponse	0 Participants	3 Participants	4 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain BRB · No seroresponse	12 Participants	9 Participants	8 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain SC2 · Seroresponse	1 Participants	4 Participants	4 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain SC2 · No seroresponse	11 Participants	8 Participants	8 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain W161 · Seroresponse	8 Participants	9 Participants	10 Participants	—
Number and Percentage of Subjects With Serum Neutralizing Antibody Seroresponse, by Rotavirus Strain W161 · No seroresponse	4 Participants	3 Participants	2 Participants	—

Adverse Events

10 μg P2-VP8

Serious events: 0 serious events

Other events: 9 other events

Deaths: 0 deaths

30 μg P2-VP8

Serious events: 0 serious events

Other events: 5 other events

Deaths: 0 deaths

60 μg P2-VP8

Serious events: 1 serious events

Other events: 7 other events

Deaths: 0 deaths

Placebo

Serious events: 0 serious events

Other events: 6 other events

Deaths: 0 deaths

Serious adverse events

Serious adverse events
Measure	10 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 participants at risk 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Musculoskeletal and connective tissue disorders Back pain	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)

Other adverse events

Additional Information

Jorge Flores

PATH

Phone: 2028220033

Email: [email protected]

Results disclosure agreements

Principal investigator is a sponsor employee
Publication restrictions are in place

Measure	10 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 10 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	30 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 30 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	60 μg P2-VP8 n=12 participants at risk 3 doses of P2-VP8 subunit rotavirus vaccine (Lot # 1746) produced in E. coli was adsorbed onto aluminum hydroxide (0.6 mg/dose) adjuvant prior to administration. Each dose contained 60 μg of active ingredient. P2-VP8 subunit rotavirus vaccine: P2-VP8 subunit rotavirus vaccine was made by inserting a codon optimized synthetic gene for the VP8 region of rotavirus VP4 fused to the P2 T-cell epitope of tetanus toxin into the Pj411 proprietary cloning vector developed by DNA 2.0, Menlo Park, CA. The vector carries a kanamycin resistance gene as a selection marker. The vector was transfected into the BL21 strain of E. coli. The fusion protein was purified from Isopropyl β-D-1-thiogalactopyranoside (IPTG)-induced and physically lysed cultures using standard column chromatographic techniques employing Q-Sepharose and Butyl 650 as resins in addition to ultrafiltration and diafiltration.	Placebo n=12 participants at risk 3 doses of placebo delivered intramuscularly. placebo: Sodium Chloride 0.9%, USP for Injection was used to dilute the active P2-VP8 vaccine to final dosing concentration and was used for the Placebo for the study.
Gastrointestinal disorders Constipation	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Gastrointestinal disorders Diarrhoea	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Infections and infestations Atypical pneumonia	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Infections and infestations Fungal infection	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Infections and infestations Gastroenteritis viral	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Infections and infestations Upper respiratory tract infection	50.0% 6/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	33.3% 4/12 • 6 months after final vaccination (224 days)
Infections and infestations Vaginitis bacterial	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Injury, poisoning and procedural complications Muscle strain	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Investigations Alanine aminotransferase increased	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Investigations Haemoglobin decreased	25.0% 3/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Investigations Neutrophil count decreased	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Investigations Platelet count decreased	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Investigations White blood cell count decreased	16.7% 2/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Investigations White blood cell count increased	8.3% 1/12 • 6 months after final vaccination (224 days)	16.7% 2/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	25.0% 3/12 • 6 months after final vaccination (224 days)
Nervous system disorders Balance disorder	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Nervous system disorders Dizziness	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	16.7% 2/12 • 6 months after final vaccination (224 days)
Nervous system disorders Headache	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Nervous system disorders Syncope	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Reproductive system and breast disorders Dysmenorrhoea	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Respiratory, thoracic and mediastinal disorders Cough	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Respiratory, thoracic and mediastinal disorders Hiccups	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Respiratory, thoracic and mediastinal disorders Oropharyngeal pain	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Respiratory, thoracic and mediastinal disorders Rhinorrhoea	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Respiratory, thoracic and mediastinal disorders Throat irritation	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)
Skin and subcutaneous tissue disorders Blood blister	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)
Skin and subcutaneous tissue disorders Dry skin	0.00% 0/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)	8.3% 1/12 • 6 months after final vaccination (224 days)	0.00% 0/12 • 6 months after final vaccination (224 days)