Introduction: The purpose of this manuscript is to provide a narrative review of the literature and the basis of ozone therapy to treat viral illnesses including COVID-19 therapy.

Methods: We performed a narrative review of 239 relevant publications and present new data not previously published from our group.

Result: Ozone, a tri-atomic oxygen molecule is a natural substance made by the human white blood cells and metabolized into hydrogen peroxide and many lipo-peroxidases. Ozone is one of the most important modulators of the human immune system. Many investigators purport multiple potential mechanisms by which ozone treats a variety of viral and other illnesses at the atomic and cellular levels. While these mechanisms are operative, they represent passive events resulting from the ozone’s impartation of resonant energy to the human energetics’ fields. We present data demonstrating that the basis of most all human disease results from specific organ energetic deficiencies. Ozone may be one of the most potent preconditioning agents yet discovered in the human body. We discuss ozone dynamics, the vascular/blood connection, ozone preconditioning, and ozone therapy for specific viral diseases. Our review of the literature uncovered a spate of case reports describing beneficial outcomes with ozone treatment in diverse viral syndromes including Human Immunodeficiency Virus (HIV), Human Papillomavirus (HPV), hepatitis C, herpes zoster, ebola, and SARS-CoV-2. Additionally, we found in vitro studies describing inactivation of herpes and HIV by ozone treatment. We reviewed our successful use of ozone as an adjuvant and/or primary agent for the successful treatment of COVID-19. We report two cases of very successful use of ozone therapy in the maternal-fetal dyads in the severely infected/affected fetuses with CMV; this has never been reported in the medical literature.

Conclusion: Ozone is an effective primary or adjuvant therapy for COVID-19 and for many other viral illnesses. Most all disease processes represent an energy deficient state and we have shown that the primary mechanism of ozone is to impart and restore energy deficiencies.

Influenza viruses and coronaviruses cause several human diseases, such as bronchitis, bronchiolitis, and pneumonia, and exacerbate bronchial asthma, chronic obstructive pulmonary disease and pulmonary fibrosis. Human airway epithelial cells infected with these viruses release progeny viruses and inflammatory cytokines, such as Interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor-α, partly through the activation of nuclear factor kappa B. Modulation of airway damage and inflammation may modulate viral infection-induced airway and lung diseases. Human tracheal and nasal epithelial cells express proteases, including Transmembrane Protease Serine S1 Member 2 (TMPRSS2), and the proteases activate influenza viruses and coronaviruses and the subsequent replication processes of these viruses. The protease inhibitors camostat and nafamostat reduced influenza virus and coronavirus replication and the amounts of cytokines released from human airway epithelial cells. Nafamostat also reduced the release of influenza virus in the lungs of mice. The development of clinically available protease inhibitors is required to treat patients infected with influenza virus or coronavirus.

The Omicron variant of SARS-COV-2 (GISAID GRA clade (B.1.1.529, BA.1 and BA.2)) is now the single dominant Variant Of Concern (VOC). The high number of mutations in the Omicron Spike (S) protein promotes humoral immunological escape. Although a third homologous boost with S, derived from the ancestral strain, was able to increase neutralizing antibody titers and breadth including to Omicron, the magnitude of virus neutralization could benefit from further optimization. Moreover, combining SARS-COV-2 strains as additional valences may address the current antigenicity range occupied by VOCs.

Using Trimer-TagTM platform we have previously demonstrated phase 3 efficacy and safety of a prototypic vaccine SCB-2019 in the SPECTRA trial and have submitted applications for licensure. Here, we successfully generated a bivalent vaccine candidate including both Ancestor and Omicron variant S-proteins. Preclinical studies demonstrate this SARS-CoV-2 bivalent S-Trimer™ subunit vaccine elicits high titers of neutralizing antibodies against all VOCs, with markedly enhanced Omicron specific neutralizing antibody responses.

The Omicron variant of Severe Acute Respiratory Syndrome Corona Virus-2 (SARS-CoV-2) is threatening our global efforts fighting the COVID-19 pandemic. As the vast majority of current countermeasures against SARS-CoV-2 are completely or markedly losing their effectiveness, including most of the clinically approved neutralizing antibodies, better and more efficacious novel agents are urgently needed. We have developed a bispecific antibody, BAT2022, which bonds simultaneously with high affinity to two non-overlapping epitopes on the Receptor-Binding Domain (RBD), competitively blocks the binding of RBD to human Angiotensin-Converting Enzyme2 (ACE2) and potently neutralizes SARS-CoV-2 and all of the variants tested. The IC50 values in the pseudovirus assay for Omicron (BA.1) and Delta are 30 pM and 50 pM, respectively. A mouse model of SARS-CoV-2, BAT2022 showed strong prophylactic and therapeutic effects. Prophylactically, a single administration of BAT2022 completely protected mice from bodyweight loss, as compared with up to 20% loss of body weight in placebo- treated mice, reduced the lung viral titers to undetectable in all mice treated with BAT2022 either prophylactically or therapeutically, as compared with around 1 × 10⁵ pfu/g lung tissue in placebo-treated mice. Overall, Bispecific Antibody BAT2022 showed potent binding and neutralizing activity across a variety of SARS- CoV-2 variants and could be an attractive weapon to combat the ongoing waves of the Omicron pandemic.

Detecting and diagnosing small changes occurred by gene alteration via PCR is globally considered as an effective and precise method for handling various diseases. From the internal factors which change the normal body to develop disorders such as cancer to mutated external sources of infection that alters the intact human body, mainly bacteria and viruses. Among these, SARS-CoV-2 has emerged to be one of the most concerning threats to public health due to its rapid mutation. From the initial outbreak in 2019, research groups continuously identified various subtypes, some of which branched out as a novel variant of the original form that caused global health crisis among the population. While the standard PCR method of real-time quantitative reverse transcription is considered effective in detection and prevention, as more SARS-CoV-2 variants emerged, the overall process of identifying each variant one by one has deteriorated the standard procedure to be tiresome and time-consuming. To overcome the inefficiency, a novel PCR method using oligonucleotides (STexS) was introduced to significantly increase the specificity in detecting single nucleotide polymorphisms. Furthermore, the implementation of simultaneous multi-target PCR successfully isolated both Delta and Omicron variants within a single PCR trial, condensing the excessively long PCR procedure to be overall efficient and precise at the same time. The improvements will provide crucial insight in SARS-CoV-2 detection and diagnosis.

Importance: The SARS-CoV-2 non-spike structural proteins of Nucleocapsid (N), Membrane (M) and Envelope (E) are critical in the host cell interferon response and memory T-cell immunity and have been grossly overlooked in the development of COVID vaccines.

Objective: To determine the safety and immunogenicity of UB-612, a multitope vaccine containing S1-RBD-sFc protein and rationally-designed promiscuous peptides representing sequence-conserved Th and CTL epitopes on the Sarbecovirus Nucleocapsid (N), Membrane (M) and Spike (S2) proteins.

Design, setting and participants: UB-612 booster vaccination was conducted in Taiwan. A UB-612 booster dose was administered 6-8 months post-2nd dose in 1,478 vaccines from 3,844 healthy participants (aged 18 years to 85 years) who completed a prior placebo (saline)-controlled, randomized, observer-blind, multi-center Phase-2 primary 2-dose series (100 μg per dose; 28 days apart) of UB-612. The interim safety and immunogenicity were evaluated until 14 days post-booster.

Exposure: Vaccination with a booster 3rd dose (100 μg) of UB-612 vaccine.

Main outcomes and measures: Solicited local and systemic AEs were recorded for seven days in the e-diaries of study participants, while skin allergic reactions were recorded for fourteen days. The primary immunogenicity endpoints included viral-neutralizing antibodies against live SARS-CoV-2 Wild-Type (WT, Wuhan strain) and live Delta variant (VNT50), and against pseudovirus WT anbnd Omicron variant (pVNT50). The secondary immunogenicity endpoints included anti-S1- RBD IgG antibody, S1-RBD: ACE2 binding inhibition, and T-cell responses by ELISpot and Intracellular Staining.

Results: No post-booster vaccine-related serious adverse events were recorded. The most common solicited adverse events were injection site pain and fatigue, mostly mild and transient. The UB-612 booster prompted a striking upsurge of neutralizing antibodies against live WT Wuhan strain (VNT50, 1,711) associated with unusually high cross-neutralization against Delta variant (VNT50, 1,282); and similarly with a strong effect against pseudo virus WT (pVNT50, 6,245) and Omicron variant (pVNT50, 1,196). Upon boosting, the lower VNT50 and pVNT50 titers of the elderly in the primary series were uplifted to the same levels as those of the young adults. The UB-612 also induced robust, durable VoC antigen-specific Th1-oriented (IFN-γ+ -) responses along with CD8+ T-cell (CD107a+ -Granzyme B+ ) cytotoxicity.

Conclusions and relevance: With a pronounced cross-reactive booster effect on B and T-cell immunity, UB-612 may serve as a universal vaccine booster for comprehensive immunity enhancement against emergent VoCs.