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Mass Cytometry Analysis of High-Dimensional Single-Cell Immune Profiles in ZF2001-Vaccinated Patients Infected with
Xin Zhang1,2, Li Li3,4, Yongmei Liu5
1Department of Infectious Disease, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei, People's Republic of China.
Insights
One year after three doses of the ZF2001 COVID-19 vaccine, immune cell profiles showed significant changes in vaccinated patients. These alterations indicate the vaccine effectively modulated the immune response against SARS-CoV-2 infection.
Area of Science:
- Immunology
- Virology
- Vaccinology
Background:
- Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, prompted global health responses.
- The ZF2001 protein subunit vaccine targets the SARS-CoV-2 receptor-binding domain (RBD).
Purpose of the Study:
- To investigate peripheral immune cell profiles one year post-vaccination with ZF2001.
- To characterize the immune response and evaluate ZF2001's mechanism against chronic disease and mortality.
Main Methods:
- Single-cell mass spectrometry flow cytometry (CyTOF) was used to analyze peripheral blood mononuclear cells (PBMCs).
- A 41-antibody panel was employed on PBMCs from 16 vaccinated COVID-19 patients and 8 healthy donors.
- Data analysis involved FlowJo gating, Seurat/Harmony batch correction, PhenoGraph clustering, and t-SNE visualization.
Main Results:
- Significant differences in immune cell populations were observed between infected and healthy individuals one year after three ZF2001 doses.
- Elevated CD8+ cytotoxic T cells (expressing Ki67 and CD138), expanded B cells, and reduced CD16+ monocytes were noted.
- Depletion of CD4+ T cells and T cell differentiation were also observed, suggesting vaccine-induced immune modulation.
Conclusions:
- The study provides a detailed single-cell immune profile following SARS-CoV-2 infection in ZF2001-vaccinated individuals.
- Observed changes in immune cell kinetics indicate an active innate and adaptive immune response.
- ZF2001 vaccination appears to modulate the immune system effectively one year post-immunization.
Introduction:
Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was declared a public health emergency of international concern (PHEIC) by the WHO. ZF2001, a protein subunit vaccine targeting the RBD, was utilized to evaluate its impact on the immune system of COVID-19 patients. This study aimed to investigate peripheral cell profiles one year after three doses of ZF2001 vaccine using single cell mass spectrometry flow cytometry (CyTOF), a technique that allows detailed characterization of the immune response against SARS-COV-2 infection and further evaluation of ZF2001 mechanisms as a prophylactic against chronic disease and reducing mortality.
Methods:
This study profiled peripheral blood mononuclear cells (PBMCs) from 16 vaccinated COVID-19 patients (Omicron 5.2) and 8 hDs using CyTOF with a 41-antibody panel. PBMCs isolated via Lymphoprep density gradient underwent metal-tagged antibody staining. Data analysis included FlowJo gating, Seurat/Harmony batch correction, PhenoGraph clustering (k=45), and t-SNE visualization. Statistical assessments employed Wilcoxon tests and Spearman correlation.
Results:
Our findings revealed significant differences between infected and healthy individuals one year after three doses of ZF2001. Specifically, infected individuals exhibited: significant elevation of cytotoxic T cells expressing CD8 with a proliferation marker antigen-Kiel 67 (Ki67) and an adhesion molecule (CD138), expansion of B cells and reduction of monocytes expressing CD16, as well as depletion of CD4+ T cells and differentiation of T cells 1 year after the vaccine. These changes suggested that the vaccine was effectively modulating the immune response.
Discussion:
Our results provided a detailed single-cell profile of the immune response to SARS-CoV-2 infection in vaccinated patients, highlighting significant changes in immune cell kinetics indicative of an active innate and adaptive immune cell response.
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