Tambahan Data Pra-OPB Vaksin

ABO1020-DP kepada BPOM

Jawaban Pertanyaan Nomor 2

Pertanyaan

2. Mengingat efikasi vaksin prototipe AWCorna terhadap varian Omicron sangat rendah (cross
protection 28%), maka untuk mendukung data efikasivaksin mRNA COVID-19 BA.4/5 masih
diperlukan beberapa dokumen:

2.1 Agar dikirimkan data hasil uji tantang (challenge study) menggunakan vaksin mRNA
COVID-19 BA.4/5 yang dibandingkan dengan vaksin dengan platform yang sama atau
dengan pelarut.

2.2 Agar dilengkapi dengan data immunological markers sesuai yang tercantum dalam
Pedoman WHO Annex 3: ADDENDUM to Considerations for Evaluation of COVID-19
Vaccines for Prequalification or Emergency Use Listing version 12 March 2021.

Jawaban:

2.1 To confirm that ABO1020 vaccine will confer protection against virus challenge, we
performed a mouse challenge study with two groups of mice (5 mice/group) that received
intramuscularly two doses of either ABO1020 vaccine or placebo PBS. 42 days after the first
immunization, both groups of mice were intranasally infected with the maximal dose of Omicron
BA.4 virus and the mice were sacrificed three days after infection to detect lung and nasal virus
load by PCR and to analyze lung pathological change by HE (Hematoxylin and Eosin) staining.
PCR analysis showed high levels of viral RNA (>10 5 copies in the lung and >107 copies in the
nasal per g) in the PBS control group. In contrast, virus load in the lung and in the nasal from
vaccinated group is significantly reduced to almost baseline level. (Figure 2-1)The data
demonstrate that ABO1020 vaccine have a potent protection efficacy against BA.4 virus and
indicate it will have a high protection against many other Omicron VOCs since the vaccine
induced high level of cross-reactive neutralizing antibody titer.

Since the translation from Chinese to English is still underway, the full report of this challenge
study is expected to be available at the end November 2022.
Figure 2-1 Virus titer in the Lung and nasal turbinate tissues of mice challenged with BA.4 variant

2.2 According to the WHO guideline, some studies should be conducted in the case of modified
COVID-19 vaccine are presented as follows:

Table 1. The test items based on the WHO guidelines

No. Test items Availability

1 Levels of neutralizing and non-neutralizing antibodies Presented
2 Antibody avidity (the binding strength between antigen and N/A
antibodies)
3 T-cell response profile dan examination of CD8+ and CD4+ T cell Presented
response
4 Characterization of lung histopathology Presented

Levels of neutralizing and non-neutralizing antibodies

The preclinical studies of ABO1020 vaccine showed that the BA.4/5 RBD specific IgG titer in
mice after 2 dose injection is 390499, and the neutralization titer against BA.4/5 pseudovirus is
21870. The ratio between neutralizing Ab titer and IgG titer is 0.056. The detailed data is
presented at Appendix number 3.

Antibody avidity

As to antibody avidity, we did not perform such analysis since we do not think it is essential to
do so. Antibody avidity has been sometimes used to assess the quality of antibody induced by a
vaccine. However, the assays used by different groups are largely different and showed great
variation in the results. In our studies, the neutralizing antibodies from mice immunized with
ABO1020 were evaluated using pseudovirus system and the data is highly consistent and clearly
showed that this vaccine could induce high neutralizing antibody titer with GMT value of 21,870
against COVID -19 BA.4 variant, indicating a strong protection efficacy.

T-cell response profile dan examination of CD8+ and CD4+ T cell response

A mouse study was performed to demonstrate the T cell responses of ABO1020. In the study,
spleen cells from mice immunized with ABO1020 or with AWcorna were ex vivo stimulated
with peptide pools covering the specific RBD region of VOCs, and the T cells expressing
cytokines IFNγ, IL-2 and IL-4 were enumerated. The result showed both ABO1020 and
AWcorna induced typical Th1 T cell response by inducing high level of IFNγ secretion, with
ABO1020 having a trend of higher response than AWcorna and low IL-4-secreting T cell
numbers. The detailed data is presented at Appendix number 4, while the full report of this study
will be available at the end of November 2022 due to translation progress still underway.

Table 2-1 Data of cytokines secretion by splenic T cells in AWcorna/ABO1020 immunized

mice

Groups
Peptides pool Cytokines
AWCorna ABO1020 PBS
Omicron ABO-RBD.529 peptide Pool IFN-γ 332±31 1360±47 1±1
(It’s also BA.1 RBD peptides) IL-2 156±11 384±43 5±2
（Mean±SEM, n=5） IL-4 12±3 10±4 6±1
IFN-γ 506±41 1828±66 4±1
Omicron ABO-BA.2 peptide Pool
IL-2 368±94 629±38 16±2
（Mean±SEM, n=5）
IL-4 81±20 51±3 48±8
Omicron ABO-BA.4 peptide Pool IFN-γ 445±42 2041±84 2±1
 IL-2 207±29 1011±48 17±3
（Mean±SEM, n=5）
IL-4 50±10 96±13 44±16

Characterization of lung histopathology

The study for lung histopathology characterization was inferred from the previous virus
challenge study for AWcorna because challenge study data is replaced by the immunogenicity
based on previous explanation. According In that study, the mice were immunized twice with 2
µg and 10 µg of AWcorna and the neutralizing Ab titer were measured at Day 35 after the first
immunization. 40 days after the first immunization, mice were challenged with a mouse adaptive
COVID-19 virus and the lung virus load and the lung histopathology analysis were performed.

The study showed that the mice were well protected from virus challenge reflected by complete
clearance of lung virus titer and reduced lung pathological change (Figure 2-1 & 2-2).

The following paragraphs are brief summary data from AWcorna challenge protection study.

Pathological analysis of lung tissues from SARS-CoV-2 virus challenged mice received
AWcorna or placebo

The mice were immunized twice with 2 µg or 10 µg of AWcorna and were challenged 40 days
after the first immunization with a mouse adaptive COVID-19 virus. The lung virus load and the
lung histopathology analysis were performed after 5 days of virus challenge. Typical pulmonary
tissue injury such as epithelial degeneration, alveolar septal thickening and inflammatory cell
infiltration were noted in control mice, while the pulmonary structure is intact without obvious
histology abnormal changes in mice treated with twice AWcorna.

Since the lung histopathology study is a part of challenge study, the result of ABO1020 lung
histopathology will be available at the end of November 2022 the same when challenge study is
completed to be translated to English.
PBS control AWcorna 2 µg AWcorna 10 µg
 Figure 2-2 Lung pathological analysis from vaccinated and placebo mice challenged with
mouse adapted SARS-CoV-2 virus

Figure 2-3 Scoring of lung tissues with HE staining for pathological changes upon SARS-CoV-2
virus infection in vaccinate mice and placebo mice