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The establishment of CMV antigen-specific enzyme-linked immunospot assay among Chinese: A pilot study
Yutong Liu1, Susu Ye1, Baotong Zhou2
1Department of Infectious Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Insights
Cytomegalovirus antigen-specific enzyme-linked immunospot assay effectively distinguishes infection states. Phosphoprotein 65 stimulation yielded a stronger cellular immune response than immediate early protein-1 in patients.
Area of Science:
- Immunology
- Virology
Background:
- Cytomegalovirus (CMV) infection is prevalent in China, posing significant health risks.
- Cellular immunity plays a crucial role in controlling CMV.
- The diagnostic performance of CMV antigen-specific enzyme-linked immunospot assay (ELISpot) in different infection states was investigated.
Purpose of the Study:
- To evaluate the efficacy of CMV antigen-specific ELISpot assay.
- To assess cellular immune responses in individuals with varying CMV infection statuses.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) were isolated from 28 patients.
- PBMCs were stimulated with CMV phosphoprotein 65 (pp65) or immediate early protein-1 (IE1) antigens.
- Spot-forming cells (SFCs) were quantified to measure antigen-specific cellular immunity.
Main Results:
- ELISpot assay demonstrated clear spot formation with a clean background.
- Significant differences in SFC counts were observed between uninfected (0 SFCs), latently infected (426 SFCs), and actively infected (230 SFCs) individuals.
- CMV pp65 antigen elicited a significantly higher SFC response compared to IE1 antigen.
Conclusions:
- CMV antigen-specific ELISpot assay effectively differentiates between CMV infection states.
- CMV pp65 is a more potent stimulator of cellular immunity than IE1 in this context.
Background:
The prevalence of cytomegalovirus in China is high and the infection could result in disastrous consequences. Cellular immunity is the main mechanism for virus controlling. We explored the performance of cytomegalovirus antigen-specific enzyme-linked immunospot assay in patients with different infection states in endemic area.
Methods:
Twenty-eight patients with reliable results were included in statistical analysis. Peripheral blood mononuclear cells were extracted and were stimulated by cytomegalovirus phosphoprotein 65 or immediate early protein-1 antigen respectively. Spot forming cells (SFCs) were counted to evaluate the cellular immune response elicited by antigens.
Results:
Spots could be clearly displayed, and evenly dispersed with a clean background. The numbers of SFCs were 0 [0-0], 426 [210-601] and 230 [57-513] for uninfected individuals (n = 7), latently infected patients (n = 11) and actively infected patients (n = 10) respectively, which were statistically different. The number of SFCs stimulated by phosphoprotein 65 was significantly higher than that by immediate early protein-1.
Conclusions:
The number of SFCs was significantly different among patients with different infection state. The stimulatory effect of phosphoprotein 65 was better than that of immediate early protein-1.
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