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Article ; Online: Circulating antibody-secreting cells are a biomarker for early diagnosis in patients with Lyme disease.

Haddad, Natalie S / Nozick, Sophia / Ohanian, Shant / Smith, Robert / Elias, Susan / Auwaerter, Paul G / Lee, F Eun-Hyung / Daiss, John L

PloS one

2023 Volume 18, Issue 11, Page(s) e0293203

Abstract: Background: Diagnostic immunoassays for Lyme disease have several limitations including: 1) not all patients seroconvert; 2) seroconversion occurs later than symptom onset; and 3) serum antibody levels remain elevated long after resolution of the ... ...

Abstract	Background: Diagnostic immunoassays for Lyme disease have several limitations including: 1) not all patients seroconvert; 2) seroconversion occurs later than symptom onset; and 3) serum antibody levels remain elevated long after resolution of the infection. Introduction: MENSA (Medium Enriched for Newly Synthesized Antibodies) is a novel diagnostic fluid that contains antibodies produced in vitro by circulating antibody-secreting cells (ASC). It enables measurement of the active humoral immune response. Methods: In this observational, case-control study, we developed the MicroB-plex Anti-C6/Anti-pepC10 Immunoassay to measure antibodies specific for the Borrelia burgdorferi peptide antigens C6 and pepC10 and validated it using a CDC serum sample collection. Then we examined serum and MENSA samples from 36 uninfected Control subjects and 12 Newly Diagnosed Lyme Disease Patients. Results: Among the CDC samples, antibodies against C6 and/or pepC10 were detected in all seropositive Lyme patients (8/8), but not in sera from seronegative patients or healthy controls (0/24). Serum antibodies against C6 and pepC10 were detected in one of 36 uninfected control subjects (1/36); none were detected in the corresponding MENSA samples (0/36). In samples from newly diagnosed patients, serum antibodies identified 8/12 patients; MENSA antibodies also detected 8/12 patients. The two measures agreed on six positive individuals and differed on four others. In combination, the serum and MENSA tests identified 10/12 early Lyme patients. Typically, serum antibodies persisted 80 days or longer while MENSA antibodies declined to baseline within 40 days of successful treatment. Discussion: MENSA-based immunoassays present a promising complement to serum immunoassays for diagnosis and tracking therapeutic success in Lyme infections.
MeSH term(s)	Humans ; Case-Control Studies ; Antigens, Bacterial ; Immunoglobulin G ; Lyme Disease ; Antibodies, Bacterial ; Biomarkers ; Antibody-Producing Cells ; Early Diagnosis ; Borrelia burgdorferi
Chemical Substances	Antigens, Bacterial ; Immunoglobulin G ; Antibodies, Bacterial ; Biomarkers
Language	English
Publishing date	2023-11-03
Publishing country	United States
Document type	Observational Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0293203
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: The Majority of SARS-CoV-2 Plasma Cells are Excluded from the Bone Marrow Long-Lived Compartment 33 Months after mRNA Vaccination.

Nguyen, Doan C / Hentenaar, Ian T / Morrison-Porter, Andrea / Solano, David / Haddad, Natalie S / Castrillon, Carlos / Lamothe, Pedro A / Andrews, Joel / Roberts, Danielle / Lonial, Sagar / Sanz, Ignacio / Lee, F Eun-Hyung

medRxiv : the preprint server for health sciences

2024

Abstract: The goal of any vaccine is to induce long-lived plasma cells (LLPC) to provide life-long protection. Natural infection by influenza, measles, or mumps viruses generates bone marrow (BM) LLPC similar to tetanus vaccination which affords safeguards for ... ...

Abstract	The goal of any vaccine is to induce long-lived plasma cells (LLPC) to provide life-long protection. Natural infection by influenza, measles, or mumps viruses generates bone marrow (BM) LLPC similar to tetanus vaccination which affords safeguards for decades. Although the SARS-CoV-2 mRNA vaccines protect from severe disease, the serologic half-life is short-lived even though SARS-CoV-2-specific plasma cells can be found in the BM. To better understand this paradox, we enrolled 19 healthy adults at 1.5-33 months after SARS-CoV-2 mRNA vaccine and measured influenza-, tetanus-, or SARS-CoV-2-specific antibody secreting cells (ASC) in LLPC (CD19
Language	English
Publishing date	2024-03-05
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.03.02.24303242
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Article ; Online: The Majority of SARS-CoV-2 Plasma Cells are Excluded from the Bone Marrow Long-Lived Compartment 33 Months after mRNA Vaccination

Nguyen, Doan C. / Hentenaar, Ian T. / Morrison-Porter, Andrea / Solano, David / Haddad, Natalie S. / Castrillon, Carlos / Lamothe, Pedro A. / Andrews, Joel / Roberts, Danielle / Lonial, Sagar / Sanz, Ignacio / Lee, F. Eun-Hyung

medRxiv

Abstract	The goal of any vaccine is to induce long-lived plasma cells (LLPC) to provide life-long protection. Natural infection by influenza, measles, or mumps viruses generates bone marrow (BM) LLPC similar to tetanus vaccination which affords safeguards for decades. Although the SARS-CoV-2 mRNA vaccines protect from severe disease, the serologic half-life is short-lived even though SARS-CoV-2-specific plasma cells can be found in the BM. To better understand this paradox, we enrolled 19 healthy adults at 1.5-33 months after SARS-CoV-2 mRNA vaccine and measured influenza-, tetanus-, or SARS-CoV-2-specific antibody secreting cells (ASC) in LLPC (CD19-) and non-LLPC (CD19+) subsets within the BM. All individuals had IgG ASC specific for influenza, tetanus, and SARS-CoV-2 in at least one BM ASC compartment. However, only influenza- and tetanus-specific ASC were readily detected in the LLPC whereas SARS-CoV-2 specificities were mostly excluded. The ratios of non-LLPC:LLPC for influenza, tetanus, and SARS-CoV-2 were 0.61, 0.44, and 29.07, respectively. Even in five patients with known PCR-proven history of infection and vaccination, SARS-CoV-2-specific ASC were mostly excluded from the LLPC. These specificities were further validated by using multiplex bead binding assays of secreted antibodies in the supernatants of cultured ASC. Similarly, the IgG ratios of non-LLPC:LLPC for influenza, tetanus, and SARS-CoV-2 were 0.66, 0.44, and 23.26, respectively. In all, our studies demonstrate that rapid waning of serum antibodies is accounted for by the inability of mRNA vaccines to induce BM LLPC.
Keywords	covid19
Language	English
Publishing date	2024-03-05
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2024.03.02.24303242
Database	COVID19

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Article ; Online: Reduced COVID-19 Vaccine Response in Patients Treated with Biologic Therapies for Asthma.

Runnstrom, Martin C / Morrison-Porter, Andrea / Ravindran, Mayuran / Quehl, Hannah / Ramonell, Richard P / Woodruff, Matthew / Patel, Rahulkumar / Kim, Caroline / Haddad, Natalie S / Lee, F Eun-Hyung

American journal of respiratory and critical care medicine

2022 Volume 205, Issue 10, Page(s) 1243–1245

MeSH term(s)	Anti-Asthmatic Agents/therapeutic use ; Asthma/drug therapy ; Biological Therapy ; COVID-19 ; COVID-19 Vaccines/therapeutic use ; Humans
Chemical Substances	Anti-Asthmatic Agents ; COVID-19 Vaccines
Language	English
Publishing date	2022-03-11
Publishing country	United States
Document type	Letter ; Research Support, N.I.H., Extramural
ZDB-ID	1180953-x
ISSN	1535-4970 ; 0003-0805 ; 1073-449X
ISSN (online)	1535-4970
ISSN	0003-0805 ; 1073-449X
DOI	10.1164/rccm.202111-2496LE
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Article: Novel immunoassay for diagnosis of ongoing Clostridioides difficile infections using serum and medium enriched for newly synthesized antibodies (MENSA)

Haddad, Natalie S / Nozick, Sophia / Kim, Geena / Ohanian, Shant / Kraft, Colleen / Rebolledo, Paulina A / Wang, Yun / Wu, Hao / Bressler, Adam / Le, Sang Nguyet Thi / Kuruvilla, Merin / Cannon, L. Edward / Lee, F. Eun-Hyung / Daiss, John L

Elsevier B.V. Journal of immunological methods. 2021 May, v. 492

2021

Abstract: Clostridioides difficile infections (CDI) have been a challenging and increasingly serious concern in recent years. While early and accurate diagnosis is crucial, available assays have frustrating limitations.Develop a simple, blood-based immunoassay to ... ...

Abstract	Clostridioides difficile infections (CDI) have been a challenging and increasingly serious concern in recent years. While early and accurate diagnosis is crucial, available assays have frustrating limitations.Develop a simple, blood-based immunoassay to accurately diagnose patients suffering from active CDI.Uninfected controls (N = 95) and CDI patients (N = 167) were recruited from Atlanta area hospitals. Blood samples were collected from patients within twelve days of a positive CDI test and processed to yield serum and PBMCs cultured to yield medium enriched for newly synthesized antibodies (MENSA). Multiplex immunoassays measured Ig responses to ten recombinant C. difficile antigens.Sixty-six percent of CDI patients produced measurable responses to C. difficile antigens in their serum or MENSA within twelve days of a positive CDI test. Fifty-two of the 167 CDI patients (31%) were detectable in both serum and MENSA, but 32/167 (19%) were detectable only in MENSA, and 27/167 (16%) were detectable only in serum.We describe the results of a multiplex immunoassay for the diagnosis of ongoing CDI in hospitalized patients. Our assay resolved patients into four categories: MENSA-positive only, serum-positive only, MENSA- and serum-positive, and MENSA- and serum-negative. The 30% of patients who were MENSA-positive only may be accounted for by nascent antibody secretion prior to seroconversion. Conversely, the serum-positive only subset may have been more advanced in their disease course. Immunocompromise and misdiagnosis may have contributed to the 34% of CDI patients who were not identified using MENSA or serum immunoassays.While there was considerable overlap between patients identified through MENSA and serum, each method detected a distinctive patient group. The combined use of both MENSA and serum to detect CDI patients resulted in the greatest identification of CDI patients. Together, longitudinal analysis of MENSA and serum will provide a more accurate evaluation of successful host humoral immune responses in CDI patients.
Keywords	Clostridium difficile ; antibodies ; blood serum ; disease course ; immunoassays ; longitudinal studies ; patients ; secretion ; seroconversion
Language	English
Dates of publication	2021-05
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2020.112932
Database	NAL-Catalogue (AGRICOLA)

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Article: Poor immunogenicity upon SARS-CoV-2 mRNA vaccinations in autoimmune SLE patients is associated with pronounced EF-mediated responses and anti-BAFF/Belimumab treatment.

medRxiv : the preprint server for health sciences

2023

Abstract: Novel mRNA vaccines have resulted in a reduced number of SARS-CoV-2 infections and hospitalizations. Yet, there is a paucity of studies regarding their effectiveness on immunocompromised autoimmune subjects. In this study, we enrolled subjects naïve to ... ...

Abstract	Novel mRNA vaccines have resulted in a reduced number of SARS-CoV-2 infections and hospitalizations. Yet, there is a paucity of studies regarding their effectiveness on immunocompromised autoimmune subjects. In this study, we enrolled subjects naïve to SARS-CoV-2 infections from two cohorts of healthy donors (HD, n=56) and systemic lupus erythematosus (SLE, n=69). Serological assessments of their circulating antibodies revealed a significant reduction of potency and breadth of neutralization in the SLE group, only partially rescued by a 3
Language	English
Publishing date	2023-06-12
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.06.08.23291159
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Article ; Online: Detection of Newly Secreted Antibodies Predicts Nonrecurrence in Primary Clostridioides difficile Infection.

Haddad, Natalie S / Nozick, Sophia / Kim, Geena / Ohanian, Shant / Kraft, Colleen S / Rebolledo, Paulina A / Wang, Yun / Wu, Hao / Bressler, Adam / Le, Sang Nguyet Thi / Kuruvilla, Merin / Runnstrom, Martin C / Ramonell, Richard P / Cannon, L Edward / Lee, F Eun-Hyung / Daiss, John L

Journal of clinical microbiology

2022 Volume 60, Issue 3, Page(s) e0220121

Abstract: Within 8 weeks of primary Clostridioides difficile infection (CDI), as many as 30% of patients develop recurrent disease with the associated risks of multiple relapses, morbidity, and economic burden. There are no clear clinical correlates or validated ... ...

Abstract	Within 8 weeks of primary Clostridioides difficile infection (CDI), as many as 30% of patients develop recurrent disease with the associated risks of multiple relapses, morbidity, and economic burden. There are no clear clinical correlates or validated biomarkers that can predict recurrence during primary infection. This study demonstrated the potential of a simple test for identifying hospitalized CDI patients at low risk for disease recurrence. Forty-six hospitalized CDI patients were enrolled at Emory University Hospitals. Samples of serum and a novel matrix from circulating plasmablasts called "medium-enriched for newly synthesized antibodies" (MENSA) were collected during weeks 1, 2, and 4. Antibodies specific for 10 C. difficile antigens were measured in each sample. Among the 46 C. difficile-infected patients, 9 (19.5%) experienced recurrence within 8 weeks of primary infection. Among the 37 nonrecurrent patients, 23 (62%; 23/37) had anti-C. difficile MENSA antibodies specific for any of the three toxin antigens: TcdB-CROP, TcdBvir-CROP, and/or CDTb. Positive MENSA responses occurred early (within the first 12 days post-symptom onset), including six patients who never seroconverted. A similar trend was observed in serum responses, but they peaked later and identified fewer patients (51%; 19/37). In contrast, none (0%; 0/9) of the patients who subsequently recurred after hospitalization produced antibodies specific for any of the three C. difficile toxin antigens. Thus, patients with a negative early MENSA response against all three C. difficile toxin antigens had a 19-fold greater relative risk of recurrence. MENSA and serum levels of immunoglobulin A (IgA) and/or IgG antibodies for three C. difficile toxins have prognostic potential. These immunoassays measure nascent immune responses that reduce the likelihood of recurrence thereby providing a biomarker of protection from recurrent CDI. Patients who are positive by this immunoassay are unlikely to suffer a recurrence. Early identification of patients at risk for recurrence by negative MENSA creates opportunities for targeted prophylactic strategies that can reduce the incidence, cost, and morbidity due to recurrent CDI.
MeSH term(s)	Bacterial Toxins ; Biomarkers ; Clostridioides difficile ; Clostridium Infections/epidemiology ; Culture Media ; Humans ; Immunoglobulin A ; Immunoglobulin G ; Recurrence
Chemical Substances	Bacterial Toxins ; Biomarkers ; Culture Media ; Immunoglobulin A ; Immunoglobulin G
Language	English
Publishing date	2022-03-16
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	390499-4
ISSN	1098-660X ; 0095-1137
ISSN (online)	1098-660X
ISSN	0095-1137
DOI	10.1128/jcm.02201-21
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Article ; Online: Inactivation of SARS-CoV-2 and COVID-19 Patient Samples for Contemporary Immunology and Metabolomics Studies.

Eddins, Devon J / Bassit, Leda C / Chandler, Joshua D / Haddad, Natalie S / Musall, Kathryn L / Yang, Junkai / Kosters, Astrid / Dobosh, Brian S / Hernández, Mindy R / Ramonell, Richard P / Tirouvanziam, Rabindra M / Lee, F Eun-Hyung / Zandi, Keivan / Schinazi, Raymond F / Ghosn, Eliver E B

ImmunoHorizons

2022 Volume 6, Issue 2, Page(s) 144–155

Abstract: Due to the severity of COVID-19 disease, the U.S. Centers for Disease Control and Prevention and World Health Organization recommend that manipulation of active viral cultures of SARS-CoV-2 and respiratory secretions from COVID-19 patients be performed ... ...

Abstract	Due to the severity of COVID-19 disease, the U.S. Centers for Disease Control and Prevention and World Health Organization recommend that manipulation of active viral cultures of SARS-CoV-2 and respiratory secretions from COVID-19 patients be performed in biosafety level (BSL)3 laboratories. Therefore, it is imperative to develop viral inactivation procedures that permit samples to be transferred to lower containment levels (BSL2), while maintaining the fidelity of complex downstream assays to expedite the development of medical countermeasures. In this study, we demonstrate optimal conditions for complete viral inactivation following fixation of infected cells with commonly used reagents for flow cytometry, UVC inactivation in sera and respiratory secretions for protein and Ab detection, heat inactivation following cDNA amplification for droplet-based single-cell mRNA sequencing, and extraction with an organic solvent for metabolomic studies. Thus, we provide a suite of viral inactivation protocols for downstream contemporary assays that facilitate sample transfer to BSL2, providing a conceptual framework for rapid initiation of high-fidelity research as the COVID-19 pandemic continues.
MeSH term(s)	COVID-19/prevention & control ; Hot Temperature ; Humans ; Metabolomics/methods ; Pandemics/prevention & control ; SARS-CoV-2 ; Specimen Handling/methods ; Ultraviolet Rays ; Virus Inactivation
Language	English
Publishing date	2022-02-16
Publishing country	United States
Document type	Journal Article
ISSN	2573-7732
ISSN (online)	2573-7732
DOI	10.4049/immunohorizons.2200005
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Article ; Online: Novel immunoassay for diagnosis of ongoing Clostridioides difficile infections using serum and medium enriched for newly synthesized antibodies (MENSA).

Journal of immunological methods

2020 Volume 492, Page(s) 112932

Abstract: Background: Clostridioides difficile infections (CDI) have been a challenging and increasingly serious concern in recent years. While early and accurate diagnosis is crucial, available assays have frustrating limitations.: Objective: Develop a simple, ...

Abstract	Background: Clostridioides difficile infections (CDI) have been a challenging and increasingly serious concern in recent years. While early and accurate diagnosis is crucial, available assays have frustrating limitations. Objective: Develop a simple, blood-based immunoassay to accurately diagnose patients suffering from active CDI. Materials and methods: Uninfected controls (N = 95) and CDI patients (N = 167) were recruited from Atlanta area hospitals. Blood samples were collected from patients within twelve days of a positive CDI test and processed to yield serum and PBMCs cultured to yield medium enriched for newly synthesized antibodies (MENSA). Multiplex immunoassays measured Ig responses to ten recombinant C. difficile antigens. Results: Sixty-six percent of CDI patients produced measurable responses to C. difficile antigens in their serum or MENSA within twelve days of a positive CDI test. Fifty-two of the 167 CDI patients (31%) were detectable in both serum and MENSA, but 32/167 (19%) were detectable only in MENSA, and 27/167 (16%) were detectable only in serum. Discussion: We describe the results of a multiplex immunoassay for the diagnosis of ongoing CDI in hospitalized patients. Our assay resolved patients into four categories: MENSA-positive only, serum-positive only, MENSA- and serum-positive, and MENSA- and serum-negative. The 30% of patients who were MENSA-positive only may be accounted for by nascent antibody secretion prior to seroconversion. Conversely, the serum-positive only subset may have been more advanced in their disease course. Immunocompromise and misdiagnosis may have contributed to the 34% of CDI patients who were not identified using MENSA or serum immunoassays. Importance: While there was considerable overlap between patients identified through MENSA and serum, each method detected a distinctive patient group. The combined use of both MENSA and serum to detect CDI patients resulted in the greatest identification of CDI patients. Together, longitudinal analysis of MENSA and serum will provide a more accurate evaluation of successful host humoral immune responses in CDI patients.
MeSH term(s)	Antibodies, Bacterial/analysis ; Antibodies, Bacterial/immunology ; Antigens, Bacterial/immunology ; Antigens, Bacterial/metabolism ; Case-Control Studies ; Cell Culture Techniques ; Clostridioides difficile/immunology ; Clostridioides difficile/isolation & purification ; Clostridium Infections/blood ; Clostridium Infections/diagnosis ; Clostridium Infections/microbiology ; Culture Media/metabolism ; Female ; Humans ; Leukocytes, Mononuclear/immunology ; Leukocytes, Mononuclear/metabolism ; Male ; Middle Aged ; Recombinant Proteins/immunology ; Recombinant Proteins/metabolism ; Serologic Tests/methods
Chemical Substances	Antibodies, Bacterial ; Antigens, Bacterial ; Culture Media ; Recombinant Proteins
Language	English
Publishing date	2020-11-19
Publishing country	Netherlands
Document type	Journal Article ; Observational Study ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	120142-6
ISSN	1872-7905 ; 0022-1759
ISSN (online)	1872-7905
ISSN	0022-1759
DOI	10.1016/j.jim.2020.112932
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Article: Relaxed peripheral tolerance drives broad

Woodruff, Matthew C / Ramonell, Richard P / Saini, Ankur Singh / Haddad, Natalie S / Anam, Fabliha A / Rudolph, Mark E / Bugrovsky, Regina / Hom, Jennifer / Cashman, Kevin S / Nguyen, Doan C / Kyu, Shuya / Piazza, Michael / Tipton, Christopher M / Jenks, Scott A / Lee, F Eun-Hyung / Sanz, Ignacio

medRxiv : the preprint server for health sciences

2021

Abstract: An emerging feature of COVID-19 is the identification of autoreactivity in patients with severe disease that may contribute to disease pathology, however the origin and resolution of these responses remain unclear. Previously, we identified strong ... ...

Abstract	An emerging feature of COVID-19 is the identification of autoreactivity in patients with severe disease that may contribute to disease pathology, however the origin and resolution of these responses remain unclear. Previously, we identified strong extrafollicular B cell activation as a shared immune response feature between both severe COVID-19 and patients with advanced rheumatic disease. In autoimmune settings, this pathway is associated with relaxed peripheral tolerance in the antibody secreting cell compartment and the generation of
Keywords	covid19
Language	English
Publishing date	2021-07-27
Publishing country	United States
Document type	Preprint
DOI	10.1101/2020.10.21.20216192
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