Prevention and reversal of experimental autoimmune myasthenia gravis by a monoclonal antibody against acetylcholine receptor-specific T cells.

We have recently described an algorithm to design, among others, peptides with complementarity contour to autoimmune epitopes. Immunization with one such peptide resulted in a monoclonal antibody (mAb), termed CTCR8, that specifically recognized Vbeta15 containing TCR on acetylcholine receptor (AChR) alpha-chain residue 100-116-specific T cells. CTCR8 was found to label the cell surface of AChR100-116-specific T cell lines and clones, immunoprecipitate the TCR from such cells, and block their proliferative responses to AChRalpha100-116. In the present report, we have found that there is a marked reduction in IFN-gamma and no effect on IL-10 production in a CTCR8-treated AChRalpha100-116-specific T cell line. Interestingly, when AChR100-116-primed, primary T cells were stimulated with peptide and treated with CTCR8, there was once again inhibition of IFN-gamma but also marked stimulation of IL-10 production. The change in the Th1/Th2 cytokine profile was paralleled by a reduction in AChR-specific IgG2a and IgM with no effect on IgG1. Remarkably, the most profoundly inhibited Ab population was that which causes experimental autoimmune myasthenia gravis (EAMG) by reaction with the main immunogenic region (alpha61-76) of the AChR. Based on these results, CTCR8 was tested for prophylactic and therapeutic effects in EAMG. EAMG induced by immunization with purified native Torpedo AChR was both inhibited and reversed by CTCR8. These findings suggest a means to produce therapeutic mAb for the treatment of autoimmune diseases.

[Manifestation of primary biliary cirrhosis and Basedow's disease caused by exposure to carbon monoxide in a patient with HTLV-1 associated myelopathy].

A 55-year-old woman with HTLV-1 associated myelopathy (HAM) was discovered in a rentan kotatsu (Japanese foot warmer with a frame and a coverlet by burning briquet) with conscious disorder and admitted to an emergency hospital. Her conscious disturbance waned the 3rd day after admission with gradual improvement of communication and food intake. However, on the 18th day after admission, her orientation was poor again and she was unable to take food for herself and keep sitting. She was diagnosed as suffering from an interval form of acute carbon monoxide (CMO) poisoning and transferred to our hospital for the purpose of hyperbaric oxygen therapy on the 20th day after exposure to CMO. In the course of treatment she recovered but showed jaundice, pruritus, liver dysfunction and elevation of antimitochondrial antibody. She received liver biopsy and was found to have primary biliary cirrhosis (PBC). On the 150th day, she manifested perspiration and hypertension. The clinical and immunological feature revealed her Basedow's disease. The relationship between HAM and PBC due to the autoimmune process has been predicted by investigators. The implication of autoimmune disease and HLA haplotype is a main focus of attention. Our case supports their hypothesis, and suggested that the complication occurred with immunological and genetic correlation. Anti-HTLV-1 antibody was positive at a titer of 1:8192 before exposure to CMO. On transferring to us, it was negative and revealed excessive positive at a titer of 1:20,480 on the 80th day. Immunoglobulin analysis was normal on admission and increased during hospitalization. It was reported that prenatal exposure to relatively mild concentrations of CMO in rats reduces splenic macrophage phagocytosis and killing ability as well as macrophage respiratory burst. These data suggested that PBC and Basedow's disease were manifested by exposure to carbon monoxide.

A peptide vaccine that prevents experimental autoimmune myasthenia gravis by specifically blocking T cell help.

Myasthenia gravis (MG) and its animal model, experimental autoimmune (EA) MG, are caused by T cell-dependent autoantibodies that react with the nicotinic acetylcholine receptor (AChR) on muscle and interfere with neuromuscular transmission. Thus, selective inactivation of CD4(+) AChR-specific T helper cells should lower AChR Ab levels and ameliorate disease. In the Lewis rat model of EAMG, alpha chain residues 100-116 of the AChR represent the dominant T cell epitope, which is important in helping Ab responses to this autoantigen. In the present report, we have applied a new design technique that requires no knowledge of Ag receptor sequences on errant T cells in order to develop a synthetic peptide vaccine against T cells reactive with the aforementioned T cell epitope. Immunization with the peptide 1) induced polyclonal and monoclonal Ab, which inhibited AChR 100-116 stimulation of AChR-sensitized lymphocytes and recognized Vbeta15 containing T cell receptors on AChR 100-116-specific T cell lines and clones; 2) lowered AChR Ab levels; 3) reduced the loss of muscle AChR; and 4) lessened the incidence and severity of EAMG. These findings suggest a new strategy for the functional abrogation of epitope-specific T cells that could have potential application to human autoimmune diseases.

A complementary peptide vaccine that induces T cell anergy and prevents experimental allergic neuritis in Lewis rats.

We have developed and described a new method of altering T cell-mediated autoimmune diseases by immunization with the complementary peptide against T cell epitopes. The complementary peptide (denoted NAE 07-06) to the bovine P2 protein, residues 60-70 (denoted EAN 60-70), was tested in the Lewis rat model of experimental allergic neuritis (EAN). Immunization with NAE 07-06 induced polyclonal and monoclonal Abs that inhibited the proliferation of the P2-specific T cell line, stimulated with EAN 60-70, and recognized Vbeta, but not Valpha, of TCRs. Proliferation of T cells treated with anti-NAE 07-06 Abs could be partially restored by treatment with rIL-2, in accordance with an anergy model. A homologous sequence was found between NAE 07-06 and the VDJ junction of the TCR beta-chain from an EAN 60-70-specific T cell line. Rats preimmunized with NAE 07-06 in vivo before EAN induction showed less disease severity clinically and histologically. These data suggest a new therapeutic approach for T cell-mediated autoimmune disorders through the induction of anti-TCR Abs with complementary peptide Ags.

Plasma lactic acid and pyruvic acid levels in migraine and tension-type headache.

We examined the lactic and pyruvic acid levels in the plasma of 14 patients with migraine, 17 patients with tension-type headache, and 12 normal controls. The lactic and pyruvic acid levels in the plasma of the migraine patients were significantly higher than those of the normal controls (9.6 +/- 5.0 mg/dL and 0.51 +/- 0.30 mg/dL versus 3.3 +/- 1.9 mg/dL and 0.26 +/- 0.20 mg/dL, respectively). There were no significant differences in the levels between the patients with tension-type headache and normal controls. Our results suggest that migraine patients may have functional abnormalities in mitochondrial energy metabolism.

[Neurologic signs and symptoms in viral infections--disease involving meninges, central nervous system or peripheral nerve system].

Knowledge about the roles of virus in the neurological infection has been expanded because of the recent advancement of viral detection techniques such as enzyme immunoassay and polymerase chain reaction: However, detection of neurological signs and symptoms can still indicate a specific virologic diagnosis. In addition, neurological symptoms that are less commonly seen or newly described are reviewed, including opsoclonus myoclonus syndrome, Rasmussen's encephalitis and slow viral infections.

Prevention of experimental autoimmune myasthenia gravis by a monoclonal antibody to a complementary peptide for the main immunogenic region of the acetylcholine receptors.

We have previously reported that a complementary peptide (denoted RhCA 67-16), encoded by RNA complementary to that of the Torpedo acetylcholine receptor (AChR) main immunogenic region (MIR), AChR residues alpha 61-76, induces polyclonal and monoclonal Ab reactive with Ig against the AChR MIR. RhCA 67-16 vaccination also protected against the development of experimental autoimmune myasthenia gravis (EAMG) in Lewis rats. In the present report, we found that a mAb (denoted TCM 240, IgG1 kappa) against RhCA 67-16 recognized three different idiotypic Ab (mAb 6, mAb 35, and mAb 198), which were previously reported by others to recognize the AChR MIR and to cause EAMG. Based on these results, TCM 240 was tested for prophylactic effects in EAMG. EAMG induced passively by mAb 35 was inhibited by simultaneous injection with TCM 240. The disease severity was inversely paralleled by the ratio of mAb 35 to TCM 240. EAMG induced by immunization with purified native Torpedo AChR was also inhibited by TCM 240, but not a control mAb. The inhibitory effect of TCM 240 on actively induced EAMG occurred without significantly lowering the overall AChR Ab levels, which indicates a limited repertoire of disease-causing Ab in EAMG and perhaps MG. Such findings suggest the existence of an EAMG-associated Id and also support the concept of an MIR. In a more general sense, these results demonstrate that prophylactic and perhaps diagnostic mAb for autoimmune diseases can be produced by immunization with complementary peptides for disease-associated epitopes.

The two-color analysis of the T-lymphocyte subsets in experimental allergic neuritis in rats.

We examined the T-cell subsets in the peripheral blood, spleen and lymph nodes during the course of experimental allergic neuritis (EAN) by using two-color analysis. In the acute phase, the percentages of CD4+ major histocompatibility complex (MHC)-II+ cells and CD8+ MHC-II+ cells in the lymph nodes of EAN rats were significantly higher than in the control rats. In the recovery phase, the percentage of CD4+ CD45RC+ cells and CD8+ CD45RC+ cells in the lymph nodes in EAN rats were significantly higher than in the control rats. However, significant changes of the T-cell subsets were not detected in either the spleen or the peripheral blood during the course of EAN. These results suggest that CD4+ MHC-II+ cells, CD8+ MHC-II+ cells, CD4+ CD45RC+ cells and CD8+ CD45RC+ cells may play a role in the course of EAN. The relationship between these double staining cells and EAN is discussed.

The CD5+ B cells and myasthenia gravis.

A high frequency of CD5+ B lymphocytes in the peripheral blood of patients with myasthenia gravis (MG) has been reported recently. These results seem to indicate an attractive linkage between CD5+ B lymphocytes and autoantibodies against Acetylcholine receptor in MG. We examined the frequency of CD5+ B cells in 20 patients with MG and 21 normal healthy controls by two-color flow cytometry. However, there were no significant differences in the percentages of CD5+ B lymphocytes between the two groups. We also examined the frequency of CD5+ B lymphocytes in the resected thymus of patients. The frequency of CD5+ B lymphocytes in the thymus was low and similar pattern to that in the peripheral blood. We checked the antibody (Ab) production against the human acetylcholine receptor in either CD5+ B or CD5- B lymphocytes using B lymphoblastoid cell line generated from the lymphocytes of 11 patients with anti-AChR Abs in the sera. Abs against the AChR in the human were mostly produced by CD5- B, not CD5+ B lymphocytes. The anti-AChR Abs (IgG) production of CD5+ B cells and CD5- B cells (mean +/- SD) were 6.8 +/- 2.4 fmol/ml and 18.5 +/- 17.6 fmol/ml, respectively. These results suggest that in MG, the frequencies of the CD5+ B lymphocytes in PBL may be genetic background and that there may be no strong linages between AChR Ab production and CD5+ B lymphocytes.

Use of complementary peptides and their antibodies in B-cell-mediated autoimmune disease: prevention of experimental autoimmune myasthenia gravis with a peptide vaccine.

We have developed and describe a new method of altering B-cell-mediated autoimmune diseases by induction of anti-idiotypic (Id) antibodies (Abs) by immunization with complementary peptides. Specifically, a peptide denoted RhCA 67-16 encoded by RNA complementary to RNA for the Torpedo acetylcholine receptor (AChR) main immunogenic region, AChR 61-76, was tested in the Lewis rat model of experimental autoimmune myasthenia gravis (EAMG). Immunization with RhCA 67-16 induced monoclonal and polyclonal anti-Id Ab against Abs to Torpedo AChR 61-76. RhCA 67-16 antisera inhibited AChR binding by AChR-specific Abs. In addition, a mAb to RhCA 67-16 (denoted TCM 240) recognized two well known EAMG-causing mAbs, 6 and 35. TCM 240, but not a control mAb F28C, inhibited mAb 6 binding to Torpedo AChR 67-76 peptide. mAb 35 binding to TCM 240 was inhibited by native Torpedo AChR as well as by RhCA 67-16. In in vivo experiments, immunization with a RhCA 67-16 keyhole limpet hemacyanin (KLH) conjugate blocked the development of EAMG after challenge with native Torpedo AChR (25% disease incidence versus 90% in the controls). This new approach may provide a novel therapy for MG and perhaps other B-cell-mediated autoimmune disorders through the induction of anti-Id Abs with complementary peptide antigens.

Prevention of experimental autoimmune myasthenia gravis by manipulation of the immune network with a complementary peptide for the acetylcholine receptor.

Myasthenia gravis (MG) and experimental autoimmune myasthenia gravis (EAMG) are caused, in part, by the production of autoantibodies against the main immunogenic region, amino acids 61-76, of the alpha chain of the acetylcholine receptor (AChR). Theoretically, induction of anti-idiotypic (Id) antibodies (Abs) should be a highly specific treatment for the disease by virtue of their potential ability to neutralize Abs to the AChR. We have tested this idea by attempting to evoke such anti-Id Abs by immunization with a peptide (termed RhCA 67-16) encoded by RNA complementary to the Torpedo AChR main immunogenic region and determining whether such treatment will prevent the development of EAMG. Immunization with RhCA 67-16, but not a control peptide termed PBM 9-1, was found to elicit the production of anti-Id Abs that blocked recognition of native Torpedo AChR by its Ab. This anti-Id Ab activity was ablated by incubation of the anti-RhCA 67-16 serum with RhCA 67-16, but PBM 9-1, prior to the assay for Ab binding to AChR. The anti-Id Ab-inducing activity of RhCA 67-16 was confirmed by the ability to produce a rat monoclonal Ab to RhCA 67-16 that showed anti-Id activity for polyclonal rat Ab reactive with AChR residues 67-76. Most importantly, RhCA 67-16 immunization also prevented the development of EAMG in Lewis rats challenged with Torpedo AChR (25% incidence versus 90% in the controls) and diminished the AChR Ab levels in animals injected with low doses of AChR. Our results suggest a therapy for MG and perhaps other autoimmune diseases through the induction of anti-Id Abs by peptide immunogens.

Immunoglobulin kappa/lambda ratios in migraine and tension-type headache.

The possibility of immunological mechanisms causing headaches has been proposed in the past. To investigate the immunological system activation in patients with chronic headaches, we evaluated the kappa/lambda ratios of immunoglobulins in 40 patients with migraine and 49 patients with tension-type headache. Nineteen healthy volunteers composed the control group. The serum kappa and lambda levels of immunoglobulins were determined by using the enzyme-linked immunosorbent assay (ELISA) techniques. The kappa/lambda ratios of IgG in the patients with tension-type headache were significantly higher than those in the controls. The kappa/lambda ratios of IgA and IgM in the patients with headaches were higher than those in the controls, but they were not statistically significant. The total concentrations of IgG, IgA and IgM were significantly higher in the patients with migraine. In the patients with tension-type headache, the total concentrations of IgG and IgA were significantly higher than those in the controls. The high levels of kappa/lambda ratios of IgG in the patients with tension-type headache and the increase in the total concentrations of immunoglobulins in the patients with migraine and tension-type headache, observed in this study, suggest that the humoral immunological system activation might exist, and it might be related to the etiology of tension-type headache and migraine.

Recurrent herpes zoster myelitis treated with human interferon alpha: a case report.

Recurrent herpes zoster myelitis is very rare. However, a case was recently observed in our hospital. A 43-year-old woman developed myelitis 2 weeks after development of shingles. Her condition was improved by methylprednisolone. Seven months later, she developed myelitis after development of shingles again. Antibody against varicella-zoster (VZV), increased in the serum, but was negative in the cerebrospinal fluid. Methylprednisolone was not sufficiently effective against this attack. The refractory sensory disturbance was improved by human interferon alpha (IFN-alpha). Natural killer cell activity, the helper T-cell/suppressor T-cell ratio and the kappa/lambda ratio of B-cells increased with clinical improvement. In this case, delayed-type hypersensitivity after VZV infection played a role in the occurrence of myelopathy and clinical improvement resulted from the immunosuppressive effects of IFN-alpha.

Immunosuppressive effect of FK506 on experimental allergic neuritis in Lewis rats: change of T cell subsets.

The effect of a new immunosuppressant, FK506, on experimental allergic neuritis (EAN) was examined in Lewis rats. EAN was induced by inoculation with bovine peripheral myelin. The EAN rats were divided into two groups. FK506-treated EAN rats were prophylactically administered FK506 by injection into the peritoneal cavity at a dosage of 5.0 mg/kg/day for 13 days beginning the day after inoculation. The control EAN rats were injected with only saline solution. FK506 prevented the development of EAN, histologically and clinically. In FK506-treated EAN rats, flow cytometric analysis of T cell subsets of the lymph nodes showed a significant decrease in W3/13 positive T cells on the 14th and 21st day and a decrease in W3/25 positive T cells on the 21st day after inoculation when compared with the control EAN rats. The percentage of OX-8 positive T cells were not significantly different between the two groups. Our results suggest that FK506 prevented the development of EAN by decreasing W3/13 and W3/25 positive T cells.

Interleukin-6 levels in the cerebrospinal fluid of patients with multiple sclerosis.

We developed the competitive enzyme-linked immunosorbent assay for interleukin-6 (IL-6). The detection limit was 30 pg per ml. Using this method, we examined the IL-6 levels in the cerebrospinal fluid of 10 patients with multiple sclerosis (MS) and 12 age-matched normal controls. IL-6 was detected in 6 out of 10 patients with MS. There was no significant correlation between the IL-6 levels and other parameters, including IgG, IgG index, cell counts, total protein and albumin in the CSF. Our results suggest that IL-6 detected in the MS-CSF may have no correlation to the immunological processes.

Reduced natural killer cell activity in patients with dementia of the Alzheimer type.

We examined the natural killer (NK) cell activity in 50 patients with dementia of the Alzheimer type (DAT) and 37 age-matched normal controls. The NK cell activity in DAT was significantly lower than in the normal controls. The NK cell activity induced by either interferon-alpha (IFN-alpha) or interleukin-2 (IL-2) in DAT was also significantly lower than in the normal controls. There were no significant differences in the level of serum IL-2 and IFN-alpha between the two groups. As regards NK cell subsets, two-color flow cytometric analysis showed no significant differences between the percentages of Leu-11+ Leu-7- cells, Leu-11+ Leu-7+ cells and Leu-11- Leu-7+ cells in the two groups. Our results suggest that NK cells in DAT may have functional abnormalities and may provide important clues to fundamental cellular and molecular aberrations in DAT.

Serum interleukin-2 levels in patients with dementia of the Alzheimer type.

We examined serum interleukin 2 (IL-2) levels in 127 normal subjects and 22 patients with dementia of the Alzheimer type (DAT) using the enzyme-linked immunosorbent assay technique. There was no significant correlation between serum IL-2 levels and age. However, serum IL-2 levels in elderly normal subjects (older than 70 years of age) were significantly lower than in the younger normal subjects (younger than 30 years of age) (p less than 0.05). There were no significant differences in serum IL-2 levels between the patients with DAT and the age-matched normal controls. These results suggest that the changes of serum IL-2 levels in patients with DAT may be natural in the normal aging process.