Quantification of Immune Cell Subsets

Quantification of T cells, B cells and plasmablasts was performed on 100 µl heparinized whole blood exactly as described before⁴ (link)
^,¹⁹ using monoclonal antibodies towards CD3 (clone SK7), CD19 (clone HIB19), CD27 (clone L128), CD38 (clone HB7), and IgD (clone IA6-2). Cells were analyzed using flow cytometry (FACS-Canto-II or FACSLyric) and FACS-Diva-V6.1.3 or FlowJo software (BD Biosciences). Among total lymphocytes, T and B cells were identified by expression of CD3 and CD19, respectively. Plasmablasts were gated as CD38 positive cells among IgD-CD27+ CD19 positive switched-memory B cells as described before.⁴ (link) Differential blood counts were used to calculate absolute lymphocyte numbers.

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Schmidt T., Klemis V., Schub D., Schneitler S., Reichert M.C., Wilkens H., Sester U., Sester M, & Mihm J. (2022). Cellular immunity predominates over humoral immunity after homologous and heterologous mRNA and vector-based COVID-19 vaccine regimens in solid organ transplant recipients. American Journal of Transplantation, 21(12), 3990-4002.

Publication 2021

FACS-Canto-II FACSLyric FACS-Diva-V6.1.3

B cells Blood Cells Clone Flow cytometry Lymphocyte numbers Lymphocytes Memory b cells Monoclonal antibodies T cells

Corresponding Organization : Saarland University

Other organizations : Institute of Medical Microbiology and Hygiene

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Variable analysis

independent variables

Monoclonal antibodies towards CD3 (clone SK7), CD19 (clone HIB19), CD27 (clone L128), CD38 (clone HB7), and IgD (clone IA6-2)

dependent variables

Quantification of T cells, B cells and plasmablasts

control variables

100 µl heparinized whole blood
Flow cytometry (FACS-Canto-II or FACSLyric) and FACS-Diva-V6.1.3 or FlowJo software (BD Biosciences)
Differential blood counts

controls

No positive or negative controls were explicitly mentioned in the input.

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