Dataset Information

Preclinical study of formulated recombinant nucleocapsid protein, the receptor binding domain of the spike protein, and truncated spike (S1) protein as vaccine candidates against COVID-19 in animal models.

ABSTRACT:

Background

In this pre-clinical study, we designed a candidate vaccine based on severe acute respiratory syndrome-related -coronavirus 2 (SARS-CoV-2) antigens and evaluated its safety and immunogenicity.

Methods

SARS-CoV-2 recombinant protein antigens, including truncated spike protein (SS1, lacking the N-terminal domain of S1), receptor-binding domain (RBD), and nucleoprotein (N) were used. Immunization program was performed via injection of RBD, SS1 +RBD, and SS1 +N along with different adjuvants, Alum, AS03, and Montanide at doses of 0, 40, 80, and 120 μg at three-time points in mice, rabbits, and primates. The humoral and cellular immunity were analyzed by ELISA, VNT, splenocyte cytokine assay, and flow cytometry.

Results

The candidate vaccine produced strong IgG antibody titers at doses of 80 and 120 μg on days 35 and 42. Even though AS03 and Montanide produced high-titer antibodies compared to Alum adjuvant, these sera did not neutralize the virus. Strong virus neutralization was recorded during immunization with SS1 +RBD and RBD with Alum. AS03 and Montanide showed a strong humoral and cellular immunity; however, Alum showed mild to moderate cellular responses. Ultimately, no cytotoxicity and pathologic change were observed.

Conclusion

These findings strongly suggest that RBD with Alum adjuvant is highly immunogenic as a potential vaccine.

SUBMITTER: Nazarian S

PROVIDER: S-EPMC9222294 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Publications

Preclinical study of formulated recombinant nucleocapsid protein, the receptor binding domain of the spike protein, and truncated spike (S1) protein as vaccine candidates against COVID-19 in animal models.

Nazarian Shahram S Olad Gholamreza G Abdolhamidi Raziyeh R Motamedi Mohammad Javad MJ Kazemi Rouhollah R Kordbacheh Emad E Felagari Alireza A Olad Hanieh H Ahmadi Ali A Bahiraee Alireza A Farahani Parisa P Haghighi Leila L Hassani Faezeh F Hajhassan Vahideh V Nadi Mona M Sheikhi Abdolkarim A Salimian Jafar J Amani Jafar J

Molecular immunology 20220623

<h4>Background</h4>In this pre-clinical study, we designed a candidate vaccine based on severe acute respiratory syndrome-related -coronavirus 2 (SARS-CoV-2) antigens and evaluated its safety and immunogenicity.<h4>Methods</h4>SARS-CoV-2 recombinant protein antigens, including truncated spike protein (SS1, lacking the N-terminal domain of S1), receptor-binding domain (RBD), and nucleoprotein (N) were used. Immunization program was performed via injection of RBD, SS1 +RBD, and SS1 +N along with d ...[more]

PMID: 35802999

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Project description:BackgroundThe intricacies of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causing acute lung injury (ALI) and modulating inflammatory factor dynamics in vivo remain poorly elucidated. The present study endeavors to explore the impact of the recombinant SARS-CoV-2 spike protein S1 subunit (S1SP) on ALI and inflammatory factor profiles in mice, aiming to uncover potential therapeutic targets and intervention strategies for the prevention and management of Coronavirus Disease 2019 (COVID-19).MethodsTo mimic COVID-19 infection, K18-hACE2 transgenic mice were intratracheally instilled with S1SP, while C57BL/6 mice were administered LPS to form a positive control group. This setup facilitated the examination of lung injury severity, inflammatory factor levels, and alterations in signaling pathways in mice mimicking COVID-19 infection. Histopathological assessment through HE staining, along with analysis of lung wet/dry ratio and ultrasound imaging, revealed severe lung injury.ResultsAfter molding, K18-hACE2 mice exhibited a pronounced reduction in body weight and showed more significant lung injury (P < 0.05). Notably, there was a significant elevation in vascular permeability, total protein, and total white blood cells in bronchoalveolar lavage fluid (BALF) (P < 0.05), indicative of tissue damage. Additionally, the tight junction of lung tissue was compromised (P < 0.05), accompanied by intense oxidative stress marked by decreased SOD activity and elevated MDA content (P < 0.05). Cytokine levels, including IL-6, IL-1β, TNF-α, and MIG, were significantly upregulated in both BALF and serum of S1SP + K18 mice (P < 0.05). Furthermore, S1SP prominently augmented the expression of p-p65/P65 and attenuated IκBα expression in the NF-κB signaling pathway of humanized mice (P < 0.05), corroborating a heightened inflammatory response at the tissue level (P < 0.05).ConclusionThe administration of S1SP to K18-hACE2 mice resulted in severe lung injury, enhanced vascular permeability, and compromised epithelial barrier function in vivo. This was accompanied by disruption of lung tight junctions, the manifestation of severe oxidative stress and a cytokine storm, as well as the activation of the NF-κB signaling pathway, highlighting key pathological processes underlying COVID-19-induced lung injury.

Project description:The current lack of a straightforward and convenient modeling approach to simulate the onset of acute lung injury (ALI) has impeded fundamental research and hindered the screening of therapeutic drugs in coronavirus disease 2019 (COVID-19). The co-cultured human pulmonary alveolar epithelial cells (HPAEpics) and alveolar macrophages (AMs) were exposed to the complete medium, three concentrations of recombinant spike S1 protein (0.1, 1, and 10 μg/mL), or lipopolysaccharide (LPS) (10 μg/mL). The cells were harvested at 1, 2, and 3 days post-exposure. Lactate dehydrogenase (LDH) release, and IL-6, TNF-ɑ, and malondialdehyde (MDA) production were quantified and compared. Compared to those exposed to medium, co-cultures of HPAEpics and AMs exposed to a concentration of S1 protein at 10 μg/mL demonstrated significantly increased levels of LDH release (22.9% vs. 9.1%, and 25.7%), IL-6 (129 vs. 74, and 110 pg/mg of protein), and TNF-ɑ (75 vs. 51, and 86 pg/mg of protein) production, and similar to those exposed to LPS. However, no statistically significant differences were observed in MDA production. Compared to those harvested at 1 or 2 days post-exposure, co-cultured cells harvested at 3 days post-exposure exhibited increased levels of LDH release (23.4% vs. 14.9%, or 16.7%), IL-6 (127 vs. 81, or 97 pg/mg of protein) and MDA (5.6 vs. 3.2, or 3.8 nmol/mg of protein) production, but exhibited lower TNF-ɑ (58 vs. 79 pg/mg of protein) production than those harvested at 2 days post-exposure. After 3 days of exposure, co-cultures of HPAEpics and AMs showed significantly increased levels of LDH release (25.3% vs. 18.4%), and MDA production (5.5 vs. 4.3 nmol/mg of protein) compared to HPAEpics monocultures, and increased levels of LDH release (25.3% vs. 13.8%), IL-6 (139 vs. 98 pg/mg of protein) and MDA (5.5 vs. 4.7 nmol/mg of protein) production, and decreased TNF-ɑ (59 vs. 95 pg/mg of protein) production compared to AMs monocultures. Conclusions: The exposure to a concentration of S1 protein at 10 μg/mL in co-cultures of HPAEpics and AMs induced significant injury and inflammation three days post-exposure. This methodology for establishing a COVID-19-associated ALI model may have promising potential applications and value.

Dataset Information

Preclinical study of formulated recombinant nucleocapsid protein, the receptor binding domain of the spike protein, and truncated spike (S1) protein as vaccine candidates against COVID-19 in animal models.

Background

Methods

Results

Conclusion

Publications

Preclinical study of formulated recombinant nucleocapsid protein, the receptor binding domain of the spike protein, and truncated spike (S1) protein as vaccine candidates against COVID-19 in animal models.

Similar Datasets

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