scholarly journals Antibody Responses to MAP 1B and Other Cowdria ruminantium Antigens Are Down Regulated in Cattle Challenged with Tick-Transmitted Heartwater

2001 ◽  
Vol 8 (2) ◽  
pp. 388-396 ◽  
Author(s):  
S. M. Semu ◽  
T. F. Peter ◽  
D. Mukwedeya ◽  
A. F. Barbet ◽  
F. Jongejan ◽  
...  
Keyword(s):  
Tick Infestation ◽  
Repeated Exposure ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Cattle ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Cowdria Ruminantium ◽  
Infection Pressure ◽  
Specific Igg

ABSTRACT Serological diagnosis of heartwater or Cowdria ruminantium infection has been hampered by severe cross-reactions with antibody responses to related ehrlichial agents. A MAP 1B indirect enzyme-linked immunosorbent assay that has an improved specificity and sensitivity for detection of immunoglobulin G (IgG) antibodies has been developed to overcome this constraint (A. H. M. van Vliet, B. A. M. Van der Zeijst, E. Camus, S. M. Mahan, D. Martinez, and F. Jongejan, J. Clin. Microbiol. 33:2405–2410, 1995). When sera were tested from cattle in areas of endemic heartwater infection in Zimbabwe, only 33% of the samples tested positive in this assay despite a high infection pressure (S. M. Mahan, S. M. Samu, T. F. Peter, and F. Jongejan, Ann. N.Y. Acad. Sci 849:85–87, 1998). To determine underlying causes for this observation, the kinetics of MAP 1B-specific IgG antibodies in cattle after tick-transmitted C. ruminantium infection and following recovery were investigated. Sera collected weekly over a period of 52 weeks from 37 cattle, which were naturally or experimentally infected with C. ruminantium via Amblyomma hebraeum ticks, were analyzed. MAP 1B-specific IgG antibody responses developed with similar kinetics in both field- and laboratory-infected cattle. IgG levels peaked at 4 to 9 weeks after tick infestation and declined to baseline levels between 14 and 33 weeks, despite repeated exposure to infected ticks and the establishment of a carrier state as demonstrated by PCR and xenodiagnosis. Some of the serum samples from laboratory, and field-infected cattle were also analyzed by immunoblotting and an indirect fluorescent-antibody test (IFAT) to determine whether this observed seroreversion was specific to the MAP 1B antigen. Reciprocal IFAT and immunoblot MAP 1-specific antibody titres peaked at 5 to 9 weeks after tick infestation but also declined between 30 and 45 weeks. This suggests that MAP 1B-specific IgG antibody responses and antibody responses to other C. ruminantium antigens are down regulated in cattle despite repeated exposure to C. ruminantium via ticks. Significantly, serological responses to the MAP 1B antigen may not be a reliable indicator of C. ruminantium exposure in cattle in areas of endemic heartwater infection.

scholarly journals Detection of Genotype-Specific Antibody Responses to Glycoproteins B and H in Primary and Non-Primary Human Cytomegalovirus Infections by Peptide-Based ELISA

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 399
Author(s):  
Federica Zavaglio ◽  
Loretta Fiorina ◽  
Nicolás M. Suárez ◽  
Chiara Fornara ◽  
Marica De Cicco ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Primary Infection ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Background Strain ◽  
Specific Igg ◽  
Cytomegalovirus Infections ◽  
Specific Igg Antibodies

Background: Strain-specific antibodies to human cytomegalovirus (HCMV) glycoproteins B and H (gB and gH) have been proposed as a potential diagnostic tool for identifying reinfection. We investigated genotype-specific IgG antibody responses in parallel with defining the gB and gH genotypes of the infecting viral strains. Methods: Subjects with primary (n = 20) or non-primary (n = 25) HCMV infection were studied. The seven gB (gB1-7) and two gH (gH1-2) genotypes were determined by real-time PCR and whole viral genome sequencing, and genotype-specific IgG antibodies were measured by a peptide-based enzyme-linked immunosorbent assay (ELISA). Results: Among subjects with primary infection, 73% (n = 8) infected by gB1-HCMV and 63% (n = 5) infected by gB2/3-HCMV had genotype-specific IgG antibodies to gB (gB2 and gB3 are similar in the region tested). Peptides from the rarer gB4-gB7 genotypes had nonspecific antibody responses. All subjects infected by gH1-HCMV and 86% (n = 6) infected by gH2-HCMV developed genotype-specific responses. Among women with non-primary infection, gB and gH genotype-specific IgG antibodies were detected in 40% (n = 10) and 80% (n = 20) of subjects, respectively. Conclusions: Peptide-based ELISA is capable of detecting primary genotype-specific IgG responses to HCMV gB and gH, and could be adopted for identifying reinfections. However, about half of the subjects did not have genotype-specific IgG antibodies to gB.

scholarly journals A comparison of non-magnetic and magnetic beads for measuring IgG antibodies against Plasmodium vivax antigens in a multiplexed bead-based assay using Luminex technology (Bio-Plex 200 or MAGPIX)

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0238010
Author(s):  
Ramin Mazhari ◽  
Jessica Brewster ◽  
Rich Fong ◽  
Caitlin Bourke ◽  
Zoe S. J. Liu ◽  
...  
Keyword(s):  
Plasmodium Vivax ◽  
Magnetic Beads ◽  
Antibody Responses ◽  
Strongly Correlated ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Standard Curve ◽  
Luminex Technology ◽  
Specific Igg ◽  
Comparability Of Results

Multiplexed bead-based assays that use Luminex® xMAP® technology have become popular for measuring antibodies against proteins of interest in many fields, including malaria and more recently SARS-CoV-2/COVID-19. There are currently two formats that are widely used: non-magnetic beads or magnetic beads. Data are lacking regarding the comparability of results obtained using these two types of beads, and for assays run on different instruments. Whilst non-magnetic beads can only be run on flow-based instruments (such as the Luminex® 100/200™ or Bio-Plex® 200), magnetic beads can be run on both these and the newer MAGPIX® instruments. In this study we utilized a panel of purified recombinant Plasmodium vivax proteins and samples from malaria-endemic areas to measure P. vivax-specific IgG responses using different combinations of beads and instruments. We directly compared: i) non-magnetic versus magnetic beads run on a Bio-Plex® 200, ii) magnetic beads run on the Bio-Plex® 200 versus MAGPIX® and iii) non-magnetic beads run on a Bio-Plex® 200 versus magnetic beads run on the MAGPIX®. We also performed an external comparison of our optimized assay. We observed that IgG antibody responses, measured against our panel of P. vivax proteins, were moderately-strongly correlated in all three of our comparisons (pearson r>0.5 for 18/19 proteins), however higher amounts of protein were required for coupling to magnetic beads. Our external comparison indicated that results generated in different laboratories using the same coupled beads are also highly comparable (pearson r>0.7), particularly if a reference standard curve is used.

scholarly journals Systemic and Mucosal Immunizations with Fibronectin-Binding Protein FBP54 Induce Protective Immune Responses against Streptococcus pyogenes Challenge in Mice

2001 ◽  
Vol 69 (2) ◽  
pp. 924-930 ◽  
Author(s):  
Shigetada Kawabata ◽  
Eiji Kunitomo ◽  
Yutaka Terao ◽  
Ichiro Nakagawa ◽  
Ken Kikuchi ◽  
...  
Keyword(s):  
Streptococcus Pyogenes ◽  
Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Antibody Titers ◽  
Fibronectin Binding Protein ◽  
Specific Igg ◽  
Fibronectin Binding ◽  
Specific Igg Antibodies

ABSTRACT The purpose of this study was to examine the suitability of fibronectin-binding protein FBP54 as a putative vaccine forStreptococcus pyogenes infections. When the distribution of the fbp54 gene among the clinical isolates representing various M serotypes was tested by PCR and Southern blot assays, it was found that all of the strains possess this gene. Furthermore, a significant increase in immunoglobulin G (IgG) antibody titers against FBP54 was observed in sera from patients with S. pyogenesinfections compared with those from healthy volunteers (P < 0.005). Mice were immunized with FBP54 subcutaneously, orally, or nasally. An enzyme-linked immunosorbent assay revealed that antigen-specific IgG antibodies were induced in the sera of immunized mice, while high salivary levels of IgA antibodies were detected after oral and nasal immunizations. Mice subcutaneously or orally immunized with FBP54 survived significantly longer following the challenge with S. pyogenes than did nonimmunized mice (P < 0.001). These results indicate that FBP54 is a promising vaccine for the prevention of S. pyogenesinfections.

scholarly journals Influence of Exogenous Reproductive Hormones on Specific Antibody Production in Genital Secretions after Vaginal Vaccination with Recombinant Cholera Toxin B Subunit in Humans

2006 ◽  
Vol 13 (2) ◽  
pp. 202-207 ◽  
Author(s):  
Lotta Wassen ◽  
Marianne Jertborn
Keyword(s):  
Cholera Toxin ◽  
Specific Antibody ◽  
Large Scale ◽  
Reproductive Hormones ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cholera Toxin B Subunit ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Predictive Values

ABSTRACT The objective of this study was to investigate the influence of exogenous reproductive hormones on the local and systemic production of specific immunoglobulin A (IgA) and IgG antibodies after vaginal vaccination with recombinant cholera toxin subunit B (CTB). Three groups of women using either progesterone-containing intrauterine devices (n = 9), oral contraceptives (n = 8), or no hormonal contraceptive methods (n = 9) were vaginally immunized twice, 2 weeks apart. Cervical secretions, vaginal fluids, and serum were collected before and after vaccination. Total and CTB-specific IgA and IgG antibodies in genital secretions and serum were analyzed by enzyme-linked immunosorbent assay. A majority of the women presented strong CTB-specific IgA and IgG antibody responses in cervicovaginal secretions after vaccination, whereas the antitoxin responses in serum were weaker. Exogenously administered steroid hormones did not seem to have any impact on the production of specific antibodies. Both the frequencies and the magnitudes of IgA and IgG antitoxin responses in genital secretions were comparable among the three immunization groups. An association, in particular for IgA, was found between the magnitudes of the CTB-specific antibody responses in cervical secretions and vaginal fluids after vaccination. The sensitivities and positive predictive values of vaginal antibody analyses to reflect responses in cervical secretions were also high, suggesting that vaginal fluids alone might be used for evaluation of genital immune responses in large-scale vaccination studies in the future.

scholarly journals A comparison of non-magnetic and magnetic beads for measuring IgG antibodies against P. vivax antigens in a multiplexed bead-based assay using Luminex® technology (Bio-Plex® 200 or MAGPIX®)

2020 ◽  
Author(s):  
Ramin Mazhari ◽  
Jessica Brewster ◽  
Rich Fong ◽  
Caitlin Bourke ◽  
Zoe SJ Liu ◽  
...  
Keyword(s):  
Magnetic Beads ◽  
External Validation ◽  
Antibody Responses ◽  
Strongly Correlated ◽  
Reference Standard ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Standard Curve ◽  
Specific Igg ◽  
Comparability Of Results

AbstractMultiplexed bead-based assays that use Luminex xMAP® technology have become popular for measuring antibodies against proteins of interest in many fields, including malaria and more recently SARS-CoV-2/COVID-19. There are currently two formats that are widely used: non-magnetic beads or magnetic beads. Data is lacking regarding the comparability of results obtained using these two types of beads, and for assays run on different instruments. Whilst non-magnetic beads can only be run on flow-based instruments (such as the Luminex® 100/200™ or Bio-Plex® 200), magnetic beads can be run on both these and the newer MAGPIX® instruments. In this study we utilized a panel of purified recombinant Plasmodium vivax proteins and samples from malaria-endemic areas to measure P. vivax-specific IgG responses using different combinations of beads and instruments. We directly compared: i) non-magnetic versus magnetic beads run on a Bio-Plex® 200, ii) magnetic beads run on the Bio-Plex® 200 versus MAGPIX® and iii) non-magnetic beads run on a Bio-Plex® 200 versus magnetic beads run on the MAGPIX®. We also performed an external validation of our optimized assay. We observed that IgG antibody responses, measured against our panel of P. vivax proteins, were strongly correlated in all three of our comparisons, however higher amounts of protein were required for coupling to magnetic beads. Our external validation indicated that results generated in different laboratories using the same coupled beads are also highly comparable, particularly if a reference standard curve is used.

scholarly journals Evaluation of cysticercus-specific IgG (total and subclasses) and IgE antibody responses in cerebrospinal fluid samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies

2013 ◽  
Vol 71 (2) ◽  
pp. 106-109 ◽  
Author(s):  
Lisandra Akemi Suzuki ◽  
Cláudio Lúcio Rossi
Keyword(s):  
Cerebrospinal Fluid ◽  
Neurological Disorders ◽  
Taenia Solium ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Ige Antibodies ◽  
Future Studies ◽  
Specific Igg ◽  
Specific Igg Antibodies

In the present study, an enzyme-linked immunosorbent assay (ELISA) standardized with vesicular fluid of Taenia solium cysticerci was used to screen for IgG (total and subclasses) and IgE antibodies in cerebrospinal fluid (CSF) samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies and patients with other neurological disorders. The following results were obtained: IgG-ELISA: 100% sensitivity (median of the ELISA absorbances (MEA)=1.17) and 100% specificity; IgG1-ELISA: 72.7% sensitivity (MEA=0.49) and 100% specificity; IgG2-ELISA: 81.8% sensitivity (MEA=0.46) and 100% specificity; IgG3-ELISA: 63.6% sensitivity (MEA=0.12) and 100% specificity; IgG4-ELISA: 90.9% sensitivity (MEA=0.85) and 100% specificity; IgE-ELISA 93.8% sensitivity (MEA=0.60) and 100% specificity. There were no significant differences between the sensitivities and specificities in the detection of IgG-ELISA and IgE-ELISA, although in CSF samples from patients with neurocysticercosis the MEA of the IgG-ELISA was significantly higher than that of the IgE-ELISA. The sensitivity and MEA values of the IgG4-ELISA were higher than the corresponding values for the other IgG subclasses. Future studies should address the contribution of IgG4 and IgE antibodies to the physiopathology of neurocysticercosis.

Antibody responses to fluke cysteine proteinases in Paragonimus- and Fasciola-infected rats

1998 ◽  
Vol 72 (3) ◽  
pp. 187-191 ◽  
Author(s):  
T. Ikeda
Keyword(s):  
Cross Reactivity ◽  
Antibody Responses ◽  
Cysteine Proteinases ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Capture Elisa ◽  
Igm Antibody ◽  
Specific Igg ◽  
Secretory Products ◽  
Specific Igg Antibodies

AbstractIgG and IgM antibody responses to fluke cysteine proteinases in Paragonimus ohirai- and Fasciola sp.-infected rats were followed by means of cystatin capture ELISA using fluke excretory-secretory products for 10 weeks after infection. The specific IgG antibodies were detectable at week 2 postinfection in all P. ohirai-infected and some Fasciola-infected rats. Levels of specific IgG antibodies increased rapidly between week 2 and 6, and slightly thereafter, in both infected groups. From week 3, specific IgG antibody levels were higher in Fasciola-infected than P. ohirai-infected rats. Sera from infected rats did not react with heterologous cysteine proteinases throughout the infection periods. In both infected groups, the kinetic patterns of specific IgM antibody responses were similar to those of specific IgG antibody responses although the ELISA levels of the IgM antibody responses were much lower. In abnormal infections with P. ohirai metacercariae x-irradiated at 2 krad, the specific IgG antibodies were detectable at week 2 postinfection with similar ELISA values to normal P. ohirai infection, but thereafter increased little. In infections with P. westermani, for which the rat is not a suitable host, even stunted worms induced a comparable specific IgG antibody response, although the response was lower than in normal infections with P. ohirai. These results indicate that cystatin capture ELISA can distinguish clearly between Paragonimus and Fasciola infections which show immunodiagnostic cross-reactivity and is useful even in the early stages of the infection and in the infection of unsuitable hosts.

scholarly journals Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

mSphere ◽  
2018 ◽  
Vol 3 (4) ◽  
pp. e00128-18 ◽  
Author(s):  
Danka Pavliakova ◽  
Peter C. Giardina ◽  
Soraya Moghazeh ◽  
Shite Sebastian ◽  
Maya Koster ◽  
...  
Keyword(s):  
Human Serum ◽  
Lower Limit ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Correlate Of Protection ◽  
Immune Correlate ◽  
Relative Standard ◽  
Limit Of Quantitation

ABSTRACT A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysaccharide (PnPS) serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. The assay uses poly-l-lysine (PLL)-conjugated PnPS, chemically coupled to spectrally distinct Luminex microspheres. Assay validation experiments were performed using residual human serum samples obtained from 13-valent pneumococcal conjugate vaccine (13vPnC) clinical studies. Assay results are expressed as IgG antibody concentrations in micrograms per milliliter using the international reference serum, 007sp. The lower limit of quantitation (LLOQ) for all serotypes covered in the 13-plex dLIA fell within the range of 0.002 to 0.038 µg/ml serum IgG. The difference between the lower limit and upper limit of the assay range was >500-fold for all serotypes, and assay variability was <20% relative standard deviation (RSD) for all serotypes. IgG antibody measurements were shown to be serotype-specific (some cross-reactivity was observed only between the structurally related serotypes 6A and 6B as well as 19A and 19F), and no interference was observed between the serotypes when the assay was performed in the 13-plex format compared to the singleplex assays. The 13-plex dLIA platform developed by Pfizer Inc. generates up to 143 test results in a single 96-well plate and is a suitable replacement of the enzyme-linked immunosorbent assay (ELISA) platform for evaluating vaccine clinical trials. IMPORTANCE The pneumococcal enzyme-linked immunosorbent assay (ELISA) measures IgG antibodies in human serum, and it is an important assay that supports licensure of pneumococcal vaccines. The immune correlate of protection, 0.35 µg/ml of IgG antibodies, was determined by the ELISA method. Pfizer has developed a new Luminex-based assay platform to replace the ELISA. These papers describe the important work of (i) validating the Luminex-based assay and (ii) bridging the immune correlate of protection (0.35 µg/ml IgG) to equivalent values reported by the Luminex platform.

scholarly journals Antibodies to Protein but Not Glycolipid Structures Are Important for Host Defense againstMycoplasma pneumoniae

2018 ◽  
Vol 87 (2) ◽  
Author(s):  
Patrick M. Meyer Sauteur ◽  
Adrianus C. J. M. de Bruijn ◽  
Catarina Graça ◽  
Anne P. Tio-Gillen ◽  
Silvia C. Estevão ◽  
...  
Keyword(s):  
Neurological Disorders ◽  
Antibody Formation ◽  
Antibody Responses ◽  
Igg Antibodies ◽  
Wild Type ◽  
Content Type ◽  
Bruton Tyrosine Kinase ◽  
Specific Igg ◽  
Deficient Mice ◽  
Specific Igg Antibodies

ABSTRACTAntibody responses toMycoplasma pneumoniaecorrelate with pulmonaryM. pneumoniaeclearance. However,M. pneumoniae-specific IgG antibodies can cross-react with the myelin glycolipid galactocerebroside (GalC) and cause neurological disorders. We assessed whether antiglycolipid antibody formation is part of the physiological immune response toM. pneumoniae. We show that antibodies againstM. pneumoniaeproteins and glycolipids arise in serum ofM. pneumoniae-infected children and mice. Although antibodies toM. pneumoniaeglycolipids were mainly IgG, anti-GalC antibodies were only IgM. B-1a cells, shown to aid in protection against pathogen-derived glycolipids, are lacking in Bruton tyrosine kinase (Btk)-deficient mice.M. pneumoniae-infected Btk-deficient mice developedM. pneumoniae-specific IgG responses toM. pneumoniaeproteins but not toM. pneumoniaeglycolipids, including GalC. The equal recovery fromM. pneumoniaeinfection in Btk-deficient and wild-type mice suggests that pulmonaryM. pneumoniaeclearance is predominantly mediated by IgG reactive withM. pneumoniaeproteins and thatM. pneumoniaeglycolipid-specific IgG or IgM is not essential. These data will guide the development ofM. pneumoniae-targeting vaccines that avoid the induction of neurotoxic antibodies.

scholarly journals Detection of antibodies to recombinant truncated flagellin and sonicated whole cell antigen of Burkholderia pseudomallei in acute melioidosis and in healthy Bangladeshi individuals

2020 ◽  
Vol 14 (1) ◽  
pp. 47-52
Author(s):  
Md Shariful Alam Jilani ◽  
Tang Thean Hock ◽  
Sraboni Mazumder ◽  
Fahmida Rahman ◽  
Md Mohiuddin ◽  
...  
Keyword(s):  
Rural Areas ◽  
Burkholderia Pseudomallei ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Healthy Individuals ◽  
Serum Samples ◽  
Igg Antibody ◽  
Whole Cell ◽  
Cell Antigen ◽  
The Mean

Background and objectives: Several types of Burkholderia pseudomallei antigens have been used to determine the antibody response in acute and asymptomatic cases. In the present study, we have detected immunoglobulin G (IgG) antibody to recombinant truncated flagellin antigen (RTFA) of B. pseudomallei in the sera of acute melioidosis cases and healthy individuals from melioidosis endemic areas of Bangladesh by indirect enzyme-linked immunosorbent assay (ELISA). In parallel, IgG antibody to sonicated whole cell antigen (SWCA) of B. pseudomallei was determined to compare with anti-RTFA antibody. Methodology: Serum samples from culture confirmed melioidosis cases and from healthy individuals aged 21 years and above residing in melioidosis endemic rural areas were included in the study. Serum IgG antibody to RTFA and SWCA of B. pseudomallei was determined by indirect ELISA. Results: Out of 8 culture confirmed acute melioidosis cases, 7 (87.5%) and 8 (100%) were positive for anti-B. pseudomallei IgG antibodies by RTFA and SWCA methods respectively. Among 361 healthy individuals, the rate of seropositivity by RTFA-ELISA was significantly less than that of SWCA-ELISA (16.1% versus 26.8%; p = 0.001). The mean optical density (OD) of RTFA-ELISA of positive cases was significantly less than that of SWCA-ELISA in both melioidosis and healthy individuals (0.79±0.11 versus 2.4±0.08, p = 0.0001; 0.67±0.01 versus 1.27±0.02, p = 0.0001). The sensitivity and specificity of RTFA-ELISA were 88.9% and 100% respectively. Conclusion: Findings of the study suggest that multiple or combination of antigens should be used to study the seroprevalence of B. pseudomallei infection in a community. Also, prospective study is necessary to find out the duration of persistence of antibodies to different antigenic components of B. pseudomallei after exposure. Ibrahim Med. Coll. J. 2020; 14(1): 47-52

Sign in / Sign up

Export Citation Format

Share Document