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Journal of Clinical Laboratory Analysis Dec 2022We described a patient who exhibited a gradual increase in carbohydrate antigen 19-9 (CA19-9) concentrations for 4 years at three hospitals, with no associated...
BACKGROUND
We described a patient who exhibited a gradual increase in carbohydrate antigen 19-9 (CA19-9) concentrations for 4 years at three hospitals, with no associated clinical manifestations; however, we were unable to define the cause of this increase, forcing us to consider whether it was a false-positive result.
METHODS
Given the potential for interference, this study used multiple system detection, gradient dilution, Polyethylene glycol (PEG) precipitation and heterophilic antibody blocking assay to evaluate the reliability of CA19-9 concentration increase.
RESULTS
Analysis of the patient sample using multiple systems indicated that CA19-9 concentrations showed an obvious increase (154.0, and 889.2 IU/ml, respectively) using the Cobas E602 and Advia Centaur XP systems, and were within the reference ranges (<10 IU/ml) on other modules. PEG precipitation on the Cobas E602 and Advia Centaur XP systems reduced the CA19-9 concentration, as did heterophilic blocking tube (HBT-6, HBT-1) blockade.
CONCLUSION
CA19-9 was incorrectly identified to increase due to the presence of heterophilic antibodies. We recommend that heterophilic antibodies should be evaluated in cases with elevated CA19-9 level but no associated clinical manifestations to prevent false positives.
Topics: Humans; CA-19-9 Antigen; Reproducibility of Results; Follow-Up Studies; Antibodies, Heterophile; Carbohydrates
PubMed: 36447406
DOI: 10.1002/jcla.24792 -
Frontiers in Veterinary Science 2022The genome contributes to the uniqueness of an individual breed, and enables distinctive characteristics to be passed from one generation to the next. The allelic... (Review)
Review
The genome contributes to the uniqueness of an individual breed, and enables distinctive characteristics to be passed from one generation to the next. The allelic heterogeneity of a certain breed results in a different response to a pathogen with different genomic expression. Disease resistance in chicken is a polygenic trait that involves different genes that confer resistance against pathogens. Such resistance also involves major histocompatibility (MHC) molecules, immunoglobulins, cytokines, interleukins, T and B cells, and CD4+ and CD8+ T lymphocytes, which are involved in host protection. The MHC is associated with antigen presentation, antibody production, and cytokine stimulation, which highlight its role in disease resistance. The natural resistance-associated macrophage protein 1 (Nramp-1), interferon (IFN), myxovirus-resistance gene, myeloid differentiation primary response 88 (MyD88), receptor-interacting serine/threonine kinase 2 (RIP2), and heterophile cells are involved in disease resistance and susceptibility of chicken. Studies related to disease resistance genetics, epigenetics, and quantitative trait loci would enable the identification of resistance markers and the development of disease resistance breeds. Microbial infections are responsible for significant outbreaks and have blighted the poultry industry. Breeding disease-resistant chicken strains may be helpful in tackling pathogens and increasing the current understanding on host genetics in the fight against communicable diseases. Advanced technologies, such as the CRISPR/Cas9 system, whole genome sequencing, RNA sequencing, and high-density single nucleotide polymorphism (SNP) genotyping, aid the development of resistant breeds, which would significantly decrease the use of antibiotics and vaccination in poultry. In this review, we aimed to reveal the recent genetic basis of infection and genomic modification that increase resistance against different pathogens in chickens.
PubMed: 36439341
DOI: 10.3389/fvets.2022.1032983 -
Toxins Sep 2022Ustilaginoidins are a class of bis-naphtho-γ-pyrone mycotoxins produced by the pathogen of rice false smut, which has recently become one of the most devastating...
Ustilaginoidins are a class of bis-naphtho-γ-pyrone mycotoxins produced by the pathogen of rice false smut, which has recently become one of the most devastating diseases in rice-growing regions worldwide. In this research, the nanobody phage display library was established after an alpaca was immunized with the hemiustilaginoidin F-hapten coupled with bovine serum albumin (BSA). Heterologous antigen selection and combing trypsin with competition alternant elution methods were performed for nanobody screening. Two nanobodies, namely, Nb-B15 and Nb-C21, were selected for the establishment of indirect competitive enzyme-linked immunosorbent assays (ic-ELISAs). For Nb-B15 and Nb-C21, their IC values were 11.86 μg/mL and 11.22 μg/mL, and the detection ranges were at 3.41-19.98 μg/mL and 1.17-32.13 μg/mL, respectively. Two nanobodies had a broad spectrum to quantify the contents of total ustilaginoidins in rice samples according to cross-reactivity. The recognition mechanisms of Nb-B15 and Nb-C21 against ustilaginoidin A were elucidated by molecular modeling and docking. The key amino acid sites for the binding of Nb-B15 or Nb-C21 to ustilaginoidin A were mainly located in the FR1 and CDR1 regions. As Nb-B15 was superior to Nb-C21 in the aspects of protein expression, ELISA titer, and tolerance to organic solvents, it was selected for application in the detection of actual contaminated rice samples. The total ustilaginoidin contents of rice samples were analyzed by Nb-B15-based ic-ELISA and HPLC-DAD, between which the results were found to be consistent. The developed immunoassay based on the nanobody from the alpaca can be employed as a rapid and effective method for detection of total utilaginoidins in contaminated rice samples.
Topics: Animals; Oryza; Pyrones; Single-Domain Antibodies; Camelids, New World; Serum Albumin, Bovine; Trypsin; Mycotoxins; Immunoassay; Enzyme-Linked Immunosorbent Assay; Solvents; Haptens; Amino Acids; Antigens, Heterophile
PubMed: 36287930
DOI: 10.3390/toxins14100659 -
Ugeskrift For Laeger Sep 2022Heterophile antibodies can cause falsely elevated levels of prostate-specific antigen (PSA), which is illustrated in this case report with two patient cases. In the...
Heterophile antibodies can cause falsely elevated levels of prostate-specific antigen (PSA), which is illustrated in this case report with two patient cases. In the first case, the falsely elevated PSA resulted in thorough and unnecessary examinations with multiple transrectal biopsies causing psychological distress and a risk of infection. In the second case, a patient was diagnosed with prostate cancer, and falsely rising levels of PSA possibly resulted in prolonged treatment with medical castration. These cases underline the importance of suspecting interference, when clinical findings and PSA levels do not match.
Topics: Antibodies, Heterophile; Biopsy; Humans; Male; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms
PubMed: 36178179
DOI: No ID Found -
Frontiers in Immunology 2022We assessed if immune responses are enhanced in CD-1 mice by heterologous vaccination with two different nucleic acid-based COVID-19 vaccines: a next-generation human...
We assessed if immune responses are enhanced in CD-1 mice by heterologous vaccination with two different nucleic acid-based COVID-19 vaccines: a next-generation human adenovirus serotype 5 (hAd5)-vectored dual-antigen spike (S) and nucleocapsid (N) vaccine (AdS+N) and a self-amplifying and -adjuvanted S RNA vaccine (AAHI-SC2) delivered by a nanostructured lipid carrier. The AdS+N vaccine encodes S modified with a fusion motif to increase cell-surface expression and an N antigen modified with an Enhanced T-cell Stimulation Domain (N-ETSD) to direct N to the endosomal/lysosomal compartment and increase MHC class I and II stimulation potential. The S sequence in the AAHI-SC2 vaccine comprises the D614G mutation, two prolines to stabilize S in the prefusion conformation, and 3 glutamines in the furin cleavage region to confer protease resistance. CD-1 mice received vaccination by homologous and heterologous prime > boost combinations. Humoral responses to S were the highest with any regimen that included the AAHI-SC2 vaccine, and IgG bound to wild type and Delta (B.1.617.2) variant S1 at similar levels. An AAHI-SC2 prime followed by an AdS+N boost particularly enhanced CD4+ and CD8+ T-cell responses to both wild type and Delta S peptides relative to all other vaccine regimens. Sera from mice receiving AAHI-SC2 homologous or heterologous vaccination were found to be highly neutralizing for all pseudovirus strains tested: Wuhan, Beta, Delta, and Omicron strains. The findings here, taken in consideration with the availability of both vaccines in thermostable formulations, support the testing of heterologous vaccination by an AAHI-SC2 > AdS+N regimen in animal models of SARS-CoV-2 infection to assess its potential to provide increased protection against emerging SARS-CoV-2 variants particularly in regions of the world where the need for cold-chain storage has limited the distribution of other vaccines.
Topics: Animals; Antibodies, Neutralizing; Antigens, Heterophile; COVID-19; COVID-19 Vaccines; DNA; Humans; Mice; SARS-CoV-2; Vaccination; Vaccines, Synthetic; Viral Vaccines; mRNA Vaccines
PubMed: 35911728
DOI: 10.3389/fimmu.2022.910136 -
Life (Basel, Switzerland) Jul 2022Serum levels of cardiac troponins can be increased both with myocardial damage and in the absence of myocardial damage. In the second case, this is due to the influence... (Review)
Review
Serum levels of cardiac troponins can be increased both with myocardial damage and in the absence of myocardial damage. In the second case, this is due to the influence of false-positive factors, among which heterophilic antibodies play a significant role. Understanding the causes of the formation of heterophilic antibodies, the features and mechanisms of their effect on serum levels of cardiac troponins, is an important condition for interpreting a false-positive result due to the influence of heterophilic antibodies. This brief, descriptive review presents the causes of heterophilic-antibodies formation and discusses their effect on serum levels of cardiac troponins.
PubMed: 35892916
DOI: 10.3390/life12081114 -
Cureus Jun 2022Other than acute coronary syndrome (ACS), many clinical conditions are associated with increased cardiac troponin I (cTnI) levels. Conditions such as pulmonary embolism,...
Other than acute coronary syndrome (ACS), many clinical conditions are associated with increased cardiac troponin I (cTnI) levels. Conditions such as pulmonary embolism, acute heart failure, myocarditis, sepsis, and renal failure are commonly reported as underlying causes. Analytical interference with the cTnI assay can also lead to falsely elevated troponin I levels. That can happen due to multiple causes such as fibrin clots, heterophile antibodies, microparticles contained in the sample, rheumatoid factor, interference by bilirubin, hemolysis, and elevated alkaline phosphatase activity. Herein, we present the case of a 66-year-old female who presented with pleuritic chest pain and had a cTnI of 35.5 ng/mL post-transfusion of three units of packed red blood cells. The patient had a complete ischemic workup for ACS, including coronary angiography, which was negative for coronary artery disease.
PubMed: 35891818
DOI: 10.7759/cureus.26193 -
Frontiers in Immunology 2022Porcine islets surviving the acute injury caused by humoral rejection and IBMIR will be subjected to cellular xenograft rejection, which is predominately mediated by CD4... (Review)
Review
Porcine islets surviving the acute injury caused by humoral rejection and IBMIR will be subjected to cellular xenograft rejection, which is predominately mediated by CD4 T cells and is characterised by significant infiltration of macrophages, B cells and T cells (CD4 and CD8). Overall, the response is different compared to the alloimmune response and more difficult to suppress. Activation of CD4 T cells is both by direct and indirect antigen presentation. After activation they recruit macrophages and direct B cell responses. Although they are less important than CD4 T cells in islet xenograft rejection, macrophages are believed to be a major effector cell in this response. Rodent studies have shown that xenoantigen-primed and CD4 T cell-activated macrophages were capable of recognition and rejection of pancreatic islet xenografts, and they destroyed a graft the secretion of various proinflammatory mediators, including TNF-α, reactive oxygen and nitrogen species, and complement factors. B cells are an important mediator of islet xenograft rejection xenoantigen presentation, priming effector T cells and producing xenospecific antibodies. Depletion and/or inhibition of B cells combined with suppressing T cells has been suggested as a promising strategy for induction of xeno-donor-specific T- and B-cell tolerance in islet xenotransplantation. Thus, strategies that expand the influence of regulatory T cells and inhibit and/or reduce macrophage and B cell responses are required for use in combination with clinical applicable immunosuppressive agents to achieve effective suppression of the T cell-initiated xenograft response.
Topics: Animals; Antigens, Heterophile; Graft Rejection; Heterografts; Humans; Immunity, Cellular; Islets of Langerhans Transplantation; Swine; Transplantation, Heterologous
PubMed: 35874735
DOI: 10.3389/fimmu.2022.893985 -
Journal of Applied Microbiology Oct 2022This study aimed to provide a safe, stable and efficient SARS-CoV-2 oral vaccine development strategy based on the type III secretion system of attenuated Salmonella and...
AIMS
This study aimed to provide a safe, stable and efficient SARS-CoV-2 oral vaccine development strategy based on the type III secretion system of attenuated Salmonella and a reference for the development of a SARS-CoV-2 vaccine.
METHODS AND RESULTS
The attenuated Salmonella mutant ΔhtrA-VNP was used as a vector to secrete the antigen SARS-CoV-2 based on the type III secretion system (T3SS). The Salmonella pathogenicity island 2 (SPI-2)-encoded T3SS promoter (sifB) was screened to express heterologous antigens (RBD, NTD, S2), and the SPI-2-encoded secretion system (sseJ) was employed to secrete this molecule (psifB-sseJ-antigen, abbreviated BJ-antigen). Both immunoblotting and fluorescence microscopy revealed effective expression and secretion of the antigen into the cytosol of macrophages in vitro. The mixture of the three strains (BJ-RBD/NTD/S2, named AisVax) elicited a marked increase in the induction of IgA or IgG S-protein Abs after oral gavage, intraperitoneal and subcutaneous administration. Flow cytometric analysis proved that AisVax caused T-cell activation, as shown by a significant increase in CD44 and CD69 expression. Significant production of IgA or IgG N-protein Abs was also detected by using psifB-sseJ-N(FL), indicating the universality of this strategy.
CONCLUSIONS
Delivery of multiple SARS-CoV-2 antigens using the type III secretion system of attenuated Salmonella ΔhtrA-VNP is a potential COVID-19 vaccine strategy.
SIGNIFICANCE AND IMPACT OF THE STUDY
The attenuated Salmonella strain ΔhtrA-VNP showed excellent performance as a vaccine vector. The Salmonella SPI-2-encoded T3SS showed highly efficient delivery of SARS-COV-2 antigens. Anti-loss elements integrated into the plasmid stabilized the phenotype of the vaccine strain. Mixed administration of antigen-expressing strains improved antibody induction.
Topics: Antigens, Heterophile; Bacterial Proteins; COVID-19; COVID-19 Vaccines; Humans; Immunoglobulin A; Immunoglobulin G; SARS-CoV-2; Salmonella typhimurium; Type III Secretion Systems; Vaccine Development
PubMed: 35858677
DOI: 10.1111/jam.15720 -
BMC Genomic Data Jul 2022Previous studies have identified the carbohydrate epitope Galα1-3Galβ1-4GlcNAc-R (termed the α-galactosyl epitope), known as the α-Gal antigen as the primary...
BACKGROUND
Previous studies have identified the carbohydrate epitope Galα1-3Galβ1-4GlcNAc-R (termed the α-galactosyl epitope), known as the α-Gal antigen as the primary xenoantigen recognized by the human immune system. The α-Gal antigen is regulated by galactosyltransferase (GGTA1), and α-Gal antigen-deficient mice have been widely used in xenoimmunological studies, as well as for the immunogenic risk evaluation of animal-derived medical devices. The objective of this study was to develop α-Gal antigen-deficient rabbits by GGTA1 gene editing with the CRISPR/Cas9 system.
RESULTS
The mutation efficiency of GGTA1 gene-editing in rabbits was as high as 92.3% in F0 pups. Phenotype analysis showed that the α-Gal antigen expression in the major organs of F0 rabbits was decreased by more than 99.96% compared with that in wild-type (WT) rabbits, and the specific anti-Gal IgG and IgM antibody levels in F1 rabbits increased with increasing age, peaking at approximately 5 or 6 months. Further study showed that GGTA1 gene expression in F2-edited rabbits was dramatically reduced compared to that in WT rabbits.
CONCLUSIONS
α-Gal antigen-deficient rabbits were successfully generated by GGTA1 gene editing via the CRISPR/Cas9 system in this study. The feasibility of using these α-Gal antigen-deficient rabbits for the in situ implantation and residual immunogenic risk evaluation of animal tissue-derived medical devices was also preliminarily confirmed.
Topics: Animals; Antigens, Heterophile; CRISPR-Cas Systems; Gene Editing; Humans; Infant; Mice; Rabbits
PubMed: 35820824
DOI: 10.1186/s12863-022-01068-4