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1.
J Infect Dis ; 224(5): 771-776, 2021 09 01.
Article in English | MEDLINE | ID: covidwho-1410005

ABSTRACT

We aimed to characterize presence of culturable virus in clinical specimens during acute illness, and antibody kinetics up to 6 months after symptom onset, among 14 early patients with coronavirus disease 2019 in the United States. We isolated viable severe acute respiratory syndrome coronavirus 2 from real-time reverse-transcription polymerase chain reaction-positive respiratory specimens collected during days 0-8 after onset, but not after. All 13 patients with 2 or more serum specimens developed anti-spike antibodies; 12 developed detectable neutralizing antibodies. We did not isolate virus after detection of neutralizing antibodies. Eight participants provided serum at 6 months after onset; all retained detectable anti-spike immunoglobulin G, and half had detectable neutralizing antibodies. Two participants reported not feeling fully recovered at 6 months.


Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Antibody Formation/immunology , COVID-19/immunology , Seroconversion/physiology , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , COVID-19/blood , COVID-19/virology , Follow-Up Studies , Humans , Immunoglobulin G/blood , Immunoglobulin G/immunology , Spike Glycoprotein, Coronavirus/immunology , United States
3.
J Med Virol ; 94(1): 287-290, 2022 01.
Article in English | MEDLINE | ID: covidwho-1391642

ABSTRACT

In the 10th month of the pandemic, coronavirus disease 2019 (COVID-19) vaccination was given first to healthcare workers in Turkey after receiving emergency use approval from the Ministry of Health. This study, which was performed at the COVID-19 reference center in Ankara (the capital of Turkey) aimed to evaluate the seroconversion rate of the CoronaVac vaccine. The anti-spike immunoglobulin G response to the two-dose vaccination was retrospectively examined in healthcare workers who had no previous history of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The postvaccine seroconversion rate was investigated by measuring the antibody levels of healthcare workers who had received CoronaVac. Vaccination was administered as 600 SU in 28-day intervals. The healthcare workers' anti-SARS-CoV-2 immunoglobulin G levels were used to determine the seroconversion rate 2 months after the second dose of the vaccine. Of the healthcare workers, 22.9% (n = 155) were seronegative. The younger the age of the participant, the higher the level of anti-SARS-CoV-2 immunoglobulin G. Furthermore, anti-SARS-CoV-2 immunoglobulin G levels were much higher in women than men.


Subject(s)
Antibodies, Neutralizing/blood , Antibodies, Viral/blood , COVID-19 Vaccines/immunology , COVID-19/prevention & control , Immunization, Secondary/methods , SARS-CoV-2/immunology , Adult , Aged , COVID-19/immunology , Coronavirus Nucleocapsid Proteins/immunology , Female , Health Personnel/statistics & numerical data , Humans , Immunization Schedule , Immunoglobulin G/blood , Male , Middle Aged , Phosphoproteins/immunology , Retrospective Studies , Seroconversion/physiology , Turkey , Vaccines, Inactivated/immunology , Vaccines, Synthetic/immunology , Young Adult
4.
Genes (Basel) ; 12(7)2021 06 25.
Article in English | MEDLINE | ID: covidwho-1295802

ABSTRACT

Peripheral blood transcriptome is a highly promising area for biomarker development. However, transcript abundances (TA) in these cell mixture samples are confounded by proportions of the component leukocyte subpopulations. This poses a challenge to clinical applications, as the cell of origin of any change in TA is not known without prior cell separation procedure. We developed a framework to develop a cell-type informative TA biomarkers which enable determination of TA of a single cell-type (B lymphocytes) directly in cell mixture samples of peripheral blood (e.g., peripheral blood mononuclear cells, PBMC) without the need for subpopulation separation. It is applicable to a panel of genes called B cell informative genes. Then a ratio of two B cell informative genes (a target gene and a stably expressed reference gene) obtained in PBMC was used as a new biomarker to represent the target gene expression in purified B lymphocytes. This approach, which eliminates the tedious procedure of cell separation and directly determines TA of a leukocyte subpopulation in peripheral blood samples, is called the Direct LS-TA method. This method is applied to gene expression datasets collected in influenza vaccination trials as early predictive biomarkers of seroconversion. By using TNFRSF17 or TXNDC5 as the target genes and TNFRSF13C or FCRLA as the reference genes, the Direct LS-TA B cell biomarkers were determined directly in the PBMC transcriptome data and were highly correlated with TA of the corresponding target genes in purified B lymphocytes. Vaccination responders had almost a 2-fold higher Direct LS-TA biomarker level of TNFRSF17 (log 2 SMD = 0.84, 95% CI = 0.47-1.21) on day 7 after vaccination. The sensitivity of these Direct LS-TA biomarkers in the prediction of seroconversion was greater than 0.7 and area-under curves (AUC) were over 0.8 in many datasets. In this paper, we report a straightforward approach to directly estimate B lymphocyte gene expression in PBMC, which could be used in a routine clinical setting. Moreover, the method enables the practice of precision medicine in the prediction of vaccination response. More importantly, seroconversion could now be predicted as early as day 7. As the acquired immunology pathway is common to vaccination against influenza and COVID-19, these biomarkers could also be useful to predict seroconversion for the new COVID-19 vaccines.


Subject(s)
B-Lymphocytes/physiology , Gene Expression , Influenza Vaccines/immunology , Seroconversion/genetics , B-Cell Activation Factor Receptor/genetics , Biomarkers/analysis , COVID-19 Vaccines/immunology , Computational Biology/methods , Databases, Genetic , Humans , Leukocytes, Mononuclear/physiology , Models, Theoretical , Network Meta-Analysis , Protein Disulfide-Isomerases/genetics , ROC Curve , Receptors, Fc/genetics , Seroconversion/physiology
5.
PLoS One ; 16(4): e0250319, 2021.
Article in English | MEDLINE | ID: covidwho-1206197

ABSTRACT

Projections of the stage of the Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) pandemic and local, regional and national public health policies to limit coronavirus spread as well as "reopen" cities and states, are best informed by serum neutralizing antibody titers measured by reproducible, high throughput, and statically credible antibody (Ab) assays. To date, a myriad of Ab tests, both available and FDA authorized for emergency, has led to confusion rather than insight per se. The present study reports the results of a rapid, point-in-time 1,000-person cohort study using serial blood donors in the New York City metropolitan area (NYC) using multiple serological tests, including enzyme-linked immunosorbent assays (ELISAs) and high throughput serological assays (HTSAs). These were then tested and associated with assays for neutralizing Ab (NAb). Of the 1,000 NYC blood donor samples in late June and early July 2020, 12.1% and 10.9% were seropositive using the Ortho Total Ig and the Abbott IgG HTSA assays, respectively. These serological assays correlated with neutralization activity specific to SARS-CoV-2. The data reported herein suggest that seroconversion in this population occurred in approximately 1 in 8 blood donors from the beginning of the pandemic in NYC (considered March 1, 2020). These findings deviate with an earlier seroprevalence study in NYC showing 13.7% positivity. Collectively however, these data demonstrate that a low number of individuals have serologic evidence of infection during this "first wave" and suggest that the notion of "herd immunity" at rates of ~60% or higher are not near. Furthermore, the data presented herein show that the nature of the Ab-based immunity is not invariably associated with the development of NAb. While the blood donor population may not mimic precisely the NYC population as a whole, rapid assessment of seroprevalence in this cohort and serial reassessment could aid public health decision making.


Subject(s)
COVID-19/epidemiology , SARS-CoV-2/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Neutralizing/blood , Antibodies, Viral/immunology , Blood Donors , COVID-19/immunology , Cohort Studies , Enzyme-Linked Immunosorbent Assay/methods , Female , Humans , Immunoglobulin G/blood , Male , Middle Aged , New York City/epidemiology , SARS-CoV-2/pathogenicity , Sensitivity and Specificity , Seroconversion/physiology , Seroepidemiologic Studies , Serologic Tests/methods , Spike Glycoprotein, Coronavirus/immunology
6.
Sci Rep ; 11(1): 3461, 2021 02 10.
Article in English | MEDLINE | ID: covidwho-1078603

ABSTRACT

Despite ongoing efforts to characterize the host response toward SARS-CoV-2, a major gap in our knowledge still exists regarding the magnitude and duration of the humoral response. Analysis of the antibody response in mild versus moderate/severe patients, using our new developed quantitative electrochemiluminescent assay for detecting IgM/IgA/IgG antibodies toward SARS-CoV-2 antigens, revealed a rapid onset of IgG/IgA antibodies, specifically in moderate/severe patients. IgM antibodies against the viral receptor binding domain, but not against nucleocapsid protein, were detected at early stages of the disease. Furthermore, we observed a marked reduction in IgM/IgA antibodies over-time. Adapting our assay for ACE2 binding-competition, demonstrated that the presence of potentially neutralizing antibodies is corelated with IgG/IgA. Finally, analysis of the cytokine profile in COVID-19 patients revealed unique correlation of an IL-12p70/IL33 and IgG seroconversion, which correlated with disease severity. In summary, our comprehensive analysis has major implications on the understanding and monitoring of SARS-CoV-2 infections.


Subject(s)
COVID-19/immunology , Immunoglobulin G/immunology , Interleukin-12/blood , Interleukin-33/blood , Seroconversion/physiology , Antibody Formation , COVID-19/blood , COVID-19/diagnosis , Humans , Severity of Illness Index
7.
Nat Biomed Eng ; 4(12): 1180-1187, 2020 12.
Article in English | MEDLINE | ID: covidwho-780007

ABSTRACT

Sensitive assays are essential for the accurate identification of individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we report a multiplexed assay for the fluorescence-based detection of seroconversion in infected individuals from less than 1 µl of blood, and as early as the day of the first positive nucleic acid test after symptom onset. The assay uses dye-encoded antigen-coated beads to quantify the levels of immunoglobulin G (IgG), IgM and IgA antibodies against four SARS-CoV-2 antigens. A logistic regression model trained using samples collected during the pandemic and samples collected from healthy individuals and patients with respiratory infections before the first outbreak of coronavirus disease 2019 (COVID-19) was 99% accurate in the detection of seroconversion in a blinded validation cohort of samples collected before the pandemic and from patients with COVID-19 five or more days after a positive nasopharyngeal test by PCR with reverse transcription. The high-throughput serological profiling of patients with COVID-19 allows for the interrogation of interactions between antibody isotypes and viral proteins, and should help us to understand the heterogeneity of clinical presentations.


Subject(s)
COVID-19/immunology , Immunoassay/methods , Seroconversion/physiology , Aged , Aged, 80 and over , Antibodies/immunology , Humans , Immunoglobulin G/immunology , Male , Middle Aged , Pandemics/prevention & control , SARS-CoV-2/immunology , Sensitivity and Specificity
8.
Obstet Gynecol ; 136(4): 838-841, 2020 10.
Article in English | MEDLINE | ID: covidwho-671131

ABSTRACT

BACKGROUND: There is a growing understanding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19) in the general population. The unique immunology of pregnancy may result in variations from the reported course of disease. CASE: A 27-year-old primigravid woman presented with mild COVID-19 symptoms at 28 2/7 weeks of gestation, testing positive for SARS-CoV-2 infection by nasopharyngeal swab reverse transcription-polymerase chain reaction (RT-PCR). Antibody seroconversion was detected at 36 6/7 weeks of gestation. She presented for delivery at 38 1/7 weeks of gestation, and her SARS-CoV-2 RT-PCR test result was positive. Severe acute respiratory syndrome coronavirus 2 RNA remained detectable 34 days postpartum and 104 days from her initial positive test. CONCLUSION: Prolonged viral shedding of SARS-CoV RNA may occur in the pregnant patient. If prevalent, this complicates the interpretation of a positive SARS-CoV-2 RT-PCR test result in the asymptomatic gravid patient.


Subject(s)
Antibodies, Viral/blood , Betacoronavirus , Clinical Laboratory Techniques/methods , Coronavirus Infections , Pandemics , Pneumonia, Viral , Pregnancy Complications, Infectious , Seroconversion/physiology , Virus Shedding/immunology , Adult , Betacoronavirus/immunology , Betacoronavirus/isolation & purification , COVID-19 , COVID-19 Testing , Coronavirus Infections/diagnosis , Coronavirus Infections/immunology , Female , Gestational Age , Humans , Monitoring, Immunologic/methods , Pneumonia, Viral/diagnosis , Pneumonia, Viral/immunology , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Pregnancy Complications, Infectious/immunology , SARS-CoV-2 , Time Factors
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