A novel assay to measure B cell responses to keyhole limpet haemocyanin vaccination in healthy volunteers and subjects with systemic lupus erythematosus

The aim of the study was to characterize performance of a complementary set of assays to measure antigen‐specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet...

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Published in	British journal of clinical pharmacology Vol. 76; no. 2; pp. 188 - 202
Main Authors	Ferbas, John, Belouski, Shelley S., Horner, Michelle, Kaliyaperumal, Arunan, Chen, Li, Boyce, Malcolm, Colaço, C. Bernie, McHugh, Neil, Quick, Vanessa, Nicholl, Richard J., Siu, Gerald, Chung, James
Format	Journal Article
Language	English
Published	England Blackwell Science Inc 01.08.2013
Subjects	Adjuvants, Immunologic - administration & dosage B-Lymphocytes - drug effects B-Lymphocytes - immunology ELISPOT immunoassay Enzyme-Linked Immunosorbent Assay flow cytometric bead immunoassay Flow Cytometry Hemocyanins - administration & dosage Hemocyanins - immunology Humans Immunoglobulin G - blood keyhole limpet haemocyanin Lupus Erythematosus, Systemic - immunology Lupus Erythematosus, Systemic - prevention & control Reviews systemic lupus erythematosus Vaccination vaccine response systemic lupus erythematosus keyhole limpet haemocyanin flow cytometric bead immunoassay vaccine response ELISPOT immunoassay
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ISSN	0306-5251 1365-2125 1365-2125
DOI	10.1111/bcp.12172

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Abstract	The aim of the study was to characterize performance of a complementary set of assays to measure antigen‐specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet haemocyanin (KLH) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array (CBA) that multiplexed the KLH response alongside pre‐existing anti‐tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT. These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti‐KLH IgG responses rose to a mean of 65–93‐fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260‐170‐fold above baseline. High levels of anti‐tetanus IgG were detected in pre‐immunization samples and their levels did not change over the course of study. Anti‐KLH IgG1‐4 subclasses were characterized by a predominant IgG1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti‐KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT, with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti‐KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune‐therapeutics.
AbstractList	The aim of the study was to characterize performance of a complementary set of assays to measure antigen‐specific immune responses in subjects immunized with a neoantigen. Healthy volunteers ( HV ) ( n = 8) and patients with systemic lupus erythematosus ( SLE ) ( n = 6) were immunized with keyhole limpet haemocyanin ( KLH ) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array ( CBA ) that multiplexed the KLH response alongside pre‐existing anti‐tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT . These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti‐ KLH IgG responses rose to a mean of 65–93‐fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260‐170‐fold above baseline. High levels of anti‐tetanus IgG were detected in pre‐immunization samples and their levels did not change over the course of study. Anti‐ KLH IgG 1‐4 subclasses were characterized by a predominant IgG 1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti‐ KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT , with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti‐ KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune‐therapeutics. The aim of the study was to characterize performance of a complementary set of assays to measure antigen-specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet haemocyanin (KLH) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array (CBA) that multiplexed the KLH response alongside pre-existing anti-tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT. These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti-KLH IgG responses rose to a mean of 65-93-fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260-170-fold above baseline. High levels of anti-tetanus IgG were detected in pre-immunization samples and their levels did not change over the course of study. Anti-KLH IgG1-4 subclasses were characterized by a predominant IgG1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti-KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT, with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti-KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune-therapeutics.The aim of the study was to characterize performance of a complementary set of assays to measure antigen-specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet haemocyanin (KLH) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array (CBA) that multiplexed the KLH response alongside pre-existing anti-tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT. These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti-KLH IgG responses rose to a mean of 65-93-fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260-170-fold above baseline. High levels of anti-tetanus IgG were detected in pre-immunization samples and their levels did not change over the course of study. Anti-KLH IgG1-4 subclasses were characterized by a predominant IgG1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti-KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT, with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti-KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune-therapeutics. The aim of the study was to characterize performance of a complementary set of assays to measure antigen-specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet haemocyanin (KLH) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array (CBA) that multiplexed the KLH response alongside pre-existing anti-tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT. These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti-KLH IgG responses rose to a mean of 65-93-fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260-170-fold above baseline. High levels of anti-tetanus IgG were detected in pre-immunization samples and their levels did not change over the course of study. Anti-KLH IgG1-4 subclasses were characterized by a predominant IgG1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti-KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT, with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti-KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune-therapeutics.
Author	Chung, James Horner, Michelle Kaliyaperumal, Arunan Belouski, Shelley S. Nicholl, Richard J. Colaço, C. Bernie Siu, Gerald Chen, Li Boyce, Malcolm Quick, Vanessa Ferbas, John McHugh, Neil
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CitedBy_id	crossref_primary_10_1002_cyto_b_21371 crossref_primary_10_1002_cpt_2539 crossref_primary_10_1111_bcp_14588 crossref_primary_10_1111_bcp_12202 crossref_primary_10_1111_bcp_12422 crossref_primary_10_1111_cts_13457 crossref_primary_10_3389_fddsv_2022_992087 crossref_primary_10_3389_fimmu_2022_1009304 crossref_primary_10_1080_1547691X_2019_1635234 crossref_primary_10_1126_scitranslmed_aar6584 crossref_primary_10_1136_lupus_2016_000146 crossref_primary_10_1016_j_medmic_2023_100088 crossref_primary_10_1038_srep43486 crossref_primary_10_1017_cts_2020_493
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Snippet	The aim of the study was to characterize performance of a complementary set of assays to measure antigen‐specific immune responses in subjects immunized with a... The aim of the study was to characterize performance of a complementary set of assays to measure antigen-specific immune responses in subjects immunized with a...
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SubjectTerms	Adjuvants, Immunologic - administration & dosage B-Lymphocytes - drug effects B-Lymphocytes - immunology ELISPOT immunoassay Enzyme-Linked Immunosorbent Assay flow cytometric bead immunoassay Flow Cytometry Hemocyanins - administration & dosage Hemocyanins - immunology Humans Immunoglobulin G - blood keyhole limpet haemocyanin Lupus Erythematosus, Systemic - immunology Lupus Erythematosus, Systemic - prevention & control Reviews systemic lupus erythematosus Vaccination vaccine response
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Title	A novel assay to measure B cell responses to keyhole limpet haemocyanin vaccination in healthy volunteers and subjects with systemic lupus erythematosus
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