Interplaying factors that effect multiple sclerosis causation and sustenance

Abstract

The author hypothesized that multiple sclerosis (MS) is a humoral autoimmune disease, caused by faulty interplay between myelin-specific, dimeric IgE, specifically competing non-IgE antibodies and IgE-triggered degranulating mast cells. The principal fault was believed to be insufficient quantity of protective, specific non-IgE antibodies. Also conjectured was the possibility of an unexpected and adverse immune suppression caused by none-MS pharmaceuticals being consumed by patients for their MS or for other conditions. To test both hypotheses, a mimotopic, peptide antigen-based, serum immunoassay was developed to measure dimer-bound IgE excess among MS patients, wherein the IgE specifically complexes with two or more myelin surface epitopes at an interval of 40-100 Angstroms, a separation critical for mast cell degranulation and cell damaging effect. MS test sensitivity and specificity, when analyzing five previously untreated patients for dimeric IgE presence, was 100%. In direct comparison, twenty age- and gender-matched female and male control subjects were test negative. Analysis of 35 multiple sclerosis patients, who were concomitantly being treated with potentially immunosuppressive pharmaceuticals, appeared to show the substances' negative effect upon MS causation, progression, or specific immunoassay performance. Therefore, MS is likely an autoimmune disease caused by IgE-mediated mast cell degranulation possibly in conjunction with immunosuppressive agents.

Figure 1

Check description in Section 2.

Figure 2

A schematic illustration of myelin proteolipid protein Isoform 1 is shown. Depicted are: (a) amino acid sequence portions that are net hydrophilic and located on the myelin protein (oligodendrocyte) surface (blue-green highlight); (b) portions that are net hydrophobic and project inwardly within the myelin glycolipid layer (uncolored); and (c) portions that are hydrophilic and intracellular (yellow highlighted).

Figure 3

A schematic drawing is shown wherein the location of a myelin-specific epitope, ADARM, is illustrated by performing a pictorially functional readjustment of Figure 1(a), removing rows 1–38, 48-49, 50–116, 155–188, 198-199, and 209–277 to visualize the 3 hydrophilic surface platforms, susceptible to autoantibody binding. The individual platforms encompass amino acids number 39−49, 189–197, and 200–208. Two intracellular, hydrophilic platforms encompass amino acids number, 117–141 and 145–154. The unique, corresponding amino acid hydrophilic indices, −0.5, 3, −0.5, 3, and −1.3 depicted in column 6.

Figure 4

Displayed is the measured distance between two IgE autoantibodies if each was to bind a potential epitopic dimer site (QAPEY and VTLRI) with each site incorporating five, uniquely sequenced, contiguous amino acids flanked on either end by a nonreactive, normally present amino acid thus making a 7 amino acid, antibody binding footprint. Each intervening amino acid between epitopes is estimated to be 10.6 Ångströms in width. When the interfootprint dimer distance analysis is performed, the potential dimer between QAPEY and VTLRI is inadequate for mast cell degranulation because there are 13 intervening amino acids between the two epitopes, and this is equivalent to a distance of 103 Ångströms, which is 3 Ångströms above the mandated upper limit of 100 Ångströms. HI: peptide hydrophilic index.

Figure 5

A potentially functional dimer site with an interval distance of 95 Ångströms between the epitopes VTLRI and HSYQE is illustrated.

Figure 6

A potentially functional dimer site with an interval distance of 56 Ångströms between the epitopes RNVRF and HSYQE is illustrated.

Figure 7

A potentially non-functional dimer site with an interval distance of 103 Ångströms between the epitopes RNVRF and AAMEL is illustrated.

Figure 8

A potentially functional dimer site with an interval distance of 95 Ångströms between the intracellular MOG epitopes IENLH and KTGQF is illustrated. The epitopes' complexing with specific IgE antibodies and mast cells likely hinges upon disruption of the overhanging oligodendrocyte membrane surface.

Figure 9

Illustrated are three potentially functional dimer sites with interval distances of 80, 71, and 64 Ångströms between the interlayer MOG epitopes NLHRT and KTGQF, LHRTF and KTGQF, and HRTFE and KTGQF. Dimeric IgE complexing hinges upon disruption of the overhanging oligodendrocyte membrane surface and facilitated intracellular antibody inflow. For serum antibody immunoassay purposes, the longer, inclusive peptide NLHRTFE can be used together with KTGQF as both peptides are sufficiently hydrophilic when coupled with the peptide-solubilizing, amino-ADOOA-ADOOA linker.

Figure 10

Seven structurally unique epitopes located on the outer surface of myelin basic protein (MBP) Isoform 1 are illustrated. The dimer group 1 combinations encompass the overlapping, epitopic pentamers DNEVF, NEVFG, EVFGE, and VFGEA coupled to QDTAV. The dimer group 2 combinations encompass the pentamers DNEVF, NEVFG, EVFGE, and VFGEA coupled to DTAVT. The dimer group 3 combinations encompass the overlapping, epitopic pentamers QDTAV and DTAVT individually coupled to PKNAW. Dimer group 1 displays epitope intervals that are 95, 87, 80, and 72 Ångströms. Dimer group 2 displays epitope intervals that are 88, 80, 72, and 64 Ångströms. Dimer group 3 displays epitope intervals that are 48 and 40 Ångströms. Dimeric IgE complexing hinges upon disruption of the overhanging oligodendrocyte membrane surface and facilitated intracellular autoantibody inflow. For serum antibody immunoassay purposes, the longer, inclusive peptide DNEVFGEA can be used together with QDTAVT and QDTAVT used together with PKNAW as all three peptides are sufficiently hydrophilic when coupled with the peptide-solubilizing construct, amino-ADOOA-ADOOA linker.

Figure 11

A second set of potentially functional intracellular dimer sites on are displayed on MBP Isoform 1. The inclusive epitope pairs are: DNTFK, LQTIQ and DNTFK, QTIQE and NTFKD, LQTIQ and NTFKD, QTIQE with respective interval distances of 40 and 48 Ångströms. Dimeric IgE complexing hinges upon disruption of the overhanging oligodendrocyte membrane surface and facilitated intracellular autoantibody inflow. For serum antibody immunoassay purposes, the longer, inclusive peptide DNTFKD can be used together with LQTIQE as both peptides are sufficiently hydrophilic when coupled with the peptide-solubilizing, amino-ADOOA-ADOOA linker.

Figure 12

A third set of potentially functional intracellular dimer sites on MBP Isoform 1 is displayed incorporating the epitopes KDSHH, DSHHP, and SHHPA individually coupled to HGRTQ. The dimer epitopes' complexing with specific IgE antibodies likely hinges upon disruption of the overhanging myelin surface and specific antibody inflow. For serum antibody immunoassay purposes, the longer, inclusive peptide KDSHHPA can be used together with HGRTQ as both solubilize readily with the amino-ADOOA-ADOOA linker.

Figure 13

Potentially functional dimer sites on MBP Isoform 2 and conditions match the dimersets on MBP Isoform 1 displayed in Figure 10.

Figure 14

Potentially functional dimer sites and conditions on MBP Isoform 2 match a dimer set on MBP Isoform 1 as displayed in Figure 11.

Figure 15

Potentially functional intracellular dimer sites and conditions on MBP Isoform 3 which are similar to the third dimer set on MBP Isoform 1 as displayed in Figure 12.

Figure 16

MS test microplate layout of mimotopic peptide antigens covalently coupled to individual test wells. The test plate format was employed for both specific IgE and non-IgE serum antibody determinations.

Figure 17

MS test results for ten female control serum samples, ages 20–66, are displayed. Tested samples were from Caucasian and African-American donors who did not have multiple sclerosis. Specific IgE/(kappa + lambda)-positive results are confined to single, nondimer participating epitope.

Figure 18

MS test results for ten male control serum samples, ages 20–66, are displayed. Tested samples were from Caucasian and African-American donors who did not have multiple sclerosis. Specific IgE/(kappa + lambda)-positive results are confined to single, non-dimer participating epitope.

Figure 19

MS test results for serum samples obtained from multiple sclerosis patients (4 Caucasians and 1 African American) who had not yet received pharmacotherapy. Individual epitope- positive results are highlighted with yellow plus signs. To be dimer test positive, MS patients had to be ADARM-specific, IgE/(kappa + lambda)-positive and/or IgE/(kappa + lambda)-positive for the dimer pairs HSYQE and VTLRI, HSYQE and RNVRF, IENLH and KTGQF, and/or NLHRTFE and KTGQF.

Figure 20

MS test results for serum samples obtained from multiple sclerosis patients only treated with interferon or Copaxone. Individual epitope-positive results (all against the PLP epitope ADARM) are highlighted with a yellow plus sign. Being test-positive to PLP indicates dimer-positive presence because of the PLP monomers' high myelin surface prevalence and adequate intermolecular monomer-to-monomer separation (65–71 Ångströms).

Figure 21

MS test results for serum samples obtained from multiple sclerosis patients treated with interferon plus psychotropic pharmaceuticals and/or other potentially immunosuppressive agents. Individual dimer-positive results (just one) are highlighted with a yellow plus sign. The immunosuppressive substances are identified by yellow-highlighted numbers at the bottom of columnar, individual patient test results and referenced in literary citations provided in Table 3 that are listed at the end of the paper.

Figure 22

MS test results for serum samples obtained from multiple sclerosis patients only treated with psychotropic pharmaceuticals or other immunosuppressive agents. Individual epitope-positive test results (4 ADARM positives) are highlighted with a yellow plus sign. The immunosuppressive substances are identified by yellow-highlighted numbers at the bottom of the columnar, individual patient test results and referenced in literary citations provided in Table 3 that are listed at the end of the paper.

Figure 23

MS test results for serum samples obtained from multiple sclerosis patients treated with Copaxone plus psychotropic pharmaceuticals or other immunosuppressive agents. Individual epitope-positive results (just one tested individual) are highlighted with yellow plus signs. The immunosuppressive substances are identified by yellow-highlighted numbers at the bottom of columnar, individual patient test results and referenced in literary citations provided in Table 3 that are listed at the end of the paper.