ng well recombinant chikv e2 protein  (Sino Biological)

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: NHPs were vaccinated with a low (LD) or high doses (HD) of EILV/CHIKV, CHIKV 181/25 or PBS (mock). At day 350, all animals were implanted with electronic data loggers as temperature sensor and challenged subcutaneously with 1.0 × 10 5 PFU of WT La Reunion strain of CHIKV at day 370. A Study design and vaccination timeline [Created with BioRender.com Wang, T. (2023) BioRender.com/o29z240]. B Body temperature changes were recorded every 15 min and reported as mean ± standard error of the mean (SEM) starting 7 days before to 14 days after infection with WT CHIKV strain La Réunion. C Viremia were measured by plaque assays at indicated days post-challenge (PC). UD: Undetectable. Data are presented as means ± SEM.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques: Infection

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: A – E Transcriptional changes of PBMCs of vaccinated macaques and mock controls at day 4 PC. A Principal component analysis (PCA) of all normalized transcripts to visualize the relatedness of samples via dimensional reduction. Each dot represents an individual RNA sample for mock (pink; n = 3), EILV/CHIKV (gray; n = 3), and CHIKV 181/25 (aqua; n = 3) groups. Samples that cluster are considered overall more similar, whereas distant samples are considered overall more distinct. B Heatmap of the most differentially expressed mRNAs in each group. Transcripts with a Benjamini–Hochberg adjusted p -value < 0.05 were sorted by log fold change values for EILV/CHIKV-vaccinated compared to CHIKV 181/25-immunized subjects. Red indicates increased expression; blue indicates decreased expression. C Bar plot of the most significant signaling pathways in EILV/CHIKV-vaccinated compared to mock-immunized subjects. Any differentially expressed transcripts with a Benjamini–Hochberg corrected P < 0.1 were deemed significant for enrichment. Pathways were sorted by statistical significance (a higher -log ( P ) indicates higher significance). Red indicates increased expression; blue indicates decreased expression. D Heatmap of the most upregulated pathways in each group; samples were sorted by z-score for EILV/CHIKV-vaccinated compared to CHIKV 181/25-immunized subjects. Yellow/green indicates increased expression, blue/purple indicates decreased expression. E Dot plots depicting predicted cell type trend scores for mock (pink; n = 3), EILV/CHIKV (gray; n = 3), and CHIKV 181/25 (aqua; n = 3) groups. Values were derived from transcriptional signatures indicative for a particular cell subset. F , G PBMCs of day 350 vaccinated NHPs were cultured ex vivo with CHIKV capsid, E3, E2 and E1 peptide pools for 6 h, and stained for IFN-γ, CD3, CD4, or CD8. Total number of IFN-γ + CD4 + and CD8 + T cells is shown. *** P < 0.001, ** P < 0.01, or * P < 0.05 compared to mock. #### P < 0.001, ## P < 0.01, or # P < 0.05 compared to CHIKV 181/25.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques: Expressing, Derivative Assay, Cell Culture, Ex Vivo, Staining

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: A – C CHIKV-specific MBC responses by ELISPOT analysis. PBMCs of day 30 ( A ) and day 350 ( B - C ) vaccinated NHPs were stimulated for 5 d with R848 plus rIL-2 and seeded onto ELISPOT plates coated with CHIKV capsid, E3, E2 and E1 peptide pools ( A ), total IgG or CHIKV recombinant E2 protein ( B , C ). A – C Frequencies of CHIKV-specific antibody secreting cells (ASC)s per 10 6 input cells in MBC cultures from the subject. B Images of total IgG-ASCs, CHIKV peptide-specific or CHIKV E2 protein- specific MBCs, are shown. D Serum neutralizing activity against CHIKV 181/25 was measured by a plaque reduction neutralization test (PRNT). E CHIKV E2 binding IgG responses at indicate time points by ELISA. F , G Passive immunization study. Pooled sera collected at day 350 PV of macaques were diluted 1:2 in PBS and transferred to 6 week-old AB6 mice 24 h before infection with a LD100 dose of WT CHIKV. Mice were monitored daily for morbidity. F Survival rate. G Percent weight loss compared to prior infection. *** P < 0.001, ** P < 0.01, or * P < 0.05 compared to mock. #### P < 0.001, ## P < 0.01, or # P < 0.05 compared to CHIKV 181/25.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques: Enzyme-linked Immunospot, Recombinant, Activity Assay, Plaque Reduction Neutralization Test, Binding Assay, Enzyme-linked Immunosorbent Assay, Infection

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: NHPs were implanted with electronic data loggers as temperature sensor and bled 14 days before vaccination. NHPs were vaccinated with 1.3 × 10 8 PFU EILV/CHIKV ( n = 3), 5 × 10 5 PFU inactivated WT CHIKV ( n = 3) or PBS (mock, n = 2). At day 6 PV, NHPs were challenged subcutaneously with 10 5 PFU of WT La Reunion strain of CHIKV. A Study design and vaccination timeline [Created with BioRender.com Wang, T. (2023) BioRender.com/o29z240]. B Body temperature changes were recorded every 15 min and reported as hourly mean ± SEM starting 8 days before to 14 days after the challenge with WT CHIKV. C , D Viremia were measured by plaque assays ( C ) or Q-PCR ( D ) at indicated days PC. Undetectable: UD. ** P < 0.01 compared to mock group. Unpaired, 2-tailed Student’s t -test was used to determine the differences. Data are presented as means ± SEM.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques:

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: A – D Transcriptional changes of PBMCs in EILV/CHIKV- vaccinated macaques and mock controls at day 7 PV. A PCA of all normalized transcripts to visualize the relatedness of samples via dimensional reduction. Each dot represents an individual RNA sample for mock (pink; n = 3) and EILV/CHIKV (gray; n = 3) groups. Samples that cluster are considered overall more similar, whereas distant samples are considered overall more distinct. B Heatmap of the most differentially expressed mRNAs in EILV/CHIKV-vaccinated compared to the mock-immunized subjects; sorted by statistical significance (Benjamini–Hochberg adjusted P < 0.05). Red indicates increased expression, white indicates no change in expression, blue indicates decreased expression. C Bar plot of the most significant signaling pathways in EILV/CHIKV-vaccinated compared to mock-immunized subjects. Any differentially expressed transcripts with a Benjamini–Hochberg corrected P < 0.1 were deemed significant for enrichment purposes. Pathways are sorted by statistical significance (a higher -log( p -value) indicates higher significance). Red indicates increased expression, blue indicates decreased expression. D Dot plots depicting predicted cell type trend scores for mock (pink; n = 3) and EILV/CHIKV (gray; n = 3) groups. Values were derived from transcriptional signatures indicative for a particular cell subset. E – G Sera cytokines were measured by nonhuman primate inflammation 13-plex kits at indicated time points PC. Data are presented as fold increases compared to the mock group.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques: Expressing, Derivative Assay

Journal: NPJ Vaccines

Article Title: A safe insect-based chikungunya fever vaccine affords rapid and durable protection in cynomolgus macaques

doi: 10.1038/s41541-024-01047-z

Figure Lengend Snippet: A , B NHPs were vaccinated with EILV/CHIKV, inactivated WT CHIKV or PBS (mock). At day 6 PV, NHPs were challenged subcutaneously with 10 5 PFU of the WT La Reunion strain of CHIKV. At day 4 PC, cells expressing NK cell markers were analyzed. Percent positive ( A ) or total number ( B ) of NK or NK T cells in PBMCs are shown. C Serum neutralizing activity against CHIKV 181/25 was measured by PRNT. D CHIKV E2 binding IgG responses at indicate time points by ELISA. E . ELISPOT quantification of peripheral T cell responses. PBMCs of macaques collected at day 4 PC were stimulated with CHIKV capsid, E3, E2 and E1 peptide pools for 24 h. SFCs were measured by IFN-γ ELISPOT. Data are shown as # of SFC per 10 6 cells. ** P < 0.01, or * P < 0.05 compared to mock. ## P < 0.01 compared to CHIKV 181/25.

Article Snippet: Millipore ELISPOT plates (Millipore Ltd, Darmstadt, Germany) were coated with CHIKV peptide pools (15 µg/ml), or CHIKV E2 recombinant protein (SinoBiological, USA, 15 ug/ml), EILV/CHIKV (1 × 10 8 PFU/ well), CHIKV VLP (The Native Antigen Company, Oxford, UK, 15 µg/ml), or anti-human Ig capture Ab (Mabtech).

Techniques: Expressing, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Yonsei Medical Journal

Article Title: Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

doi: 10.3349/ymj.2008.49.5.828

Figure Lengend Snippet: The seroreactivity of the recombinant CHIKV E1 and E2 envelope proteins using indirect IgM ELISA. Forty anti-CHIKV positive serum samples were used to evaluate the seroreactivity of the recombinant CHIKV E1 (•) and E2 (•) envelope proteins. The absorbance was read at 450 nm. IgM capture ELISA data (○) were supplied from Laboratoire Marcel Merieux. CHIKV, chikungunya virus.

Article Snippet: The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Virus

Journal: Yonsei Medical Journal

Article Title: Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

doi: 10.3349/ymj.2008.49.5.828

Figure Lengend Snippet: Analysis of expression of CHIKV E1 (A) and E2 (B) envelope protein in Sf9 cells. Sf9 cell lysates were analysed by SDS-PAGE, and the gel was stained with Coomassie blue (lanes 1 to 3) or analysed by Western blot with pooled anti-CHIKV positive serum (lane 4). M; Prestained protein marker, lane 1; lysate of Sf9 cells infected with mock-vector, lane 2; lysate of Sf9 cells infected with recombinant virus containing CHIKV E1 (A) or E2 (B) envelope protein gene, lane 3; purified recombinant CHIKV E1 (A) or E2 (B) envelope protein, lane 4; Western blot analysis with pooled anti-CHIKV positive serum. CHIKV, chikungunya virus.

Article Snippet: The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA.

Techniques: Expressing, SDS Page, Staining, Western Blot, Marker, Infection, Plasmid Preparation, Recombinant, Virus, Purification

Journal: Yonsei Medical Journal

Article Title: Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

doi: 10.3349/ymj.2008.49.5.828

Figure Lengend Snippet: Detection of anti-CHIKV IgM antibodies by recombinant CHIKV E1 (A) and E2 (B) envelope protein-based ELISA. Each well of a microtiter plate was coated with various amounts of recombinant CHIKV E1 and E2 envelope protein and evaluated for their reactivity with pooled anti-CHIKV positive or negative serum. Absorbance was read at 450 nm. The P/N ratio is A450 of positive serum/A450 of negative serum. CHIKV, chikungunya virus.

Article Snippet: The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Virus

Journal: Yonsei Medical Journal

Article Title: Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

doi: 10.3349/ymj.2008.49.5.828

Figure Lengend Snippet: Agreement, Sensitivity, and Specificity of Anti-CHIKV IgM Indirect ELISA Using the Recombinant CHIKV E1 and E2 Envelope Proteins with Anti-CHIKV IgM Capture ELISA from LMM

Article Snippet: The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA.

Techniques: Indirect ELISA, Recombinant, Enzyme-linked Immunosorbent Assay