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Investigating the Antigen Specificity of Multiple Sclerosis Central Nervous System-Derived Immunoglobulins.

Willis SN, Stathopoulos P, Chastre A, Compton SD, Hafler DA, O'Connor KC - Front Immunol (2015)

Bottom Line: This infiltrate often includes B cells that are found in multiple locations throughout the CNS, including the cerebrospinal fluid (CSF), parenchyma, and the meninges, frequently forming tertiary lymphoid structures in the latter.However, the antigen(s) driving this response have yet to be conclusively defined.We conclude that while MS CNS resident B cells display the characteristics of an antigen-driven B cell response, the antigen(s) driving this response remain at large.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Yale School of Medicine , New Haven, CT , USA ; Walter and Eliza Hall Institute of Medical Research , Parkville, VIC , Australia ; Department of Medical Biology, University of Melbourne , Parkville, VIC , Australia.

ABSTRACT
The central nervous system (CNS) of patients with multiple sclerosis (MS) is the site where disease pathology is evident. Damaged CNS tissue is commonly associated with immune cell infiltration. This infiltrate often includes B cells that are found in multiple locations throughout the CNS, including the cerebrospinal fluid (CSF), parenchyma, and the meninges, frequently forming tertiary lymphoid structures in the latter. Several groups, including our own, have shown that B cells from distinct locations within the MS CNS are clonally related and display the characteristics of an antigen-driven response. However, the antigen(s) driving this response have yet to be conclusively defined. To explore the antigen specificity of the MS B cell response, we produced recombinant human immunoglobulin (rIgG) from a series of expanded B cell clones that we isolated from the CNS tissue of six MS brains. The specificity of these MS-derived rIgG and control rIgG derived from non-MS tissues was then examined using multiple methodologies that included testing individual candidate antigens, screening with high-throughput antigen arrays and evaluating binding to CNS-derived cell lines. We report that while several MS-derived rIgG recognized particular antigens, including neurofilament light and a protocadherin isoform, none were unique to MS, as non-MS-derived rIgG used as controls invariably displayed similar binding specificities. We conclude that while MS CNS resident B cells display the characteristics of an antigen-driven B cell response, the antigen(s) driving this response remain at large.

No MeSH data available.


Related in: MedlinePlus

MS and control-derived rIgG binding to MOG detected with a cell-based assay. Representative binding of MS (MS-B1) or germinoma control-derived (GCT-A3) rIgG to Jurkat cells transfected with MOG-GFP (left column) or GFP alone (right column). Histograms show the MFI of transfected cells gated on those that were positive for both GFP and a florescent anti-human secondary antibody (red). The blue histograms show secondary antibody alone. A humanized monoclonal antibody, h8-18c5, specific for human MOG served as a positive control for the Jurkat–MOG–GFP binding. FACS data for additional rIgGs from the MS and control groups are shown in the Supplementary Material.
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Figure 2: MS and control-derived rIgG binding to MOG detected with a cell-based assay. Representative binding of MS (MS-B1) or germinoma control-derived (GCT-A3) rIgG to Jurkat cells transfected with MOG-GFP (left column) or GFP alone (right column). Histograms show the MFI of transfected cells gated on those that were positive for both GFP and a florescent anti-human secondary antibody (red). The blue histograms show secondary antibody alone. A humanized monoclonal antibody, h8-18c5, specific for human MOG served as a positive control for the Jurkat–MOG–GFP binding. FACS data for additional rIgGs from the MS and control groups are shown in the Supplementary Material.

Mentions: We also examined binding to MOG; autoantibodies to MOG have recently been described in a small subset of MS patients (13), in pediatric MS (14) and in NMO (39). MOG binding was evaluated using a cell-based assay that preserves conformational epitopes and, accordingly, has become a widely accepted approach for detection of such antibodies (13). Robust binding by the humanized monoclonal anti-MOG monoclonal antibody (h8-18C5) was recorded (Figure 2). The clear recognition of MOG by our humanized h8-18C5 demonstrates that our recombinant expression system produces fully functional whole human IgG and did not introduce any artifacts that might confound native specificity. Applying this approach to the MS- and germinoma-derived rIgG demonstrated that none of these antibodies recognized MOG expressed on the surface of the cells (Figure 2; Figure S1 in Supplementary Material).


Investigating the Antigen Specificity of Multiple Sclerosis Central Nervous System-Derived Immunoglobulins.

Willis SN, Stathopoulos P, Chastre A, Compton SD, Hafler DA, O'Connor KC - Front Immunol (2015)

MS and control-derived rIgG binding to MOG detected with a cell-based assay. Representative binding of MS (MS-B1) or germinoma control-derived (GCT-A3) rIgG to Jurkat cells transfected with MOG-GFP (left column) or GFP alone (right column). Histograms show the MFI of transfected cells gated on those that were positive for both GFP and a florescent anti-human secondary antibody (red). The blue histograms show secondary antibody alone. A humanized monoclonal antibody, h8-18c5, specific for human MOG served as a positive control for the Jurkat–MOG–GFP binding. FACS data for additional rIgGs from the MS and control groups are shown in the Supplementary Material.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663633&req=5

Figure 2: MS and control-derived rIgG binding to MOG detected with a cell-based assay. Representative binding of MS (MS-B1) or germinoma control-derived (GCT-A3) rIgG to Jurkat cells transfected with MOG-GFP (left column) or GFP alone (right column). Histograms show the MFI of transfected cells gated on those that were positive for both GFP and a florescent anti-human secondary antibody (red). The blue histograms show secondary antibody alone. A humanized monoclonal antibody, h8-18c5, specific for human MOG served as a positive control for the Jurkat–MOG–GFP binding. FACS data for additional rIgGs from the MS and control groups are shown in the Supplementary Material.
Mentions: We also examined binding to MOG; autoantibodies to MOG have recently been described in a small subset of MS patients (13), in pediatric MS (14) and in NMO (39). MOG binding was evaluated using a cell-based assay that preserves conformational epitopes and, accordingly, has become a widely accepted approach for detection of such antibodies (13). Robust binding by the humanized monoclonal anti-MOG monoclonal antibody (h8-18C5) was recorded (Figure 2). The clear recognition of MOG by our humanized h8-18C5 demonstrates that our recombinant expression system produces fully functional whole human IgG and did not introduce any artifacts that might confound native specificity. Applying this approach to the MS- and germinoma-derived rIgG demonstrated that none of these antibodies recognized MOG expressed on the surface of the cells (Figure 2; Figure S1 in Supplementary Material).

Bottom Line: This infiltrate often includes B cells that are found in multiple locations throughout the CNS, including the cerebrospinal fluid (CSF), parenchyma, and the meninges, frequently forming tertiary lymphoid structures in the latter.However, the antigen(s) driving this response have yet to be conclusively defined.We conclude that while MS CNS resident B cells display the characteristics of an antigen-driven B cell response, the antigen(s) driving this response remain at large.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Yale School of Medicine , New Haven, CT , USA ; Walter and Eliza Hall Institute of Medical Research , Parkville, VIC , Australia ; Department of Medical Biology, University of Melbourne , Parkville, VIC , Australia.

ABSTRACT
The central nervous system (CNS) of patients with multiple sclerosis (MS) is the site where disease pathology is evident. Damaged CNS tissue is commonly associated with immune cell infiltration. This infiltrate often includes B cells that are found in multiple locations throughout the CNS, including the cerebrospinal fluid (CSF), parenchyma, and the meninges, frequently forming tertiary lymphoid structures in the latter. Several groups, including our own, have shown that B cells from distinct locations within the MS CNS are clonally related and display the characteristics of an antigen-driven response. However, the antigen(s) driving this response have yet to be conclusively defined. To explore the antigen specificity of the MS B cell response, we produced recombinant human immunoglobulin (rIgG) from a series of expanded B cell clones that we isolated from the CNS tissue of six MS brains. The specificity of these MS-derived rIgG and control rIgG derived from non-MS tissues was then examined using multiple methodologies that included testing individual candidate antigens, screening with high-throughput antigen arrays and evaluating binding to CNS-derived cell lines. We report that while several MS-derived rIgG recognized particular antigens, including neurofilament light and a protocadherin isoform, none were unique to MS, as non-MS-derived rIgG used as controls invariably displayed similar binding specificities. We conclude that while MS CNS resident B cells display the characteristics of an antigen-driven B cell response, the antigen(s) driving this response remain at large.

No MeSH data available.


Related in: MedlinePlus