This study examined 328 CFS sera in a study with 17 CCFP, 8 Gulf War Veterans (GWV), 24 Prostate Cancer (PC), and 52 normal sera in the modified Membrane Immunobead Assay (MIA) procedure for CTX. Three hundred and twenty-eight CFS patients' sera were examined by the modified MIA with purified MAb-CTX and 91.2% gave a titre > or =1:40. 76% of the 17 CCFP sera samples and 100% of the 8 GWV sera samples also had a titre > or =1:40. 92.3% of 52 normal sera showed titres of 1:20 or less, while 4 gave titres of > or =1:40. In addition, 41 sera were examined for Anti-Cardiolipin (aCL) by a commercial ELISA procedure with 87.8% demonstrating IgM, IgM+IgA, or IgM+IgG aCL antibodies. These results showed mostly the IgM aCL antibody alone in the sera samples. In addition, 41 serum samples were examined for aCL, with 37 showing positive for aCL, representing 90.2% positive for the three disease categories examined: CFS, CCFP and GWV. Examination for antiMitochondrial-M2 autoantibody (aM-M2) in 28 patients (CFS (18), CCFP (5), and GWV (5)) was negative for aM-M2. Inhibition analysis with antigens, CTX, CFS "Acute Phase Lipids", commercial Cardiolipin (CL) and 1,2-Dipalmitoyl-sn-Glycero-3-[Phospho-L-Serine] (PS) and antibodies, MAb-CTX and aCL from patients' serum show that the phospholipids in CL and CTX are antigenically indistinguishable with antibodies MAb-CTX and CFS-aCL. Preliminary chemical analyses have shown the lipids to be phospholipids associated with CL of the mitochondria. We designate this "Acute Phase Lipid" comparable to "Acute Phase Proteins" (C-reactive protein (CRP) and Serum Amyloid A (SAA)) in inflammatory conditions.
Examination of anticardiolipin antibodies (ACAs) in the sera of patients clinically diagnosed with chronic fatigue syndrome (CFS) using an enzyme-linked immunoassay procedure demonstrated the presence of immunoglobulin M isotypes in 95% of CFS serum samples tested. The presence of immunoglobulin G and immunoglobulin A isotypes were also detected in a subset of the samples. Future studies will focus on elucidating whether alterations to mitochondrial inner membranes and/or metabolic functions play a possible role in the expression of ACAs.
A plaque assay was employed to quantify the number of peripheral K cells in autoimmune thyroid diseases. The percentages of peripheral K cells determined were 3.1 +/- 2.2% and 3.3 +/- 1.3% (mean +/- SD) in forty-one patients with Graves' disease and twenty-two patients with Hashimoto's thyroiditis, respectively, which were significantly lower than 5.3 +/- 2.5% in 22 normal subjects. Circulating immune complexes (ICs) were measured by three different methods. The positivity of these ICs was investigated in relation to the level of peripheral K cells. The percentage of peripheral K cells in the patients positive for ICs was found to be lower than that in the patients negative for any ICs. Moreover, it was observed that the number of detectable K cells from a normal subject was significantly decreased by incubating the K cells with sera of patients positive for ICs. These results suggest that the decrease of peripheral K cells in the patients with autoimmune thyroid diseases may be due to saturation of K cell Fc receptors by binding ICs.
Acute phase phospholipids isolated from the sera of patients with clinically diagnosed chronic fatigue syndrome (CFS) were examined for effects on the sodium channel of mouse neuroblastoma cells in tissue culture after studies suggested a structural correlation between CFS lipids and the lipophilic marine toxin, ciguatoxin. The neuroblastoma cell bioassay is a directed cytotoxicity assay that records sensitive changes in the sodium channel of neuroblastoma cells in the presence of sodium channel activators or inhibitors. The ciguatoxins are characterized by their ability to cause the persistent activation of voltage‐sensitive sodium channels, leading to increased cell Na+ permeability. Ciguatera fish poisoning in humans is characterized by neurological, gastrointestinal, and cardiovascular manifestations that are similar to the symptoms of CFS.Several lipids were examined for sodium channel activity: ciguatoxin, cholesterol, cholesterol myristate, and the phospholipids isolated from CFS patients as well as antagonists gluconate, ursodeoxycholic acid, d‐ribose, and anti‐CTX antibody. Results indicate that cholesterol and cholesterol myristate have weak but observable interactions at the sodium channel as activators, and that CTX and CFS lipids both have strong activity as sodium channel activators. However, studies revealed that none of the potential antagonists evaluated were effective at blocking the activity of the lipids at the sodium channel.[Supported in part by National CFIDS Foundation, and the Hokama‐Yagawa Fund]
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