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The Chikungunya Virus Capsid Protein Contains Linear B Cell Epitopes in the N- and C-Terminal Regions that are Dependent on an Intact C-Terminus for Antibody Recognition.

Goh LY, Hobson-Peters J, Prow NA, Baker K, Piyasena TB, Taylor CT, Rana A, Hastie ML, Gorman JJ, Hall RA - Viruses (2015)

Bottom Line: Competitive binding also revealed that five of the CP-specific mAbs recognized a series of overlapping epitopes in the latter domain.Our data also provides evidence that the C-terminus of CP is required for authentic antigenic structure of CP.This study shows that these anti-CP mAbs will be valuable research tools for further investigating the structure and function of the CHIKV CP.

View Article: PubMed Central - PubMed

Affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Queensland 4072, Australia. l.goh1@uq.edu.au.

ABSTRACT
Chikungunya virus (CHIKV) is an arthropod-borne agent that causes severe arthritic disease in humans and is considered a serious health threat in areas where competent mosquito vectors are prevalent. CHIKV has recently been responsible for several millions of cases of disease, involving over 40 countries. The recent re-emergence of CHIKV and its potential threat to human health has stimulated interest in better understanding of the biology and pathogenesis of the virus, and requirement for improved treatment, prevention and control measures. In this study, we mapped the binding sites of a panel of eleven monoclonal antibodies (mAbs) previously generated towards the capsid protein (CP) of CHIKV. Using N- and C-terminally truncated recombinant forms of the CHIKV CP, two putative binding regions, between residues 1-35 and 140-210, were identified. Competitive binding also revealed that five of the CP-specific mAbs recognized a series of overlapping epitopes in the latter domain. We also identified a smaller, N-terminally truncated product of native CP that may represent an alternative translation product of the CHIKV 26S RNA and have potential functional significance during CHIKV replication. Our data also provides evidence that the C-terminus of CP is required for authentic antigenic structure of CP. This study shows that these anti-CP mAbs will be valuable research tools for further investigating the structure and function of the CHIKV CP.

No MeSH data available.


Related in: MedlinePlus

Reactivity of anti-CP mAbs with full length and truncated versions of native and recombinant CP. Reactivity of mAbs 1.7B2 (lanes A1–3), 5.2H7 (lanes A4–6), cocktail of group 2 mAbs (lanes B1–3) and anti-V5 mAb (lanes B4–6) in Western blot against boiled and reduced lysates of COS-7L cells expressing rCap (lanes 1 and 4), mock-transfected COS-7L cells (lanes 2 and 5), and CHIKV-infected C6/36 cells (lanes 3 and 6).
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viruses-07-02754-f006: Reactivity of anti-CP mAbs with full length and truncated versions of native and recombinant CP. Reactivity of mAbs 1.7B2 (lanes A1–3), 5.2H7 (lanes A4–6), cocktail of group 2 mAbs (lanes B1–3) and anti-V5 mAb (lanes B4–6) in Western blot against boiled and reduced lysates of COS-7L cells expressing rCap (lanes 1 and 4), mock-transfected COS-7L cells (lanes 2 and 5), and CHIKV-infected C6/36 cells (lanes 3 and 6).

Mentions: IFA and Western blot analysis of N-terminally truncated recombinant CP indicated that group 1 mAbs recognized a region in the C-terminal half of the CHIKV CP while group 2 mAbs bound the N-terminal region. We also observed that, in addition to detecting the rCap in lysates of CHIKV-infected cells (~35 kDa), group 1 mAbs also detected smaller, truncated versions of native CP (sCP) ranging from ~15–30 kDa (Figure 6A). In contrast, antibodies from group 2 only recognized full-length CP in these lysates (Figure 6B, Figure S4).


The Chikungunya Virus Capsid Protein Contains Linear B Cell Epitopes in the N- and C-Terminal Regions that are Dependent on an Intact C-Terminus for Antibody Recognition.

Goh LY, Hobson-Peters J, Prow NA, Baker K, Piyasena TB, Taylor CT, Rana A, Hastie ML, Gorman JJ, Hall RA - Viruses (2015)

Reactivity of anti-CP mAbs with full length and truncated versions of native and recombinant CP. Reactivity of mAbs 1.7B2 (lanes A1–3), 5.2H7 (lanes A4–6), cocktail of group 2 mAbs (lanes B1–3) and anti-V5 mAb (lanes B4–6) in Western blot against boiled and reduced lysates of COS-7L cells expressing rCap (lanes 1 and 4), mock-transfected COS-7L cells (lanes 2 and 5), and CHIKV-infected C6/36 cells (lanes 3 and 6).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488721&req=5

viruses-07-02754-f006: Reactivity of anti-CP mAbs with full length and truncated versions of native and recombinant CP. Reactivity of mAbs 1.7B2 (lanes A1–3), 5.2H7 (lanes A4–6), cocktail of group 2 mAbs (lanes B1–3) and anti-V5 mAb (lanes B4–6) in Western blot against boiled and reduced lysates of COS-7L cells expressing rCap (lanes 1 and 4), mock-transfected COS-7L cells (lanes 2 and 5), and CHIKV-infected C6/36 cells (lanes 3 and 6).
Mentions: IFA and Western blot analysis of N-terminally truncated recombinant CP indicated that group 1 mAbs recognized a region in the C-terminal half of the CHIKV CP while group 2 mAbs bound the N-terminal region. We also observed that, in addition to detecting the rCap in lysates of CHIKV-infected cells (~35 kDa), group 1 mAbs also detected smaller, truncated versions of native CP (sCP) ranging from ~15–30 kDa (Figure 6A). In contrast, antibodies from group 2 only recognized full-length CP in these lysates (Figure 6B, Figure S4).

Bottom Line: Competitive binding also revealed that five of the CP-specific mAbs recognized a series of overlapping epitopes in the latter domain.Our data also provides evidence that the C-terminus of CP is required for authentic antigenic structure of CP.This study shows that these anti-CP mAbs will be valuable research tools for further investigating the structure and function of the CHIKV CP.

View Article: PubMed Central - PubMed

Affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Queensland 4072, Australia. l.goh1@uq.edu.au.

ABSTRACT
Chikungunya virus (CHIKV) is an arthropod-borne agent that causes severe arthritic disease in humans and is considered a serious health threat in areas where competent mosquito vectors are prevalent. CHIKV has recently been responsible for several millions of cases of disease, involving over 40 countries. The recent re-emergence of CHIKV and its potential threat to human health has stimulated interest in better understanding of the biology and pathogenesis of the virus, and requirement for improved treatment, prevention and control measures. In this study, we mapped the binding sites of a panel of eleven monoclonal antibodies (mAbs) previously generated towards the capsid protein (CP) of CHIKV. Using N- and C-terminally truncated recombinant forms of the CHIKV CP, two putative binding regions, between residues 1-35 and 140-210, were identified. Competitive binding also revealed that five of the CP-specific mAbs recognized a series of overlapping epitopes in the latter domain. We also identified a smaller, N-terminally truncated product of native CP that may represent an alternative translation product of the CHIKV 26S RNA and have potential functional significance during CHIKV replication. Our data also provides evidence that the C-terminus of CP is required for authentic antigenic structure of CP. This study shows that these anti-CP mAbs will be valuable research tools for further investigating the structure and function of the CHIKV CP.

No MeSH data available.


Related in: MedlinePlus