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 ◽  
...  

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.

2001 ◽  
Vol 8 (2) ◽  
pp. 388-396 ◽  
Author(s):  
S. M. Semu ◽  
T. F. Peter ◽  
D. Mukwedeya ◽  
A. F. Barbet ◽  
F. Jongejan ◽  
...  

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.


2001 ◽  
Vol 69 (2) ◽  
pp. 924-930 ◽  
Author(s):  
Shigetada Kawabata ◽  
Eiji Kunitomo ◽  
Yutaka Terao ◽  
Ichiro Nakagawa ◽  
Ken Kikuchi ◽  
...  

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.


2013 ◽  
Vol 71 (2) ◽  
pp. 106-109 ◽  
Author(s):  
Lisandra Akemi Suzuki ◽  
Cláudio Lúcio Rossi

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.


1998 ◽  
Vol 72 (3) ◽  
pp. 187-191 ◽  
Author(s):  
T. Ikeda

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.


Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 516
Author(s):  
Shuyi Yang ◽  
Keith R. Jerome ◽  
Alexander L. Greninger ◽  
Joshua T. Schiffer ◽  
Ashish Goyal

While SARS-CoV-2 specific neutralizing antibodies have been developed for therapeutic purposes, the specific viral triggers that drive the generation of SARS-CoV-2 specific IgG and IgM antibodies remain only partially characterized. Moreover, it is unknown whether endogenously derived antibodies drive viral clearance that might result in mitigation of clinical severity during natural infection. We developed a series of non-linear mathematical models to investigate whether SARS-CoV-2 viral and antibody kinetics are coupled or governed by separate processes. Patients with severe disease had a higher production rate of IgG but not IgM antibodies. Maximal levels of both isotypes were governed by their production rate rather than different saturation levels between people. Our results suggest that an exponential surge in IgG levels occurs approximately 5–10 days after symptom onset with no requirement for continual antigenic stimulation. SARS-CoV-2 specific IgG antibodies appear to have limited to no effect on viral dynamics but may enhance viral clearance late during primary infection resulting from the binding effect of antibody to virus, rather than neutralization. In conclusion, SARS-CoV-2 specific IgG antibodies may play only a limited role in clearing infection from the nasal passages despite providing long-term immunity against infection following vaccination or prior infection.


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 ◽  
...  

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.


2021 ◽  
Vol 12 ◽  
Author(s):  
Jun Wu ◽  
Bo-Yun Liang ◽  
Yao-Hui Fang ◽  
Hua Wang ◽  
Xiao-Li Yang ◽  
...  

Approximately half of the SARS-CoV-2 infections occur without apparent symptoms, raising questions regarding long-term humoral immunity in asymptomatic individuals. Plasma levels of immunoglobulin G (IgG) and M (IgM) against the viral spike or nucleoprotein were determined for 25,091 individuals enrolled in a surveillance program in Wuhan, China. We compared 405 asymptomatic individuals who mounted a detectable antibody response with 459 symptomatic COVID-19 patients. The well-defined duration of the SARS-CoV-2 endemic in Wuhan allowed a side-by-side comparison of antibody responses following symptomatic and asymptomatic infections without subsequent antigen re-exposure. IgM responses rapidly declined in both groups. However, both the prevalence and durability of IgG responses and neutralizing capacities correlated positively with symptoms. Regardless of sex, age, and body weight, asymptomatic individuals lost their SARS-CoV-2-specific IgG antibodies more often and rapidly than symptomatic patients did. These findings have important implications for immunity and favour immunization programs including individuals after asymptomatic infections.


PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0238010
Author(s):  
Ramin Mazhari ◽  
Jessica Brewster ◽  
Rich Fong ◽  
Caitlin Bourke ◽  
Zoe S. J. Liu ◽  
...  

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.


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