Neutralizing antibodies to canine distemper and measles virus in multiple sclerosis.

Serum neutralizing antibody to canine distemper virus (CDV) was measured in 142 patients with multiple sclerosis (MS), an equal number of age and sex-matched normal controls, and 75 patients with other neurological disorders. An elevated antibody titer was found in the MS population compared to controls. Measles-neutralizing antibody was also found to be elevated in 128 patients with MS. The mean ratio of measles to CDV antibody was similar in the control and MS groups, but a wide range of measles to CDV ratios was found in individual patients and controls. No significant correlation was found between IgG levels and neutralizing antibody titers in MS sera. While the increased titer of measles and CDV antibody may represent a non-specific immunological response, these findings are also consistent with the hypothesis that CDV or a closely related measles virus may stimulate the increased level of measles and CDV antibodies found in MS sera.     

Complement dependent cytotoxic antibody activity against measles virus in multiple sclerosis

Summary The presence of measles cytotoxic (CT) and hemagglutination inhibiton (HI) antibodies in 195 multiple sclerosis (MS) patients and 251 controls was tested. The measles virus Lu carrier cells labeled with ⁵¹Cr were exposed to serum specimens in the presence of complement in order to test the presence of CT antibody. The analysis of complement dependent CT antibodies against measles virus revealed significantly (P<0.01) higher titers in MS patients than in the control group. However, the measles HI test failed to show this difference. Measles CT titers 1:32 among MS patients occured in 54.9% and in 35.5% among the controls. In comparison with this the HI method revealed measles titers 1:128 more often in the control group than in MS cases (27.9 and 17.9%, respectively). The presence of CT antibodies against measles virus in MS proves that these patients have a functional defence mechanism to eliminate virus infected cells. The high measles antibody titer among MS patients could be due to recurrent antigenic stimulation caused by measles virus persistency. Whether this virus persistency plays a role in MS can not be decided on the available data.Supported by Deutsche Forschungsgemeinschaft (Schwerpunkt Ätiologie und Pathogenese der multiplen Sklerose und verwandter Erkrankungen)     

Incidence of CSF abnormalities in siblings of multiple sclerosis patients and unrelated controls

We found that 19% (9/47) of healthy siblings of patients with clinically definite multiple sclerosis had an intrathecal immunological reaction with two or more 2 CSF-enriched oligoclonal bands (OCBs), in contrast to (4%) (2/50) unrelated healthy controls. Furthermore, in this group of nine healthy sibs the measles CSF IgG antibody titers were higher than that of the other sibs and that of controls. There were also differences in the serum titers for measles IgG antibody, which were higher in the group of all healthy sibs than in healthy volunteers, and (as with CSF titers) higher in the subgroup of healthy sibs with two or more 2 CSF-enriched OCBs than the other sibs. Thus a significant proportion of healthy siblings to MS patients have a partially hyperimmune condition similar to that occurring in MS, which in 19% manifested itself as an OCB reaction, in 9% as increased CSF measles IgG antibody titers, and in 21% as increased serum measles IgG antibody titers, these phenomena tending to occur in the same individuals. This condition is characterized by CSF-enriched OCBs with undefined specificity, although some increased antiviral reactivity is found both in the serum and CSF. While it needs further characterization, a genetic trait interacting with common infections is suggested. The recurrence risk of this condition is approximately five times higher than the 3-4% recurrence risk for manifest MS reported for sibs.     

Risk factors in multiple sclerosis: a population-based case-control study in Hautes-Pyrénées, France

A population-based study of the prevalence and risk factors of multiple sclerosis (MS) was conducted in the Hautes-Pyrénées, the southwestern region of France. The prevalence rate per 100,000 was equal to 40. Data on the past medical history of 63 MS patients and matched controls were collected. The frequency and age at occurrence of common childhood infections were similar for both the MS cases and controls. There was no difference between the frequency of vaccination for MS patients and for controls. However, the age at which MS patients were immunized against poliomyelitis was significantly higher than the corresponding age for controls (15.8 years versus 8.9 years, P less than 0.01). Antibody titers for various viruses were measured. The mumps antibody titer was significantly higher in the MS patients than in the controls. Also, MS patients tended to have higher titers for measles antibodies.     

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.     

Comprehensive viral immunology of multiple sclerosis. II. Analysis of serum and CSF antibodies by standard serologic methods

Sera and CSFs of 85 patients with multiple sclerosis (MS), 49 patients with probable MS, and 165 control patients with other neurologic diseases were assayed for antibodies to rubella, mumps, measles, parainfluenza I (strain 6/94), herpes simplex, cytomegalovirus, varicella-zoster, and vaccinia viruses. Methods included complement fixation (CF), hemagglutination inhibition (HI), and complement-dependent plaque reduction (CPR). Significant differences between the groups with MS and the control groups were higher serum antibody titers to measles virus in the groups with MS, higher proportion of patients with MS with CSF antibodies to measles, rubella, and vaccinia viruses, and greater percentage of patients with MS with more than one CSF viral antibody. Duration and severity of disease in the patients with MS were associated with presence of multiple CSF antibodies. Presence of CSF antibody was positively correlated with the height of the correspnding serum titer, yet a high serum titer did not ensure the presence of CSF antibody. Oligoclonal bands were present in the CSFs of equal proportions of patients with MS with and without CSF viral antibody. Our data support the hypothesis of local antibody synthesis within the CNS. However, we favor the view that preprogrammed antibody-forming lymphocytes enter the CNS and then produce antibody either because of nonspecific polyclonal activation in situ or because of failure of normal regulation.     

Lymphocyte blast transformation responses and viral antibodies in relation to HLA antigens in multiple sclerosis

Fifty four clinically stable multiple sclerosis (MS) patients and 54 age- and sex-matched control subjects were HLA-typed, and their responses to herpes simplex, measles, mumps and rubella antigens were examined by the lymphocyte blast transformation test and by serum antibody titrations. Blast transformation response to purified tuberculin (PPD), mitogen phytohaemagglutinin (PHA), pokeweed mitogen (PWN) and concanavalin A (Con A) and spontaneous proliferation of lymphocytes were also studied. MS patients differed from controls by higher antibody levels to measles and rubella viruses and by lower specific blast transformation responses to rubella and measles antigens. When the relative strength of transformation responses was measured, mumps and herpes simplex responses were also lower in MS patients than in controls. In addition, spontaneous lymphocyte proliferation of MS patients in 6-day cultures was lower than that of control lymphocytes. In mitogen stimulations there were no differences between whole groups, but the oldest patients had lower responses to PHA and Con A than their matched controls. The frequency of HLA-Dw2 was 56.6% in MS patients and 32.1% in controls. The patients with and without Dw2 differed from each other only by a lower specific response to PPD in the Dw2-positive group. The immunological response of Dw2-positive controls resembled that of MS patients: low transformation response to viral antigens, low spontaneous proliferation and elevated measles antibodies. This finding supports the function of a genetically determined type of immune responsiveness with low cell-mediated immunity and high levels of certain viral antibodies as one susceptibility factor in multiple sclerosis.     

Optic neuritis and distribution of genetic markers of the HLA system

Thirty patients with optic neuritis (ON), in which neither multiple sclerosis (MS) nor any other etiology could be discriminated, were reexamined after a mean observeation period of 5 years. Eleven patients revealed oligoclonal IgG in the CSF and in five of them a measles virus antibody response within the CNS was demonstrable. the remaining 19 patients did not display oligoclonal CSF IgG and no local antibody production was detectable.
The occurrence of the HLA antigens A3 and B7 in ON did not correlate to the presence of oligoclonal IgG in CSF. the frequencies did not differ from those found in controls. the HLA-B7 linked lymphocyte defined antigen HLA-Dw2 occurred in ON at increased frequency, which was intermediate to that observed in MS and controls. an association was found in ON between oligoclonal IgG in CSF and Dw2. This association was of the same magnitude as in 22 MS patients who had ON as their first symptom of MS. In ON without oligoclonal CSF IgG the frequency of Dw2 was similar to that of controls.
No association was observed between the occurrence of the HLA antigens A3, B7 and Dw2, and increased measles antibody titers in serum or a measles virus antibody response in the CNS.
The occurrence of oligoclonal IgG in CSF in patients with ON may be assumed to increase the risk of developing MS.     

The effect of live, attenuated measles vaccine and measles infection on measles antibody levels in serum and CSF of patients with multiple sclerosis or clinically isolated syndrome

High occurrence of measles, rubella and varicella zoster antibodies has been used as a biomarker for MS (the MRZ test). We analyzed measles antibody titres with respect to measles infection/measles vaccination status in 166 patients with MS or clinically isolated syndrome. Fifty blood donors served as controls. Measles vaccination yielded CSF measles antibodies in fewer patients (62%) than measles infection did (87%, p=0.001) and yielded lower measles titres in both serum and CSF (p<0.001). Controls had lower CSF measles titres than patients with measles vaccination alone (p<0.001). Childhood vaccinations probably reduce the sensitivity of the MRZ diagnostic test for MS.     

OLIGOCLONAL IgG AND MEASLES ANTIBODY IN CSF OF MULTIPLE SCLEROSIS PATIENTS

Cerebrospinal fluid (CSF) was obtained from 50 neurological patients, 26 of whom suffered from multiple sclerosis (MS). Agar gel electrophoresis and determination of anti-measles antibody titre were performed on the CSF samples. The frequency of elevated measles antibody titre was higher among the MS patients than among the patients with other neurological diseases. Furthermore, oligoclonal IgG in CSF occurred more often among the MS patients than among the other neurological patients. An association between high measles antibody titre and the presence of oligoclonal IgG was observed in CSF from both the MS patients and the patients with other neurological diseases. However, the nature of the correlation between occurrence of oligoclonal IgG and high measles antibody titre remains to be explained.     

Measles virus-specific immunoglobulin D antibody in cerebrospinal fluid and serum from patients with subacute sclerosing panencephalitis and multiple sclerosis

Quantitation of measles-specific immunoglobulin D (IgD) antibody was carried out in cerebrospinal fluid (CSF) and serum samples from 18 patients with subacute sclerosing panencephalitis (SSPE), 12 patients with multiple sclerosis (MS) and seven normal controls with high measles antibody titers in serum, using polyclonal and monoclonal antibodies specific for human IgD and enzyme-linked immunosorbent assay. Measles-specific IgD activity was significantly higher in CSF and serum from SSPE patients compared to that found in patients with MS or normal controls. The IgD antibody to measles virus was not due to high levels of measles-specific IgG since significant measles IgD activity was found after eluting IgG from SSPE serum. The increased level of measles-specific IgD found in SSPE sera is consistent with the levels observed in patients with acute and chronic viral infections.     

VIRUS ANTIBODY LEVELS IN THE CEREBROSPINAL FLUID FROM PATIENTS WITH OPTIC NEURITIS

Virus antibody levels were studied in the cerebrospinal fluid (CSF) of 58 patients with optic neuritis and 58 control patients with no indication of multiple sclerosis (MS) or infectious disorders of the central nervous system (CNS). The specimens were tested against three different structural components of measles virus with measles hemagglutination inhibition (HI), measles hemolysis inhibition (HLI) and gel precipitation (GP) tests. Measles antibodies occurred in 62 per cent of CSF specimens from patients with optic neuritis, and 21 per cent of the controls. In the specimens from patients with optic neuritis, the positive rate figures were: for rubella HI test 35, parainfluenza-1 HI 16, and Epstein-Barr virus immunofuorescence (IF) 53 per cent. The frequencies in the control group were 10, 10 and 26 per cent, respectively. Serum/CSF antibody ratios below 80 occurred in measles tests in 45 per cent of patients with optic neuritis and 16 per cent of the control group. Some patients with optic neuritis (but none from the control group) had a reduced serum/CSF antibody ratio in more than one measles antibody test, The patients with optic neuritis had a higher frequency of low serum/CSF albumin ratios indicating blood brain barrier damage, There were, however, several patients with a normal serum/CSF albumin ratio but low serum/CSF immunoglobulin G and measles antibody ratios. This supports the hypothesis that local production of measles antibodies takes place in CNS in some patients with optic neuritis as well as in MS patients. The CSF specimens were further tested against 12 other viruses and mycoplasma pneumoniae complement fixation, but there were no positive specimens. New CSF specimens were taken from five patients during optic neuritis, and from seven patients later on during the follow-up because of the appearance of new neurological symptoms. There were no changes in virus antibody levels, except for two patients with an increase of measles virus antibody titres.     

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.     

Pet ownership,distemper antibodies and multiple sclerosis

The incidence of cat or dog ownership was similar in 64 multiple sclerosis (MS) patients in south-east England and age- and sex-matched controls. 86% of both groups had owned a cat or dog before the date of onset of MS in the index case so that it would be difficult to detect an association between pet ownership and MS. The titres of neutralising antibody to canine distemper virus were higher in the MS patients (mean 48 ± 68) than the controls (32 ± 35), a significant difference (P < 0.05). This difference is not necessarily explained by a cross-reaction with measles antibody of which the titre was much higher in MS (610 ± 560) than in the controls (320 ± 330, P < 0.001).     

Virus antibodies in the cerebrospinal fluid of multiple sclerosis patients detected with ELISA tests

The enzyme-linked immunosorbent assay (ELISA) was used to determine levels of specific IgG antibodies against measles, rubella, vaccinia, corona (OC43) and mumps viruses in cerebrospinal fluid (CSF) and serum of 18 patients with clinically definite multiple sclerosis (MS), 8 patients with optic neuritis (ON), 27 patients with other neurological disease (OND), and 88 control subjects without central nervous system disease. Serum antibody levels were not significantly different between the four groups. Differences in the frequency and levels of CSF antibodies between the four groups were observed. Control patients had serum/CSF antibody ratios from 2.0 to 3.0 (log) with an average of 2.5 corresponding to a 320-fold difference between serum and CSF antibody levels. MS patients had ratios from 1.1 to 2.1 with an average of 1.6. The average was 2.0 for the ON patients. The average for the OND patients was similar to the controls. The altered serum/CSF ratios for several viruses within an individual patient was similar. These results suggest that nonspecific immunostimulation is responsible for the increased levels of CSF virus antibodies.     

MEASLES ANTIBODIES DETECTED WITH VARIOUS TECHNIQUES IN SERA OF PATIENTS WITH MULTIPLE SCLEROIS

Hemagglutination inhibition (HI) antibody titers to measles, rubella, mumps, and La Crosse virus in 137 patients with multiple sclerosis and 137 controls matched by date of birth and place of residence were measured and are reported according to age. The measles HI titers were significantly higher in serum specimens of patients with multiple sclerosis (geometric mean 1/82) than in the controls (geometric mean 1/43). In addition, the measles HI titers decreased significantly in the older age groups of the controls, whereas a similar decrease was not observed in the MS patients. No significant differences in the HI titers of rubella, mumps, and La Crosse virus were found between the MS group and the control group. Measles antibodies were further analyzed in a representative smaller series of sera from 49 MS patients and 49 controls by inhibition of salt-dependent hemagglutinin, gel precipitation, and platelet aggregation. With each of the techniques used, higher levels of measles antibody titers or stronger reactions were observed in the MS group. However, when measles antibodies detected with various techniques were compared in individual serum specimens of MS patients, no clear correlation between the high titers or strong reactions was discernible. The nature of the antibody response to measles virus in MS patients and the light thrown by the results obtained on the etiopathogenesis of multiple sclerosis is discussed.     

Genetic basis of multiple sclerosis: HLA antigens, disease progression, and oligoclonal IgG in CSF

The HLA antigens B7 and Dw2 occurred at elevated frequencies in 105 multiple sclerosis (MS) patients (49 and 47%, respectively), compared to healthy controls (29 and 30%), especially in MS patients with oligoclonal CSF-IgG (51 and 50%), in cases with CSF-IgG index values above 1.5 (64 and 64%), and in those with the most malignant course of the disease (47 and 59%). Normal or only slightly elevated frequencies of B7 and Dw2 were found in MS patients without oligoclonal CSF IgG (35 and 29%), normal CSF-IgG index (43 and 39%), and the most benign course (42 and 37%). No correlation was found between the HLA type and measles virus antibody titers in serum or a measles virus antibody response within the CNS.     

Comprehensive viral immunology of multiple sclerosis. III. Analysis of CSF antibodies by radioimmunoassay

The CSF from 279 patients with multiple sclerosis (MS), probable MS, or controls was examined by radioimmunoassay (RIA) for antibodies to measles, rubella, mumps, parainfluenza 1 (Sendai) (strain 6/94), herpes simplex (HSV), varicella, and vaccinia viruses. Significantly more patients with MS than noninflammatory control patients had antibody to measles, rubella and varicella viruses, of which antibody to measles was the most prevalent. The percentage of patients with MS with two or more CSF antibodies was significantly greater than that in the controls. There was no tendency for certain antibodies to be associated. There was a general relationship between presence of CSF antibodies and severity of MS. The data support the hypothesis of local CNS antibody synthesis of several viral antibodies; however, such local synthesis may be a random event, possibly dependent on the number and specificity of peripheral virus antibody-forming lymphocytes available for ingress into the CNS.     

Viral antibodies in twins with multiple sclerosis

Viral antibodies to measles, rubella, corona, vaccinia, and mumps viruses in serum and CSF (and to Epstein-Barr virus in serum only) were studied in 24 twin pairs, both discordant and concordant for clinical MS. In pairs, CSF antibody titers for rubella in MS monozygotic and dizygotic twins and for vaccinia in dizygotic twins were higher than for unaffected twins. Increased CSF titers among MS twins existed for measles, rubella, and vaccinia when pairing was ignored. Among MS twins, serum rubella and measles and CSF measles antibody titers, and CSF:serum ratios for measles virus, were higher in those who were DW2 positive.     

VIRUS ANTIBODY LEVELS IN SERUM SPECIMENS FROM PATIENTS WITH OPTIC NEURITIS AND FROM MATCHED CONTROLS

Virus antibody levels in serum specimens taken in acute and convalescent phases from 77 patients with optic neuritits were tested by measles hamagglutination inhibition (HI), measles hemolysis inhibition (HLI), rubella HI, parainfluenza-1 HI, Epstein-Barr immunofluorescence (IF), and against 11 other viruses and mycoplasma pneumoniae with the complement fixation (CF) technique. The virus antibody levels were indicated to be usually very stable, and a fourfold change in virus antibody levels was demonstrated in only eight patients. The virus antibody levels were compared with specimens from two carefully selected control groups. The first control group consisted of 71 healthy persons matched in age, sex and place of residence with the patients with optic neuritis. The other control group consisted of 58 patients with various neurological diseases other than multiple sclerosis (MS) or infectious diseases of the central nervous system. The patients with optic neuritis had significantly higher measles antibody titres than the two control groups in both measles HI and measles HLI tests. Also in 33 patients with optic neuritis of unknown cause, the measles antibody levels were higher than in the control groups. On the other hand, various other antibody tests showed no statistically significant differences between patients with optic neuritis and the control group.