il 4 elispot plus kit  (Mabtech Inc)

Journal: Nature Communications

Article Title: Chemically modified tRNA enhances the translation capacity of mRNA rich in cognate codons

doi: 10.1038/s41467-025-62981-7

Figure Lengend Snippet: a Left, schematic diagram showing that LNP formulation consisted of tRNA, Spike-mRNA, ionizable lipid, PEG2000-DMG, DSPC and cholesterol. Right, abbreviation of LNPs and tRNAs. T1 (Lst-UUA) was used as the benchmark vaccine. Placebo (blank LNP) was used as a negative control. Figure was generated using BioRender. b , c Dynamic changes of RNA levels and antigen concentrations of LNP formulations transfected in HEK293T cells ( n = 3 biological replicates). d Schematic diagram of the mRNA-tRNA vaccination, antibody detection, and splenic lymphocytes detection in BALB/c mice. Figure was generated using BioRender. e Measurement of the IgG antibody endpoint GMTs elicited by the mRNA-tRNA vaccines using ELISA ( n = 8 mice in each group). f Quantification of the NT50 of mRNA-tRNA immunized mouse sera by Spike B.1.1.529 pseudovirus neutralizing assay. The sera were collected 2 weeks after the booster injection (T1 ~ T7, n = 8 mice in each group; placebo, n = 3 mice). g ELISpot assay measurement of the SARS-CoV-2 Spike-specific IFN-γ, TNF-α, IL-2, and IL-4 responses of splenocytes from immunized mice. Mouse spleens were isolated and dissociated 5 weeks post boost-injection (T1 ~ T7, n = 8 mice in each group; placebo, n = 3 mice). h FACS analysis results showing the percentages and cell counts of CD4 + Tem cells producing IL-2, and IL-4 after stimulation with Spike-peptide pool ( n = 8 mice in each group). i FACS analysis results showing the percentages and cell counts of CD8 + Tem cells producing IFN-γ and TNF-α after stimulation ( n = 8 mice in each group). Statistical significance was analysed using one-way ANOVA with Dunnett’s correlation ( e – i ) or two-way ANOVA with Dunnett’s correlation ( c ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns, not significant. Data are presented as mean ± s.d. Source data are provided as a Source Data file.

Article Snippet: ELISpot assay of splenocytes was performed using the mouse IFN-γ, TNF-α, IL-2, and IL-4 ELISpot PLUS Kit (MabTech) according to the manufacturer’s instructions.

Techniques: Formulation, Negative Control, Generated, Transfection, Vaccines, Enzyme-linked Immunosorbent Assay, Neutralizing Assay, Injection, Enzyme-linked Immunospot, Isolation

Journal: iScience

Article Title: Preclinical evaluation of PHH-1V vaccine candidate against SARS-CoV-2 in non-human primates

doi: 10.1016/j.isci.2023.107224

Figure Lengend Snippet: Cellular immune responses in cynomolgus macaques on PHH-1V vaccination (A and B) PBMCs were isolated 7 days post-immunization (dpi) (D28) in PHH-1V vaccinated (n = 6) and PBS-mock vaccinated control (n = 3 or 4) group and then they were stimulated with an RBD peptide pool from Alpha and Beta variants (A) or Omicron (BA.1) variant (B) IFNɣ + - and IL-4 + -expressing cells were determined by ELISpot assay. Each data point represents an individual animal with bars indicating the mean and the SD. Arcsine square root-transformed percentage data were analyzed using generalized least squares models. (C and D) PBMC from vaccinated (PHH-1V, n = 6) and control (PBS, n = 4) cynomolgus macaques were isolated 7 dpi (D28). PBMCs were stimulated with an RBD peptide pool from Alpha and Beta variants and analyzed by ICS. Frequencies of cytokine expressing CD4 + T cells (C) and CD8 + T cells (D) are shown. Frequencies of CD4 + and CD8 + T cells expressing Th1-like cytokines (IFNɣ, TNFα and/or IL-2) are also depicted. Arcsine square root-transformed percentage data were analyzed using generalized least squares models. Basal expression of cytokines stimulated with media was considered the background value and was subtracted from the peptide-specific response. Two animals from the control group were excluded in both assays (ELISpot and ICS) because one showed a consistent RBD-specific response and the other showed a higher percentage of dead cells in their PBMCs. In the ELISpot assay stimulated with Omicron (BA.1) peptides, another one was excluded for not having enough cells to perform the analysis. Each data point represents an individual animal with bars indicating the mean and the SD. Data were analyzed using Welch’s permutation t-tests or Mann-Whitney U-tests. (∗p < 0.05, ∗∗p < 0.01).

Article Snippet: The NHP IFNɣ and IL-4 ELISpot Plus kits were used following manufacturer’s instructions (Mabtech).

Techniques: Isolation, Control, Variant Assay, Expressing, Enzyme-linked Immunospot, Transformation Assay, MANN-WHITNEY

Journal: iScience

Article Title: Preclinical evaluation of PHH-1V vaccine candidate against SARS-CoV-2 in non-human primates

doi: 10.1016/j.isci.2023.107224

Figure Lengend Snippet:

Article Snippet: The NHP IFNɣ and IL-4 ELISpot Plus kits were used following manufacturer’s instructions (Mabtech).

Techniques: Virus, Variant Assay, Expressing, Luciferase, Recombinant, Saline, Staining, Enzyme-linked Immunospot, Software, Sterility, Real-time Polymerase Chain Reaction, Microscopy, Flow Cytometry

Journal: iScience

Article Title: Preclinical evaluation of a COVID-19 vaccine candidate based on a recombinant RBD fusion heterodimer of SARS-CoV-2

doi: 10.1016/j.isci.2023.106126

Figure Lengend Snippet:

Article Snippet: ELISpot assays were performed with mouse IFN-γ and IL-4 ELISpot PLUS kits according to the manufacturer’s instructions (3321-4HPT-10 and 3311-4HPW-10, Mabtech, Herndon, VA, USA).

Techniques: Virus, Recombinant, Luciferase, Modification, Expressing, Transfection, Cell Viability Assay, Transgenic Assay, Software, Luminex