Herpesviruses and parkinsonism. Herpes simplex virus types 1 and 2, and cytomegalovirus antibodies in serum and CSF

Antibodies against herpes simplex virus (HSV) types 1 and 2 and cytomegalovirus (CMV) were assayed with a microindirect hemagglutination (IHA) test in the serum of 67 pairs of patients with Parkinson's disease and controls. Cerebrospinal fluid from 30 pairs was assayed. All patient and control serum was tested with a radioimmunoassay (RIA) for antibodies against HSV type 1 subunit antigens. Serum IHA antibody level against HSV type 1 was increased in patients with Parkinson's disease and RIA antibody levels against the same viral antigen were significantly higher in the patients than controls. Herpes simplex virus type 2 and CMV serum antibodies were equal in the patient and control groups. Most of the CSF samples tested negatively for IHA; small and comparable numbers of the patients and controls had low antibody levels against HSV and CMV antigens.     

Herpes simplex virus subunit antibodies in patients with Parkinson's disease.

Serum IgG antibodies against herpes simplex virus (HSV) type 1 capsid, envelope, and excreted antigens in 52 patients with idiopathic Parkinson's disease, and in their age- and sex-matched controls, were assayed with a solid-phase radioimmunoassay. When compared with the controls, patients with Parkinson's disease were found to have a substantially increased antibody response against each of the HSV subunit antigens tested. The increased antibody response in patients with Parkinson's disease was not associated with the occurrence of recurrent HSV infections, since the difference in antibody levels was most evident when comparing patients without recurrent HSV infections with their respective control group. Consequently, the increased HSV antibody response in patient with Parkinson's disease might depend on some antigenic stimulation other than ordinary recurrent HSV infections, or alternatively, on the generally enhanced immunological reaction of the patients against HSV.     

Virus antibodies in Parkinson's disease: Herpes simplex and measles virus antibodies in serum and CSF and their relation to HLA types

Serum and cerebrospinal fluid (CSF) immunoglobulin G (IgG) antibodies to herpes simplex (HSV) and measles viruses were assayed with a radioimmunoassay in 56 patients with idiopathic Parkinson's disease and in a similar number of age- and sex-matched controls with other neurological diseases. As a group, the patients with Parkinson's disease had a significantly increased serum antibody level against HSV, but measles virus antibody levels were similar in both groups. Both in the Parkinson's group and in the control group, the levels of the total IgG in CSF were within normal limits and the CSF antibodies to HSV and measles virus paralleled the serum antibody titers relative to the total IgG serum-to-CSF ratios. This indicates no increased intrathecal antibody production in either group. In 48 patients with Parkinson's disease who were HLA-typed, no association of viral antibody levels with particular HLA antigens were noted.The findings suggest that HSV is not present within the central nervous system of the patients with Parkinson's disease. The increased HSV antibody level seen in Parkinson's disease patients may reflect a more general disturbance of the patients' immune functions.     

Multiple sclerosis: cellular and humoral immune responses to several viruses.

One hundred and eight multiple sclerosis (MS) patients and 108 matched controls were studied for antibody levels and cellular immune responses to several viruses. There were significant increases in the mean titers of complement fixation (CF) or hemagglutination inhibition (HI), and complement-mediated cytotoxicity (CMC) tests for measles antibodies in MS patients; there was no increase in antibody titers to herpesviruses 1 and 2, or cytomegalovirus (CMV). The direct migration inhibition (DMI) tests showed no difference between MS patients and controls for measles, CMV, herpesviruses 1 and 2, or vaccinia virus antigens. Lymphocyte-mediated cytotoxicity (LMC) tests showed no difference between patients and controls, using cultures infected with measles and CMV viruses. In a study of stimulation or blocking of the LMC response by serum or cerebrospinal fluid (CSF), no effect was found. Therefore, increased levels of measles antibody in serum were again demonstrated in MS patients, but there was no difference in these patients' cellular immunity to measles virus versus that of the controls, and there was no abnormality of cellular immunity against the other viruses tested.     

Herpes simplex virus antibodies in patients with Parkinson's disease

Sera from 40 patients with idiopathic Parkinson's disease and their age- and sex-matched controls were assayed for immunoglobulin G (IgG) antibodies against herpes simplex virus (HSV) type 1-induced cell surface antigens with the indirect immunofluorescent test. An increased level of HSV antibodies was found among the patients with Parkinson's disease when the distribution of titres (P less than 0.001), the mean titres (P less than 0.001), or the mean paired titre difference (P less than 0.001) was compared with the controls. This may suggest a part of HSV in the pathogenesis of Parkinson's disease. Alternatively, the increased HSV antibody response might be an epiphenomenon indirectly associated with Parkinson's disease.     

HERPES SIMPLEX VIRUS-IgM SPECIFIC ANTIBODIES IN GUILLAIN-BARRÉ SYNDROME AND ENCEPHALITIS

Herpes simplex virus (HSV) has veen associated with a variety of inflammatory neurologic disorders. Recently we studied a patient with Guillain-Barré syndrome (GBS) following acute herpes vaginalis infection. Since IgM virus-specific antibody is thought to be a reliable indicator of acute viral infection, we employed a 2-h serologic assay for serum IgM antibodies to HSV using an indirect immunofluorescent technique. This patient demonstrated high serum IgM titers to HSV type 2 during the acute phase of her neurologic syndrome. The titer dropped substantially as convalescence progressed. A search for similar elevations in HSV-IgM specific antibody was made on sera from more than 70 other GBS patients. No other significant IgM antibody titers to either HSV type 1 or type 2 were found in this GBS series and a large number of neurologic controls. However, sera from two patients with a presumptive diagnosis of acute herpes encephalitis based on clinical and cerebrospinal fluid findings were positive, showing high titers in our test. The results of this study suggest that associated acute HSV infection is uncommon in GBS and an immunofluorescent seroassay of the type reported here may be a valuable noninvasive technique enabling the clinical laboratory to rapid confirm a diagnosis of herpes encephalitis.     

Virus antibodies in serum specimens from patients with multiple sclerosis, from siblings, and matched controls. A final report

Interviews were conducted with 229 patients with multiple sclerosis, 172 of their siblings and 219 matched controls from the general population. Serum specimens were taken and tested with 15 different antigens. An increased antibody response was observed in the patients and to a lesser extent in the siblings, but this seems to be directed almost selectively against measles virus antigens. The antibody levels to measles were highest in all study groups in that area in Finland for which the population has the highest relative risk to contract multiple sclerosis. No correlation could be found between the antibody levels and different clinical variables. Reports on increased antibody titers against different viruses in various chronic diseases, as well as the validity of serological results indicating an immunopathological process in the central nervous system, are discussed.     

Antibody and cell-mediated immunity to herpes simplex virus in psychotic depression.

The incidence and antibody titers to herpes simplex virus (HSV) were found significantly higher in patients with psychotic depression as compared to normal controls. Furthermore, the cell-mediated immunity (CMI) to HSV in psychotic depression was similar to that observed after acute HSV infection or recurrence. The results suggest therefore an association between HSV infection and psychotic depression.     

Increased titers of antibodies against streptococcal M12 and M19 proteins in patients with Tourette's syndrome

It has been suggested that a post-streptococcal autoimmune process may be involved in the pathogenesis of a subgroup of children with tics and obsessive-compulsive symptoms (PANDAS). Elevated antibody titers against streptococcal antigens have also been described in adult patients suffering from Tourette's syndrome (TS). In order to characterise further streptococcal antigens, we focussed on M proteins. M proteins are a major virulence factor of group A streptococci and known to evoke an immunologic cross-reaction with diverse epitopes of human tissue including brain tissue. Therefore, antibodies against M proteins may play a role in the pathophysiology of at least a subgroup of TS patients. Antibodies against M proteins were studied in 25 adult patients suffering from TS and 25 healthy controls after careful medical examination. The antibody titers against the peptides M1, M4, M6, M12 and M19 were estimated by ELISA. Our results show increased titers of antibodies against the streptococcal M12 and M19 proteins in TS patients as compared with controls, while antibody titers against M1, M4 and M6 did not differ between the TS and control groups. Elevated serum titers of antibodies against M12 and M19 proteins support the view that a streptococcus-induced autoimmune process may be involved in TS. The finding of a possible autoimmune origin of TS has implications for both pathophysiology and future therapeutic strategies.     

Serological diagnostic trial of the causative virus of Bell's palsy by anti-herpes virus antibodies in the paired sera]

Substantial evidence obtained through polymerase chain reaction techniques strongly supports that reactivation of latently infected herpes simplex virus (HSV) in geniculate ganglia is the main cause of Bell's palsy. However, serum antibody titers to HSV rarely increase in patients with this disease. This discrepancy may result from the difficulty in detecting a small increase in antibody titers by conventional serological analysis. To detect such a small increase in antibody titer, we defined the significant increase in IgG antibody titers more precisely, and examined the association of HSV or varicella-zoster virus (VZV) with Bell's palsy. From 40 patients with Bell's palsy, paired sera were obtained within the 4th disease day and 2 weeks later. IgG antibodies to HSV, VZV, cytomegalovirus (CMV), or measles virus (MsV) were measured with solid-phase enzyme immunoassay (EIA). IgM antibodies were measured with captured EIA. Each antibody titer was expressed as an EIA-value, which was quantitatively measured by using a calibration curve prepared from the samples containing known titers to the virus in proportion to the logarithm of its titer. An EIA-value ratio (EVR) between paired sera and a corrected EVR was calculated according to the formulae: EVR = [EIA-value in the second serum]/[EIA-value in the first serum]; corrected EVR = [EVR to HSV, VZV or CMV]/[EVR to MsV]. The association with a virus was determined to be positive when the corrected EVR of the virus was beyond the normal range (the logarithmical mean +/- 3 SD of corrected EVRs among 24 healthy controls) while the corrected EVR of the other viruses were within it. Using the corrected EVR, 6 patients were positive: 4 for HSV, 2 for VZV. On the other hand, the conventional serological analysis, which confirms positivity by a 4-fold increase in IgG antibody titer or a demonstration of IgM antibody, disclosed only 2 positive patients (1 for HSV, 1 for VZV). EIA is a very sensitive method of detecting antibodies. Corrected EVRs can exclude a decrease in antibody titers induced by corticosteroids, which are generally used for therapy of Bell's palsy. Moreover, the normal range of corrected EVRs can be defined more precisely than in conventional serological analyses. Our results indicate that some sensitive analysis, such as the corrected EVR method, may make it possible to serologically detect the causative virus of Bell's palsy.     

Histocompatibility types and measles antibodies in multiple sclerosis

The relationship between HLA antigens and measles antibody titers was investigated in 105 multiple sclerosis (MS) patients. HLA antigens were determined serologically by a micro-lymphocytotoxicity test and measles antibody titers were determined by the hemagglutination inhibition test. There was an increased frequency of HLA-B7 and HLA-Bw35 antigens and a decreased frequency of HLA-A2, HLA-B12, and HLA-Bw40 antigens in MS. Measles antibody titers were significantly higher in MS than in control cases. In the MS patients, the chi-square test for homogeneity did not show any significant difference between the presence or absence of antigens HLA-A3, HLA-B7, HLA-B8 HLA-A2 or HLA-B12 when compared with measles antibody titers. The chi-square test for association did not show any significant difference between the presence or absence of these antigens compared with measles antibody titers above 1:64 or 1:128 or 1:256. However, there were significantly higher measles antibody geometric mean titers in MS patients with HLA-A3 antigen or lacking HLA-A2 antigen. In the case of HLA-A3, this was reflected in the female but not in the male patients. The increased measles HI antibody titer in MS may be related to the increased frequency of HLA-A3, or the decreased frequency of HLA-A2, or both.     

Measles and canine distemper virus antibodies in patients with multiple sclerosis determined by radioimmunoassay

Antibodies against measles virus (MV) and canine distemper virus (CDV) were measured by solid-phase radioimmunoassay (RIA) of sera and cere-brospinal fluid (CSF) from 28 patients with multiple sclerosis (MS) and matched neurological controls. When the groups were compared for MV antibody titers and CDV antibody titers of sera and MV/CDV serum antibody titer ratios, no significant difference was found. The CDV antibody titers and the MV antibody titers were in good correlation. CDV antibodies showed RIA titration curves typical of low avidity antibodies. In tests for MV antibodies in CSF, 82% of the MS patients and 19% of the controls were positive, whereas 36% of the MS patients and 4% of the controls were positive in CDV RIA. The correlation between MV and CDV antibody levels, the low avidity of CDV antibodies and the fact that absorption of the specimens with MV antigen abolished all CDV antibody activity suggest that the CDV antibodies are MV antibodies cross-reacting with CDV. It is concluded that canine distemper virus is unlikely to be involved in the etiology of multiple sclerosis.     

Detection of antibody classes and subpopulations in Myasthenia gravis patients using a new nonradioactive enzyme immunoassay

To investigate the presence of antibodies in myasthenia gravis (MG) patients, we have developed a new reproducible and sensitive enzyme immunoassay (EIA-AChR), in which a β subunit-specific monoclonal antibody (mAb 73) immobilizes fetal calf acetylcholine receptors (AChRs). We tested 92 MG patients (42 with positive and 50 with negative antibody titers), 60 healthy controls, and 40 controls with other autoimmune diseases. EIA-AChR detected immunoglobulin G (IgG) titers in all of the seropositive samples, with a significant correlation between these and those obtained using the traditional immunoprecipitation method. Moreover, 5 seronegative patients at immunoprecipitation assay were positive at EIA-AChR. EIA-AChR was also useful in revealing: (1) a seropositive patient subpopulation with generalized MG who had Abs directed against α-Bungarotoxin binding sites; and (2) patients with IgM directed against fetal calf AChR (detected in 13 seronegative and 16 seropositive MG patients, and in 6 of the patients with other autoimmune diseases). © 1997 John Wiley & Sons, Inc. Muscle Nerve 20: 800–808, 1997     

Anti-glycolipid antibodies and their immune complexes in multiple sclerosis

Antibody titers against myelin constituents in sera and CSF of patients with multiple sclerosis (MS) were examined by a solid-phase radioimmunoassay. Anti-GM4 and anti-galactocerebroside antibody titers were significantly elevated in the CSF of MS patients, but not anti-GM1 and anti-myelin basic protein antibodies. In sera of MS patients, the titers of antibodies against these myelin constituents were not elevated. Total IgG level was also significantly elevated in the CSF, but not in the sera of MS patients. Immune complexes from the CSF of MS patients were dissociated by acid-ultrafiltration and assayed for antibodies to GM4, GM1, and galactocerebroside. Anti-GM4 and antigalactocerebroside antibody titers were significantly enhanced after acid dissociation and ultrafiltration. These data suggest that antibodies of the IgG class against GM4 and galactocerebroside are present in CSF of MS patients, and a significant number of them exist as immune complexes with their corresponding glycolipid antigens.     

Isolated cranial neuropathy associated with anti-glycolipid antibodies

We describe seven patients with isolated cranial neuropathy in whom serum anti-glycolipid antibodies were detected. Trigeminal sensory neuropathy was found in four patients, who had exhibited symptoms for 2 months to 4 years. The other three patients showed facial nerve palsy with or without ophthalmoparesis. Temporal profile analysis of anti-glycolipid antibodies revealed that titers of anti-glycolipid IgM antibodies against GM2 and LM1 gradually decreased in patients having chronic trigeminal sensory neuropathy. In patients with acute trigeminal sensory neuropathy, elevation of anti-LM1 antibody titers continued over 12 months although anti-GalNAc-GD1a antibody disappeared. On the other hand, titers of anti-glycolipid antibodies rapidly decreased in patients with acute facial nerve palsy with or without ophthalmoparesis. We conclude that anti-glycolipid antibodies may play an important role in the development of isolated cranial neuropathy in some patients.     

Studies of cellular immunity, serum interferon titers, and natural killer cell activity in schizophrenic patients

In one of the hypotheses on the causes of schizophrenia it is proposed that viruses may play a role in the etiology and pathogenesis of at least some forms of schizophrenia. In our study, 30 healthy adults and 30 schizophrenic patients were investigated for humoral and cellular immunity against herpes simplex virus (HSV) and cytomegalovirus (CMV). A whole blood test system with several advantages over commonly used procedures was used to study cell-mediated immunity to CMV and HSV. Our data showed no difference between serum antibody levels or cellular immunity of the schizophrenic patients and control individuals as far as HSV and CMV were tested. Serum interferon may be indicative for the presence of a yet unidentified virus. Therefore, we tested interferon levels in the serum of patients. No interferon was detected in any of the samples tested. Further, we tested the activity of natural killer (NK) cells in the blood of schizophrenic patients and controls. NK-activity of the patients' blood cells was not different from that of normal donors.     

Fine specificities of antibodies in sera and cerebrospinal fluid in herpes virus infections of the central nervous system as detected by the antigen variable immunoblot technique

Applying the immunoblot technique a sensitive and specific method was developed for the detection of intrathecally synthesized antibodies against individual specific proteins that are antigens of various infectious agents causing encephalitis. Paired serum and cerebrospinal fluid (CSF) samples from five patients with herpes virus infections of the central nervous system (CNS) (three herpes simplex virus encephalitis, one varicella zoster virus encephalitis, one zoster ganglionitis) were investigated for the presence of locally produced IgG against the electrophoretically separated antigens of herpes simplex virus (HSV), varicella zoster virus (VZV) and human cytomegalovirus (HCMV), as well as for IgM antibodies in one case of HSV encephalitis. In two cases (HSV encephalitis and VZV encephalitis) four and one antibody, respectively, were found that were synthesized intrathecally only. In the other cases the patterns of sera and CSF antibodies were similar, the CSF antibodies showing an all-over stronger reaction, at identical IgG concentrations. In contrast to the conception of a 'limited heterogeneity' of intrathecal antibody synthesis in encephalitis, we thus found an 'expanded heterogeneity' of the intrathecally synthesized antibodies in comparison to the corresponding serum antibodies.     

Serum autoantibodies measured by immunofluorescence confirm a failure to differentiate PANDAS and Tourette syndrome from controls

PANDAS and some cases of Tourette syndrome (TS) have been proposed to be post-streptococcal movement disorders in which antibodies produced against group A beta-hemolytic streptococcus cross react against brain epitopes. Attempts to identify disease specific anti-striatal antibodies in the serum of affected patients have focused on the use of Western immunoblotting and ELISA methodologies. In this study, immunohistochemical techniques were used to identify serum anti-striatal antibody reactivity. In positive samples, double staining with anti-GFAP (glial) and anti-MAP2 (neuronal) was used to establish localization of the immunofluorescence. No significant differences in immunofluorescence or localization were identified in patients with PANDAS (n=30) and TS (n=30) as compared to controls (n=30). IF reactivity did not correlate with tic severity or elevated titers of antistreptococcal antibodies. Further comparisons showed no correlation between autoreactivity determined by immunofluorescence and the presence of previously measured immunoblot reactivity against human caudate or putative antigens (pyruvate kinase M1 and aldolase C). These results confirm an inability to distinguish patient populations by antibody measurements and raise further concerns about the presence of an autoimmune mechanism in PANDAS and TS.     

Effect of ara-A on the experimental herpes simplex virus encephalitis--pathology and fluctuation of the cerebrospinal fluid and serum antibody levels

The effect of Ara-A treatment on the experimentally induced herpes simplex virus (HSV) encephalitis in rabbits was examined. Simultaneously practical value of enzyme-linked immunosorbent assay (ELISA) for rapid diagnosis was tested to find an early rise of antibody in cerebrospinal fluid (CSF) and serum in connection with an adoption of Ara-A administration. HSV encephalitis was produced by inoculation with type-1 HSV into the bilateral corneas of the eyes. A large amount of pus and typical neurological signs characteristic to encephalitis appeared after several days of infection. Ara-A therapy begun at the 3rd day of infection clearly increased the survival rate (90%) and improved the clinical signs, but the start of the drug at the 6th day could only increase the survival rate (60%) without cure of the clinical symptoms. ELISA could catch the early rise of IgG antibody in CSF from 3-4 days and in serum from 4-6 days after infection. Ara-A didn't interfere the antibody production. Viral antigens were found in the corneal epithelia and propria, neurons and Schwann's cells of trigeminal ganglia, neurons and glial cells of pons and cerebrum, especially in hippocampal nuclei. Typical Cowdry A and full type intranuclear inclusion bodies in the neurons were observed in the above tissues with infiltration of large amount of small round cells. In conclusion, Ara-A therapy in HSV encephalitis at the early stage of infection was quite effective for cure and the ELISA method is a useful tool for detection of early antibody rise in CSF and serum.     

Autoreactive IgG to intracellular proteins in sera of MS patients.

IgG binding to multiple protein constituents in lysates of Jurkat cells was detected by Western blot in sera of patients with multiple sclerosis (MS) and systemic lupus erythematosus (SLE). The distribution patterns of bands with sera tested against protein lysates from normal Jurkat cells or from Jurkat cells exposed to apoptosis or oxidative stress inducing conditions were similar in most patients, but with inter-individual differences. The number of bands with sera of both patient populations far exceeded those (0 or 2 bands) detected with sera of healthy controls. Proteinase K, RNase and DNase pre-treatment of cell lysates suggested a protein nature for all of the antigens and a ribonucleoprotein (RNP) nature for some of the antigens recognized by serum IgG of MS and SLE patients. Only two MS patients had positive anti-nuclear antibody (ANA) titers, while all of them had positive Western blots. In addition to similarities, dissimilarities were also recognized between the humoral immune responses in MS and SLE. No IgG molecules were detected against phosphorylated proteins in the sera of MS patients, while multiple phosphoproteins were recognized by IgG molecules of SLE patients in immunoprecipitation experiments. These data suggest that in addition to ANA, the sera of MS patients contain autoantibodies directed against multiple intracellular proteins. The protein recognition patterns of immunoglobulins in MS share similarities, but also have distinct features when compared to those in SLE. The biological significance of these autoantibodies in MS remains to be understood.