Anti-CHIKV Monoclonal antibody, Clone F37E0.13

Introduction Longitudinal data and trends about chikungunya virus (CHIKV) are critical for its control, however in Mozambique very few studies were conducted over 5 decades, between 1957 and 2013. In this study, we retrospectively investigated the occurrence, geographical distribution and trend of anti-CHIKV antibodies between 2009 and 2015 in Mozambique using serum samples from febrile patients. Methods A total of 895 serum samples collected from febrile patients for measles and rubella surveillance between 2009 and 2015 in 127 districts of Mozambique were retrospectively tested for IgM and IgG antibodies against CHIKV using a commercially available ELISA. Results The median age of patients was 2 years (IQR: 1-5 years) and 44.2% (395/895) of them were female. We found that 54 (6.0%) of samples were positive for anti-IgM chikungunya, and 160 (17.9%) were positive for anti-CHIKV IgG. Antibodies against CHIKV (IgM and IgG) were identified in serum throughout 2009 to 2015. While frequency of IgG antibodies was significantly higher in 2015 as compared to other years, frequency of IgM antibodies was homogeneous between 2009 and 2015. Antibodies against CHIKV were reported in all provinces and in 84 (66.1%) of the districts studied. Frequency of IgM and IgG antibodies was not significantly similar between age groups. Conclusion This is the largest and longest serological screening of antibodies against CHIKV in febrile patients in Mozambique and findings from this study suggest that Mozambicans from all over the country have been silently exposed to CHIKV for several years.

Alphaviruses and flaviviruses are important human pathogens that include Chikungunya virus (CHIKV), Dengue virus (DENV), and Zika virus (ZIKV), which can cause diseases in humans ranging from arthralgia to hemorrhagic fevers and microcephaly. It was previously shown that treatment with surface layer (S-layer) protein, present on the bacterial cell-envelope of Lactobacillus acidophilus, is able to inhibit viral and bacterial infections by blocking the pathogen's interaction with DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN), a trans-membrane protein that is a C-type calcium-dependent lectin. DC-SIGN is known to act as an attachment factor for several viruses including alphaviruses and flaviviruses. In the present study, we used alphaviruses as a model system to dissect the mechanism of S-layer inhibition. We first evaluated the protective effect of S-layer using 3T3 cells, either wild type or stably expressing DC-SIGN, and infecting with the alphaviruses Semliki Forest virus (SFV) and CHIKV and the flaviviruses ZIKV and DENV. DC-SIGN expression significantly enhanced infection by all four viruses. Treatment of the cells with S-layer prior to infection decreased infectivity of all viruses only in cells expressing DC-SIGN. In vitro ELISA experiments showed a direct interaction between S-layer and DC-SIGN; however, confocal microscopy and flow cytometry demonstrated that S-layer binding to the cells was independent of DC-SIGN expression. S-layer protein prevented SFV binding and internalization in DC-SIGN-expressing cells but had no effect on virus binding to DC-SIGN-negative cells. Inhibition of virus binding occurred in a time-dependent manner, with a significant reduction of infection requiring at least a 30-min pre-incubation of S-layer with DC-SIGN-expressing cells. These results suggest that S-layer has a different mechanism of action compared to mannan, a common DC-SIGN-binding compound that has an immediate effect in blocking viral infection. This difference could reflect slower kinetics of S-layer binding to the DC-SIGN present at the plasma membrane (PM). Alternatively, the S-layer/DC-SIGN interaction may trigger the activation of signaling pathways that are required for the inhibition of viral infection. Together our results add important information relevant to the potential use of L. acidophilus S-layer protein as an antiviral therapy.