scholarly journals Human Milk from Previously COVID-19-Infected Mothers: The Effect of Pasteurization on Specific Antibodies and Neutralization Capacity

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1645
Author(s):  
Britt J. van Keulen ◽  
Michelle Romijn ◽  
Albert Bondt ◽  
Kelly A. Dingess ◽  
Eva Kontopodi ◽  
...  
Keyword(s):  
Human Milk ◽  
Passive Immunization ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Neutralization Capacity ◽  
N Protein ◽  
Unpasteurized Milk ◽  
Lactating Mothers ◽  
Neutralizing Capacity ◽  
Antibody Levels

Background: Since the outbreak of coronavirus disease 2019 (COVID-19), many put their hopes in the rapid availability of effective immunizations. Human milk, containing antibodies against syndrome coronavirus 2 (SARS-CoV-2), may serve as means of protection through passive immunization. We aimed to determine the presence and pseudovirus neutralization capacity of SARS-CoV-2 specific IgA in human milk of mothers who recovered from COVID-19, and the effect of pasteurization on these antibodies. Methods: This prospective case control study included lactating mothers, recovered from (suspected) COVID-19 and healthy controls. Human milk and serum samples were collected. To assess the presence of SARS-CoV-2 antibodies we used multiple complementary assays, namely ELISA with the SARS-CoV-2 spike protein (specific for IgA and IgG), receptor binding domain (RBD) and nucleocapsid (N) protein for IgG in serum, and bridging ELISA with the SARS-CoV-2 RBD and N protein for specific Ig (IgG, IgM and IgA in human milk and serum). To assess the effect of pasteurization, human milk was exposed to Holder (HoP) and High Pressure Pasteurization (HPP). Results: Human milk contained abundant SARS-CoV-2 antibodies in 83% of the proven cases and in 67% of the suspected cases. Unpasteurized milk with and without these antibodies was found to be capable of neutralizing a pseudovirus of SARS-CoV-2 in (97% and 85% of the samples respectively). After pasteurization, total IgA antibody levels were affected by HoP, while SARS-CoV-2 specific antibody levels were affected by HPP. Pseudovirus neutralizing capacity of the human milk samples was only retained with the HPP approach. No correlation was observed between milk antibody levels and neutralization capacity. Conclusions: Human milk from recovered COVID-19-infected mothers contains SARS-CoV-2 specific antibodies which maintained neutralization capacity after HPP. All together this may represent a safe and effective immunization strategy after HPP.

scholarly journals Breastmilk; a source of SARS-CoV-2 specific IgA antibodies

2020 ◽  
Author(s):  
Britt J van Keulen ◽  
Michelle Romijn ◽  
Albert Bondt ◽  
Kelly A Dingess ◽  
Eva Kontopodi ◽  
...  
Keyword(s):  
Rapid Development ◽  
Neutralization Capacity ◽  
N Protein ◽  
Lactating Mothers ◽  
Neutralizing Capacity ◽  
Before And After ◽  
Additional Protection ◽  
Good Hygiene ◽  
Control Study

Background: Since the outbreak of COVID-19, many put their hopes in the rapid development of effective immunizations. For now patient isolation, physical distancing and good hygiene are the sole measures for prevention. Processed breast milk with antibodies against SaRS-CoV-2 may serve as additional protection. We aimed to determine the presence and neutralization capacity of antibodies against SaRS-CoV-2 in breastmilk of mothers who have recovered from COVID-19. Methods: This prospective case control study included lactating mothers, recovered from (suspected) COVID-19 and healthy controls. Serum and breastmilk was collected. To assess the presence of antibodies in breastmilk and serum, we used multiple complementary assays, namely ELISA with the SARS-CoV-2 spike protein, SARS-CoV-2 receptor binding domain (RBD) and with the SARS-CoV-2 nucleocapsid (N) protein for IgG and bridging ELISA with the SARS-CoV-2 RBD and N protein for total Ig. To assess the effect of pasteurization breastmilk was exposed to Holder Pasteurization and High Pressure Pasteurization. Results: Breastmilk contained antibodies against SARS-CoV-2 using any of the assays in 24 out of 29 (83%) proven cases, in six out of nine (67%) suspected cases and in none of the 13 controls. In vitro neutralization of SARS-CoV-2 clinical isolate virus strain was successful in a subset of serum (13%) and milk samples (26%). Although after pasteurization of the milk SARS-CoV-2 antibodies were detected with both methods of pasteurization, virus neutralizing capacity of those antibodies was only retained with the HPP approach. Conclusion: Breastmilk of mothers who recovered from COVID-19 contains significant amounts of IgA against SARS-CoV-2, both before and after pasteurization.

scholarly journals Seven-month kinetics of SARS-CoV-2 antibodies and protective role of pre-existing antibodies to seasonal human coronaviruses on COVID-19

2021 ◽  
Author(s):  
Natalia Ortega ◽  
Marta Ribes ◽  
Marta Vidal ◽  
Rocío Rubio ◽  
Ruth Aguilar ◽  
...  
Keyword(s):  
Protective Role ◽  
Neutralization Capacity ◽  
Care Workers ◽  
Neutralizing Capacity ◽  
Human Coronaviruses ◽  
Antibody Levels ◽  
The Impact ◽  
Kinetics Of ◽  
Over Time

AbstractUnraveling the long-term kinetics of antibodies to SARS-CoV-2 and its determinants, including the impact of pre-existing antibodies to human coronaviruses causing common cold (HCoVs), is essential to understand protective immunity to COVID-19 and devise effective surveillance strategies. IgM, IgA and IgG levels against six SARS-CoV-2 and four HCoV antigens were quantified by Luminex, and antibody neutralization capacity was assessed by flow cytometry, in a cohort of health care workers followed-up for 6 months. Seroprevalence increased over time from 13.5% (month 0) and 15.6% (month 1) to 16.4% (month 6). Levels of antibodies, including those with neutralizing capacity, were stable over time, except IgG to nucleocapsid antigen and IgM levels that waned. After the peak response, anti-spike antibody levels increased from ∼150 days post-symptom onset in all individuals (73% for IgG), in the absence of any evidence of re-exposure. Pre-existing antibodies to alpha-HCoV were lower in individuals who subsequently seroconverted for SARS-CoV-2. IgG and IgA to HCoV were significantly higher in asymptomatic than symptomatic seropositive individuals. Thus, pre-existing cross-reactive HCoVs antibodies could have a protective effect against SARS-CoV-2 infection and COVID-19 disease.

scholarly journals Sensitive detection of SARS-CoV-2-specific-antibodies in dried blood spot samples

2020 ◽  
Author(s):  
Gabriella L. Morley ◽  
Stephen Taylor ◽  
Sian Jossi ◽  
Marisol Perez-Toledo ◽  
Sian E. Faustini ◽  
...  
Keyword(s):  
Sampling Method ◽  
Dried Blood Spot ◽  
Serological Testing ◽  
Spike Glycoprotein ◽  
Serum Samples ◽  
Perfect Agreement ◽  
Specific Antibodies ◽  
Antibody Levels ◽  
Matched Samples ◽  
Blood Spot

AbstractImportancePopulation-wide serological testing is an essential component in understanding the COVID-19 pandemic. The logistical challenges of undertaking widespread serological testing could be eased through use of a reliable dried blood spot (DBS) sampling method.ObjectiveTo validate the use of dried blood spot sampling for the detection of SARS-CoV-2-specific antibodies.Design, setting and participantsEighty-seven matched DBS and serum samples were obtained from eighty individuals, including thirty-one who were previously PCR-positive for SARS-CoV-2. DBS eluates and sera were used in an ELISA to detect antibodies to the viral spike protein.ResultsSpecific anti-SARS-Cov-2 spike glycoprotein antibodies were detectable in both serum and DBS eluate and there was a significant correlation between the antibody levels detected in matched samples (r = 0.96, p<0.0001). Using serum as the gold standard in the assay, matched DBS samples achieved a Cohen’s kappa coefficient of 0.975 (near-perfect agreement), a sensitivity of 98.1% and specificity of 100%, for detecting anti-spike glycoprotein antibodies.Conclusions and relevanceEluates from DBS samples are a reliable and reproducible source of antibodies to be used for the detection of SARS-CoV-2-specific antibodies. The use of DBS sampling could complement the use of venepuncture in the immunosurveillance of COVID-19 in both low and high income settings.

scholarly journals Enzyme-linked immunosorbent assays for the measurement of specific antibodies in experimentally induced ovine toxoplasmosis

1988 ◽  
Vol 100 (2) ◽  
pp. 205-212 ◽  
Author(s):  
R. A. Payne ◽  
D. H. M. Joynson ◽  
A. J. Wilsmore
Keyword(s):  
Toxoplasma Gondii ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Capture Assay ◽  
Previous Infection ◽  
Specific Igg ◽  
Immunosorbent Assays ◽  
Antibody Class ◽  
Antibody Levels ◽  
Indirect Assay

SUMMARYTachyzoitcs of the RH strain of Toxoplasma gondii were inoculated intravenously into sheep following which serum samples were collected at approximately weekly intervals for 9 months. The sera were examined by the toxoplasma dye test and two enzymc-linkcd immunosorbent assays (ELISA) specifically developed for investigations of ovine toxoplasmosis. One was an antibody class capture assay for the detection of anti-toxoplasma specific IgM, the other an indirect assay which detected anti-toxoplasma IgG.Some of the sheep had antibodies to toxoplasma prior to inoculation but none had specific IgM. Sera collected 17 days after inoculation showed that all had raised specific antibody levels but the only sheep that produced specific antitoxoplasma IgM were those that were initially without any antibody. Specific IgM could be detected in all these particular sheep for at least 1 month after infection and up to 3 months in some. Specific IgG persisted at high levels for at least 3 months and could still be detected at moderate levels for at least 9 months. The ELISA methods described are simple to perform and could clearly distinguish between previous infection and this experimental infection with Toxoplasma gondii.

scholarly journals Titers, Prevalence, and Duration of SARS-CoV-2 Antibodies in a Local COVID-19 Outbreak and Following Vaccination

Vaccines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 587
Author(s):  
Jodi F. Hedges ◽  
Macy A. Thompson ◽  
Deann T. Snyder ◽  
Amanda Robison ◽  
Matthew P. Taylor ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Natural Infection ◽  
Herd Immunity ◽  
Antibody Responses ◽  
Receptor Binding Domain ◽  
Specific Antibodies ◽  
Neutralization Capacity ◽  
Protein Receptor ◽  
Asymptomatic Infections ◽  
Antibody Levels

Information concerning the development of neutralizing antibodies and their duration will be critical to establishing herd immunity for COVID-19. We sought to evaluate SARS-CoV-2 spike protein receptor-binding domain (RBD)-specific antibodies, their duration, and capacity for SARS-CoV-2 neutralization in volunteers while the pandemic spread within our community starting in March 2020. Those participants with the highest starting titers had the longest-lasting response, up to 12 months post-diagnosis. SARS-CoV-2 neutralization capacity was correlated with anti-RBD antibody levels. The majority of our participants with confirmed COVID-19 diagnosis had very mild or asymptomatic infections. We also detected low and largely non-neutralizing anti-RBD IgG titers in a few participants with no known COVID-19 diagnosis. Finally, we found that antibody responses induced by vaccination were significantly higher than those induced by natural infection. Thus, our study suggests that vaccination is still critical even for those naturally infected or diagnosed with COVID-19.

scholarly journals Reduced serum neutralization capacity against SARS-CoV-2 variants in a multiplex ACE2 RBD competition assay

2021 ◽  
Author(s):  
Daniel Junker ◽  
Alex Dulovic ◽  
Matthias Becker ◽  
Teresa R Wagner ◽  
Philipp D Kaiser ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Protective Immunity ◽  
Competition Assay ◽  
Serum Samples ◽  
Who Grade ◽  
Live Virus ◽  
Neutralization Capacity ◽  
Neutralizing Capacity ◽  
Single Well ◽  
Vaccination Campaigns

As global vaccination campaigns against SARS-CoV-2 proceed, there is emerging interest in the longevity of immune protection, especially with regard to increasingly infectious virus variants. Neutralizing antibodies (Nabs) targeting the receptor binding domain (RBD) of SARS-CoV-2 are promising correlates of protective immunity and have been successfully used for prevention and therapy. To assess neutralizing capacity, we developed a bead-based multiplex ACE2 RBD competition assay as a large scalable, time-, cost-, and material-saving alternative to infectious live-virus neutralization tests. By mimicking the interaction between ACE2 and RBD, this assay detects the presence of Nabs against SARS-CoV2 in serum. Using this multiplex approach allows the simultaneous analysis of Nabs against all SARS-CoV-2 variants of concern and variants of interest in a single well. Following validation, we analyzed 325 serum samples from 186 COVID-19 patients of varying severity. Neutralization capacity was reduced for all variants examined compared to wild-type, especially for those displaying the E484K mutation. The neutralizing immune response itself, while highly individualistic, positively correlates with IgG levels. Neutralization capacity also correlated with disease severity up to WHO grade 7, after which it reduced.

scholarly journals Codominant IgG and IgA expression with minimal vaccine mRNA in milk of BNT162b2 vaccinees

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Jia Ming Low ◽  
Yue Gu ◽  
Melissa Shu Feng Ng ◽  
Zubair Amin ◽  
Le Ye Lee ◽  
...  
Keyword(s):  
Human Milk ◽  
Protective Immunity ◽  
Post Dose ◽  
Post Vaccination ◽  
Lactating Women ◽  
Vaccination Programs ◽  
Specific Antibodies ◽  
Lactating Mothers ◽  
Protective Antibodies ◽  
Specific Igg

AbstractLactating women can produce protective antibodies in their milk after vaccination, which has informed antenatal vaccination programs for diseases such as influenza and pertussis. However, whether SARS-CoV-2-specific antibodies are produced in human milk as a result of COVID-19 vaccination is still unclear. In this study, we show that lactating mothers who received the BNT162b2 vaccine secreted SARS-CoV-2-specific IgA and IgG antibodies into milk, with the most significant increase at 3–7 days post-dose 2. Virus-specific IgG titers were stable out to 4–6 weeks after dose 2. In contrast, SARS-CoV-2-specific IgA levels showed substantial decay. Vaccine mRNA was detected in few milk samples (maximum of 2 ng/ml), indicative of minimal transfer. Additionally, infants who consumed post-vaccination human milk had no reported adverse effects up to 28 days post-ingestion. Our results define the safety and efficacy profiles of the vaccine in this demographic and provide initial evidence for protective immunity conferred by milk-borne SARS-CoV-2-specific antibodies. Taken together, our study supports recommendations for uninterrupted breastfeeding subsequent to mRNA vaccination against COVID-19.

scholarly journals Comparison of Different Assays To Assess Human Papillomavirus (HPV) Type 16- and 18-Specific Antibodies after HPV Infection and Vaccination

2013 ◽  
Vol 20 (8) ◽  
pp. 1329-1332 ◽  
Author(s):  
Mirte Scherpenisse ◽  
Rutger M. Schepp ◽  
Madelief Mollers ◽  
Sofie H. Mooij ◽  
Chris J. L. M. Meijer ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Serum Antibody ◽  
Hpv Infection ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Multiplex Immunoassay ◽  
Specific Serum ◽  
Virus Like Particle ◽  
Large Panel ◽  
Antibody Levels

ABSTRACTWe compared the measurement of human papillomavirus (HPV)-specific serum antibody levels with the virus-like-particle multiplex immunoassay (VLP-MIA), competitive Luminex immunoassay (cLIA), and glutathioneS-transferase (GST) L1-based MIA. Using a large panel of serum samples, these assays showed mutually good correlations for both naturally induced and vaccine-derived HPV-specific antibody levels. However, an adaptation of the GST L1-based MIA resulted in an improved correlation with both cLIA and VLP-MIA.

scholarly journals Determinants of early antibody responses to COVID-19 mRNA vaccines in exposed and naive healthcare workers

2021 ◽  
Author(s):  
Gemma Moncunill ◽  
Ruth Aguilar ◽  
Marta Ribes ◽  
Natalia Ortega ◽  
Rocío Rubio ◽  
...  
Keyword(s):  
Health Care Workers ◽  
Healthcare Workers ◽  
Healthy Individuals ◽  
Wild Type ◽  
Post Vaccination ◽  
Neutralization Capacity ◽  
Care Workers ◽  
Neutralizing Capacity ◽  
One Year ◽  
Antibody Levels

Background Two doses of mRNA vaccination have shown >94% efficacy at preventing COVID-19 mostly in naive adults, but it is not clear if the second dose is needed to maximize effectiveness in those previously exposed to SARS-CoV-2 and what other factors affect responsiveness. Methods We measured IgA, IgG and IgM levels against SARS-CoV-2 spike (S) and nucleocapsid (N) antigens from the wild-type and S from the Alpha, Beta and Gamma variants of concern, after BNT162b2 (Pfizer/BioNTech) or mRNA-1273 (Moderna) vaccination in a cohort of health care workers (N=578). Neutralizing capacity and antibody avidity were evaluated. Data were analyzed in relation to COVID-19 history, comorbidities, vaccine doses, brand and adverse events. Findings Vaccination induced robust IgA and IgG levels against all S antigens. Neutralization capacity and S IgA and IgG levels were higher in mRNA-1273 vaccinees, previously SARS-CoV-2 exposed, particularly if symptomatic, and in those experiencing systemic adverse effects. A second dose in pre-exposed did not increase antibody levels. Smoking and comorbidities were associated with lower neutralization and antibody levels. Among fully vaccinated, 6.3% breakthroughs were detected up to 189 days post-vaccination. Among pre-exposed non-vaccinated, 90% were IgG seropositive more than 300 days post-infection. Interpretation Our data support administering a single-dose in pre-exposed healthy individuals. However, heterogeneity of responses suggests that personalized recommendations may be necessary depending on COVID-19 history and life-style. Higher mRNA-1273 immunogenicity would be beneficial for those expected to respond worse to vaccination. Persistence of antibody levels in pre-exposed unvaccinated indicates maintenance of immunity up to one year.

The level of protective post-vaccination antibodies in NIPH-NIH employees after administration of Pfizer vaccine against COVID-19

2021 ◽  
pp. 3-13
Author(s):  
Waldemar Rastawicki ◽  
Kaludia Płaza
Keyword(s):  
Effective Vaccine ◽  
Elisa Assay ◽  
Igg Antibodies ◽  
Human Immune System ◽  
Serum Samples ◽  
S Protein ◽  
N Protein ◽  
Global Pandemic ◽  
Incidence And Mortality ◽  
Antibody Levels

INTRODUCTION. The new SARS-CoV-2 coronavirus, first recognized in China in 2019, within a few months caused a global pandemic of a disease called COVID-19. The high incidence and mortality of COVID-19 was the reason for the beginning of intensive work on the development of an effective vaccine. In Poland, mass vaccinations against this disease began at the end of December 2020. OBJECTIVES. The aim of the presented study was to determine the effectiveness of stimulating the production of specific antibodies for SARS-CoV-2 by the Pfizer vaccine. MATERIAL AND METHODS. The presence of IgA and IgG antibodies to the spike (S protein) of SARSCoV-2 was tested by the ELISA/Euroimmun in serum samples obtained from 140 the employees of NIPH-NIH (137 were vaccinated). In addition, the presence of IgG antibodies to S protein, nucleoprotein, and mixture of both in selected serum samples was tested by the newly developed in NIPH-NIH in-house ELISA assay. RESULTS. IgA and IgG antibodies to the S protein of the SARS-CoV-2 were detected by ELISA/Euroimmun, respectively in 136 and in all 137 vaccinated persons. There were no statistically significant differences in the level of antibodies depending on the sex and age of the vaccinated persons. Slightly higher levels of antibodies have been demonstrated in vaccinated subjects with documented preexisting SARS-CoV-2 immunity compared to subjects without COVID-19 history. The presence of IgA and IgG antibodies was found in respectively, 18 (45.0%) and all 40 (100.0%) tested vaccinated persons by the in-house ELISA with mixture antigen. The study showed that ELISA assay with N protein as an antigen may enable the distinction between antibodies acquired after infection and after vaccination. CONCLUSIONS. The results obtained in the presented study clearly demonstrate the high effectiveness of the Pfizer vaccine in stimulation of the human immune system to produce antibodies specific for the S protein of the SARS-CoV-2. It is necessary to continue testing vaccine antibody levels at various times after vaccination to determine the potential duration of humoral immunity.

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