Limits...
Early appearance of neutralizing immunoglobulin G3 antibodies is associated with chikungunya virus clearance and long-term clinical protection.

Kam YW, Simarmata D, Chow A, Her Z, Teng TS, Ong EK, Rénia L, Leo YS, Ng LF - J. Infect. Dis. (2012)

Bottom Line: Although these patients endured a more severe disease progression during the acute viremic phase, they cleared the virus faster and did not experience persistent arthralgia.However, significant persistent arthralgia was observed in patients with low viremia who developed IgG3 at a later stage.Absence of early CHIKV-specific IgG3 may therefore serve as a specific marker of patients with increased risk of disease.

View Article: PubMed Central - PubMed

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

ABSTRACT

Background: Chikungunya virus (CHIKV) and related arboviruses have been responsible for large epidemic outbreaks with serious economic and social impact. Although infected individuals clear the virus from the blood, some develop debilitating and prolonged arthralgia.

Methods: We investigated specificity and strength of antibody responses in a longitudinal study on CHIKV-infected patients and analyzed their association with viral load, cytokine profile, and severity.

Results: We found that CHIKV-specific response is dominated by immunoglobulin G3 (IgG3) antibodies. The antibodies were neutralizing, and patients with high viremia rapidly developed high levels of anti-CHIKV antibodies of this specific isotype. Although these patients endured a more severe disease progression during the acute viremic phase, they cleared the virus faster and did not experience persistent arthralgia. However, significant persistent arthralgia was observed in patients with low viremia who developed IgG3 at a later stage.

Conclusions: Absence of early CHIKV-specific IgG3 may therefore serve as a specific marker of patients with increased risk of disease.

Show MeSH

Related in: MedlinePlus

Role of anti–chikungunya virus (CHIKV) immunoglobulin G3 (IgG3) in in vitro neutralizing activity against CHIKV from plasma samples of early and late IgG3 responders. A, Pooled plasma samples collected at median 10 d post–illness onset (PIO) (early IgG3, n = 16; late IgG3, n = 14) were added to plates precoated with purified Chikungunya virion for depletion of anti-CHIKV antibodies. Depleted samples were subjected to anti-CHIKV IgG3 antibody detection with virion-based enzyme-linked immunosorbent assay (ELISA). B, Depleted samples were subjected to in vitro neutralizing activity detection with a seroneutralization assay. C, IgG3 antibodies from pooled plasma samples collected at a median of 10 d PIO (early IgG3, n = 16; late IgG3, n = 14) were depleted as described in the “Methods” section and measured for anti-CHIKV IgG3 antibodies with virion-based ELISA. D, Depleted samples were subjected to in vitro neutralizing detection in a seroneutralization assay. All samples assayed were performed at 1:500 dilution (n = 3). Plasma from healthy donors was used as negative controls. Data are presented as mean ± SD. Data are representative of 3 independent experiments. Statistical significance was measured using Mann–Whitney U test. *P < .05. **P < .01.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3295607&req=5

fig3: Role of anti–chikungunya virus (CHIKV) immunoglobulin G3 (IgG3) in in vitro neutralizing activity against CHIKV from plasma samples of early and late IgG3 responders. A, Pooled plasma samples collected at median 10 d post–illness onset (PIO) (early IgG3, n = 16; late IgG3, n = 14) were added to plates precoated with purified Chikungunya virion for depletion of anti-CHIKV antibodies. Depleted samples were subjected to anti-CHIKV IgG3 antibody detection with virion-based enzyme-linked immunosorbent assay (ELISA). B, Depleted samples were subjected to in vitro neutralizing activity detection with a seroneutralization assay. C, IgG3 antibodies from pooled plasma samples collected at a median of 10 d PIO (early IgG3, n = 16; late IgG3, n = 14) were depleted as described in the “Methods” section and measured for anti-CHIKV IgG3 antibodies with virion-based ELISA. D, Depleted samples were subjected to in vitro neutralizing detection in a seroneutralization assay. All samples assayed were performed at 1:500 dilution (n = 3). Plasma from healthy donors was used as negative controls. Data are presented as mean ± SD. Data are representative of 3 independent experiments. Statistical significance was measured using Mann–Whitney U test. *P < .05. **P < .01.

Mentions: To determine if the antibodies also have protective capacity, in vitro infections of HEK 293T cells with CHIKV were carried out in the presence of plasma from patients or healthy donors (Figure 2). The experiments revealed that plasma samples collected at a median of 10 days PIO effectively inhibited CHIKV infection (Figure 2A). Preincubation of CHIKV with plasma samples induced a clear and dose-dependent reduction in the detection of CHIKV antigens (Figure 2A and 2B). In line with the observed differences in IgG3 titer, plasma from early IgG3 responders showed a higher neutralizing activity than plasma from the late IgG3 responders (Figure 2B). To confirm the protective role of anti-CHIKV IgG3 antibodies, CHIKV-infected patient plasma samples were depleted of antibodies against the purified Chikungunya virion (Figure 3A). Removal of anti-CHIKV IgG3 antibodies led to a marked decrease in neutralization for both early and late IgG3 responders (Figure 3B). In addition, the partial removal of IgG3 from the plasma of CHIKV patients by plate-bound anti-IgG3 reduced the IgG3 titer by 70%–80% (Figure 3C) and led to a marked decrease in neutralization for both early and late IgG3 responders (Figure 3D), confirming the importance of IgG3 antibodies in virus neutralization.


Early appearance of neutralizing immunoglobulin G3 antibodies is associated with chikungunya virus clearance and long-term clinical protection.

Kam YW, Simarmata D, Chow A, Her Z, Teng TS, Ong EK, Rénia L, Leo YS, Ng LF - J. Infect. Dis. (2012)

Role of anti–chikungunya virus (CHIKV) immunoglobulin G3 (IgG3) in in vitro neutralizing activity against CHIKV from plasma samples of early and late IgG3 responders. A, Pooled plasma samples collected at median 10 d post–illness onset (PIO) (early IgG3, n = 16; late IgG3, n = 14) were added to plates precoated with purified Chikungunya virion for depletion of anti-CHIKV antibodies. Depleted samples were subjected to anti-CHIKV IgG3 antibody detection with virion-based enzyme-linked immunosorbent assay (ELISA). B, Depleted samples were subjected to in vitro neutralizing activity detection with a seroneutralization assay. C, IgG3 antibodies from pooled plasma samples collected at a median of 10 d PIO (early IgG3, n = 16; late IgG3, n = 14) were depleted as described in the “Methods” section and measured for anti-CHIKV IgG3 antibodies with virion-based ELISA. D, Depleted samples were subjected to in vitro neutralizing detection in a seroneutralization assay. All samples assayed were performed at 1:500 dilution (n = 3). Plasma from healthy donors was used as negative controls. Data are presented as mean ± SD. Data are representative of 3 independent experiments. Statistical significance was measured using Mann–Whitney U test. *P < .05. **P < .01.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3295607&req=5

fig3: Role of anti–chikungunya virus (CHIKV) immunoglobulin G3 (IgG3) in in vitro neutralizing activity against CHIKV from plasma samples of early and late IgG3 responders. A, Pooled plasma samples collected at median 10 d post–illness onset (PIO) (early IgG3, n = 16; late IgG3, n = 14) were added to plates precoated with purified Chikungunya virion for depletion of anti-CHIKV antibodies. Depleted samples were subjected to anti-CHIKV IgG3 antibody detection with virion-based enzyme-linked immunosorbent assay (ELISA). B, Depleted samples were subjected to in vitro neutralizing activity detection with a seroneutralization assay. C, IgG3 antibodies from pooled plasma samples collected at a median of 10 d PIO (early IgG3, n = 16; late IgG3, n = 14) were depleted as described in the “Methods” section and measured for anti-CHIKV IgG3 antibodies with virion-based ELISA. D, Depleted samples were subjected to in vitro neutralizing detection in a seroneutralization assay. All samples assayed were performed at 1:500 dilution (n = 3). Plasma from healthy donors was used as negative controls. Data are presented as mean ± SD. Data are representative of 3 independent experiments. Statistical significance was measured using Mann–Whitney U test. *P < .05. **P < .01.
Mentions: To determine if the antibodies also have protective capacity, in vitro infections of HEK 293T cells with CHIKV were carried out in the presence of plasma from patients or healthy donors (Figure 2). The experiments revealed that plasma samples collected at a median of 10 days PIO effectively inhibited CHIKV infection (Figure 2A). Preincubation of CHIKV with plasma samples induced a clear and dose-dependent reduction in the detection of CHIKV antigens (Figure 2A and 2B). In line with the observed differences in IgG3 titer, plasma from early IgG3 responders showed a higher neutralizing activity than plasma from the late IgG3 responders (Figure 2B). To confirm the protective role of anti-CHIKV IgG3 antibodies, CHIKV-infected patient plasma samples were depleted of antibodies against the purified Chikungunya virion (Figure 3A). Removal of anti-CHIKV IgG3 antibodies led to a marked decrease in neutralization for both early and late IgG3 responders (Figure 3B). In addition, the partial removal of IgG3 from the plasma of CHIKV patients by plate-bound anti-IgG3 reduced the IgG3 titer by 70%–80% (Figure 3C) and led to a marked decrease in neutralization for both early and late IgG3 responders (Figure 3D), confirming the importance of IgG3 antibodies in virus neutralization.

Bottom Line: Although these patients endured a more severe disease progression during the acute viremic phase, they cleared the virus faster and did not experience persistent arthralgia.However, significant persistent arthralgia was observed in patients with low viremia who developed IgG3 at a later stage.Absence of early CHIKV-specific IgG3 may therefore serve as a specific marker of patients with increased risk of disease.

View Article: PubMed Central - PubMed

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

ABSTRACT

Background: Chikungunya virus (CHIKV) and related arboviruses have been responsible for large epidemic outbreaks with serious economic and social impact. Although infected individuals clear the virus from the blood, some develop debilitating and prolonged arthralgia.

Methods: We investigated specificity and strength of antibody responses in a longitudinal study on CHIKV-infected patients and analyzed their association with viral load, cytokine profile, and severity.

Results: We found that CHIKV-specific response is dominated by immunoglobulin G3 (IgG3) antibodies. The antibodies were neutralizing, and patients with high viremia rapidly developed high levels of anti-CHIKV antibodies of this specific isotype. Although these patients endured a more severe disease progression during the acute viremic phase, they cleared the virus faster and did not experience persistent arthralgia. However, significant persistent arthralgia was observed in patients with low viremia who developed IgG3 at a later stage.

Conclusions: Absence of early CHIKV-specific IgG3 may therefore serve as a specific marker of patients with increased risk of disease.

Show MeSH
Related in: MedlinePlus