-
FEMS Microbiology Ecology Jun 2016Mycobacterium ulcerans(MU) is the causative agent of Buruli ulcer, an emerging human infectious disease. However, both the ecology and life cycle of MU are poorly...
Mycobacterium ulcerans(MU) is the causative agent of Buruli ulcer, an emerging human infectious disease. However, both the ecology and life cycle of MU are poorly understood. The occurrence of MU has been linked to the aquatic environment, notably water bodies affected by human activities. It has been hypothesized that one or a combination of environmental factor(s) connected to human activities could favour growth of MU in aquatic systems. Here, we testedin vitrothe growth effect of two ubiquitous polysaccharides and five chemical components on MU at concentration ranges shown to occur in endemic regions. Real-time PCR showed that chitin increased MU growth significantly providing a nutrient source or environmental support for thebacillus, thereby, providing a focus on the association between MU and aquatic arthropods. Aquatic environments with elevated population of arthropods provide increased chitin availability and, thereby, enhanced multiplication of MU. If calcium very slightly enhanced MU growth, iron, zinc, sulphate and phosphate did not stimulate MU growth, and at the concentration ranges of this study would limit MU population in natural ecosystems.
Topics: Buruli Ulcer; Calcium; Chitin; Ecosystem; Humans; Iron; Mycobacterium ulcerans; Phosphates; Real-Time Polymerase Chain Reaction; Zinc
PubMed: 27020062
DOI: 10.1093/femsec/fiw067 -
Journal of Clinical Laboratory Analysis Feb 2023Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) inhibits infection-induced inflammation and multiorgan injury through several methods. The present study aimed to...
BACKGROUND
Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) inhibits infection-induced inflammation and multiorgan injury through several methods. The present study aimed to estimate the association of serum ITIH4 with inflammatory cytokines, multiorgan injury, and death risk in sepsis patients.
METHODS
Serum samples were collected to detect ITIH4 by enzyme-linked immunosorbent assay in 127 sepsis patients at admission (baseline), day (D)1, D3, and D7 after admission, as well as in 30 healthy controls (HCs). Additionally, 28-day mortality was recorded in sepsis patients.
RESULTS
ITIH4 was reduced in sepsis patients versus HCs (median [interquartile range]: 147.9 [78.2-208.8] vs. 318.8 [237.2-511.4] ng/ml) (p < 0.001). In sepsis patients, ITIH4 was associated with the absence of cardiovascular and cerebrovascular disease history (p = 0.021). Additionally, ITIH4 was negatively correlated with tumor necrosis factor-α (p < 0.001), interleukin (IL)-1β (p < 0.001), IL-6 (p = 0.019), IL-17A (p = 0.002), and C-reactive protein (p = 0.001), but positively related to IL-10 (p = 0.007). Moreover, ITIH4 was also inversely associated with Acute Physiology and Chronic Health Evaluation II score (p = 0.002), Sequential Organ Failure Assessment (SOFA) score (p < 0.001), SOFA-respiratory system score (p = 0.023), and SOFA-renal system score (p = 0.007). Interestingly, ITIH4 gradually increased from baseline to D7 (p < 0.001); besides, ITIH4 at baseline (p = 0.009), D1 (p = 0.002), D3 (p < 0.001), and D7 (p = 0.015) were all decreased in sepsis deaths versus sepsis survivors.
CONCLUSION
Serum ITIH4 is raised from baseline to D7 after disease onset, and it reflects the reduction of systemic inflammation, disease severity, and 28-day mortality for sepsis. However, further verification is required.
Topics: Humans; Alpha-Globulins; Cytokines; Inflammation; Multiple Organ Failure; Prognosis; Sepsis
PubMed: 36725250
DOI: 10.1002/jcla.24834 -
Animal Biotechnology Jan 2019Foot-and-mouth disease (FMD) is an acute, highly contagious, and economically devastating viral disease of domestic and wildlife species. For effective implementation of...
Foot-and-mouth disease (FMD) is an acute, highly contagious, and economically devastating viral disease of domestic and wildlife species. For effective implementation of FMD control program, there is an imperative need for developing a rapid, sensitive, and specific diagnostics which help in the identification of serotypes involved in the outbreaks. The humoral immune response of the Camelidae is unique since in these animals 75% of circulating antibodies are constituted by heavy-chain antibodies and 25% are conventional immunoglobulin with two identical heavy chains. In the present study, we developed and characterized FMD virus-specific single-domain heavy-chain antibodies (VHHs) against inactivated whole-virus antigens of FMDV serotypes O (INDR2/1975), A (IND40/2000), and Asia 1 (IND63/1972) vaccine strains. After six rounds of panning and enrichment, these VHHs were stably expressed in Escherichia coli cells. The VHHs directed against outer capsid proteins of FMD virus were successfully utilized as the capture antibody in liquid-phase blocking ELISA (LPBE) thus replacing rabbit coating antibodies. Our study demonstrated the utility of FMD virus-specific VHHs as potential candidates in FMD research and diagnostic application.
Topics: Animals; Antibodies, Viral; Antibody Specificity; Camelus; Capsid Proteins; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Foot-and-Mouth Disease; Foot-and-Mouth Disease Virus; Male; Single-Domain Antibodies; Species Specificity
PubMed: 29527970
DOI: 10.1080/10495398.2018.1433191 -
Clinical Lymphoma, Myeloma & Leukemia Apr 2018Heavy/light chain (HLC) assay can quantify involved as well as uninvolved immunoglobulin pairs and is used to detect monoclonal proteins.
BACKGROUND
Heavy/light chain (HLC) assay can quantify involved as well as uninvolved immunoglobulin pairs and is used to detect monoclonal proteins.
PATIENTS AND METHODS
We compared the sensitivity between HLC assay and serum protein electrophoresis, serum immunofixation electrophoresis (IFE), and free light chain (FLC) assay in patients with symptomatic multiple myeloma (n = 111) whose responses were stable disease or better.
RESULTS
Among patients with negative IFE and normal FLC ratios, 84.4% (38 of 45) and 80% (36 of 45) exhibited normal HLC ratios and no pair suppression, respectively (13.3% [6 of 45], moderate pair suppression and 6.7% [3 of 45], severe pair suppression). The lower the monoclonal protein levels, the more the possibility that the patients had normal HLC ratios and no matched pair suppression (both P < .000001). HLC ratios or pair suppression combined with IFE results and FLC ratios were more sensitive for detecting monoclonal proteins than were IFE results and FLC ratios alone (P = .016 and .0039, respectively). A combination of all 4 methods (IFE, FLC, HLC, and pair suppression) was far more sensitive than were IFE findings plus FLC ratios alone (P = .00024).
CONCLUSION
Abnormal HLC ratios and HLC-matched pair suppression can increase the sensitivity for detecting residual disease in patients with multiple myeloma with deep responses.
Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Blood Protein Electrophoresis; Female; Humans; Immunoglobulin Heavy Chains; Immunoglobulin Light Chains; Male; Middle Aged; Multiple Myeloma; Myeloma Proteins; Neoplasm, Residual; Prospective Studies; Sensitivity and Specificity
PubMed: 29472112
DOI: 10.1016/j.clml.2018.01.008 -
Circulation Research Aug 2022
Topics: Animals; Heart; Mice; Myocardium; Myosin Heavy Chains; Organogenesis
PubMed: 35862119
DOI: 10.1161/CIRCRESAHA.122.321461 -
Arteriosclerosis, Thrombosis, and... Jan 2017The importance of maintaining contractile function in aortic smooth muscle cells (SMCs) is evident by the fact that heterozygous mutations in the major structural... (Review)
Review
The importance of maintaining contractile function in aortic smooth muscle cells (SMCs) is evident by the fact that heterozygous mutations in the major structural proteins or kinases controlling contraction lead to the formation of aneurysms of the ascending thoracic aorta that predispose to life-threatening aortic dissections. Force generation by SMC requires ATP-dependent cyclic interactions between filaments composed of SMC-specific isoforms of α-actin (encoded by ACTA2) and myosin heavy chain (MYH11). ACTA2 and MYH11 mutations are predicted or have been shown to disrupt this cyclic interaction predispose to thoracic aortic disease. Movement of the myosin motor domain is controlled by phosphorylation of the regulatory light chain on the myosin filament, and loss-of-function mutations in the dedicated kinase for this phosphorylation, myosin light chain kinase (MYLK) also predispose to thoracic aortic disease. Finally, a mutation in the cGMP-activated protein kinase (PRKG1) results in constitutive activation of the kinase in the absence of cGMP, thus driving SMC relaxation in part through increased dephosphorylation of the regulatory light chain and predisposes to thoracic aortic disease. Furthermore, SMCs cannot generate force without connections to the extracellular matrix through focal adhesions, and mutations in the major protein in the extracellular matrix, fibrillin-1, linking SMCs to the matrix also cause thoracic aortic disease in individuals with Marfan syndrome. Thus, disruption of the ability of the aortic SMC to generate force through the elastin-contractile units in response to pulsatile blood flow may be a primary driver for thoracic aortic aneurysms and dissections.
Topics: Actins; Aortic Dissection; Animals; Aortic Aneurysm, Thoracic; Calcium-Binding Proteins; Cyclic GMP-Dependent Protein Kinase Type I; Dilatation, Pathologic; Elastin; Genetic Markers; Genetic Testing; Heredity; Humans; Mechanotransduction, Cellular; Muscle, Smooth, Vascular; Mutation; Myocytes, Smooth Muscle; Myosin Heavy Chains; Myosin-Light-Chain Kinase; Phenotype; Pulsatile Flow; Vasoconstriction
PubMed: 27879251
DOI: 10.1161/ATVBAHA.116.303229 -
The Journal of Allergy and Clinical... Mar 2019Predominantly antibody deficiencies (PADs) are the most common primary immunodeficiencies, characterized by hypogammaglobulinemia and inability to generate effective... (Comparative Study)
Comparative Study
BACKGROUND
Predominantly antibody deficiencies (PADs) are the most common primary immunodeficiencies, characterized by hypogammaglobulinemia and inability to generate effective antibody responses.
OBJECTIVE
We intended to report most common monogenic PADs and to investigate how patients with PAD who were primarily diagnosed as suffering from agammaglobulinemia, hyper-IgM (HIgM) syndrome, and common variable immunodeficiency (CVID) have different clinical and immunological findings.
METHODS
Stepwise next-generation sequencing and Sanger sequencing were performed for confirmation of the mutations in the patients clinically diagnosed as suffering from agammaglobulinemia, HIgM syndrome, and CVID.
RESULTS
Among 550 registered patients, the predominant genetic defects associated with agammaglobulinemia (48 Bruton's tyrosine kinase [BTK] and 6 μ heavy chain deficiencies), HIgM syndrome (21 CD40 ligand and 7 activation-induced cytidine deaminase deficiencies), and CVID (17 lipopolysaccharides-responsive beige-like anchor deficiency and 12 atypical Immunodeficiency, Centromeric instability, and Facial dysmorphism syndromes) were identified. Clinical disease severity was significantly higher in patients with μ heavy chain and CD40 ligand mutations compared with patients with BTK (P = .003) and activation-induced cytidine deaminase (P = .009) mutations. Paralysis following live polio vaccination was considerably higher in patients with μ heavy chain deficiency compared with BTK deficiency (P < .001). We found a genotype-phenotype correlation among patients with BTK mutations regarding clinical manifestation of meningitis and chronic diarrhea. Surprisingly, we noticed that first presentations in most patients with Immunodeficiency, Centromeric instability, and Facial dysmorphism were respiratory complications (P = .008), whereas first presentations in patients with lipopolysaccharides-responsive beige-like anchor deficiency were nonrespiratory complications (P = .008).
CONCLUSIONS
This study highlights similarities and differences in the clinical and genetic spectrum of the most common PAD-associated gene defects. This comprehensive comparison will facilitate clinical decision making, and improve prognosis and targeted treatment.
Topics: Adolescent; Adult; Agammaglobulinaemia Tyrosine Kinase; Agammaglobulinemia; CD40 Ligand; Child; Child, Preschool; Common Variable Immunodeficiency; Diarrhea; Female; Genetic Association Studies; Humans; Hyper-IgM Immunodeficiency Syndrome; Immunoglobulin mu-Chains; Male; Meningitis; Mutation; Poliomyelitis; Severity of Illness Index; Young Adult
PubMed: 30240888
DOI: 10.1016/j.jaip.2018.09.004 -
Kidney International Jul 2021Heavy chain/light chain (HLC) antibodies target conformational epitopes at the junctions of the heavy chain and light chain constant regions (CH1 and CL) of serum IgGκ,...
Immunofluorescence staining for immunoglobulin heavy chain/light chain on kidney biopsies is a valuable ancillary technique for the diagnosis of monoclonal gammopathy-associated kidney diseases.
Heavy chain/light chain (HLC) antibodies target conformational epitopes at the junctions of the heavy chain and light chain constant regions (CH1 and CL) of serum IgGκ, IgGλ, IgAκ, IgAλ, IgMκ, and IgMλ to provide quantitation of intact HLC pairs. Here, we developed an HLC tissue immunofluorescence protocol to test if it can complement conventional immunofluorescence in the diagnosis of monoclonal gammopathy-associated kidney diseases. HLC immunofluorescence was performed on archived frozen tissue of 104 kidney biopsies. The sensitivity and specificity of HLC immunofluorescence was confirmed by testing cases of lupus nephritis, other polyclonal immunoglobulin nephropathies, and light chain nephropathies (light chain amyloidosis and deposition disease). Testing of ten cases of the IgG variant of proliferative glomerulonephritis with monoclonal immunoglobulin deposits excluded monoclonal deposits in two by revealing positivity for IgGκ and IgGλ. Testing of 12 cases of monotypic IgA nephropathy excluded monoclonal deposits in six by revealing staining for IgAκ and IgAλ. Testing of six cases of monotypic fibrillary glomerulonephritis excluded monoclonal deposits in three by revealing positivity for IgGκ and IgGλ. None of 14 cases of glomerulonephritis in which HLC immunofluorescence unmasked polytypic deposits were associated with a serum or urine monoclonal immunoglobulins matching the conventional immunofluorescence results. HLC immunofluorescence outperformed paraffin immunofluorescence and IgG subclass staining in 10/13 (77%) of cases. Testing of 18 cases of cryoglobulinemic glomerulonephritis showed better correlation with serum cryoprecipitate immunofixation than conventional immunofluorescence with regards to the type of cryoglobulin in 47% of cases. Thus, HLC immunofluorescence is a valuable ancillary technique in kidney pathology for the diagnosis of monoclonal gammopathy-associated nephropathies, and could be utilized to confirm or exclude the monoclonal nature of deposits.
Topics: Biopsy; Fluorescent Antibody Technique; Glomerulonephritis; Humans; Immunoglobulin Heavy Chains; Immunoglobulin Light Chains; Kidney; Paraproteinemias; Staining and Labeling
PubMed: 33774084
DOI: 10.1016/j.kint.2021.02.038 -
Biochimie Apr 2015The deposition of misfolded peptides and proteins in the form of amyloid fibrils is the hallmark of nearly fifty medical disorders, including Alzheimer's disease,... (Review)
Review
The deposition of misfolded peptides and proteins in the form of amyloid fibrils is the hallmark of nearly fifty medical disorders, including Alzheimer's disease, Parkinson's disease, prion diseases and type II diabetes. These disorders, referred to as amyloidoses, generally become apparent late in life. Their psycho-sociological and economic incidence in western societies will be therefore considerable in the coming decades due to the ageing of the population. Neither preventing nor curative treatments are available yet. These disorders constitute therefore a medical challenge of great importance. Thus, an extensive research is being carried out to understand, at the molecular level, (i) how amyloidogenic proteins misfold and convert from their soluble form into amyloid fibrils, and (ii) how these aggregates or some of their oligomeric precursor species are toxic. The formation of amyloid fibrils proceeds through a complex nucleation/polymerisation mechanism with the formation of various species, including small oligomers. In this review, we focus on how VHHs or nanobodies, the antigen-binding domains of camelid heavy-chain antibodies, are being increasingly used to characterise each of the species formed on the pathway of fibril formation in terms of structure, stability, kinetics of formation and toxicity. We first introduce the characteristic features of nanobodies compared to those of conventional antibody fragments. Thereafter, we discuss how nanobodies, due to their unique properties, are used as probes to dissect the molecular mechanisms of misfolding and aggregation of six proteins associated with diseases, i.e. human lysozyme, β2-microglobulin, α-synuclein, prion, polyadenylate binding protein nuclear 1 and amyloid β-peptide. A brief general presentation of each disease and the associated peptide/protein is also provided. In addition, we discuss how nanobodies could be used as early diagnostic tools and as novel strategies to treat diseases associated with protein misfolding and aggregation.
Topics: Animals; Camelids, New World; Humans; Immunoglobulin Heavy Chains; Protein Aggregation, Pathological; Proteostasis Deficiencies; Single-Domain Antibodies
PubMed: 25656912
DOI: 10.1016/j.biochi.2015.01.012 -
Developmental and Comparative Immunology Mar 2021In salmonids, stress responses increase cortisol levels and disease susceptibility, including to Flavobacterium psychrophilum (Fp), the causative agent of BCWD. A...
In salmonids, stress responses increase cortisol levels and disease susceptibility, including to Flavobacterium psychrophilum (Fp), the causative agent of BCWD. A BCWD-resistant line (R-line) of rainbow trout was used here to investigate potential differences in immunoglobulin response after a combined treatment of cortisol and Fp, as compared to a susceptible (S-line) control line. Expression of membrane and secreted immunoglobulin heavy chains mu and tau were determined by RT-qPCR in spleen and anterior kidney. Cortisol treatment did not affect B cell development in the anterior kidney, but delayed IgM responses at the early stage of infection in the spleen of both lines. An earlier IgM response was a determining factor in differential disease progression between resistant- and susceptible fish after Fp-challenge. S-line fish had a delayed and exacerbated IgM response after cortisol implant indicative of a detrimental cycle of sustained IgM responses and high pathogen loads. In contrast, R-line fish had delayed but milder, and protective IgM responses and cleared pathogen successfully. Fp challenge after cortisol implant increased serum cortisol levels on days 3 and 5 compared to mock treatments in S-line fish, but only on day 3 in R-line. Hence, combined cortisol treatment and Fp challenge differentially modulated B cell activation and Fp loads in BCWD-resistant and susceptible lines of rainbow trout. We propose that under conditions of increased stress, BCWD-resistant fish may remain immunologically better equipped to respond to infections compared to BCWD susceptible fish.
Topics: Animals; Bacterial Load; Disease Resistance; Fish Diseases; Flavobacteriaceae Infections; Flavobacterium; Hydrocortisone; Immunoglobulin M; Kidney; Leukocytes; Oncorhynchus mykiss; Spleen
PubMed: 33212092
DOI: 10.1016/j.dci.2020.103921