Use of BCG Vaccine as a Preventive Measure for COVID-19 in Health Care Workers
Phase: PHASE2
Conditions: COVID 19 Vaccine
Sponsor: Universidade Federal do Rio de Janeiro

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Use of BCG Vaccine as a 
Preventive Measure for COVID-19 
in Health Care Workers 
 
 
VERSION: 14.0 / September 30, 2021 
 
HISTORIC: 
Version 1.0 / May 5, 2020 
Version 2.0 / June 7, 2020 
Version 3.0 / August 14, 2020 
Version 4.0 / September 9, 2020 
Version 5.0 / October 11, 2020 
Version 6.0 / October 22, 2020 
Version 7.0 / November 16, 2020 
Version 8.0 / December 11, 2020 
Version 9.0 / February 2, 2021 
Version 10.0 / April 24, 2021 
Version 11.0 / May 11, 2021 
Version 12.0 / July 29, 2021 
Version 13.0 / August 18, 2021 
 
ACRONYM: ProBCG 
 
NTC number: 04659941 
 
 

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SUMMARY: 
RESEARCHERS 
4 
PARTICIPATING INSTITUTIONS 
4 
SUBSCRIPTION PAGE (RESEARCHER - SPONSOR) 
5 
STATEMENT BY THE PRINCIPAL INVESTIGATOR 
6 
SUMMARY 
7 
INTRODUCTION 
8 
JUSTIFICATION 
12 
GOALS 
13 
Primary Objectives 
13 
Secondary Objectives 
13 
METHODOLOGY 
15 
Study Definitions 
15 
Study Design 
15 
Study site 
16 
Eligibility criteria 
16 
   Inclusion criteria 
16 
   Exclusion criteria 
16 
Intervention Plan 
17 
   Intervention 
17 
   Comparator 
18 
   Allocation and randomization 
18 
Study visits 
18 
   Storage of specimens during study visits 
22 
BCG or placebo vaccine application procedure 
22 
   BCG Vaccine 
22 
   Placebo 
23 
   Blinding 
23 
   Reaction to BCG vaccine and placebo 
23 
   Adverse events 
24 

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   Serious adverse events 
26 
Risks 
26 
Benefits 
27 
LABORATORY PROCEDURES 
28 
Blood collection 
28 
Polymerase chain reaction with real-time reverse transcription 
28 
Serological test for COVID-19 
28 
Immunoassays 
29 
Inflammatory profile and population characterization of neutrophils 
and T lymphocytes 
29 
Transcriptoma 
30 
Laboratory procedures in each research center 
30 
DATA COLLECTION AND MONITORING PLAN 
31 
Sample storage plan 
33 
Statistical Analysis Plan 
34 
Sample size 
34 
OUTCOMES 
36 
Primary Outcome 
36 
INTERIM ANALYSIS PLAN 
38 
Independent Data Monitoring and Security Committee 
38 
   CMDS Activity Plan 
39 
Plan for monitoring and analysis of adverse events 
40 
Criteria for discontinuation of the study for safety reasons 
41 
Criteria for discontinuation of experimental treatment in 
participants 
41 
QUALITY MANAGEMENT 
42 
BUDGET 
43 
SCHEDULE 
44 
REFERENCES 
45 
    
RESEARCHERS: 

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Fernanda Carvalho of Q. Mello; MD, MSc, PhD; CPF 916357567 15 
Afrânio Lineu Kritski; MD, MSc, PhD; CPF 299576569 53 
José Roberto Lapa e Silva; MD, MSc, PhD; CPF 127025017 53 
Elisangela Costa da Silva; MSc. PhD; CPF 016548077 70 
Rafael Galliez; MD, MSc. PhD; CPF 082542837 88 
Maria Armanda Monteiro da Silva Vieira; MD, MSc, PhD; CPF 707915627 00 
Ana Paula Santos; MD, MSc, PhD; CPF 047498367 61 
Antonio Guilherme Fonseca Pacheco; MD, MSc, PhD; CPF 012999877 02 
Martha Maria Oliveira; MSc, PhD; CPF 042874657 84 
Guilherme Loureiro Werneck; MD, MSc, PhD, CPF 770037137 72 
Rogério Lopes Rufino Alves; MD, MSc, PhD; CPF 821459627 00 
Luís Cristóvão de Moraes Sobrino Porto, MD, MSc, PhD; CPF: 429488357 04  
Hanna Silva Condelo, MSc; CPF: 135268117 09 
Walter Costa; MD; CPF 547574877 20 
Robson de Souza Leo; MSc, PhD; CPF 034374517 82 
Luciana Silva Rodrigues; MSc, PhD; CPF 075691427 21 
Janaina Aparecida of Medeiros Leung; MSc; CPF 160954918 00 
Fabio Silva Aguiar; MD; MD, MSc, PhD; CPF 077792197 98 
Paulo Fernando Guimarães Morando Marzocchi Tierno; MD; CPF 293080448 33 
Roberto de Andrade Medronho; MD, MSc, PhD; CPF 508401427 49 
Terezinha Marta Pereira Pinto Castineiras; MD, MSc, PhD; CPF 982013527 34 
 
 
PARTICIPATING INSTITUTIONS: 
1. Hospital Complex: Clementino Fraga Filho University Hospital - Institute of Chest 
Diseases, Federal University of Rio de Janeiro - Rio de Janeiro/RJ 
2. Pedro Ernesto University Hospital - Piquet Carneiro Polyclinic, State University of Rio 
de Janeiro - Rio de Janeiro/RJ 
3. Dr. Francisco Moran Municipal Hospital - Barueri/SP 
 
 

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SUBSCRIPTION PAGE (RESEARCHER - SPONSOR) 
 
 
I inform you that this Protocol has been revised and approved. 
We will supervise and coordinate the clinical trial, conducted in 2 centers in Brazil as 
described and ensure compliance with Good Clinical Practices (BPC) and Good 
Laboratory Practices (BPL), the principles described in the Helsinki Declaration and the 
applicable regulatory requirements. 
 
 
Name: Fernanda Carvalho de Queiroz Mello 
 
Institution: Institute of Chest Diseases - Federal University of Rio de Janeiro 
 
Signature: ________ 
                                                                                                                      day/month/year 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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STATEMENT BY THE PRINCIPAL INVESTIGATOR 
 
By signing this page, I agree to conduct the study in accordance with all current Brazilian 
regulations regarding applicable clinical research and international guidelines, as stated 
in the protocol and other information provided to me. 
I will ensure that the requirements for obtaining review and approval from the Research 
Ethics Committee (CEP) are met. I will promptly report to cep any and all changes in 
research activities covered by this protocol. 
I certify that the individuals involved with this study have completed training in Good 
Clinical Practices (BPC) in the last 3 years and, if applicable, training in the protection of 
human beings. 
I understand that all information obtained during the conduct of the study regarding the 
health status of individuals will be considered confidential. No individual name or 
personally identifiable information may be disclosed. All subject data will be anonymized 
and identified by numbers assigned in all Case Report Forms and laboratory samples. 
I will maintain the confidentiality of this protocol and all other related investigative 
materials. The information obtained from the study protocol cannot be disclosed or 
discussed with third parties without the express consent of the researcher – Dr. 
Fernanda Carvalho de Queiroz Mello. 
 
Full Name of the Principal Investigator: 
Fernanda Carvalho de Queiroz Mello 
Signature: ________ 
Signature date: __________ 
                                        day / month / year 
 
 
 
 
 
 
SUMMARY: 

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The disease promoted by coronavirus (COVID-19) is caused by SARS-Cov2, the first cases 
identified in December 2019 in China after exposure to the animal market in wuhan city, 
China (reviewed by Adhikari et al, 2020).  From the first case to the present day, the 
COVID-19 epidemic has been identified in 185 countries, with the notification of 
2,666,154 cases and 186,144 deaths (Source: COVID Visualizer, 23Apr2020). In Brazil, 
more than 45,757 cases and 2,906 confirmed deaths by COVID-19 have been confirmed 
(Source: COVID Visualizer, 23Abr2020). In our country, to date, testing for COVID-19 
occurs only in severe cases and few centers offer the service to health professionals, a 
population at high risk of infection. BCG is a vaccine produced from a live attenuated 
strain derived from a Mycobacterium bovis  isolate and is widely used worldwide as a 
tuberculosis (TB) vaccine, but there are studies demonstrating non-specific 
immunotherapeutic mechanisms of this vaccine that signal a possible relationship with 
the lowest morbidity and mortality associated with COVID-19 infections worldwide. The 
present study aims to analyze the role of BCG in the prevention of SARS-CoV-2 infection 
and also in the occurrence of severe forms of COVID-19 in addition to evaluating the 
immune response mediated by this vaccine in voluntary health professionals. 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
INTRODUCTION: 

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The disease promoted by coronavirus (COVID-19) is caused by SARS-Cov2, the first cases 
identified in December 2019 in China after exposure to the animal market in wuhan city, 
China (reviewed by Adhikari et al, 2020).   
From the first case to the present day, the COVID-19 epidemic has been identified in 185 
countries, with the notification of 2,666,154 cases and 186,144 deaths (Source: COVID 
Visualizer, 23Apr2020). In Brazil, more than 45,757 cases and 2,906 confirmed deaths 
by COVID-19 have been confirmed (Source: COVID Visualizer, 23Abr2020). In our 
country, to date, testing for COVID-19 occurs only in severe cases and few centers offer 
the service to health professionals, a population at high risk of infection. Thus, the data 
in Brazil present a high of underreporting. According to the UFRJ Technical Note, it is 
estimated that the cases reported in Brazil represent approximately only 11% of the 
cases (Kritski et al, 2020).  
It is known that the impact of the disease differs between countries. These differences 
are attributed to the different cultural norms, mitigation efforts, available health 
infrastructure and socio-economic level. Recently, Miller et al (2020) proposed, through 
an ecological study, that countries that continued with the use of vaccination with 
Bacillus Calmette Guérin (BCG) after birth have a lower occurrence of COVID-19.  
BCG is a vaccine produced from a live attenuated strain derived from an isolate of 
Mycobacterium bovis and is widely used worldwide as a tuberculosis (TB) vaccine, and 
countries such as Portugal, Japan and China adopt a universal BCG vaccination policy in 
newborns. Other countries such as Italy, the Netherlands, Spain, France, Switzerland and 
the United States of America have discontinued or have not adopted universal 
vaccination policies due to the low risk, when compared to countries with high 
prevalence of the disease, of developing Mycobacterium tuberculosis (MTB) infections, 
and also due to the variable efficacy in the prevention of tuberculosis in adults (Miller et 
al,  2020). 
Interestingly, shortly after its introduction in the 1920s, epidemiological studies showed 
that the BCG vaccine reduced infant mortality regardless of its effect on TB (reviewed 
by Shan et al, 2009). In several observational studies in West Africa, there was a 50% 
reduction in the overall mortality of children vaccinated with BCG, an effect too large to 
be explained by the protection against tuberculosis only (Garly et al, 2003; Roth et al, 
2005). More recently, these findings have been validated in controlled trials (Aaby et al, 

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2011). Therefore, the reduction in mortality in infants by BCG seems to be due to the 
induction of protection against unrelated infectious agents. These beneficial effects 
have been called "hemorologist" or "non-specific" effects.  
There are two basic mechanisms on the effect of BCG vaccine on herterologist 
responses: 
1) BCG induces recteral lymphocytic responses, resulting in more efficient immune 
responses in relation to secondary responses not related to infectious agents 
(Goodridge et al, 2016). For example, mice are protected by BCG vaccination against 
vaccinia virus infection by increasing the production of IFN-γ by CD4+ T cells (Mathurin 
et al, 2009). The recurrent lymphocytic responses may also involve the activation of 
CD4+ and CD8+ memory T  cells that are specific to non-target antigens, thus modulating TH1 
and TH17 responses to secondary non-mycobacterial infections. In healthy volunteer 
humans, BCG vaccination increased non-specific responses of 
TH1 and TH17 
subpopulations for at least 1 year after vaccination (Kleinnijenhuis et al, 2014). In 
addition, in patients with recurrent respiratory papillomatosis, which is caused by 
human papillomavirus in 90% of cases, adjuvant therapy with BCG restored the 
efficiency of antiviral T-cell response by stimulating th1 cytokine production and TREG cell 
induction. 
 
2) Less than a decade ago, only B and T lymphocytes were considered capable of 
constructing memory responses. However, in recent studies it has been shown that the 
functional program of innate immune cells is altered in certain infections or 
vaccinations, resulting in increased immunity when cells encounter a secondary stimulus 
(Netea et al, 2016). The induction of a non-specific memory in innate immune cells is 
known as "trained immunity" and is mediated by epigenetic and metabolic remodeling 
(Netea et al, 2016). Evidence suggests that the key mechanism by which BCG induces its 
non-specific effects is probably through the induction of immune memory in innate 
immune cells, especially in natural killer cells (NK), monocytes and macrophages, and 
not by means of adaptive immune mechanisms based on T cells and B cells.  
 
In mice with severe combined immunodeficiency, the BCG vaccine provided protection 
against a non-mycobacterial secondary challenge, confirming the importance of innate 

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immune cells in mediating this effect (Arts et al, 2018). In healthy human volunteers, 
BCG vaccination improved the production of pro-inflammatory cytokines, such as 
interleukin (IL)-1β and tumor necrosis factor (TNF)-α in peripheral blood mononuclear 
cells (PBMC) for up to 3 months after vaccination on in vitro stimulation with unrelated 
pathogens (Arts et al , 2018). This response was associated with an increase in CD11b, 
TLR4 and CD14 activation markers and epigenetic reprogramming of human monocytes 
at gene-promoting sites for pro-inflammatory cytokines. In a randomized BCG study, 
volunteer humans were vaccinated with the YFV vaccine, a yellow fever attenuated virus 
vaccine. In this study, a significant reduction in viremia was observed after 1 month or 
placebo in volunteers vaccinated with BCG when compared to volunteers who received 
placebo. And, this effect was correlated with epigenetic mutations induced by BCG in 
monocytes (Arts et al, 2018). BCG induced higher production of IL-1β during viral 
infections, rather than the production of IFN-γ by lymphocytes and NK cells. The 
protective role of IL-1β is related to recent findings suggesting a crucial role of this 
interleukin in antiviral immunity (Rathinam et al, 2010).  
Finally, several authors have discussed the role of BCG in inducing the IFN-I pathway in 
circulating monocytes, inducing the production of IFN-α and -β, cytokines known to be 
important in the control of viral replication (Moorlag et al, 2019). Modulation of a TH1 
response, with monocyte/macrophage activation, may help reduce the severe 
responses associated with COVID-19, as several authors point out that inflammatory 
responses associated with severe forms of Severe Acute Respiratory Syndrome (ARDS) 
are associated with neutrophil-mediated responses, 
TH17 profiles, pro-inflammatory 
cytokines such as IL-6,  in an event called a cytokine storm (reviewed by Jiang et al, 
2019).  
To build an antiviral response, the ininate immune system recognizes molecular 
structures that are produced by virus invasion, called pathogen-associated molecular 
patterns (PAMPs). For RNA viruses such as coronavirus, it is known that the PAMPs 
found in virus replication in the form of viral genomic ssRNA or double-taped RNA are 
recognized by the endosomal receptors of RNA TLR8 and TLR7 in ssRNA and by the 
cytosolic RNA sensor, RIG (retinid inducible gene)/MDA5 (gene 5 associated with 
melanoma differentiation) (Li et al,  2020; by Wit et al, 2016; Channappanavar et al, 
2017). This recognition leads to the activation of several signaling routes. Transcription 

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factors such as nuclear factor κB (NF‐κB), activating protein 1 (AP‐1), interferon response 
factor 3 and 7 (IRF3 and IRF7). NF‐κB and AP‐1 stimulate the expression of genes 
encoding various molecules necessary for inflammatory responses, including 
inflammatory cytokines (e.g., TNF-α and IL‐1) and chemokines (CCL2 and CXCL8). IRF3 
and IRF7 promote the production of type I interferon (IFN-α and IFN-β), important for 
innate antiviral immune responses capable of suppressing early-stage replication and 
viral dissemination (Deng et al, 2019; Yang et al, 2015). This process can cause 
complications such as exhaustion, weakness and cough in patients (Kindler et al, 2016). 
The important point in the current pandemic scenario by COVID19 is that, for SARS‐CoV‐
2, the response to ifn virus type I infection is suppressed (reviewed by Rokni et al, 2020), 
because hospitalized patients with severe COVID-19 have high levels of pro-
inflammatory cytokines, including IL‐2, IL‐7, G‐CSF, IP‐10, MCP‐1,  MIP‐1α, and TNF-α. 
This phenomenon known as "cytokine storm" or cytokine release syndrome becomes an 
important factor in the pathogenesis of COVID‐19 (Wongi et al, 2004; Conti et al, 2020). 
New strategies should be researched to analyze and improve the individual's response 
against SARS-CoV2 infection. 
Thus, it is understood that non-specific immunotherapeutic mechanisms of bcg vaccine 
in the light of current epidemiological data, indicate a possible relationship between the 
existence of universal BCG vaccine policies and the lower morbidity and mortality 
associated with COVID-19 infections worldwide. 
  
 
 
 
 
 
 
 
 
 
 
 

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JUSTIFICATION: 
Considering the above, the present project aims to study the effect of BCG vaccine on 
health professionals active in reference hospitals for the care of COVID-19. 
This project presents an innovative proposal, addressing a BCG vaccine with known 
efficacy against severe TB in childhood and reliable for the use of adjuvant therapy in 
patients with malignant bladder neoplasia. 
BCG is an easily accessible vaccine, widely used by SUS in newborn children, thus 
facilitating its incorporation into a new preventive proposal for COVID-19. In addition, 
the selection of health professionals will be based on diagnostic screening and 
confirmatory tests of COVID-19. 
It is intended to analyze the role of BCG in the prevention of infection and also in the 
occurrence of severe forms of COVID-19 in addition to evaluating the immune response 
mediated by this vaccine in voluntary health professionals. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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GOALS: 
In the period of 6 months, to analyze the effect of BCG as a protective factor for COVID-
19 in research volunteers, aiming at greater efficiency in the prevention of subjects at 
high risk of infection and evolution to severe forms of the disease, as well as to evaluate 
the specific and non-specific immune responses to SARS-CoV-2 in vaccinated and 
unvaccinated volunteers who evolved with COVID-19 or not.  
 
Primary Objectives: 
1. Assess the impact of BCG vaccine administration on the cumulative incidence of 
SARS-CoV-2 infection 
2. Assess the impact of BCG vaccine administration on the cumulative incidence of 
severe forms of COVID-19 
3. Evaluate the bcg vaccine-mediated immune response in volunteers 
 
Secondary Objectives: 
In groups receiving BCG or placebo, infected and not infected with SARS-CoV-2: 
 
- Epidemiological: 
1. Estimate whether BCG vaccination compared to placebo prolongs the time to the 
first respiratory disease proven by SARS-CoV-2; 
2. Estimate whether BCG vaccination compared with placebo reduces the proportion 
of symptoms associated with COVID-19 (fever or at least one sign/symptom of 
respiratory disease);  
3. Estimate whether BCG vaccination compared to placebo reduces absenteeism 
(missing days) in volunteers who are health professionals;  
4. Analyze the safety of BCG vaccination in adult volunteers; 
 
- Laboratories: 
5. Describe the inflammatory profile by dosing neutrophils, platelets, cit-H3, alpha 1 
antitrypsin (α1AT), galectin-2, ferritin, transferrin-R, glycated hemoglobin (HbA1c), 

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C-reactive protein (CRP), D-dummer, LDH, albumin and hemosedimentation velocity 
(ESR); 
6. Describe the profile of inflammatory cytokines identified in the BCG and placebo 
groups, comparing to the protective events to SARS-CoV-2; 
7. Describe the epigenetic alterations associated with monocytes and neutrophils 
related to the protection of COVID-19; 
8. Characterize the cell population (T, B lymphocytes, neutrophils and monocytes) in 
peripheral blood of the study groups (BCG and placebo); 
9. Relate the profile of immune response associated with mild and severe forms 
observed in the study groups (BCG and placebo); 
10. Analyze the serum concentration of antibodies and relate it to the immune 
response; 
11. Identify possible antibodies that may confer immunity to new COVID-19 infections; 
12. Perform transcriptomic analysis of neutrophilic, lymphocytic and monocytic 
responses in the study groups (BCG and placebo) involved in the modulation of 
immune responses in the study groups; 
13. Identify predictive biomarkers of infection and/or illness by SARS-CoV-2; 
14. Develop a predictive model of infection and/or illness by SARS-CoV-2; 
15.  Identify socio-demographic factors, life habits, associated diseases and previous 
vaccinations associated with immune response and the risk of COVID-19 infection. 
 
 
With a view to the approval of specific vaccines against COVID-19 for emergency use by 
ANVISA in January 2021 and incorporated by the National Immunization Program of the 
Brazilian Ministry of Health, the objectives for the subgroup of individuals who receive 
the intervention of this study with BCG or placebo and who are also immunized with 
specific vaccines against COVID-19 will be carried out in the part,  taking into account in 
the future analysis, the time between the intervention of the study and vaccination 
against COVID-19, the vaccine used and the performance of the second dose of the 
vaccine used, aiming to identify a possible adjuvant effect of bcg vaccine as a stimulant 

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of cellular immunity, which also plays a role in immunopathogeny against SARS-CoV-2, 
including action prior to humoral response. 
Furthermore, in cases of participants immunized with specific vaccines against SARS-
CoV-2, the identification of asymptomatic infection by the virus, diagnosed through the 
conversion of serologies to COVID-19 in the moments M3 and M6 after the intervention, 
will not be evaluated, considering the bias caused by the use of vaccines against viral 
disease. In these cases, in order to characterize the diagnosis of COVID-19, there will be 
a need for a report (self-reported) of characteristic signs/symptoms associated with 
detection of SARS CoV-2 by RT-PCR. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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METHODOLOGY: 
Study Definitions: 
The definitions of the Brazilian Ministry of Health (Brazil, 2020) will be used: 
 
Flu syndrome (GS): Individual with acute respiratory condition, characterized by 
at least two (2) of the following signs and symptoms: fever (even if reported), 
chills, sore throat, headache, cough, runny nose, olfactory disorders or taste 
disorders. 
 
Severe Acute Respiratory Syndrome (SARS): Individual with GS sit : 
dyspnea/respiratory distress OR persistent pressure in the chest OR O2 
saturation less than 95% in room air OR bluish coloration of the lips or face. 
 
Confirmed cases of COVID-19: 
- By clinical criterion: Case of GSs or SRAG with clinical confirmation 
associated with anosmia (olfactory dysfunction) OR ageusia (gustatory 
dysfunction) acute without other previous cause. 
- Por clinical - epidemiological criterion: Case of GSs or SRAG with a history 
of close or household contact, in the 14 days prior to the appearance of signs and 
symptoms with case confirmed for COVID-19. 
- By clinical criterion - imaging: Case of GSS  or SRAG or death from RagS 
that could not be confirmed by laboratory criterion And that presents at least one 
(1) of the following tomographic changes: 
   - Peripheral, bilateral , horizontal matte glass opacity , with or without 
consolidation or visible intralobular lines ("paving"), OR 
   - Multifocal frosted glass opacity of rounded morphology with or without 
consolidation or visible intralobular lines ("paving"),OR 
   - Reverse halo sign or other findings of organizing pneumonia (later observed 
in the disease). 
- By laboratory criterion: Case of SG or SRAG with test of: 
   - Molecular biology: detectable result for SARS-CoV-2 performed by the RT-PCR 
method in real time. 

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- Immunological:  reactive result for IgM, IgA and/or IgG performed by the following 
methods: Enzyme-Linked Immunosorbent Assay (ELISA); Immunochromatography 
(rapid test) for antibody detection; Electrochemiluminescence Immunoassay (ECLIA); 
  - Antigen Research: reagent result for SARS-CoV-2 by immunochromatography method 
for antigen detection. 
 
- By laboratory criterion in asymptomatic individual: Asymptomatic individual with test 
results:  
   - Molecular Biology: detectable result for SARS-CoV-2 performed by the RT-PCR 
method in real time; 
  - Immunological: reagent result for IgM and/or IgA performed by the following 
methods: ELISA or Immunochromatography (rapid test) for antibody detection. 
 
- Health Professional: For the purposes of this study, in addition to the definition of a 
health professional established in Resolution 218 - 97 - Regulation of health professions, 
the Brazilian Classification of Occupations will be used, and in an expanded way will be 
included undergraduate students in the area of health sciences who attend the hospital 
and are in direct contact with patients. Administrative technical professionals who 
circulate or perform activities in sectors within the hospitals participating in the study 
and who have direct contact with patients may also be included in the study, such as 
receptionists, maqueiros, security guards, among others. 
 
Study Design: 
The project proposes to monitor research participants for 6 months and thus conduct a 
controlled clinical trial, phase II-B, which compares the occurrence of SARS-CoV-2 
infection as well as the time to infection among health professionals receiving bcg 
vaccine (experimental group) compared to those receiving placebo (control group). 
Coupled with the clinical trial, participants will also be analyzed in an observational study 
of two cohorts of volunteers, vaccinated with BCG and not vaccinated. In this part of the 

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study, participants will be analyzed in relation to the inflammatory profile, considering 
biomarkers predicting infection and evolution to severe form of COVID-19. 
Study site: 
The possible participants of this research will be recruited in two hospitals in the city of 
Rio de Janeiro (Hospital Complex Institute of Chest Diseases - Clementino Fraga Filho 
University Hospital of UFRJ and Pedro Ernesto University Hospital Complex - Piquet 
Carneiro Polyclinic of UERJ) and a hospital in the city of Barueri in SP (Hospital Municipal 
Dr. Francisco Moran - Barueri/SP). 
 
 
Eligibility criteria: 
Health professionals (according to the definitions of this study), who are in the exercise 
of their activities in the units of the study or not, of both sexes over the age of 18 years 
and who agree to participate in the study by signing the Informed Consent Form (TCLE). 
 
Inclusion criteria: 
1. Individuals aged 18 ≥, male or female, not infected with SARS-CoV-2  
2. Agreement to participate in the study by signing the TCLE 
3. Not being pregnant in case of women able to become pregnant 
4. Have not received a specific vaccine against COVID-19 or, if vaccinated against SARS-
CoV-2, have received the complete immunization schedule only with vaccines 
approved by ANVISA and implemented by the National Immunization Program, 
(including the second dose) within a minimum of 15 days prior to the date of 
inclusion in the study 
5. If the participant have not received a specific vaccine against COVID-19 approved by 
ANVISA, be aware and agree to be able to receive them only 15 days after the 
intervention proposed in this study 
 
Exclusion criteria: 
1. Professionals with a history of sars-cov-2 confirmed infection through RT-PCR or who 
have already presented clinical and molecular diagnosis of COVID-19 prior to the 
study 

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2. Individuals who have not underwent confirmatory tests for COVID-19 
3. Breastfeeding 
4. Individuals who report having primary or acquired immunodeficiency 
5. Individuals who report being affected by malignant neoplasms 
6. Patients reporting treatment with high-dose corticosteroids (equivalent to the 
prednisone dose of 20 mg/day or more) for more than two weeks 
7. Patients who report being on other immunosuppressive therapies (antineoplastic 
chemotherapy, radiotherapy, among others) 
8. Individuals who report being diagnosed with autoimmune diseases 
9. Dermatological conditions at the vaccine site or generalized 
10. Individuals who report being treated for active tuberculosis 
11. Individuals who report a history of previous treated tuberculosis 
12. Individuals with febrile symptoms (body temperature ≥ 37.5ºC in the last 48h) 
13. Participation in other clinical prevention trials for COVID-19 (vaccines already 
approved by ANVISA for use by the National Immunization Program are not included 
in this item) 
14. Report of vaccination with live microorganism administered in the month prior to 
randomization 
15. Require that, if another vaccination with live microorganism is required, it is 
administered in the month following randomisation (If the other live vaccine can be 
administered on the same day, this exclusion criterion does not apply) 
16. Known anaphylactic reaction to any ingredient in bcg vaccine 
17. Adverse reaction prior to BCG vaccine [significant local reaction (abscess) or suputive 
lymphadenitis] 
18. BCG vaccine administered in the last year 
 
 
Intervention Plan: 
Intervention: 
Randomly eligible participants who have provided informed written consent will be 
recruited. Participants will then be allocated to one of the study regimens in a ratio of 
1:1. 

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1. Intervention: BCG vaccine, single dose 
2. Control regimen: placebo, single dose. 
In the BCG clinical trial, participation in the study will last 6 months: during the first 4 
months, or for the duration of the recruitment period, participants will receive the 
randomly designated BCG regimen or only control group. After the recruitment period, 
all study participants should have received BCG or placebo, and both groups will be 
monitored through clinical and laboratory examination for 6 months of follow-up. 
Biosafety care to combat COVID-19 adopted in the participating Health Units will also 
be monitored during the study period. 
 
 
Comparator: 
The natural evolution of SARS-CoV-2 infection in health professionals will be used to 
evaluate the efficacy of bcg use as preventive therapy, considering the use of specific 
vaccines against COVID-19, the time of positivity of COVID-19 tests and the evolution to 
severe forms of COVID-19. 
 
 
Specific vaccines against SARS-CoV-2: 
- Coronavac® (Sinovac® / Butantan Institute): 
The vaccine developed by the Sinovac laboratory® in partnership with the Butantan 
Institute is a vaccine containing the inactivated SARS-CoV-2 virus. Seroconversion 
studies of the Sinovac/Butantan vaccine showed results of >92% in participants who 
took both doses of the vaccine within 14 days and > 97% in participants who took both 
doses of the vaccine within 28 days. 
The efficacy of this vaccine was demonstrated in a regimen containing 2 doses with an 
interval of 2 weeks. For the prevention of symptomatic cases of COVID-19 that required 
outpatient or hospital care, efficacy was 77.96%. There were no serious cases in 
vaccinated subjects, compared to 7 severe cases in the placebo group. 
Considering the absence of co-administration studies, covid-19 vaccines are currently 
not recommended simultaneously with other vaccines. Thus, a minimum interval of 14 

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days is recommended between COVID-19 vaccines and the different vaccines of the 
National Vaccination Calendar. 
 
 
- Covishield® (AstraZeneca® / Fiocruz):  
The vaccine developed by the AstraZeneca/University of Oxford laboratory and 
produced in partnership with Fiocruz is a vaccine containing 5 × 1010 viral particles (pv) 
of the replication-deficient chimpanzee recombinant adenovirus vector (ChAdOx1), 
which expresses the Glycoprotein SARS-CoV-2 Spike (S). Produced in human embryonic 
brain cells (HEK) 293 genetically modified. After vaccination, in participants who were 
seronegative at the beginning of the study, seroconversion was demonstrated in ≥ 98% 
of participants at 28 days after the first dose and > 99% at 28 days after the second. For 
prevention of illness by COVID-19 the vaccine demonstrated efficacy of 73% 22 days 
after the first dose (in a period of at least 3 months) and with strong indication of 
increased immune response when the second dose is provided within 3 months, which 
provides an increase of about 7.5 times in levels of humoral response (antibody 
production). Exploratory analyses showed that increased immunogenicity was 
associated with a longer dose interval and efficacy is currently more certainly 
demonstrated for intervals of 8 to 12 weeks. It is notepoint that there were no severe 
cases or deaths 21 days or more after vaccination, and 10 hospitalizations for severe 
COVID-19 were observed in the placebo group, including 1 death. 
 
 
Allocation and randomization: 
The randomization process will be carried out centrally and managed by the team of the 
Ribeirão Preto School of Medicine of USP (FMRP-USP), under the coordination of 
Professor Domingos Alves. The process will be performed using the Microsoft Excel 
random number generation function®. The result will be composed of a randomization 
table with size appropriate to the N expected for the study (752 participants) and with 
a ratio of 1:1, which will be integrated to REDCap prior to the beginning of electronic 
data collection. The creation and validation of the table will be the responsibility of the 
data manager/project coordinator. 

Page 22 from 55 
 
By including a participant in the study, REDCap will consult the existing randomization 
table and excel® sequentially to allocated it to one of the groups defined in the study. It 
is noteworthy that REDCap does not, on its own, perform any form of randomization, 
which will be based exclusively on the said table in Excel®. 
 
Study visits: 
Social networks and electronic media will be used to disseminate the study through 
folders and calls about the research.  
Health professionals from other institutions besides those participating in the study may 
participate in the research. In this case, they will contact the participating centers, 
through telephone contact or email, informing their interest. All follow-up of these 
participants will be carried out by the center in which the subject was recruited. 
The first evaluation (Screening or recruitment) of the study can be initiated in person or 
through telephone contact, email or electronic media.  If the eligibility criteria are met, 
the individual will be invited to attend the study center for the signing of the Informed 
Consent and collection of exams that will confirm his or her eligibility or not. 
Over four other face-to-face visits [M0 or inclusion, M1 (10 days after the intervention), 
M3 (90 days after the intervention) and M6 (180 days after the intervention)] the status 
of Coronavirus infection will be monitored by real-time PCR, in the case of symptomatic, 
and coronavirus serology every 3 months after vaccination with BCG or placebo (chart 
below). The tests will be performed with the purpose of having the diagnosis of the 
infection either through the real-time polymerase chain reaction (RT-PCR) technique 
and/or through the qualitative identification of IgM+IgA and/or IgG antibodies in the 
detection of infections that occurred asymptomatically and that may have been 
attenuated by the intervention with the BCG vaccine. For individuals receiving specific 
immunization against SARS CoV 2, serology will not be used for diagnosis of 
asymptomatic infection.   
At the time of recruitment hiv serology and BHCG dosage will also be collected for 
women with status in  order to meet the eligibility criteria. The time between the 
examinations collected in the recruitment and the effective inclusion in the study should 
be up to 10 calendar days. If the return occurs between the third and tenth day, and the 

Page 23 from 55 
 
participant presents symptoms compatible with COVID-19 upon return, the participant 
should recollect the RT-PCR.  
The gamma interferon release assay (IGRA) will preferably be performed at the inclusion 
visit or M0  for the purpose of identifying patients previously infected with Mycobacterium 
tuberculosis and detecting gamma interferon (IFN) levels, as well as its role in preventing 
viral infection, despite knowledge about alpha and beta INFs in immunopathogeny 
against the virus. In case of non-conformities that make the quality in the performance 
of this test, in view of being an examination that requires technical details crucial for its 
performance, the participant will be invited to perform a recollection.  In case of 
participants with previous IGRA reagent and who prove through the exam report will 
not be submitted to the new collection for this purpose. The other tests will be useful in 
evaluating the inflammatory response of individuals vaccinated and not vaccinated with 
BCG and those who became ill or not by COVID-19.  
If RT-PCR tests for nasoarcheal swab coronavirus, coronavirus serology and/or HIV 
serology performed during recruitment, generate inconclusive or indeterminate results, 
participants will be invited to a new collection and confirmation of the result. 
Participants will only be included with certain negative or non-reagent results for the 
three tests, with the exception of cases that have received vaccines against COVID-19, 
when serology will not be used as a determinant for inclusion or not. In case of 
participants who present reagent results in hiv serology, they will be referred to the 
Infectious Diseases service of the study centers for evaluation and follow-up. 
In addition to face-to-face visits, participants will receive a telephone contact or a text 
message by phone via SMS or receive email, the latter two with a link for the participant 
to access and answer the questionnaire about signs and symptoms of COVID as well as 
sending a photo with the aspect of the injection site. This will occur weekly until M3, 
and then monthly seeking information on fever and/or respiratory symptoms and the 
site of vaccine application. The participant will be asked to send a photo of the vaccine 
application location in each of these contacts. If the participant signals any respiratory 
manifestation or fever, they will be instructed to seek the Worker Health Divisions of 
their respective work units or the private or public health services, according to their 
availability, for the nasoarcheal swab to perform RT-PCR for coronavirus, as well as for 
medical care directed to the condition. In case of adverse events to the vaccine or 

Page 24 from 55 
 
placebo, patients will be evaluated in person at the study centers where they were 
recruited. 
If any participant presents the diagnosis of OVID-19 they will be invited to attend the 
study for a face-to-face visit once they are asymptomatic and 14 days after the onset of 
symptoms. In this face-to-face visit, the participant will answer questions from a 
questionnaire about signs of symptoms of the disease and will be asked to present 
laboratory or radiological tests performed during COVID-19 infection. 
Individuals who show interest in participating in the research should not have been 
previously vaccinated with any specific vaccine against COVID-19 or if vaccinated, only 
those immunized with both doses of vaccines, and the second dose should be applied 
within a minimum of 15 days. 
Participants already included and submitted to the intervention, must wait up to 15 days 
after the intervention with BCG/Placebo to receive specific vaccines against COVID-19 
already approved by ANVISA for use by the National Immunization Program, according 
to the guidelines of the Brazilian Ministry of Health.  
In the case of recruited individuals who are eligible for inclusion and intervention, but 
who give up receiving the investigational product of the study, they should withdraw 
consent to participate in the ProBCG study. 
Participants who present some test results collected during the study outside the normal 
range will be evaluated by the study physician. It should be indicated in the participant's 
folder that this result has been verified and whether or not there is clinical significance. 
If any, the participant should be referred for specialist evaluation at the hospital where 
his follow-up is being carried out. 
 
 
 
 
 
 
 
 
 

Page 25 from 55 
 
 
 
 
 
 
 
 
Visits 
Evaluations 
Exams 
Procedures 
Screening or 
Recruitment 
Face-to-face assessment 
of eligibility criteria, 
application of the TCLE 
 
NASOarchyeal swab RT-
PCR, anti-SARS-Cov-2 
antibody levels, BHCG 
dosage and HIV serology 
Invitation to participate in 
the study, application of the 
TCLE, collection of clinical 
samples (nasopharine swab 
and 
blood). 
Inclusion or M0 
 
0 day of preventive 
treatment (ToT) 
(BCG or placebo) 
Face-to-face assessment 
with confirmation of 
eligibility, clinical inclusion 
visit, collection of exams 
and dose of BCG or 
placebo 
IGRA; Complete blood 
count, lipidogram, 
albumin, HbGlic, LDH, PCR, 
ESR, D-D-Dmer; and 
biomarkers*. 
Nursing and medical 
consultation. Collection of 
clinical sample (blood). 
Application of preventive 
treatment. 
S1 - S12 
 
7-90 days of 
preventive TTO 
(±3 days) 
Telephone and/or SMS 
contact, and/or weekly 
email and sending photo 
of the injection site 
No exams 
Fever questionnaires and/or 
respiratory signs and 
symptoms and photo 
submission of the injection 
site 
M1 
 
10 days of preventive 
TTO 
(±5 days) 
Face-to-face visit to the 
laboratory for blood 
collection for biomarkers 
Anti-Anti-SARS-Cov-2 
antibody levels; Complete 
blood count, lipidogram, 
albumin, HbGlic, LDH, PCR, 
ESR, D-D-Dmer; and 
biomarkers*. 
Collection of clinical sample 
(blood). 
M3 
 
90 days of preventive 
TTO (±14 days) 
 
Face-to-face clinical visit 
for follow-up and blood 
collection for serology  
Anti-Anti-SARS-Cov-2 
antibody levels 
Nursing and/or medical 
consultation. Collection of 
clinical sample (blood). 
M4/M5 
 
120/150 days of 
preventive Tom (±14 
days) 
Telephone contact and/or 
SMS, and/or weekly email 
No exams 
Fever questionnaires and/or 
respiratory signs and 
symptoms 
M6 
 
180 days of 
preventive TTO (±14 
days) 
Face-to-face clinical visit 
to close the study and for 
blood collection for 
serology 
Anti-Antibody levels anti-
SARS-Cov-2. 
Nursing consultation. 
Collection of clinical sample 
(blood). 
 
 
 
Fever and/or 
respiratory signs and 
symptoms 
Face-to-face visit to the 
Worker's Health sector of 
its study center to collect 
exams 
RT-PCR  
Clinical sample collection 
(swab and blood). 
Face-to-face clinical visit 
to the study center since 
asymptomatic and 14 – 30 
days after the onset of 
symptoms 
Anti-Anti-SARS-Cov-2 
antibody levels; Complete 
blood count, lipidogram, 
albumin, HbGlic, LDH, PCR, 
VHS, D-Dallin and 
biomarkers*. 
Nursing or medical 
consultation. Copy of the 
tests performed during 
COVID-19 infection. 
Collection of clinical sample 
(blood). 

Page 26 from 55 
 
Subtitles: M0: Moment zero; S1-S12: Weeks 1 to 12; M1: Moment 1; M3: Month 3; M4: Month 4, M5: Month 5; M6: 
Month 6; *Clinical Analyses and Biomarkers: Will be performed only in hucff-idt ufrj, HUPE-PPC UERJ centers. 
Storage of specimens during study visits: 
The nasopharine swab will be obtained for the realization of the real time – polymerase 
chain reation (RT-PCR) for COVID-19. The collection will be performed using the 
technique described in the Manual of the Ministry of Health. The collection should be 
performed with the rubbing of the swab in the posterior region of the nasal caress trying 
to obtain some of the mucosal cells. The swab will be stored in 15ml tubes containing 
DMEM + Fetal Bovine Serum 10% + L-Glutamine Solution–Penicillin–Streptomycin. 
After collection, the specimen will be packed in a refrigerator of specimens from the 
research center with temperature control between 2 - 8 °C.  
Blood collection will comply with the guidelines of the Ministry of Health in the Manual 
Techniques for Blood Collection (2001) for the performance of blood count, lipidogram, 
albumin, LDH, ESR, C-reactive protein, D-dummer and dosage of biomarkers described 
in the study protocol. The collected material will be distributed in 11 blood tubes [1 SST 
tube, 2 EDTA tubes – purple cap (4.5ml/each), 1 serum tube - red cap (4 ml/each), 1 gel 
activator tube - yellow cap (4 ml/each), 2 tubes with soric heparin - green cap (8 
ml/each) and 1 Tempus tube – blue cap (3 ml/each). During the screening or recruitment 
visit, blood will be collected for BHCG dosage and HIV serology. In the inclusion visit the 
material will also be sent for IGRA [4 tubes Kit IGRA (1ml/each)]. 
After collection, the specimen will be packed in a refrigerator of specimens from the 
research center with temperature control between 2 - 8 °C. 
All specimens will be delivered to their destination laboratories, transported in thermal 
boxes maintaining the necessary refrigeration and obeying the criteria of storage and 
maintenance of the viability of the collected material.  
 
 
Procedure for applying bcg or placebo vaccine: 
BCG vaccine: 
The application of bcg vaccine as well as its storage and the materials used for 
application will follow the standards of the Manual of Standards and Procedures for 
Vaccination of the Ministry of Health (Brazil, 2014). As standardized in this Manual, the 

Page 27 from 55 
 
vaccine will be applied intradermally to the lower insertion of the deltoid muscle of the 
right arm, except contraindications or impossibilities. Other details are described in the 
standard operating manual (MOP) of the study. After the intervention, participants will 
be under observation for 20 minutes in the study center waiting room and will be 
released after the period as long as they do not present adverse events. 
 
 
Placebo: 
A 0.1 ml of 0.9% NaCl saline solution applied intradermally in the lower insertion of the 
deltoid muscle of the right arm will be used as placebo, except contraindications or 
impossibilities. Other details are described in the standard operating manual (MOP) of 
the study. After the intervention, participants will be under observation for 20 minutes 
in the study center waiting room and will be released after the period as long as they do 
not present adverse events. 
 
 
Blinding: 
The study participants as well as the researchers, coordinators, nurses, physicians and 
digitizers of the study will be blind to the allocation in the experimental or control 
groups. 
Only nursing technicians who will apply the vaccine or placebo will be aware of which 
group the patient will be allocated to. This professional will not follow up on the clinical 
follow-up or outcome of the study participant.  
If the participant has access to the COVID-19 vaccine before 15 days after vaccination 
with the study product, the "blinding" may be performed at the participant's written 
request, and if the product applied was placebo, the same may be vaccinated against 
COVID-19 before the 15 days established by the National Immunization Program.  Other 
details are described in the standard operating manual (MOP) of the study. 
 
 
Reaction to BCG vaccine and placebo: 

Page 28 from 55 
 
The BCG vaccine known to generate a local inflammatory response that lasts about 12 
weeks and will be monitored throughout the study. The reactions are described below 
and will not be considered as adverse events: 
- From the first to the second week: reddish macula lasting from 5 mm to 15 mm in 
diameter. 
- From the third to the fourth week: pustule that forms with the softening of the center 
of the lesion, followed by the appearance of crust. 
- From the fourth to the fifth week: ulcer with 4 mm to 10 mm in diameter. 
- From the 6th to the 12th week: scar with 4 mm to 7 mm in diameter, found in about 
95% of those vaccinated. 
Although the response to placebo did not cause any local vaccine reaction, which would 
question its use, we believe that if we did not use any intervention in the control group, 
bias could occur, in view of the possible reduction of precautionary measures and care 
in the use of personal protective equipment (PPE) in the experimental group, believing 
that they were "protected" by a vaccine,  although its effectiveness has not been proven 
in this situation. On the other hand, those not vaccinated would be more careful in 
relation to hygiene measures and use of PPE.  Using placebo, we believe that we match 
the usual prevention measures to COVID-19 in the study population at least in the first 
month of follow-up, a period prior to the onset of the lesions expected by BCG 
vaccination. 
 
 
Adverse events: 
The definition for an adverse event described in the Manual for Notification of Adverse 
Events and Safety Monitoring in Clinical Trials (Anvisa, 2016) will be used as a definition 
for an adverse event: "any adverse medical occurrence in a patient or clinical trial 
participant to whom a pharmaceutical product has been administered and who does 
not necessarily have a causal relationship to treatment. As a result, an As can be any 
unfavorable and unintentional sign, symptom, or disease (including results outside the 
reference range), associated with the use of a product under investigation, whether 
related to it or not. A serious AE shall be considered to have one resulting in any adverse 
experience with medicinal products, biological products or devices occurring at any dose 

Page 29 from 55 
 
and resulting in any of the following outcomes: a) death; b) threat to life; (c) persistent 
or significant disability/disability; d) requires hospitalization or prolongs hospitalization; 
e) congenital anomaly or birth defect; f) any suspected transmission of infectious agent 
by means of a drug or; g) clinically significant event." 
Adverse events that do not fall within the definition of severe AE (definition above) will 
be graduated according to ananvisa 2016 manual: 
• Light Level: Problem present less than 25% of the time, with an intensity that a 
person can tolerate and that rarely happened in the last 30 days; 
• Moderate Level: A problem that is present less than 50% of the time, with an 
intensity that interferes with the day-to-day of people and that has occurred 
occasionally in the last 30 days. 
• Sharp Level: Problem that is present in more than 50% of the time, with an intensity 
that partially alters the day-to-day of the person and that has happened frequently 
in the last 30 days. 
• Complete commitment: It means that a problem that is present in more than 95% of 
the time, with an intensity that completely alters the day-to-day of the person and 
that has occurred every day for the last 30 days. 
• Unspecified: Means that there is not enough information to specify the intensity. 
• Not applicable: It means that it is inappropriate to use a gradation (e.g. menstrual 
functions). 
 
All EAs will be notified to the local E.R. by the study coordinator at each centre using its 
own form (Mop Annex 3.3). The EAs should also be reported to the coordination of the 
proposing institution. The EA notification form as well as proof of sending the 
notification to the local ZIP Code must be sent by e-mail to the coordinator of the 
proposing institution no later than 7 days after notification. 
In the case of adverse events related to bcg vaccine (ulcer with diameter greater than 1 
cm, cold subcutaneous abscess, hot subcutaneous abscess, granuloma, regional 
lymphadenopathy not suppurated greater than 3 cm, suppurated regional 
lymphadenopathy, cheloid scar or lupoid reaction), the coordinator of the proposing 
institution will report to the National Immunization Program of the Ministry of Health. 

Page 30 from 55 
 
With regard to placebo, a minimum volume of saline solution applied intradermally will 
be used. The saline solution itself does not have any immunogenic component and does 
not cause any adverse events. It is the same solution used for intravenous or oral 
hydration and for dilution of medicinal products used intravenously. At intradermal 
application, mild pain may occur and after a few days local redness. Local signs of 
infection are not expected and the same vaccination standards and protocols 
established by the National Immunization Program will be used. 
 
Serious adverse events: 
Serious adverse events (EAG) should be reported immediately to the coordination of the 
Proposing Institution through telephone contact (described in mop). From this 
notification, the EAG must be reported to ANVISA within a maximum of 24 hours by the 
coordinator of the proposing institution through the agency's electronic portal 
(Notification Form of Serious Adverse Events in Clinical Trials available on the Electronic 
Portal of Anvisa > Medicamentos > Clinical Research > Adverse Events > Form for 
Notification of Serious Adverse Events in Clinical Trials - Notification EC). 
All serious adverse events suspected to have been caused by the proposed experimental 
treatment, including deaths will be reported to CONEP, through notification and within 
24 hours. 
Participants will receive all care and will be guaranteed assistance. 
Both the vaccine and placebo will be applied using disposable materials and patients will 
have possible adverse events recorded in their own form and notified if they occur 
(Annex 3.3 of the MOP). In addition, participants will be guaranteed assistance if they 
present any adverse event. 
 
Adverse events related to Coronavac vaccine®: 
Adverse events are classified according to frequency using the following convention: 
Very common: ≥ 10%; Common: ≥ 1% and <10%; Uncommon: ≥ 0.1% and <1%; Rare: ≥ 
0.01% and <0.1%; Very rare: < 0.01% including isolated reports; Unknown: (cannot be 
estimated from the available data). 

Page 31 from 55 
 
Phase I/II clinical studies were conducted in 1,165 adults and elderly aged 18 years or 
older, and a total of 924 individuals received two doses of basic immunization. 
Systematic safety follow-up observation was performed within 7 days after each 
vaccination dose. Adverse events were collected through notifications of participants 
and regular follow-up of investigators within 8-30 days after each dose. No grade III 
adverse reactions were found. 
The overall incidence of adverse reactions in subjects vaccinated at the target dose was 
23.73%, of which 23.24% were mild and 1.94% moderate. The preliminary safety 
analysis of the phase III clinical study conducted with 5,051 adult (18-59 years) and 
elderly (over 60 years) clinical study who received the complete regimen with 2 doses 
of the vaccine showed that the general frequency of occurrence of requested adverse 
reactions (local and systemic) up to 7 days after administration of the second dose was 
50.8% in the group of adults and 36.4% in the elderly group. For unsolicited adverse 
reactions (local and systemic) up to 7 days after the administration of the second dose, 
the overall frequency of occurrence was 9.2% in the adult group and 8.1% in the elderly 
group. The most common reaction observed after the second dose of the vaccine in both 
groups was pain at the site of administration, which occurred in 40.1% of adults and 
27.8% of the elderly. The vast majority of adverse reactions observed were Grade 1/2 
and no serious adverse reaction occurred. 
- Adverse reactions observed from phase I/II clinical studies in Adults (18-59 years) and 
the elderly (over 60 years): 
- Very common reaction (> 1/10): Location: pain 
- Common reaction (> 1/100 and ≤ 1/10): Systemic: fatigue, fever, myalgia, diarrhoea, 
nausea, headache 
- Unusual reaction (> 1/1000 and ≤ 1/100): Systemic: vomiting, lower abdominal pain, 
bloating, dizziness, cough, loss of appetite, hypersensitivity, high blood pressure; and 
Site: abnormal staining at the site of administration, swelling, itching, erythema, local 
hypoesthesia, hardening. 

Page 32 from 55 
 
Adverse reactions observed from the phase III clinical study in Adults (18-59 years) up 
to 7 days after administration of the second dose of the vaccine: 
- Very common reaction (> 1/10): Systemic: headache, fatigue; and Location: pain 
- Common reaction (> 1/100 and ≤ 1/10): Systemic: nausea, diarrhea, myalgia, chills, loss 
of appetite, cough, arthralgia, itching, rinorrhoea, nasal congestion; and Location: 
erythema, swelling, duration, itching 
- Unusual reaction (> 1/1000 and ≤ 1/100): Systemic: vomiting, fever, rash, allergic 
reaction, orohargeal pain, odynophagia, sneezing, astherrries, asthesis, dizziness, 
abdominal pain, drowsiness, malaise, flushing, pain in the extremities, upper abdominal 
pain, back pain, vertigo, dyspnea, edema; and Location: hematoma 
Adverse reactions observed from the phase III clinical study in the Elderly (over 60 years) 
up to 7 days after the administration of the second dose of the vaccine: 
- Very common reaction (> 1/10): Location: pain 
- Common reaction (> 1/100 and ≤ 1/10): Systemic: nausea, diarrhea, headache, fatigue, 
myalgia, cough, arthralgia, itching, rinorrhoea, odinofagia, nasal congestion; and 
Location: itching, erythema, local edema, duration 
- Unusual reaction (> 1/1000 and ≤ 1/100): Systemic: vomiting, chills, decreased 
appetite, allergic reaction, asthena, dizziness, ecchymosis, hypothermia, discomfort in 
the limbs; and Location: hematoma 
In addition, there are theoretical risks that vaccinated individuals could develop a more 
serious disease, but so far there is no report that this has happened to the virus that 
causes COVID-19. This vaccine was previously tested in animals and they did not present 
this form of infection more severe. 
 
Adverse events related to Covishield Vaccine®: 

Page 33 from 55 
 
The overall safety of the Covishield® vaccine (recombinant) is based on the interim 
analysis of grouped data from four clinical studies conducted in the UK, Brazil and South 
Africa. At the time of analysis, 23,745 ≥ 18 years had been randomized and received the 
vaccine or control. Of these, 12,021 received at least one dose of the recombinant 
vaccine. Demographic characteristics were generally similar among individuals who 
received the vaccine and those who received control. Overall, among the individuals 
who received the recombinant vaccine, 90.3% were between 18 and 64 years old and 
9.7% were older than 65 years or more. Most individuals were white (75.5%), 10.1% 
were black and 3.5% Asian; 55.8% were women and 44.2% were men. The most 
frequently reported adverse reactions were injection site sensitivity (> 60%); injection 
site pain, headache, fatigue (> 50%); myalgia, malaise (> 40%); pyrexia, chills (> 30%); 
and arthralgia, nausea (> 20%). Most adverse reactions were mild to moderate in 
intensity and usually resolved within a few days after vaccination. Compared to the first 
dose, adverse reactions reported after the second dose were lighter and less frequently 
reported. 
Adverse reactions were generally lighter and less frequently reported in the elderly (≥ 
65 years of age). Analgesic and/or antipyretic medications (e.g., paracetamol-containing 
products) may be used to provide relief from adverse reactions after vaccination. 
Adverse drug reactions (ADRs) are organized by MedDRA Organ System Class (SOC). 
Within each SOC, the preferred terms are organized by decreasing frequency and then 
by decreasing severity. The frequencies of occurrence of adverse reactions are defined 
as: very common (≥ 1/10); (≥ 1/100 to < 1/10); unusual (≥ 1/1,000 to < 1/100); rare (≥ 
1/10,000 to < 1/1,000); (1< 10,000) and unknown (cannot be estimated with available 
data). 
 
Risks: 
The safety of BCG revaccination among health professionals will be evaluated in order 
to describe possible injection site reactions and other adverse events. 
In addition, we will minimize the occurrence of AS, excluding immunosuppressed 
individuals or using concomitant medications that impact immune responses. 

Page 34 from 55 
 
Local and systemic AEs will be evaluated at relevant times through medical visits, for 
example in the first 20 minutes after BCG injections and in later visits, either in person 
or by telephone contact. The EAs will be noted and graduated according to the ANVISA 
Manual. 
Adverse events to the BCG vaccine, more frequent have been abscess and 
lymphadenopathy, with higher reports for men than for women. The rates of As per 
10,000 distributed doses were from 11.6 to 15.4, respectively. Unsuppurated regional 
lymphadenopathy greater than 3 cm, suppurated regional lymphadenopathy, cheloid 
scar, or lupoid reaction may occur. The EAs will be monitored and handled throughout 
the follow-up visits. 
The research centers will also be open and available for extra views in case of adverse 
events. The manifestations will be recorded in the data collection instruments and will 
be treated and monitored in hospitals linked to the research centers. Similarly, the 
events will be notified to the National Immunization Program of the Ministry of Health, 
informing the batch of the vaccine applied as well as the event that occurred. 
Participants will receive all care and will be guaranteed assistance. 
Benefits: 
The results of this study may help define a form of prevention of COVID-19 for health 
professionals and in the future in the population as a whole. 
 
 
 
 
 
 
 
 

Page 35 from 55 
 
 
 
 
 
 
 
 
 
 
 
 
LABORATORY PROCEDURES: 
 
Blood collection: 
Conventional examinations will be called those performed by the Clinical Pathology 
laboratories contracted for the purpose of the study. 
These are: blood count, hemosedimentation velocity (ESR), lactate dehydrogenase 
(LDH), total cholesterol and fractions and triglycerides, D-dummer, C-reactive protein 
(CRP), albumin, ferritin, iron, glycated hemoglobin. For this purpose, 1 EDTA tube – 
purple cap and 1 serum tube (red cap) should be delivered for laboratory analysis. 
 
Polymerase chain reaction with real-time reverse transcription: 
The presence of SARS-CoV-2 in respiratory samples will be detected by real-time 
amplification by RT-PCR from the open reading chart SARS-CoV-2 1ab (ORF1ab), 
nucleocapsid protein (NP) gene fragments using kits provided by IDT USA. The conditions 

Page 36 from 55 
 
for amplifications will be 50 °C for 15 minutes, 95 ° C for 3 minutes, followed by 45 cycles 
of 95 ° C for 15 if 60 ° C for 30 s. When two targets (ORF1ab, NP) test positive by specific 
real-time RT-PCR, the case will be considered laboratory confirmed. A cycle threshold 
value (Ct value) less than 37 will be defined as a positive test and a Ct value of 40 or 
more has been defined as a negative test. An average load, defined as a Ct value of 37 
to less than 40, will require confirmation per new test. 
 
Serological test for COVID-19 
The Vircell serological test will be® used to define the inclusion or not of health 
professionals in the study. Vircell reveals® the presence of IgG and IgM/IgA against viral 
antigens as well as their titration, facilitating the monitoring of absolute antibody values. 
 
 
 
Immunoassays 
Seros will be obtained from the health professionals enrolled in the study. Two cohorts 
will be considered: health professionals vaccinated with BCG (n=376) and health 
professionals not vaccinated with BCG (n=376). 
The biomarkers evaluated will be cit-H3, alpha 1 AT, D-dummer, LDH and PCR. The 
analyzed data will be related to the blood count information, according to the scheme: 
M0, M1, M3 and M12. 
 
Inflammatory profile and population characterization of neutrophils and T 
lymphocytes 
A panel of surface and intracellular markers more related to exacerbated inflammation 
will be used, aiming to study the profile of T lymphocytes, monocytes and neutrophils.  
Cell profiles will be compared to the occurrence or not of SARS-CoV-2 infection in both 
cohorts.  

Page 37 from 55 
 
These analyses will be performed at the defined times of collection and evaluation of 
the study groups.  The markers to be studied in cell populations will be:  
Lymphocyte T: CD3, CD4, CD119, IL-12Rβ2, IFN-γ, TNF-α, IL-4, IL-10, IL-13, IL-21, IL-1RI, 
IL-6Rα, IL-17A, IL-17F, IL-22, FoxP3, CD25, Galectin-3, IL-35, TGF-β.  
Neutrophil: CCL22, CD169, CD68, ZAP70, CD11b, CD11c, LDH, CD64, IL-17.  
Monocytes: CD14, CD11c, HLA-DR, CD45, IP-10, MIP-1α, MIF.  
Inflammatory and metabolic profiles will be measured by luminex, using serum from 
patients for the panel: IFN-α2, IFN-γ, IL-10, -12 p40, -12 p70, -13, -15, -1 Ra, -1α, -1β, -2, 
-4, -6, -8, -23, -17, -18, -22, MIP-1 α, MIP-1β, TNF-α, VEGF-A, Adiponectin, Hepicidine, 
Insulin, MMP-1, -2, -8, -9, MIF, Glucagon, Greeline, Leptin, CD163s,  Galectin-1 and -3, 
Vitamin D protein, PCR, albumin, citrunilada histona H3, alpha1-anti-trypsin, P-selectin, 
D-dummer, PSGL-1, tPA, CD40L, PAI-1, Tissue Factor IX, CXCL8 (IL-8). 
 
 
Transcriptoma 
RNA obtained from peripheral blood cells will be used to evaluate the signaling 
pathways of monocytes and neutrophils in both study groups. RNA sequencing 
(RNASeq) will be used to evaluate the profiles, according to the manufacturer's 
instruction (Illumina). 
 
Laboratory procedures to be performed at each research center: 
Laboratory procedures will be different according to the research center where the 
participant is included, as follows: 
 

Page 38 from 55 
 
 
 
 
 
 
 
 
 
 
 
DATA COLLECTION AND MONITORING PLAN: 
The researchers involved in this protocol will use the forms as source documents (DF). 
The centers will record the data in the FDs when the data is generated (example: during 
the participant's visit or during telephone contact). The team will use the REDCap 
(Electronic Survey Data Capture) system for electronic data capture and data 
management and its templates will be called Case Report Forms (CRF).  A copy of the 
REDCap database will reside on a secure FMRP-USP server that has all physical, 
technical, and administrative controls. A second copy will be stored on the REDCap 
server for TB network data. We will use the group of data management researchers from 
the Faculty of Medicine of Ribeirão Preto-USP. Any professional of the Team of Brazil 
who is delegated to access the data system, either to access or to do quality control of 
the data, will have individual access with basic security requirements. The data will be 

Page 39 from 55 
 
retrieved from the centers so that no center has access to data from other centers. 
However, the study's lead investigator will have access to data from all centers. The 
team responsible for REDCap will manage access to the REDCap system, creating, 
suspending and exhaling user logins as needed. The centers of Brazil will be responsible 
for informing the team responsible for data management in advance when a 
professional is disconnected from the protocol, and then their respective access to the 
system will be canceled. The list of users who will have access to the protocol REDCap 
will be reviewed monthly.  
As recently, the release of the REDCap software license (https://projectredcap.org/) on 
behalf of REDE-TB was obtained, it will become part of the REDCap Consortium, a 
network of all partners that make an instance of the software available. The software 
was installed in the cloud computing environment of the University of São Paulo. 
REDCap supports multiple research projects in parallel. Therefore, once installed and 
configured, it can be used repeatedly for an indefinite period. We will use an intuitive 
and user-friendly KoBoToolbox (https://www.kobotoolbox.org/) interface as it also 
allows data collection online and offline, through computers and mobile devices. It can 
be used together with REDCap. The cloud computing environment of the University of 
São Paulo, called interNuvem (https://internuvem.usp.br), will be used to create virtual 
machines to instantiate the technological resources needed to execute the project, such 
as database servers, REDCap, Web servers, among others. As a way to manage the 
features of the REDCap and KoboToolbox software suite, the data collection forms, 
initially defined on paper, will be computerized. Offline data collection can be done 
through the mobile app (Android, iOS). In this case, the data must be transmitted to the 
server when internet connectivity is established. The application will be made in the 
Elixir programming language, which was created on top of erlang, and so it is a functional 
language. Within the Project there are three main modules are them: Redcap; XLSX 
Redcap Form Decode and Redcap.Encoder. 
The first module is responsible for reading the files used in the translation of the forms 
provided by KoboToolbox, in XLS format, to Redcap. The main operations involved in 
this process are migration of the form structure and the collected data. During the first 
operation, a function is responsible for reading the xls file and uses the XLSX Redcap 
Form Decode module, to transform each line of the file into an Elixir structure. After 

Page 40 from 55 
 
having each row schemated in these structures, it's time to call Redcap.Encoder that will 
transform this structure received from the previous module and transform into a new 
data structure by translating each of the rows into its data dictionary equivalents. When 
all lines are ready the main module uses this structure and "writes" in a data dictionary 
for Redcap. 
In the second operation everything happens in the same Redcap module. In it the data 
files are read and with the help of a file that I called "data_guide" it reorganizes the data 
the way Redcap asks for import. It also replaces the headers that are named after the 
xlsform data for the names that were registered in Redcap. This is done through 
"data_guide", which is a file that has on one side the column name in xlsform and on the 
other the name of that information on the Redcap platform. In addition to replacing 
headers and reorganizing columns, the data also goes through a normalization to be in 
accordance with what Redcap accepts. For example all "True" and "False" that are put 
in Ona for multiple selections have to be replaced by 1 and 0 respectively, otherwise 
Redcap does not accept. After these steps the final structure is written in csv for import. 
Specific protocols for electronic quality monitoring and tracking will be performed by a 
data manager routinely, in order to detect fill errors, inconsistent or missing data, thus 
ensuring quality control. 
An independent study monitor, appointed by the lead researcher, will be responsible 
for monitoring data quality by standard test operating procedures. Based on the 
monitoring plan, the field visit and audit will be carried out at different stages. All 
records of participants, DFs and other source documents of the patients recruited in this 
study will be made available for analysis by the monitors. A meeting of researchers from 
each site will take place monthly, via the web-based remote conferencing system, to 
share the progress of the study and discuss c problems encountered while conducting 
the test. During the conduct of the research, any critical modifications in the protocol 
will be informed to the principal investigator, to the study record and, if relevant, to the 
study participants. 
 
Sample storage plan: 

Page 41 from 55 
 
Nasophayngeal swab and peripheral blood specimens collected will be transported to 
specific laboratories. Specimens may be stored to facilitate the repetition of the test in 
case of indeterminate results or discordant cases in molecular laboratory tests or not 
for SARS-CoV-2. Stored clinical specimens will be labeled with a study identifier and not 
with personal identifiers and will be used only for the purposes of this study. 
According to CNS Resolution No. 441 of 2011 (Art. 1) and Ordinance MS No. 2,201 of 
2011 (Art. 3), any organized collection of human biological material collected for 
scientific research purposes, including samples intended for routine examinations in a 
clinical trial (e.g., blood count and renal function) are considered as constituents of a 
biorepository or biobank. The biological material will be used as provided for in the 
research protocol, with no further future analyses different from those foreseen. If there 
is a proposal for future use of the stored samples, approval in the CEP/CONEP system 
will be required and a new protocol and TCLE will be submitted so that each participant 
can choose whether or not to allow the use of the samples for future studies. 
 
 
 
Statistical Analysis Plan: 
The primary efficacy analysis will compare the cumulative incidence of SARS-CoV-2 
infection and the cumulative incidence of severe forms of COVID-19. 
These analyses will occur when the last enrolled research subject has completed 30 days 
and 6 months of preventive treatment and will be performed using the results of COVID-
19 diagnostic tests, as well as the data available in participating hospitals and national 
electronic notification systems of COVID-19. A modified treatment intention analysis 
(MITT) and protocol (PP) will be performed. The safety and tolerability analysis will 
consist of all patients who have been randomized and received a dose of the vaccine or 
placebo. Based on serum immune response concentrations, we will calculate certain 
immunoinflammatory variables related to the use of BCG versus placebo among 
subjects infected and not infected by COVID-19 among the categories with COVID-19 
vaccine (Coronavac® or Covishield®) and without covid-19 vaccine.  
The intervention and control groups will be compared at the time of inclusion in the 
study in relation to sociodemographic, clinical, radiological and biochemical imaging 
variables, in order to evaluate the adequacy of the randomization process. Multivariate 

Page 42 from 55 
 
analyses using logistic regression or Poisson models with robust variance will be used to 
evaluate the effect of BCG vaccine on primary and secondary outcomes. Associations 
will be expressed in the form of risk ratios and respective 95% confidence intervals. Data 
imputation may be required and the random forest system will be used. Statistical 
software R, version 10.0. 
 
Sample size: 
It is estimated that it will take 376 individuals in each of the comparison groups to ensure 
a statistical power of 80% and a type I error of 5%, considering an incidence of SARS-
CoV-2 infection of 20% in BCG vaccinated, 30% in unvaccinated, 15% in covid-19 
vaccinated. It is expected a reduction in the joint effect of BCG with the COVID-19 
vaccine between 10% and 15% and that the percentage of people vaccinated with BCG 
in the cohort is 50% and the percentage of people vaccinated for Covid-19 in the cohort 
is 80%.  
We will also use for the two outcomes (below) the sequential design the groups are 
followed up until there is a group with clear benefits or that both groups do not present 
difference. This design is used when the results need to be known very quickly. If there 
is a large difference between the treatments, then this study will be performed in a 
shorter time when compared to the parallel trial. The main ethical advantage is that if 
one treatment shows a superiority over the other, the study can already be stopped. 
However, the interim analyses will be previously planned, in such a way that the 
calculation of sample size allows the multiplicity of the tests. The meaning of p-values in 
sequential analyses also changes, because when using sequential analyses, more than 
one analysis is performed, and the typical definition of a p-value as data "at least as 
extreme" as observed needs to be reset. One solution is to sort the p-values of a series 
of sequential tests based on the downtime and at what time the test statistic was in a 
certain appearance, known as ordering. 
 
 
 
 
 
 

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OUTCOMES: 
Primary Outcome 
1. Compare the cumulative incidence of SARS-CoV-2 infection 
It is estimated that it will take 376 individuals in each of the comparison groups to ensure 
a statistical power of 80% for the detection of a difference in effects of 10%, considering 
that the control group will have an accumulated incidence of SARS-CoV-2 infection of 
30% after 6 months of follow-up, for a type I error of 5%. 
 
2. Compare the cumulative incidence of severe forms of COVID-19 
We will also use for the two outcomes the sequential design the groups are followed up 
until there is a group with clear benefits or that both groups do not present difference. 
This design is used when the results need to be known very quickly. If there is a large 
difference between the treatments, then this study will be performed in a shorter time 
when compared to the parallel trial. The main ethical advantage is that if one treatment 
shows a superiority over the other, the study can already be stopped. However, the 

Page 44 from 55 
 
interim analyses will be previously planned, in such a way that the calculation of sample 
size allows the multiplicity of the tests. The meaning of p-values in sequential analyses 
also changes, because when using sequential analyses, more than one analysis is 
performed, and the typical definition of a p-value as data "at least as extreme" as 
observed needs to be reset. One solution is to sort the p-values of a series of sequential 
tests based on the downtime and at what time the test statistic was in a certain 
appearance, known as ordering. 
3. Assess the BCG vaccine-mediated immune response in healthcare professionals 
SARS-CoV2 infection is estimated to occur in approximately 20% of health professionals 
vaccinated by BCG (experimental group), in 30% of health professionals not vaccinated 
by BCG (placebo group) and not vaccinated for COVID-19 and in 15% in COVID-19 
vaccinated without BCG vaccine. A reduction of 10% to 15% of the joint effect of bcg 
vaccine with Covid-19 vaccine is expected.  
In this scenario, with the bearing of 376 health professionals in each arm, if we assume 
that the percentage of covid-19 vaccinated is 80% and equal in each arm, SARS-CoV2 
infection is expected to be detected in 15 professionals vaccinated only with BCG, in 23 
professionals not vaccinated with BCG and COVID-19 vaccine,  45 professionals 
vaccinated for COVID-19 without bcg vaccine and 38 professionals vaccinated for COVID-
19 and BCG. For each subgroup (vaccinated and not vaccinated with BCG) it will be 
possible to identify the predictive biomarkers of infection by comparing infected 
professionals with those who were not infected with SARS-CoV2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Page 45 from 55 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
INTERIM ANALYSIS PLAN: 
Regarding the effect of BCG vaccine administration on the cumulative incidence of SARS-
CoV-2 infection or on severe forms of COVID-19, a single intermediate analysis will be 
performed when the number of volunteers included has reached half of the total sample 
scheduled to verify whether the evidence would already be conclusive and would allow 
the study to be interrupted. For type I error protection, intermediate analysis will use a 
Z value of 2.78 (Chow & Liu, 2004). 
The analyses that will be performed in the intermediate analysis will be restricted to the 
following outcomes: accumulated incidence of SARS-CoV-2 infection, cumulative 
incidence of severe forms of COVID-19 and proportion of serious adverse events. 
The following rules will be used to indicate early completion of the study after 
intermediate analysis: 
• Significant difference in the effect of BCG judged by comparing the proportions of 
patients who develop SARS-CoV-2 infection or severe forms of COVID-19 in each arm of 
the study; 

Page 46 from 55 
 
• If the effect of BCG is similar between groups and there is a significant difference in 
the cumulative proportion of serious adverse events; 
 The results will be evaluated by the Data and Safety Monitoring Committee, which will 
express the need to continue the study, based on the vaccine's performance in terms of 
the incidences of SARS-CoV-2 infection or severe forms of COVID-19 and adverse events. 
 
Independent Data Monitoring and Security Committee: 
The Data and Safety Monitoring Committee (CMDS) will be composed of three members 
not linked to participating institutions, in accordance with the Operational Guidelines 
for the Establishment and Functioning of Data Monitoring and Safety Committees of the 
Ministry of Health (2008). 
This committee will be responsible for reviewing the protocol and progress of the study 
in all centers, with real-time meetings by videoconference, as detailed below, and will 
consist of: 
 
Denise Rossato - Physician - Professor, Faculty of Medicine, Federal University of Rio 
Grande do Sul 
Ronir Raggio - Statistician - Institute of Collective Health Studies of the Federal University 
of Rio de Janeiro  
Silvana Spíndola - Physician - Professor, Faculty of Medicine, Federal University of Minas 
Gerais 
 
 
An early safety assessment will be carried out when the first 100 participants 
(approximately 50 per arm) are included. Security data will be reviewed by a CMDS 
member or someone designated by a CMDS member. In addition, the Committee will 
review the progress of the study after the inclusion of every 100 additional participants 
through real-time meetings by videoconference. 
 
CMDS activity plan: 
Time of study 
Activity 
Form 

Page 47 from 55 
 
Before the start of the study Protocol and MOP review 
Real-time 
video 
conferencing 
scheduling 
During the study 
Early safety assessment after 
inclusion of the first 100 
participants 
Real-time 
video 
conferencing 
scheduling 
Evaluation 
of 
study 
progression 
per 
100 
participants included 
Real-time 
video 
conferencing 
scheduling 
Adverse event analysis 
Real-time 
video 
conferencing 
scheduling 
every 
2 
weeks 
Analysis of serious adverse 
events 
Extraordinary meeting 
via 
real-time 
videoconferencing  
Intermediate analysis after 
the 
inclusion 
of 
500 
participants 
Real-time 
video 
conferencing 
scheduling  
At the close of the study 
Evaluation of the conduct of 
the study and its results 
Real-time 
video 
conferencing 
scheduling 
 
 
 
Plan for monitoring and analysis of adverse events: 
The definition for an adverse event described in the Manual for Notification of Adverse 
Events and Safety Monitoring in Clinical Trials (Anvisa, 2016) will be used as a definition 
for an adverse event: "any adverse medical occurrence in a patient or clinical trial 
participant to whom a pharmaceutical product has been administered and who does 
not necessarily have a causal relationship to treatment. As a result, an As can be any 
unfavorable and unintentional sign, symptom, or disease (including results outside the 
reference range), associated with the use of a product under investigation, whether 
related to it or not. A serious AE shall be considered to have one resulting in any adverse 
experience with medicinal products, biological products or devices occurring at any dose 
and resulting in any of the following outcomes: a) death; b) threat to life; (c) persistent 
or significant disability/disability; d) requires hospitalization or prolongs hospitalization; 
e) congenital anomaly or birth defect; f) any suspected transmission of infectious agent 
by means of a drug or; g) clinically significant event." 

Page 48 from 55 
 
All EAs will be notified to the local EC by the study coordinator at each centre using their 
own form. EAs shall also be reported to the coordination of the proposing institution no 
later than 7 days after notification of the EA.  
In the case of adverse events related to bcg vaccine (ulcer with diameter greater than 1 
cm, cold subcutaneous abscess, hot subcutaneous abscess, granuloma, regional 
lymphadenopathy not suppurated greater than 3 cm, suppurated regional 
lymphadenopathy, cheloid scar or lupoid reaction), the coordination of the proposing 
institution should be notified to the National Immunization Program of the Ministry of 
Health. 
Serious adverse events (EAG) should be reported immediately to the coordination of the 
Proposing Institution through telephone contact. From this notification, the EAG must 
be reported to ANVISA within a maximum of 24 hours by the coordinator of the 
proposing institution through the agency's electronic portal (Notification Form of 
Serious Adverse Events in Clinical Trials available on the Electronic Portal of Anvisa > 
Medicamentos > Clinical Research > Adverse Events > Form for Notification of Serious 
Adverse Events in Clinical Trials - Notification EC). 
All serious adverse events suspected to have been caused by the proposed experimental 
treatment, including deaths will be reported to CONEP, through notification and within 
24 hours. 
There will be a review of The EA data every 2 weeks. If adverse events are unexpected 
or not previously described, case review will be requested through a meeting of the data 
monitoring and security committee, which should take place in real time by 
videoconference. Serious adverse events will be immediately flagged to the CMDS for 
extraordinary analysis also by real-time videoconferencing. 
 
 
Criteria for discontinuation of the study for safety reasons: 
No formal rule will be adopted to interrupt the study for safety reasons. The CMDS 
may recommend stopping the study, based on the review of interval safety results. The 
study can only be canceled or discontinued after analysis of the reasons for 
discontinuity by the institution's ZIP Code or CONEP.  

Page 49 from 55 
 
 
Criteria for discontinuation of experimental treatment in the participants: 
No formal rule will be adopted to discontinuation of experimental treatment in the study 
participants. The CMDS may recommend this study interruption, based on the review of 
interval results as described above. Nevertheless, this decision can only be effected after 
evaluation and approval by the institution's ZIP Code or CONEP.  
 
 
 
 
 
 
 
QUALITY MANAGEMENT: 
All studies will be conducted in accordance with the ICH-GCP Guidelines (ICH-GCP E6 
R2), as far as possible in the research environment, as well as all national legal and 
regulatory requirements (as applicable). 
 
 
 
 
 
 
 
 
 

Page 50 from 55 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
BUDGET: 
 
Cost 
Value 
Legal entity 
R$84,000.00 
Bags 
R$160,800.00 
Inputs 
R$297,188.57 
Other 
R$43,571.43 
Capital 
R$14.440,00 
Total 
R$600,000.00 
 
 
 
 
 
 

Page 51 from 55 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
SCHEDULE: 
The schedule presented below is a schedule, and the study should be started only after 
approval of the CEP. 
 
Phase 
Beginning 
End 
Submission of amendment to CEP/CONEP 
01/08/2021 
31/08/2021 
Recruitment/Inclusion/Intervention 
30/10/2020 
31/12/2021 
Follow-up of patients 
30/10/2020 
31/06/2022 
Data analysis 
31/06/2022 
31/09/2022 
Reporting/Results 
31/09/2022 
31/12/2022 
 
 
 
 

Page 52 from 55 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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