Cytokine and T cell responses in post-chikungunya viral arthritis: A cross-sectional study.

OBJECTIVE: To define the relationship between chronic chikungunya post-viral arthritis disease severity, cytokine response and T cell subsets in order to identify potential targets for therapy. METHODS: Participants with chikungunya arthritis were recruited from Colombia from 2019-2021. Arthritis disease severity was quantified using the Disease Activity Score-28 and an Arthritis-Flare Questionnaire adapted for chikungunya arthritis. Plasma cytokine concentrations (interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, interferon-γ and tumor necrosis factor (TNF)) were measured using a Meso Scale Diagnostics assay. Peripheral blood T cell subsets were measured using flow cytometry. RESULTS: Among participants with chikungunya arthritis (N = 158), IL-2 levels and frequency of regulatory T cells (Tregs) were low. Increased arthritis disease activity was associated with higher levels of inflammatory cytokines (IL-6, TNF and CRP) and immunoregulatory cytokine IL-10 (p<0.05). Increased arthritis flare activity was associated with higher Treg frequencies (p<0.05) without affecting T effector (Teff) frequencies, Treg/Teff ratios and Treg subsets. Finally, elevated levels of IL-2 were correlated with increased Treg frequency, percent Tregs out of CD4+ T cells, and Treg subsets expressing immunosuppressive markers, while also correlating with an increased percent Teff out of live lymphocytes (p<0.05). CONCLUSION: Chikungunya arthritis is characterized by increased inflammatory cytokines and deficient IL-2 and Treg responses. Greater levels of IL-2 were associated with improved Treg numbers and immunosuppressive markers. Future research may consider targeting these pathways for therapy.

Regulatory T-cells and GARP expression are decreased in exercise-associated chikungunya viral arthritis flares.

Objective: Chikungunya virus (CHIKV) causes persistent arthritis, and our prior study showed that approximately one third of CHIKV arthritis patients had exacerbated arthritis associated with exercise. The underlying mechanism of exercise-associated chikungunya arthritis flare (EACAF) is unknown, and this analysis aimed to examine the regulatory T-cell immune response related to CHIKV arthritis flares. Methods: In our study, 124 Colombian patients with a history of CHIKV infection four years prior were enrolled and 113 cases with serologically confirmed CHIKV IgG were used in this analysis. Patient information was gathered via questionnaires, and blood samples were taken to identify total live peripheral blood mononuclear cells, CD4+ cells, T regulatory cells, and their immune markers. We compared outcomes in CHIKV patients with (n = 38) vs. without (n = 75) EACAF using t-tests to assess means and the Fisher's exact test, chi-squared to evaluate categorical variables, and Kruskal-Wallis tests in the setting of skewed distributions (SAS 9.3). Results: 33.6% of CHIKV cases reported worsening arthritis with exercise. EACAF patients reported higher global assessments of arthritis disease ranging from 0-100 (71.2 ± 19.7 vs. 59.9 ± 28.0, p=0.03). EACAF patients had lower ratios of T regulatory (Treg)/CD4+ T-cells (1.95 ± 0.73 vs. 2.4 ± 1.29, p = 0.04) and lower percentage of GARP (glycoprotein-A repetitions predominant) expression per Treg (0.13 ± 0.0.33 vs. 0.16 ± 0.24 p= 0.020). Conclusion: These findings suggest relative decreases in GARP expression may indicate a decreased level of immune suppression. Treg populations in patients with CHIKV arthritis may contribute to arthritis flares during exercise, though current research is conflicting.

Sleep Disturbances are a Significant Predictor of Chikungunya Arthritis Flare Severity.

OBJECTIVE: The primary objective of this research was to explore the link between sleep and flare pain associated with chikungunya virus (CHIKV) infection. The secondary objective was to investigate if cytokines and T regulatory (Treg) cells have an influence on this relationship. METHODS: A cross-sectional study was performed using data collected in Barranquilla, Colombia, which enrolled patients with and without chronic arthritis with a history of chikungunya infection. Flare severity was measured by a version of the Outcome Measures in Rheumatoid Arthritis Clinical Trials (OMERACT) flare questionnaire adapted for CHIKV arthritis, including metrics for pain, difficulty with physical activity, fatigue, stiffness and difficulty maintaining social activities due to arthritis that contribute to flare severity. In addition, four sleep disturbance items, five inflammatory cytokine levels, four anti-inflammatory cytokine levels, and six Treg levels were measured. Then, multivariable linear regression models were used to test the direct and indirect effects of flare-pain on sleep disturbance, and to determine whether this relationship was mediated by cytokines or Tregs. Finally, the SAS CALIS procedure was used to test path models showing possible causal effects with mediators and confounds. RESULTS: The analysis showed that sleep disturbance is positively correlated with CHIKV arthritis flare pain, and that it is a significant predictor of flare severity after adjusting for demographic variables, cytokine, and T cell levels. Further, neither T cells nor cytokines mediate the pain/sleep relationship in CHIKV arthritis. CONCLUSION: There is a strong association between sleep disturbance and arthritis flare pain and severity; however, this relationship is not mediated by cytokines or T cells. Since this study is unable to determine causation, further research is needed to determine the mechanism underlying the relationship between sleep disturbances and CHIKV arthritis flares.

Stiffness, pain, and joint counts in chronic chikungunya disease: relevance to disability and quality of life.

INTRODUCTION/OBJECTIVES: To characterize the importance of musculoskeletal stiffness in a cohort of chikungunya patients with chronic joint symptoms. METHOD: Eighty-two patients were followed up 3 years after chikungunya infection. Tender and swollen joint counts, a pain intensity scale, Health Assessment Questionnaire-Disability Index (HAQ-DI), and the EuroQol EQ-5D quality of life instrument were completed. A musculoskeletal stiffness questionnaire provided scores for overall stiffness and its components: stiffness severity, physical impact, and psychosocial impact. RESULTS: Patients had a mean age 51 ± 14 years. Sixty-seven patients were still experiencing chronic arthralgia. Musculoskeletal stiffness was reported by 43/67 patients with arthralgia and 3/15 patients without arthralgia. A physical impact of stiffness was reported by 87% patients and psychosocial impact by 71% patients. Mean tender joint count in patients reporting arthralgia was 6 ± 7, mean pain intensity 65 ± 20 out of 100, mean HAQ-DI was 0.54 ± 0.52, and mean EQ-VAS global health perception was 68 ± 62 out of 100. Stiffness severity was correlated with tender joint counts (ρ = 0.46) and pain intensity (ρ = 0.40). All three measures were equally well correlated with the EuroQol-VAS global health perception. Pain and tender joints were better correlated with the HAQ-DI (ρ = 0.68 and ρ = 0.63), but stiffness was more strongly correlated with several quality of life domains, including mobility. Swollen joints were a poor predictor of outcomes. CONCLUSIONS: Musculoskeletal stiffness following chikungunya infection is distinct from arthralgia. It does not always occur in the same patients or with a corresponding intensity. Joint pain and stiffness may be independently associated with disability and quality of life assessments.

Chronic Joint Pain 3 Years after Chikungunya Virus Infection Largely Characterized by Relapsing-remitting Symptoms.

OBJECTIVE: To determine the frequency of chronic joint pain and stiffness 3 years after infection with chikungunya virus (CHIKV) in a Latin American cohort. METHODS: A cross-sectional followup of 120 patients from an initial cohort of 500 patients who reported joint pain 2 years after infection from the Atlántico Department, Colombia. Patients were clinically diagnosed as having CHIKV during the 2014-2015 epidemic, and baseline and followup symptoms at 40 months were evaluated in serologically confirmed cases. RESULTS: Of the initial 500 patients enrolled in the study, 482 had serologically confirmed chikungunya infection. From this group, 123 patients reported joint pain 20 months after infection, and 54% of those patients reported continued joint pain 40 months after infection. Therefore, 1 out of every 8 people who tested serologically positive for CHIKV infection had persistent joint pain 3 years after infection. Participants who followed up in person were predominantly adult (mean ± SD age 51 ± 14 yrs) and female (86%). The most common type of pain reported in these patients at 40 months post-infection was pain with periods of relief and subsequent reoccurrence, and over 75% reported stiffness after immobility, with 39% experiencing morning stiffness. CONCLUSION: To our knowledge, this is the first report to describe persistent joint pain and stiffness 40 months after viral infection. The high frequency of chronic disease highlights the need to develop prevention and treatment methods. Further studies should be conducted to understand the similarities between post-chikungunya joint pain and rheumatoid arthritis.

A Dengue Vaccine: Will It be Accepted and Is It Feasible? Lessons from Barranquilla, Colombia, and Merida, Venezuela.

With one vaccine on the market and others in clinical trials, policy makers in dengue endemic regions face the decision of whether to introduce a dengue vaccine in their communities. The World Health Organization (WHO) recommends that individualized assessments be conducted before any vaccine introduction to evaluate disease burden and the strength of current vaccination programs. This study seeks to aid in that decision-making process by examining the acceptability and feasibility of dengue vaccine introduction in Barranquilla, Colombia, and Merida, Venezuela. Surveys were administered February-June of 2018 for three groups: patients (n = 351), health professionals (n = 197), and government officials (n = 26). In Barranquilla, most respondents reported dengue to be a moderate-severe problem, that a dengue vaccine would be useful in their communities, and that their current vaccination programs could handle the addition of a new vaccine. In Venezuela, respondents were less likely to view dengue as a major concern and listed multiple barriers to not just dengue vaccine introduction, but to providing current vaccines as well. Further work is needed in Colombia to more objectively assess the country's readiness as a whole for a future dengue vaccine. As political and social unrest continues in Venezuela, however, future initiatives should focus on trust and capacity building. This study can serve as a framework for future assessments of the acceptability and feasibility of a dengue vaccine in both targeted areas and on larger scales.

Authors’ response to the letter to the editor entitled: Co-circulation of dengue, chikungunya, and Zika viruses and cross-protection

To the editor: Dengue, zika, and chikungunya outbreaks in Central and South America countries have presented significant challenges related to their prevention and control. From the virologic point of view, the possibility has been raised that the co-circulation of the three viruses could generate cross-protection between the three alphaviruses. In order to discuss this hypothesis it must be taken into account that Zika, dengue (DENV) and chikungunya viruses are closely related flaviviruses, with identical urban transmission and some immune interactions (1). Also, it is known that secondary DENV infections may be more severe than primary infections due to the antibody-dependent immune response (i.e., heterotypic sub-neutralizing antibodies that increase virus entry into poorly susceptible cells) (2,3). In addition, the recent introduction of Zika and chikungunya viruses in the Americas and the large-scale exposure of a uniformly unexposed population could affect subsequent transmission of dengue virus. This hypothesis has not been tested, largely because insufficient epidemiological data are available for the affected sites. However, in Salvador, Brazil, after the zika outbreak there was a significant decrease in the frequency of dengue cases (4). A similar situation was observed in Colombia, where the decrease in dengue cases following the zika and chikungunya outbreaks went from 334.1 cases per 100 000 people in 2015 to 90.7 cases per 100 000 in 2017 and 173,1 cases per 100 000 in 2018 (5). Although temporary associations do not prove causation, the strength and consistency of the observations suggest that infections with Zika virus and chikungunya virus could induce cross-protective immunity against dengue. Prospective studies are needed to fully assess the risk of dengue infection after exposure to Zika and chikungunya viruses and to determine whether the supposed cross-protection is long-lasting. Although observations support this hypothesis, the potential direct implications of this hypothesis for epidemiological surveillance, immunological research on pathogenesis and vaccine development require additional studies.

Co-circulation of dengue, chikungunya, and Zika viruses in Colombia from 2008 to 2018.

OBJECTIVE: This study aimed to identify the co-circulation patterns of three viruses (dengue, Zika, and -chikungunya) in Colombia from 2008 to 2018 by using notification reports provided to the national surveillance system. METHODS: This cross-sectional study was conducted through a review of data for 2008 through 2018 from Colombia's Public Health Surveillance System (SIVIGILA). RESULTS: In 2015, when chikungunya was first detected, it had a higher incidence (1 359.0 cases per 100 000 persons) than did the two other diseases. In 2016, when the circulation of Zika virus was first found, the incidence was 296.4 cases per 100 000 persons; that incidence declined dramatically in the next two years. Between 2015 and 2018, there was a substantial decrease in the frequency of dengue circulation, with it going from 334.1 cases per 100 000 persons in 2015 to 90.7 cases per 100 000 in 2017 and 173.1 cases per 100 000 in 2018. CONCLUSIONS: The decrease in the number of dengue cases after co-circulation of the three viruses could indicate possible cross-protection. This finding should be further analyzed.

Co-circulation of dengue, chikungunya, and Zika viruses in Colombia from 2008 to 2018

[ABSTRACT]. Objective. This study aimed to identify the co-circulation patterns of three viruses (dengue, Zika, and chikungunya) in Colombia from 2008 to 2018 by using notification reports provided to the national surveillance system. Methods. This cross-sectional study was conducted through a review of data for 2008 through 2018 from Colombia’s Public Health Surveillance System (SIVIGILA). Results. In 2015, when chikungunya was first detected, it had a higher incidence (1 359.0 cases per 100 000 persons) than did the two other diseases. In 2016, when the circulation of Zika virus was first found, the incidence was 296.4 cases per 100 000 persons; that incidence declined dramatically in the next two years. Between 2015 and 2018, there was a substantial decrease in the frequency of dengue circulation, with it going from 334.1 cases per 100 000 persons in 2015 to 90.7 cases per 100 000 in 2017 and 173.1 cases per 100 000 in 2018. Conclusions. The decrease in the number of dengue cases after co-circulation of the three viruses could indicate possible cross-protection. This finding should be further analyzed.

[RESUMEN]. Objetivo: Establecer las características de la cocirculación de tres virus (dengue, Zika y chikunguña) en Colombia desde el 2008 hasta el 2018. Para ello, en este estudio se han utilizado los informes de notificación que se proporcionan al sistema nacional de vigilancia. Métodos: Este estudio transversal se llevó a cabo mediante el análisis de los datos correspondientes al período 2008-2018 del Sistema Nacional de Vigilancia en Salud Pública de Colombia (SIVIGILA). Resultados: En el 2015, cuando se detectó por primera vez el chikunguña, este virus tuvo una incidencia mayor (1 359,0 casos por 100 000 personas) que las otras dos enfermedades. En el 2016, cuando se detectó por primera vez la circulación del virus del Zika, la incidencia fue de 296,4 casos por 100 000 personas; el número de casos disminuyó enormemente en los siguientes dos años. Entre el 2015 y el 2018, se observó una reducción sustancial en la frecuencia de la circulación del dengue: se pasó de 334,1 casos por 100 000 personas en el 2015 a 90,7 casos por 100 000 en el 2017 y a 173,1 casos por 100 000 en el 2018. Conclusiones: La disminución en el número de casos del dengue que siguió a la cocirculación de los tres virus podría indicar una posible protección cruzada. Este resultado debe analizarse en otros estudios.

[RESUMO]. Objetivo. Identificar padrões de co-circulação de três vírus (dengue, Zika e chikungunya) na Colômbia de 2008 a 2018 mediante análise de casos notificados ao sistema nacional de vigilância. Métodos. Estudo transversal realizado através de análise de dados de 2008 a 2018 obtidos do Sistema de Vigilância em Saúde Pública da Colômbia (SIVIGILA). Resultados. Em 2015, quando o chikungunya foi detectado pela primeira vez, sua incidência foi superior à das duas outras doenças (1.359,0 casos por 100,000 habitantes). Em 2016, quando a circulação do vírus zika foi detectada pela primeira vez, a incidência foi de 296,4 casos por 100,000 pessoas, decaindo drasticamente nos dois anos seguintes. Entre 2015 e 2018, houve uma redução importante na frequência de circulação do vírus dengue, de 334,1 casos por 100,000 habitantes em 2015 a 90,7 casos por 100,000 em 2017 e 173,1 casos por 100,000 em 2018. Conclusões. A redução do número de casos de dengue após o estabelecimento da co-circulação dos três vírus pode indicar proteção cruzada. Este achado requer análise adicional.

Neutralizing Antibodies from Convalescent Chikungunya Virus Patients Can Cross-Neutralize Mayaro and Una Viruses.

Most alphaviruses are mosquito-borne and can cause severe disease in domesticated animals and humans. The most notable recent outbreak in the Americas was the 2014 chikungunya virus (CHIKV) outbreak affecting millions and producing disease highlighted by rash and arthralgia. Chikungunya virus is a member of the Semliki Forest (SF) serocomplex, and before its arrival in the Americas, two other member of the SF complex, Una (UNAV) and Mayaro (MAYV) viruses, were circulating in Central and South America. This study examined whether antibodies from convalescent CHIKV patients could cross-neutralize UNAV and MAYV. Considerable cross-neutralization of both viruses was observed, suggesting that exposure to CHIKV can produce antibodies that may mitigate infection with UNAV or MAYV. Understanding the impact of CHIKV exposure on population susceptibility to other emerging viruses may help predict outbreaks; moreover, identification of cross-reactive immune responses among alphaviruses may lead to the development of vaccines targeting multiple viruses.

Augmented Zika and Dengue Neutralizing Antibodies Are Associated With Guillain-Barré Syndrome.

The role of neutralizing antibodies in Zika-induced Guillain-Barré syndrome (GBS) has not yet been investigated. We conducted a case-control study using sera from the 2016 Zika epidemic in Colombia to determine the neutralizing antibody activity against Zika virus (ZIKV) and dengue virus serotype 2 (DENV2). We observed increased neutralizing antibody titers against DENV2 in ZIKV-infected individuals compared with uninfected controls and higher titers to both ZIKV and DENV2 in ZIKV-infected patients diagnosed with GBS compared with non-GBS ZIKV-infected controls. These data suggest that high neutralizing antibody titers to DENV and to ZIKV are associated with GBS during ZIKV infection.

Sequential Neuroimaging of the Fetus and Newborn With In Utero Zika Virus Exposure.

Importance: The evolution of fetal brain injury by Zika virus (ZIKV) infection is not well described. Objectives: To perform longitudinal neuroimaging of fetuses and infants exposed to in utero maternal ZIKV infection using concomitant magnetic resonance imaging (MRI) and ultrasonography (US), as well as to determine the duration of viremia in pregnant women with ZIKV infection and whether the duration of viremia correlated with fetal and/or infant brain abnormalities. Design, Setting, and Participants: A cohort of 82 pregnant women with clinical criteria for probable ZIKV infection in Barranquilla, Colombia, and Washington, DC, were enrolled from June 15, 2016, through June 27, 2017, with Colombian women identified by community recruitment and physician referral and travel-related cases of American women recruited from a Congenital Zika Program. Interventions and Exposures: Women received 1 or more MRI and US examinations during the second and/or third trimesters. Postnatally, infants underwent brain MRI and cranial US. Blood samples were tested for ZIKV. Main Outcomes and Measures: The neuroimaging studies were evaluated for brain injury and cerebral biometry. Results: Of the 82 women, 80 were from Colombia and 2 were from the United States. In 3 of 82 cases (4%), fetal MRI demonstrated abnormalities consistent with congenital ZIKV infection. Two cases had heterotopias and malformations in cortical development and 1 case had a parietal encephalocele, Chiari II malformation, and microcephaly. In 1 case, US results remained normal despite fetal abnormalities detected on MRI. Prolonged maternal polymerase chain reaction positivity was present in 1 case. Of the remaining 79 cases with normal results of prenatal imaging, postnatal brain MRI was acquired in 53 infants and demonstrated mild abnormalities in 7 (13%). Fifty-seven infants underwent postnatal cranial US, which detected changes of lenticulostriate vasculopathy, choroid plexus cysts, germinolytic/subependymal cysts, and/or calcification in 21 infants (37%). Conclusions and Relevance: In a cohort of pregnant women with ZIKV infection, prenatal US examination appeared to detect all but 1 abnormal fetal case. Postnatal neuroimaging in infants who had normal prenatal imaging revealed new mild abnormalities. For most patients, prenatal and postnatal US may identify ZIKV-related brain injury.

Second-trimester Ultrasound and Neuropathologic Findings in Congenital Zika Virus Infection.

Zika virus is a mosquito-borne virus that causes congenital Zika syndrome, characterized by microcephaly and other fetal brain anomalies. This case report presents a case of Zika virus-related fetal brain anomalies including pathologic evidence of cerebral neuronal apoptosis and macrophage infiltrates and intracerebral calcification, ventriculomegaly and corpus callosum dysgenesis detected by ultrasound at 18 weeks of pregnancy.

The Cytokine Profile in Acute Chikungunya Infection is Predictive of Chronic Arthritis 20 Months Post Infection.

The cytokine profile during acute chikungunya infection that predicts future chronic arthritis has not yet been investigated. We conducted a nested case-control study comparing serum cytokine concentrations during acute chikungunya infection in cases (n = 121) that reported the presence of chronic joint pain versus age- and gender-matched controls (n = 121) who reported recovery at 20 months post infection. We observed that a robust cytokine response during acute infection was correlated with a decreased incidence of chronic joint pain and that low TNFα, IL-13, IL-2, and IL-4 during acute infection was predictive of chronic joint pain. These data suggest that a robust cytokine response is necessary for viral clearance and cytokines that are related to immune tolerance during acute infection may be protective for chronic arthritis pathogenesis.

Frequency of Chronic Joint Pain Following Chikungunya Virus Infection: A Colombian Cohort Study.

OBJECTIVE: To estimate the frequency of chronic joint pain after infection with chikungunya virus in a Latin American cohort. METHODS: A cross-sectional follow-up of a prospective cohort of 500 patients from the Atlántico Department, Colombia who were clinically diagnosed as having chikungunya virus during the 2014-2015 epidemic was conducted. Baseline symptoms and follow-up symptoms at 20 months were evaluated in serologically confirmed cases. RESULTS: Among the 500 patients enrolled, 485 had serologically confirmed chikungunya virus and reported joint pain status. Patients were predominantly adults (mean ± SD age 49 ± 16 years) and female, had an education level of high school or less, and were of Mestizo ethnicity. The most commonly affected joints were the small joints, including the wrists, ankles, and fingers. The initial virus symptoms lasted a median of 4 days (interquartile range [IQR] 3-8 days). Sixteen percent of the participants reported missing school or work (median 4 days [IQR 2-7 days]). After 20 months, one-fourth of the participants had persistent joint pain. A multivariable analysis indicated that significant predictors of persistent joint pain included college graduate status, initial symptoms of headache or knee pain, missed work, normal activities affected, ≥4 days of initial symptoms, and ≥4 weeks of initial joint pain. CONCLUSION: This is the first report to describe the frequency of chikungunya virus-related arthritis in the Americas after a 20-month follow-up. The high frequency of chronic disease highlights the need for the development of prevention and treatment methods.