IgG Autoantibody to Brain Beta Tubulin III Associated with Cytokine Cluster-II Discriminate Cerebral Malaria in Central India

Abstract

BackgroundThe main processes in the pathogenesis of cerebral malaria caused by Plasmodium falciparum involved sequestration of parasitized red blood cells and immunopathological responses. Among immune factors, IgG autoantibodies to brain antigens are increased in P. falciparum infected patients and correlate with disease severity in African children. Nevertheless, their role in the pathophysiology of cerebral malaria (CM) is not fully defined. We extended our analysis to an Indian population with genetic backgrounds and endemic and environmental status different from Africa to determine if these autoantibodies could be either a biomarker or a risk factor of developing CM.Methods/Principal FindingsWe investigated the significance of these self-reactive antibodies in clinically well-defined groups of P. falciparum infected patients manifesting mild malaria (MM), severe non-cerebral malaria (SM), or cerebral malaria (CM) and in control subjects from Gondia, a malaria epidemic site in central India using quantitative immunoprinting and multivariate statistical analyses. A two-fold complete-linkage hierarchical clustering allows classifying the different patient groups and to distinguish the CM from the others on the basis of their profile of IgG reactivity to brain proteins defined by PANAMA Blot. We identified beta tubulin III (TBB3) as a novel discriminant brain antigen in the prevalence of CM. In addition, circulating IgG from CM patients highly react with recombinant TBB3. Overall, correspondence analyses based on singular value decomposition show a strong correlation between IgG anti-TBB3 and elevated concentration of cluster-II cytokine (IFNγ, IL1β, TNFα, TGFβ) previously demonstrated to be a predictor of CM in the same population.Conclusions/SignificanceCollectively, these findings validate the relationship between antibody response to brain induced by P. falciparum infection and plasma cytokine patterns with clinical outcome of malaria. They also provide significant insight into the immune mechanisms associated to CM by the identification of TBB3 as a new disease-specific marker and potential therapeutic target.

Vivax infection alters peripheral B‐cell profile and induces persistent serum IgM

“…PANAMA blot method was used to analyse autoantibody repertoire of sera samples from malaria and dengue patients and healthy participants (eight sera from each group), as described earlier . In brief, brain lysate proteins (200 μg/gel; Medley, Ozyme, France) were resolved on 10% SDS‐PAGE and transferred on nitrocellulose membrane.…”

“…The bands developed in the anti‐IgG probed membranes were too faint for meaningful analysis and were not processed further. Membranes probed with anti‐IgM antibodies were dried and scanned by a high‐resolution scanner (600 DPI) using an 8‐bit linear grayscale as described . Subsequently, the proteins transferred to membranes were stained with colloidal gold (Protogold, British‐BioCell, GB), and the stained membranes scanned again.…”

Multifaceted Role of Heme during Severe Plasmodium falciparum Infections in India

“…An anti-P. falciparum HRP-2 IgM (MPFM-55A; Santa Cruz Biotechnology, France) was used as the capture antibody, an anti-P. falciparum HRP-2 IgG1 was used as a detection antibody (MPFG-55; Santa Cruz Biotechnology, France), and a horseradish peroxidase-coupled monoclonal anti-IgG1 antibody (SB77e; Abcam, France) was used as the secondary antibody for detection. A standard curve was added in duplicate to each plate, and it was obtained through serial dilutions of a serum sample from a previously described cohort with known parasitemia (18). Samples out of range of detection were more or less diluted.…”

Identification of β-tubulin as a common immunogen in gastrointestinal malignancy by mass spectrometry of colorectal cancer proteome: implications for early disease detection

“…Despite these findings, the molecular mechanism of the IgG antibody production to the βIII during infection is still unknown. It seems that the IgG response against βIII found in the disease restrictively could be a new biomarker in the Indian population [35]. Some authors have found a correlation between disease stage and the presence of anticancer antibodies [6].…”