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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 neurofilament light (NF-L) by solid phase ELISA. MS-derived rIgG (n = 13) and control rIgG (n = 8) derived from a germinoma were tested by solid phase ELISA for binding to NF-L. The specific samples included in the assay are shown in the Supplementary Material. Each dot or square represents the binding of a single rIgG. The dashed line indicates the mean +2 SD of the control germinoma-derived cohort (0.76). Values above this line were determined to be positive (95% CI). To correct for non-specific binding, the reported ELISA signal (ΔOD) was calculated by subtracting the signal generated by binding to glyceraldehyde 3-phosphate dehydrogenase (GADPH) from that of the NF-L. The mean and SD are shown for each data set. Statistical differences are indicated when significant. Data associated with each rIgG for the MS and control groups are shown in the Supplementary Material.
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Figure 1: MS and control-derived rIgG binding to neurofilament light (NF-L) by solid phase ELISA. MS-derived rIgG (n = 13) and control rIgG (n = 8) derived from a germinoma were tested by solid phase ELISA for binding to NF-L. The specific samples included in the assay are shown in the Supplementary Material. Each dot or square represents the binding of a single rIgG. The dashed line indicates the mean +2 SD of the control germinoma-derived cohort (0.76). Values above this line were determined to be positive (95% CI). To correct for non-specific binding, the reported ELISA signal (ΔOD) was calculated by subtracting the signal generated by binding to glyceraldehyde 3-phosphate dehydrogenase (GADPH) from that of the NF-L. The mean and SD are shown for each data set. Statistical differences are indicated when significant. Data associated with each rIgG for the MS and control groups are shown in the Supplementary Material.

Mentions: To investigate the specificity of CNS-derived antibodies, we began by screening against candidate antigens that have previously been implicated in MS. A DELFIA and an ELISA assay were performed to test MBP (37) and contactin (26), respectively. Differences in binding to MBP and contactin between the MS and control rIgG were unremarkable (not shown). We also used an ELISA assay to assess binding of MS and control rIgG to the intracellular protein neurofilament light (NF-L) (38). Ten MS-derived antibodies that were tested showed modest binding to NF-L (Figure 1) while three antibodies (MS-B1, MS-C2, and MS-C4) displayed strong binding with absorbance values that exceeded the mean +2SD of the control data set (benchmark for strong positive binding). The difference between the MS and the control group was significant (p = 0.0018, Mann–Whitney test). However, binding was not restricted to MS-derived antibodies as a germinoma-derived antibody (GCT-A6) was also positive, indicating a lack of specificity for MS in the rIgG cohorts.


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 neurofilament light (NF-L) by solid phase ELISA. MS-derived rIgG (n = 13) and control rIgG (n = 8) derived from a germinoma were tested by solid phase ELISA for binding to NF-L. The specific samples included in the assay are shown in the Supplementary Material. Each dot or square represents the binding of a single rIgG. The dashed line indicates the mean +2 SD of the control germinoma-derived cohort (0.76). Values above this line were determined to be positive (95% CI). To correct for non-specific binding, the reported ELISA signal (ΔOD) was calculated by subtracting the signal generated by binding to glyceraldehyde 3-phosphate dehydrogenase (GADPH) from that of the NF-L. The mean and SD are shown for each data set. Statistical differences are indicated when significant. Data associated with each rIgG for 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 1: MS and control-derived rIgG binding to neurofilament light (NF-L) by solid phase ELISA. MS-derived rIgG (n = 13) and control rIgG (n = 8) derived from a germinoma were tested by solid phase ELISA for binding to NF-L. The specific samples included in the assay are shown in the Supplementary Material. Each dot or square represents the binding of a single rIgG. The dashed line indicates the mean +2 SD of the control germinoma-derived cohort (0.76). Values above this line were determined to be positive (95% CI). To correct for non-specific binding, the reported ELISA signal (ΔOD) was calculated by subtracting the signal generated by binding to glyceraldehyde 3-phosphate dehydrogenase (GADPH) from that of the NF-L. The mean and SD are shown for each data set. Statistical differences are indicated when significant. Data associated with each rIgG for the MS and control groups are shown in the Supplementary Material.
Mentions: To investigate the specificity of CNS-derived antibodies, we began by screening against candidate antigens that have previously been implicated in MS. A DELFIA and an ELISA assay were performed to test MBP (37) and contactin (26), respectively. Differences in binding to MBP and contactin between the MS and control rIgG were unremarkable (not shown). We also used an ELISA assay to assess binding of MS and control rIgG to the intracellular protein neurofilament light (NF-L) (38). Ten MS-derived antibodies that were tested showed modest binding to NF-L (Figure 1) while three antibodies (MS-B1, MS-C2, and MS-C4) displayed strong binding with absorbance values that exceeded the mean +2SD of the control data set (benchmark for strong positive binding). The difference between the MS and the control group was significant (p = 0.0018, Mann–Whitney test). However, binding was not restricted to MS-derived antibodies as a germinoma-derived antibody (GCT-A6) was also positive, indicating a lack of specificity for MS in the rIgG cohorts.

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