Human th1 2 cytokine kit 2

Cellular immunity was assessed by multicolor flow cytometric determination of surface markers using human monoclonal anti-CD3-fluorescein isothiocyanate (FITC), anti-CD4-phycoerythrin (PE), anti-CD8-allophycocyanin (APC), anti-CD19-PE, anti-CD16-APC, and anti-CD56-PE antibodies (BD Multitest), for determination of proportions and numbers of total T (CD3+), helper T (Th, CD3+CD4+), cytotoxic T (Tc, CD3+CD8+), Natural Killer (NK, CD3-CD16+CD56+), and B (CD3-CD19+) cell subsets. All samples were examined by a BD FACSCanto II Flow Cytometer. Data were analyzed by FlowJo v10.0.
Serum levels of C reactive protein (hypersensitive, hsCRP), immunoglobulin (IgG, IgM, IgA, and IgE), and complements (C3, C4) were detected by rate nephelometry immunoassay (N Antiserum to Human Ig Kit series, Siemens, Germany). The plasma levels of cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, and IFN-γ) were detected by Cytometric Bead Array using the human Th1/2 cytokine kit II (BD Ltd., USA). All tests were conducted according to the manufacturer's instructions.

According to the regulations of the local center for disease prevention and control, RT-PCR was used for SARS-CoV-2 nucleic acid testing [6 ]. RNA from the respiratory specimens was extracted with the High Pure Viral Nucleic Acid Kit (Zhongzhi, Wuhan, China). Lymphocyte subsets and cytokines were tested using a BD FACSCanto II flow cytometer (BD Biosciences). Lymphocytes were analyzed using a BD Multitest IMK Kit (cat. no. 340503, BD Biosciences); plasma cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ) were determined using a human Th1/2 cytokine kit II (BD Biosciences).

All patients were collected upper respiratory tract throat swab or nasal swab specimens when they were newly admitted to our hospital, and immediately stored in the virus transport medium [10 (link)]. Sputum specimens were also collected when necessary. SARS-CoV-2 virus were detected using the TaqMan one step RT-PCR kits (Shanghai Huirui Biotechnology Co., Ltd, China and Shanghai BioGerm Medical Biotechnology Co., Ltd, China) following the supplier’s protocol. The plasma cytokines (TNF-α, IL-6, IL-8, IL-1β, and IL-10) were measured using the Human Th1/2 Cytokine Kit II (BD Biosciences, Franklin Lakes, NJ, USA) following the product protocol.