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Early neutralizing IgG response to Chikungunya virus in infected patients targets a dominant linear epitope on the E2 glycoprotein.

Kam YW, Lum FM, Teo TH, Lee WW, Simarmata D, Harjanto S, Chua CL, Chan YF, Wee JK, Chow A, Lin RT, Leo YS, Le Grand R, Sam IC, Tong JC, Roques P, Wiesmüller KH, Rénia L, Rötzschke O, Ng LF - EMBO Mol Med (2012)

Bottom Line: E2EP3 is located at the N-terminus of the E2 glycoprotein and prominently exposed on the viral envelope.Screening of E2EP3 across different patient cohorts and in non-human primates demonstrated the value of this epitope as a good serology detection marker for CHIKV infection already at an early stage.Mice vaccinated by E2EP3 peptides were protected against CHIKV with reduced viremia and joint inflammation, providing a pre-clinical basis for the design of effective vaccine against arthralgia-inducing CHIKV and other alphaviruses.

View Article: PubMed Central - PubMed

Affiliation: Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.

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Sero-positivity of anti-E2EP3 IgG3 antibodies in CHIKV-infected patients from other cohortsCHIK virion-based ELISA was used to assess anti-CHIKV IgG titer in CHIKV-infected patients from another Singaporean cohort collected at median 10 days pio (n = 36). Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in the peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIK virion-based ELISA were used to assess anti-CHIKV IgG titer in 15 CHIKV-infected patients from another cohort collected in Malaysia at median 14 days pio. Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in a peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates. The same set of healthy donors' plasma comprising of donors from Singapore and Malaysia were used as controls throughout the study. The y axis is plotted in log 2 scale. Red straight line represents the median of the CHIKV-infected patients' group and black straight line represents the median of the healthy donors' group.
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fig06: Sero-positivity of anti-E2EP3 IgG3 antibodies in CHIKV-infected patients from other cohortsCHIK virion-based ELISA was used to assess anti-CHIKV IgG titer in CHIKV-infected patients from another Singaporean cohort collected at median 10 days pio (n = 36). Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in the peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIK virion-based ELISA were used to assess anti-CHIKV IgG titer in 15 CHIKV-infected patients from another cohort collected in Malaysia at median 14 days pio. Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in a peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates. The same set of healthy donors' plasma comprising of donors from Singapore and Malaysia were used as controls throughout the study. The y axis is plotted in log 2 scale. Red straight line represents the median of the CHIKV-infected patients' group and black straight line represents the median of the healthy donors' group.

Mentions: At median 10 days pio, almost all of the patients from this cohort were sero-positive for E2EP3 IgG3 antibodies (Supporting information Fig 4). To further validate the specificity and versatility of E2EP3 as a suitable early detection target, we screened plasma samples from another 36 CHIKV-infected patients collected from a separate cohort together with plasma obtained from 11 healthy donors (Fig 6). Here, plasma were again collected during the early convalescent phase (median 10 days pio) and tested for anti-E2EP3 IgG3 antibodies by ELISA (Fig 6B). Whole virus was used as a reference (Fig 6A). As in the previous cohort, specific E2EP3-binding was detected in virtually all CHIKV-infected patients with a clear segregation from the sero-negative healthy control donors (Fig 6A and B). Similar results were also obtained in a cohort from Malaysia where early convalescence samples of median 14 days pio were collected at outbreaks a few months later (Sam et al, 2009). Likewise, all of the patients screened were sero-positive for E2EP3, while no reactivity against the epitope was detected in healthy donors (Fig 6C and D). Thus, E2EP3 specific IgG3 antibodies appear to be a common early marker for CHIKV-infections at the population level.


Early neutralizing IgG response to Chikungunya virus in infected patients targets a dominant linear epitope on the E2 glycoprotein.

Kam YW, Lum FM, Teo TH, Lee WW, Simarmata D, Harjanto S, Chua CL, Chan YF, Wee JK, Chow A, Lin RT, Leo YS, Le Grand R, Sam IC, Tong JC, Roques P, Wiesmüller KH, Rénia L, Rötzschke O, Ng LF - EMBO Mol Med (2012)

Sero-positivity of anti-E2EP3 IgG3 antibodies in CHIKV-infected patients from other cohortsCHIK virion-based ELISA was used to assess anti-CHIKV IgG titer in CHIKV-infected patients from another Singaporean cohort collected at median 10 days pio (n = 36). Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in the peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIK virion-based ELISA were used to assess anti-CHIKV IgG titer in 15 CHIKV-infected patients from another cohort collected in Malaysia at median 14 days pio. Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in a peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates. The same set of healthy donors' plasma comprising of donors from Singapore and Malaysia were used as controls throughout the study. The y axis is plotted in log 2 scale. Red straight line represents the median of the CHIKV-infected patients' group and black straight line represents the median of the healthy donors' group.
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fig06: Sero-positivity of anti-E2EP3 IgG3 antibodies in CHIKV-infected patients from other cohortsCHIK virion-based ELISA was used to assess anti-CHIKV IgG titer in CHIKV-infected patients from another Singaporean cohort collected at median 10 days pio (n = 36). Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in the peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIK virion-based ELISA were used to assess anti-CHIKV IgG titer in 15 CHIKV-infected patients from another cohort collected in Malaysia at median 14 days pio. Healthy donors' plasma (n = 11) were used as controls. Individual samples were subjected to virion-based ELISA at a dilution of 1:2000, followed by secondary human anti-IgG-HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates.CHIKV-infected patients' and healthy donors' plasma were screened for IgG3 specific antibodies recognizing E2EP3 in a peptide-based ELISA. Individual samples were subjected to E2EP3 specific peptide-based ELISA at a dilution of 1:200, followed by secondary human anti-IgG3 isotype HRP. ***p < 0.001 by Mann–Whitney U test. Experiments were performed in triplicates. The same set of healthy donors' plasma comprising of donors from Singapore and Malaysia were used as controls throughout the study. The y axis is plotted in log 2 scale. Red straight line represents the median of the CHIKV-infected patients' group and black straight line represents the median of the healthy donors' group.
Mentions: At median 10 days pio, almost all of the patients from this cohort were sero-positive for E2EP3 IgG3 antibodies (Supporting information Fig 4). To further validate the specificity and versatility of E2EP3 as a suitable early detection target, we screened plasma samples from another 36 CHIKV-infected patients collected from a separate cohort together with plasma obtained from 11 healthy donors (Fig 6). Here, plasma were again collected during the early convalescent phase (median 10 days pio) and tested for anti-E2EP3 IgG3 antibodies by ELISA (Fig 6B). Whole virus was used as a reference (Fig 6A). As in the previous cohort, specific E2EP3-binding was detected in virtually all CHIKV-infected patients with a clear segregation from the sero-negative healthy control donors (Fig 6A and B). Similar results were also obtained in a cohort from Malaysia where early convalescence samples of median 14 days pio were collected at outbreaks a few months later (Sam et al, 2009). Likewise, all of the patients screened were sero-positive for E2EP3, while no reactivity against the epitope was detected in healthy donors (Fig 6C and D). Thus, E2EP3 specific IgG3 antibodies appear to be a common early marker for CHIKV-infections at the population level.

Bottom Line: E2EP3 is located at the N-terminus of the E2 glycoprotein and prominently exposed on the viral envelope.Screening of E2EP3 across different patient cohorts and in non-human primates demonstrated the value of this epitope as a good serology detection marker for CHIKV infection already at an early stage.Mice vaccinated by E2EP3 peptides were protected against CHIKV with reduced viremia and joint inflammation, providing a pre-clinical basis for the design of effective vaccine against arthralgia-inducing CHIKV and other alphaviruses.

View Article: PubMed Central - PubMed

Affiliation: Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.

Show MeSH
Related in: MedlinePlus