COVID-19 Pneumonia
Conditions
Keywords
Covid-19, Vaccine Reaction, Bacillus subtilis
Brief summary
This is a study trial to assess the effectiveness of the immune response stimulated by the genetically engineered Bacillus subtilis which express and display Spike protein of the SARS-COV2 on the spore coat.
Detailed description
Bacillus subtilis is regarded as safe organism by The Food and Drug Administration and it is presented in most food sources. Preliminary experiments have shown that the genetically engineered Bacillus subtilis can express and display receptor binding domain of spike protein of the SARS-COV2 on its spore coat, thus successfully induce the secretion of cytokines of human cells in vitro. Previous experiments also successfully demonstrated that a increased detection of neutralizing IgG and igM levels in mice after oral administrated with the Bacillus subtilis. This suggests that the transgenic spores of Bacillus subtilis have successfully activated the immune system, producing high-affinity neutralizing antibodies and memory B cells. Furthermore, no adverse effects were shown in all the mices. The engineered Bacillus subtilis will be further studied in a human trials through oral administration to test its safety and the immune effect resulted in human bodys.
Interventions
BIOLOGICALBacillus subtilis
Bacillus subtilis, a harmless intestinal commensal, has earned in recent years, great reputation as a vaccine production host and delivery vector with advantages such as low cost, safe for human consumption and straightforward administration. The technology team has succeeded engineering Bacillus subtilis with spore coat proteins resembling the proteins of the nucleus and spikes of coronal virus. This product could have a vaccine like activity within the intestinal environment.
Sponsors
DreamTec Research Limited
Study design
Allocation
NON_RANDOMIZED
Intervention model
PARALLEL
Primary purpose
BASIC_SCIENCE
Masking
NONE
Intervention model description
two parallel experimental groups were designed. 1\. the vaccinated volunteers and 2. unvaccinated groups.
Eligibility
Sex/Gender
ALL
Age
25 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
* healthy * age over 25 years * the outcome of the following examinations should be clinically insignificant: medical and surgical history (hypo-, hypertension, allergy, other diseases, major surgery, micturition, defecation, sleep, illness within the last 4 weeks prior to the start of the trial); * participant vaccinated with Sinovac over 4 months * anti-SARS CoV 2 neutralizing antibody is negative in serum.
Exclusion criteria
* pregnant women * history of COVID-19 infection or showing COVID-19 infection symptoms * having had contact to people with known COVID-19 infection in the last 14 days * having fever (\> 37.4oC in the last 24 hours), dry cough or feeling tired and having aches and pains, nasal congestion, runny nose, sore throat and diarrhea. * positive real time RT-PCR COVID-19 test. * persons with autoimmune diseases * allergic diathesis or any clinically significant allergic disease (i.e. asthma) * any condition that might impair the immune response * recent or current immunosuppressive medication * any other vaccine application 30 days before the first dose
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Serum Neutralizing Receptor Binding Domain IgG Antibody Concentration | Day 0, 27, 42 post oral administration | Concentration of neutralizing IgG antibody against receptor binding domain of spike protein in SARS-COV2 |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Lentirival Pseudovirus Neutralization Assay (Wild Type of SARS-CoV2) | Day 0, 27, 42 post oral administration | The ability of neutralization against SARS-CoV-2 was tested by an in vitro pseudo-virus neutralization assay. The lentivirus carrying a GFP gene was pseudotyped with the spike protein from a wild type of SARS-CoV-2. The pseudoviruses were then pre-incubated with serially diluted serum samples from orally vaccinated volunteers before being added to A549 lung carcinoma cells expressing human ACE2 and human TMPRSS2. The percentage of infection rate was measured with a fluorescent plate reader by counting GFP-positive cells. The results were fitted with a non-linear regression model. The dilution of the serum sample resulted in a 50% reduction of infection rate is designated as EC50. The results were presented as NA when the serum samples failed to neutralize pseudovirus infection. |
| Lentirival Pseudovirus Neutralization Assay (D614G SARS-COV2 Variant) | Day 0, 27, 42 post oral administration | The ability of neutralization against SARS-CoV-2 was tested by an in vitro pseudo-virus neutralization assay. The lentivirus carrying a GFP gene was pseudotyped with the spike protein from a D614G variant of SARS-CoV-2. The pseudoviruses were then pre-incubated with serially diluted serum samples from orally vaccinated volunteers before being added to A549 lung carcinoma cells expressing human ACE2 and human TMPRSS2. The percentage of infection rate was measured with a fluorescent plate reader by counting GFP-positive cells. The results were fitted with a non-linear regression model. The dilution of the serum sample resulted in a 50% reduction of infection rate is designated as EC50. The results were presented as NA when the serum samples failed to neutralize pseudovirus infection. |
Countries
Hong Kong
Participant flow
Recruitment details
Participants voluntary enrolled or recruitted by Genfortune Pharmaceuticals Limited
Pre-assignment details
6 unvaccinated participants were recruited and their level of serum neutralizing antibody against spike protein of SARS-CoV2 were lower than 0.003 ug/ml.
Participants by arm
| Arm | Count |
|---|---|
| Generation of Neutralizing Antibody for Unvaccinated Participants participants received vaccine 1 capsule of 1×10\^10 CFU of B. subtilis spore at day 0, 14, and 28 respectively.
Bacillus subtilis: Bacillus subtilis, a harmless intestinal commensal, has earned in recent years, great reputation as a vaccine production host and delivery vector with advantages such as low cost, safe for human consumption and straightforward administration. The technology team has succeeded engineering Bacillus subtilis with spore coat proteins resembling the proteins of the nucleus and spikes of coronal virus. This product could have a vaccine like activity within the intestinal environment. | 6 |
| Total | 6 |
Baseline characteristics
| Characteristic | Generation of Neutralizing Antibody for Unvaccinated Participants |
|---|---|
| Age, Categorical <=18 years | 0 Participants |
| Age, Categorical >=65 years | 2 Participants |
| Age, Categorical Between 18 and 65 years | 4 Participants |
| Race (NIH/OMB) American Indian or Alaska Native | 0 Participants |
| Race (NIH/OMB) Asian | 6 Participants |
| Race (NIH/OMB) Black or African American | 0 Participants |
| Race (NIH/OMB) More than one race | 0 Participants |
| Race (NIH/OMB) Native Hawaiian or Other Pacific Islander | 0 Participants |
| Race (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Race (NIH/OMB) White | 0 Participants |
| Region of Enrollment East Asia | 6 participants |
| Sex: Female, Male Female | 3 Participants |
| Sex: Female, Male Male | 3 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | 0 / 6 |
| other Total, other adverse events | 0 / 6 |
| serious Total, serious adverse events | 0 / 6 |
Outcome results
Primary
Serum Neutralizing Receptor Binding Domain IgG Antibody Concentration
Concentration of neutralizing IgG antibody against receptor binding domain of spike protein in SARS-COV2
Time frame: Day 0, 27, 42 post oral administration
Population: Participants were orally administered 5x10\^7 spores/kg person of recombinant B. subtilis spores mixed with sodium alginate in enteric coated capsules at day 1, 14, and 28. Furthermore, 5 mL of blood samples were drawn at day 0, 27, 42, and the serum samples were analyzed. The titer of neutralizing antibodies was determined with a CLIA-based assay.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Generation of Neutralizing Antibody for Unvaccinated Participants | Serum Neutralizing Receptor Binding Domain IgG Antibody Concentration | Day 0 | 0.2 ug/ml | Standard Deviation 0.1 |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Serum Neutralizing Receptor Binding Domain IgG Antibody Concentration | Day 27 | 9.3 ug/ml | Standard Deviation 5.4 |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Serum Neutralizing Receptor Binding Domain IgG Antibody Concentration | Day 42 | 35.8 ug/ml | Standard Deviation 8.8 |
Secondary
Lentirival Pseudovirus Neutralization Assay (D614G SARS-COV2 Variant)
The ability of neutralization against SARS-CoV-2 was tested by an in vitro pseudo-virus neutralization assay. The lentivirus carrying a GFP gene was pseudotyped with the spike protein from a D614G variant of SARS-CoV-2. The pseudoviruses were then pre-incubated with serially diluted serum samples from orally vaccinated volunteers before being added to A549 lung carcinoma cells expressing human ACE2 and human TMPRSS2. The percentage of infection rate was measured with a fluorescent plate reader by counting GFP-positive cells. The results were fitted with a non-linear regression model. The dilution of the serum sample resulted in a 50% reduction of infection rate is designated as EC50. The results were presented as NA when the serum samples failed to neutralize pseudovirus infection.
Time frame: Day 0, 27, 42 post oral administration
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (D614G SARS-COV2 Variant) | EC50 of Day 0 Serum diltuion in Pseudovirus inhibition assay | NA fold dilution | — |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (D614G SARS-COV2 Variant) | EC50 of Day 42 Serum diltuion in Pseudovirus inhibition assay | 82.4 fold dilution | Standard Deviation 29.2 |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (D614G SARS-COV2 Variant) | EC50 of Day 27 Serum diltuion in Pseudovirus inhibition assay | NA fold dilution | — |
Secondary
Lentirival Pseudovirus Neutralization Assay (Wild Type of SARS-CoV2)
The ability of neutralization against SARS-CoV-2 was tested by an in vitro pseudo-virus neutralization assay. The lentivirus carrying a GFP gene was pseudotyped with the spike protein from a wild type of SARS-CoV-2. The pseudoviruses were then pre-incubated with serially diluted serum samples from orally vaccinated volunteers before being added to A549 lung carcinoma cells expressing human ACE2 and human TMPRSS2. The percentage of infection rate was measured with a fluorescent plate reader by counting GFP-positive cells. The results were fitted with a non-linear regression model. The dilution of the serum sample resulted in a 50% reduction of infection rate is designated as EC50. The results were presented as NA when the serum samples failed to neutralize pseudovirus infection.
Time frame: Day 0, 27, 42 post oral administration
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (Wild Type of SARS-CoV2) | EC50 of Day 27 Serum diltuion in Pseudovirus inhibition assay | NA Fold of dilution | — |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (Wild Type of SARS-CoV2) | EC50 of Day 0 Serum diltuion in Pseudovirus inhibition assay | NA Fold of dilution | — |
| Generation of Neutralizing Antibody for Unvaccinated Participants | Lentirival Pseudovirus Neutralization Assay (Wild Type of SARS-CoV2) | EC50 of Day 42 Serum diltuion in Pseudovirus inhibition assay | 76.33 Fold of dilution | Standard Deviation 38.7 |