Interferon-gamma FlowSpot assay for the measurement of the T-cell response to cytomegalovirus

Qianyu Ye; Jiali Wang; Meijing Chen; Weijian Nie; Huanxi Zhang; Xiaojun Su; Liuting Ling; Xiangjun Liu; Longshan Liu; Changxi Wang; Yifang Gao

doi:10.1016/j.heliyon.2023.e16792

Interferon-gamma FlowSpot assay for the measurement of the T-cell response to cytomegalovirus

Heliyon. 2023 Jun 8;9(6):e16792. doi: 10.1016/j.heliyon.2023.e16792. eCollection 2023 Jun.

Authors

Qianyu Ye^{1

2

3}, Jiali Wang⁴, Meijing Chen⁵, Weijian Nie^{1

2

3}, Huanxi Zhang^{1

2

3}, Xiaojun Su^{1

2

3}, Liuting Ling^{1

2

3}, Xiangjun Liu⁶, Longshan Liu^{1

2

3}, Changxi Wang^{1

2

3}, Yifang Gao^{1

2

3}

Affiliations

¹ Organ Transplantation Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
² Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
³ Guangdong Provincial International Cooperation Base of Science and Technology (Organ Transplantation), The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
⁴ Department of Nephrology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
⁵ Guangzhou BFR Gene Dignostics, Guangzhou, China.
⁶ Beijing BFR Gene Diagnostics, Beijing, China.

Abstract

Objectives: We describe a new method, FlowSpot, to assess CMV-specific T-cell response by quantification of interferon-gamma (IFN-γ). CMV-specific, T-cell-released IFN-γ was captured by flow beads and measured via flow cytometry. In the present study, we used FlowSpot to assess CMV-specific T-cell response in healthy individuals. The FlowSpot results were compared with those of serological analysis and enzyme-linked immunospot (ELISpot) assay.

Methods: Experimental results and parameter analysis were investigated by using serological, ELISpot, and FlowSpot assays.

Results: The levels of IFN-γ, which is released from CMV-specific T-cells, were measured, and the results and parameter analysis showed a good correlation between FlowSpot and ELISpot. However, FlowSpot was more sensitive and better reflected the strength of IFN-γ secretion than did ELISpot.

Conclusions: Compared to ELISpot, FlowSpot has a high sensitivity and is cost and time effective. Thus, this method can be used in wider clinical and scientific applications.

Keywords: CMV-Specific T-cells; ELISpot; FlowSpot; IFN-γ; Serologic analysis.