Assessment of the Functional Integrity of the Humoral Immune Response: The Plaque-Forming Cell Assay and the Enzyme-Linked Immunosorbent Assay

Methods ◽  
1999 ◽  
Vol 19 (1) ◽  
pp. 3-7 ◽  
Author(s):  
Susan D. Wilson ◽  
Albert E. Munson ◽  
B.Jean Meade
Keyword(s):  
Immune Response ◽  
Immunosorbent Assay ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Humoral Immune ◽  
Functional Integrity ◽  
Cell Assay

scholarly journals Immunoblot Analysis of Humoral Immune Response to Helicobacter pylori in Children with and without Duodenal Ulcer

2000 ◽  
Vol 38 (5) ◽  
pp. 1777-1781 ◽  
Author(s):  
Gifone A. Rocha ◽  
Andreia M. R. Oliveira ◽  
Dulciene M. M. Queiroz ◽  
Anfrisina S. T. Carvalho ◽  
Ana M. M. F. Nogueira
Keyword(s):  
Immune Response ◽  
Duodenal Ulcer ◽  
Helicobacter Pylori ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Method ◽  
Predictive Values ◽  
Humoral Immune ◽  
Urease Test ◽  
H Pylori

Several studies have demonstrated that enzyme-linked immunosorbent assay is not a sensitive and specific method to diagnoseHelicobacter pylori infection in children, especially in the younger ones. Since serum immune response can also be determined by immunoblotting and it permits the detection of antibodies to virulence factors such as CagA and VacA, we evaluated the accuracy of a commercial immunoblotting test to diagnose H. pyloriinfection and to assess the humoral immune response to differentH. pylori antigens in 122 children who underwent upper gastrointestinal endoscopy. The presence of H. pylori was determined in antral biopsy specimens by culture, preformed urease test, and histological analysis. H. pylori was identified by microbiological and histopathological methods in 66 children (including all of the 21 who had duodenal ulcer). Antibodies toH. pylori were detected in 63 infected children and in 8 noninfected ones. The sensitivity, specificity, and positive and negative predictive values of the immunoblotting test were 95.5, 85.7, 88.7, and 94.1%, respectively. The number of immunoreactive bands increased with age (P = 0.003), and the bands of 35 kDa (P = 0.013); 89 kDa, the VacA antigen (P = 0.001); and 116 kDa, the CagA antigen (P = 0.00004) were more frequently observed in older children. The frequency of the bands of 89 kDa (P = 0.001) and 116 kDa (P = 0.03) was higher in children with duodenal ulcer than in H. pylori-positive children without the disease. In conclusion, the immunoblotting test appears to be useful for the diagnosis of H. pylori infection in children, even in the younger ones.

scholarly journals Characterization of the Humoral Immune Response to Porcine Reproductive and Respiratory Syndrome (PRRS) Virus Infection

1995 ◽  
Vol 7 (3) ◽  
pp. 305-312 ◽  
Author(s):  
Kyoung-Jin Yoon ◽  
Jeffrey J. Zimmerman ◽  
Sabrina L. Swenson ◽  
Michael J. McGinley ◽  
Ken A. Eernisse ◽  
...  
Keyword(s):  
Immune Response ◽  
Humoral Immune Response ◽  
Fluorescent Antibody ◽  
Viral Proteins ◽  
Immunoblot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Prrs Virus ◽  
Humoral Immune ◽  
Antibody Titers ◽  
Rate Of Decline

The development of the humoral immune response against porcine reproductive and respiratory syndrome (PRRS) virus was monitored by an indirect fluorescent antibody (IFA) test, immunoperoxidase monolayer assay (IPMA), enzyme-linked immunosorbent assay (ELISA), and serum virus neutralization (SVN) test over a 105-day period in 8 pigs experimentally infected with ATCC strain VR-2402. Specific antibodies against PRRS virus were first detected by the IFA test, IPMA, ELISA, and the SVN test 9-11, 5-9, 9-13, and 9-28 days postinoculation (PI), respectively, and reached their maximum values by 4-5, 5-6, 4-6, and 10-11 weeks PI, respectively, thereafter. After reaching maximum value, all assays showed a decline in antibody levels. Assuming a constant rate of antibody decay, it was estimated by regression analysis that the ELISA, IFA, IPMA, and SVN antibody titers would approach the lower limits of detection by approximately days 137, 158, 324, and 356 PI, respectively. In this study, the immunoperoxidase monolayer assay appeared to offer slightly better performance relative to the IFA test, ELISA, and SVN test in terms of earlier detection and slower rate of decline in antibody titers. Western immunoblot analysis revealed that antibody specific for the 15-kD viral protein was present in all pigs by 7 days PI and persisted throughout the 105-day observation period. Initial detection of antibodies to the 19-, 23-, and 26-kD proteins varied among pigs, ranging from 9 to 35 days PI. Thereafter, the antibody responses to these 3 viral proteins of PRRS virus continued to be detected throughout the 105-day study period. These results clearly indicate that the 15-kD protein is the most immunogenic of the 4 viral proteins identified and may provide the antigenic basis for the development of improved diagnostic tests for the detection of PRRS virus antibodies.

scholarly journals Dynamics of the Humoral Immune Response to a Prime-Boost Ebola Vaccine: Quantification and Sources of Variation

2019 ◽  
Vol 93 (18) ◽  
Author(s):  
Chloé Pasin ◽  
Irene Balelli ◽  
Thierry Van Effelterre ◽  
Viki Bockstal ◽  
Laura Solforosi ◽  
...  
Keyword(s):  
Immune Response ◽  
Antibody Response ◽  
Humoral Immune Response ◽  
Phase 1 ◽  
Humoral Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Phase 1 Trials ◽  
Humoral Immune ◽  
Boost Immunization

ABSTRACT The Ebola vaccine based on Ad26.ZEBOV/MVA-BN-Filo prime-boost regimens is being evaluated in multiple clinical trials. The long-term immune response to the vaccine is unknown, including factors associated with the response and variability around the response. We analyzed data from three phase 1 trials performed by the EBOVAC1 Consortium in four countries: the United Kingdom, Kenya, Tanzania, and Uganda. Participants were randomized into four groups based on the interval between prime and boost immunizations (28 or 56 days) and the sequence in which Ad26.ZEBOV and MVA-BN-Filo were administered. Consecutive enzyme-linked immunosorbent assay (ELISA) measurements of the IgG binding antibody concentrations against the Kikwit glycoprotein (GP) were available for 177 participants to assess the humoral immune response up to 1 year postprime. Using a mathematical model for the dynamics of the humoral response, from 7 days after the boost immunization up to 1 year after the prime immunization, we estimated the durability of the antibody response and the influence of different factors on the dynamics of the humoral response. Ordinary differential equations (ODEs) described the dynamics of antibody response and two populations of antibody-secreting cells (ASCs), short-lived (SL) and long-lived (LL). Parameters of the ODEs were estimated using a population approach. We estimated that half of the LL ASCs could persist for at least 5 years. The vaccine regimen significantly affected the SL ASCs and the antibody peak but not the long-term response. The LL ASC compartment dynamics differed significantly by geographic regions analyzed, with a higher long-term antibody persistence in European subjects. These differences could not be explained by the observed differences in cellular immune response. IMPORTANCE With no available licensed vaccines or therapies, the West African Ebola virus disease epidemic of 2014 to 2016 caused 11,310 deaths. Following this outbreak, the development of vaccines has been accelerated. Combining different vector-based vaccines as heterologous regimens could induce a durable immune response, assessed through antibody concentrations. Based on data from phase 1 trials in East Africa and Europe, the dynamics of the humoral immune response from 7 days after the boost immunization onwards were modeled to estimate the durability of the response and understand its variability. Antibody production is maintained by a population of long-lived cells. Estimation suggests that half of these cells can persist for at least 5 years in humans. Differences in prime-boost vaccine regimens affect only the short-term immune response. Geographical differences in long-lived cell dynamics were inferred, with higher long-term antibody concentrations induced in European participants.

Expression and Clinical Significance of Sperm-Associated Antigen 9 in Patients With Endometrial Carcinoma

2012 ◽  
Vol 22 (1) ◽  
pp. 87-93 ◽  
Author(s):  
Peng Yu ◽  
Lei Yan ◽  
Hui Zhang ◽  
Xiaoyan Lin ◽  
Xingbo Zhao
Keyword(s):  
Immune Response ◽  
Endometrial Cancer ◽  
Endometrial Carcinoma ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Humoral Immune ◽  
Significant Difference ◽  
New Type ◽  
Grade 3

ObjectiveTo investigate the expression and humoral immune response of sperm-associated antigen 9 (SPAG9) in endometri al carcinoma.MethodsSperm-associated antigen 9 gene expression levels were evaluated in endometrial carcinoma, endometrial hyperplasia, adjacent tissues, and normal endometrial tissues by reverse transcriptase-polymerase chain reaction, immunohistochemistry, and Western blot. Sperm-associated antigen 9 concentration in serum samples from 10 healthy women, 20 women with benign diseases, and 50 women with endometrial carcinoma was detected by enzyme-linked immunosorbent assay.Results(1) Sperm-associated antigen 9 antibodies were detected in approximately 72% of patients with endometrial cancer but not in healthy controls. (2) A significant difference has been found among pathological types and degrees (P < 0.05), and it was also found to be expressed in transferred lymph nodes. (3) Sperm-associated antigen 9 serum concentration (ng/mL) of patients with endometrial carcinoma is significantly higher than those of the healthy group (P < 0.05). Patients harboring grade 3 endometrial carcinoma were found to have significantly higher SPAG9 concentrations than those of grade 1/grade 2 (P = 0.003).ConclusionsSPAG9 is positively expressed in endometrial cancer, and with a high humoral immune response in patients. It may serve as a new type of endometrial cancer markers for early detection, diagnosis and treatment.

scholarly journals Generation and Characterization of Human Monoclonal scFv Antibodies against Helicobacter pylori Antigens

2002 ◽  
Vol 70 (8) ◽  
pp. 4158-4164 ◽  
Author(s):  
Nicole Reiche ◽  
Andreas Jung ◽  
Thomas Brabletz ◽  
Tanja Vater ◽  
Thomas Kirchner ◽  
...  
Keyword(s):  
Immune Response ◽  
Helicobacter Pylori ◽  
Phage Display ◽  
Humoral Immune Response ◽  
Vaccine Development ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Humoral Immune ◽  
Single Chain Fv ◽  
H Pylori

ABSTRACT Infection with Helicobacter pylori is chronic despite a vigorous cellular and humoral immune response and causes severe pathology in some patients. In this study, phage display was used as a new approach in order to investigate the role of the host's humoral immune response in the pathogenesis of H. pylori gastritis. Human monoclonal single-chain Fv (scFv) antibody fragments against H. pylori cell lysate and the H. pylori urease were isolated from an immune phage display library, constructed from peripheral blood lymphocytes of an H. pylori-infected patient. After affinity selection, 23% of the clones tested showed binding activity against a lysate of the H. pylori Sydney strain in enzyme-linked immunosorbent assay (ELISA) and 9% bound the H. pylori urease. Further characterization by PCR-fingerprint analysis and sequencing revealed that two closely related H. pylori binders and one antiurease scFv could be isolated. The selected scFvs were highly specific as analyzed by ELISA and immunoblots using various bacterial lysates and recombinant proteins. Analysis of the humoral immune response following H. pylori infection using human monoclonal antibodies might contribute to a better understanding of the pathogenesis of the disease. Moreover, using immune phage display libraries, it might be possible for relevant epitopes of H. pylori antigens to be determined, which might be of use for vaccine development.

scholarly journals Analysis of the Humoral Immune Response toChlamydia Outer Membrane Protein 2

1998 ◽  
Vol 5 (3) ◽  
pp. 313-318 ◽  
Author(s):  
Per Mygind ◽  
Gunna Christiansen ◽  
Kenneth Persson ◽  
Svend Birkelund
Keyword(s):  
Immune Response ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Humoral Immune Response ◽  
Chlamydia Pneumoniae ◽  
Chlamydia Psittaci ◽  
Enzyme Linked Immunosorbent Assay ◽  
Structural Protein ◽  
Humoral Immune

ABSTRACT The humoral immune response to Chlamydia outer membrane protein 2 (Omp2) was studied. Omp2 is a highly genus-conserved structural protein of all Chlamydia species, containing a variable N-terminal fragment. To analyze where the immunogenic parts were localized, seven highly purified truncated fusion proteins constituting different regions of the protein were produced (Chlamydia pneumoniae-Omp2aa23-aa93,Chlamydia psittaci-Omp2aa23-aa94, andChlamydia trachomatis-Omp2aa23-aa84, aa87-aa547, aa23-aa182, aa167-aa434, aa420-aa547). By an enzyme-linked immunosorbent assay with serologically defined patient sera, Omp2 was found to be a major immunogen of both C. pneumoniae andC. trachomatis infections (P ≪ 0.0001). The humoral immune responses were not confined to any particular region of the Omp2 protein, and no species-specific anti-Omp2 immunoglobulins were detected.

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