The significance of Epstein-Barr virus seropositivity in multiple sclerosis patients?

The objective of this study was to evaluate and investigate the significance of the previously found 100% seropositivity toward Epstein-Barr virus (EBV) found in multiple sclerosis (MS) patients in contrast to healthy controls. Using a commercially available ELISA-test (Biotest), which differentiates infections with EBV into previous infections, primary infections, reactivated infections and no previous infection, we found 137 of 138 MS patients and 124 of 138 healthy controls seropositive. A primary infection in 4 of the 124 EBV seropositive healthy controls in contrast to no primary infections in the MS EBV seropositive group was significant (P=0.049652, Fishers exact test). This may be suggestive of a lack of primary infections in MS patients, and thus strengthens the idea that MS patients are infected with EBV before development of MS. Further studies are in progress to analyse whether EBV infection is a prerequisite for the development of this disease.

[Hepatitis C virus (HCV) status in recipients transfused with blood from anti-HCV-positive donors]. / Hepatitis C-virus (HCV)-status hos recipienter transfunderet med blod fra anti-HCV-seropositive donorer.

Ten donors positive for antibodies to hepatitis C were discovered in the community of Aarhus after the introduction of screening of blood donors. These donors had donated blood products to 123 recipients. Of these recipients 76 were dead and 21 were not contacted for various reasons. Follow-up of anti-HCV status was performed in the remaining 26 recipients. Twenty-four (92%) of the recipients were positive in the RIBA confirmatory test, one was inconclusive and one was negative. Nine (90%) of the donors were hepatitis C virus RNA positive, while 17 (68%) of the recipients were HCV-RNA positive. Altogether (donors and recipients) 25 (76%) of the HCV-RNA positive patients had abnormal liver enzymes, while all HCV-RNA negative patients had normal enzyme levels. Eight of eleven HCV-RNA positive patients had an abnormal liver biopsy, while one patient in the HCV-RNA negative group had an abnormal liver biopsy. Three have been treated with interferon. In view of the liver damage already found only few years after transfusion, follow-up investigations in order to identify younger persons transfused with hepatitis C positive donations should be carried out and patients offered treatment if necessary. The National Board of Health has decided to recommend this strategy.

Multiple sclerosis in the Faroe Islands. VI. Studies of HLA markers.

Occurrence of clinical neurologic multiple sclerosis (CNMS) among resident Faroese began between 1943 and 1973 and comprised three epidemics. The occupation by British forces for 5 years during World War II was interpreted to have been of major importance for the occurrence of these epidemics and led us to believe that CNMS is the rare, late result of a single, widespread, systemic and specific infectious disease which we have labelled the primary MS affection (PMSA). In this study we describe the occurrence of genetic markers of the HLA system in 16 Faroese MS patients, 25 of their siblings, 30 unrelated healthy neighbors and spouses to MS patients, 18 healthy controls from areas where no MS cases have been detected, and 80 unrelated normal Faroese. These studies show no significant deviations of HLA class I antigens, whereas the class II antigens do deviate: 50% of the Faroese MS patients carry the HLA-DR2 (DQ1/DRB 15) antigen, compared to a frequency of 15-20% among the control groups. Also the group of siblings of MS patients showed an increased frequency of DR4 (72%) compared to normal frequency among MS patients and other normal controls (43-47%). However, if DR15-positive individuals were excluded, this difference was further reduced. If PMSA was widespread within this group, DR4 or some closely associated genetic marker may confer protection against PMSA developing into CNMS. The occurrence of CNMS in these epidemics seems therefore associated to HLA class II-linked genetic factors similar to those found in studies of other caucasians with MS. This observation seems important in understanding the pathogenesis of this disease.

[Transfusion-associated graft-vs-host disease in a patient with Hodgkin's disease]. / Transfusionsassocieret graft-versus-host disease hos en patient med Hodgkin's sygdom.

Transfusion-associated graft-versus-host disease (TA-GVHD) is a serious complication of blood transfusion which is caused by immunocompetent donor lymphocytes reacting against recipient antigens. We report a case of TA-GVHD in a male patient with Hodgkin's disease who had received several units of non-irradiated blood components. TA-GVHD was diagnosed by histological examination of affected skin and demonstration of engrafted lymphocytes of female phenotype by in situ hybridization using a Y-chromosome specific probe. The need to irradiate blood components given to patients in defined risk-groups is stressed.

[HIV-positive persons in a Danish provincial region 1985-1990]. / HIV-antistofpositive i et dansk provinsområde 1985-1990.

From January 1985 to July 1990 a total of 128 HIV-infected persons were examined in the Clinics for Infectious Diseases and Dermato-Venereology at Odense University Hospital which covers a provincial region of Denmark. Clinical and demographic data were recorded regarding their initial contact with the hospital. About 1/2 (47%) were homosexual men and these were seen primarily during the first and last years of the observation period. Approximately 1/4 (27%) were intravenous drug users (15% of these were women), and about 1/5 (18%) had become infected by heterosexual relations. Half of these were women and these were seen mostly during the latter half of the observation period. The majority (81%) were clinically healthy at the time of the initial contact with the hospital. Eight patients (6%) had developed AIDS. It is noteworthy that during the last year of the observation period, there was a great increase in the number of homosexuals with compromised health and low CD4-cell count. This tendency could not be demonstrated for the other groups. In general, at the beginning of the observation period, a very uniform group of homosexual men was observed. Most of these had become HIV-infected in Denmark. They were healthy and with good immunological function and in the age group 25-49 years. During the subsequent years, a more varied group of patients was observed. The risk behaviour was different, the percentage of women greater and there was greater variation in age, health status and CD4-count.(ABSTRACT TRUNCATED AT 250 WORDS)

HLA or HLA-linked genes reduce birthweight in families affected by idiopathic recurrent abortion.

HLA typing was performed in 49 families, each containing at least one woman with a history of unexplained recurrent spontaneous abortion (RSA), and reproductive histories were recorded for the siblings. Abortion rate in sisters sharing two HLA haplotypes with the proband was significantly (p less than 0.01) increased, whereas abortion rates of brothers' wives seemed independent of the brothers' degree of HLA sharing with the proband. Birthweights of offspring of both the sisters and the brothers decreased with increasing HLA haplotype identity between the sibling and the proband (p = 0.05). The mean birthweight of infants of siblings sharing both parental haplotypes with the proband was 3158 g which was significantly (p less than 0.02) less than the average birthweight in Denmark (3417 g). Significantly (p less than 0.05) decreased mean birthweight in infants of brothers who were HLA-identical with the proband suggested that the disposition to growth retardation could be inherited with HLA also through the male gametes. It has been shown that, in their few successful pregnancies, women with RSA bear infants with a birthweight which is approximately 300 g less than normal. The present study demonstrated that the two obstetrical conditions, RSA and retarded fetal growth, share common genetic markers: HLA. It is suggested that HLA or HLA-linked genes causing growth retardation in fetuses are part of the putative complex of genes involved in the pathogenesis of recurrent abortion.

Association of maternal HLA haplotypes with recurrent spontaneous abortions.

Histocompatibility typing was done on 63 women with idiopathic, recurrent abortions (greater than or equal to 3 consecutive, spontaneous abortions; summarized abortion rate: 92.8%), 112 of their full siblings and 101 parents. The distribution of full sisters sharing 2, 1 and 0 of the probands' haplotypes diverged from expected Mendelian segregation (p less than 0.05). Sisters sharing both the probands' haplotypes had an abortion rate of 59.1%, haploidentical sisters had an abortion rate of 25.0% whereas 6.3% of the pregnancies of the 0 haploidentical sisters had ended in miscarriage. The probands' HLA haplotypes included a significant (p less than 0.02) excess of HLA-A, B haplotypes which have previously been shown to exhibit features of positive linkage disequilibrium in Danes. Analysis of abortion rates among the probands and their siblings indicate that the idiopathic recurrent abortion syndrome and probably also the tendency to some sporadic abortions are inherited conditions determined by genes in the HLA region. The presented data may be compatible with an additive polygenic mode of inheritance with exclusive or predominant phenotypic expression in females controlled by an HLA linked spontaneous abortion susceptibility region (SASR).

No increased histocompatibility antigen-sharing in couples with idiopathic habitual abortions.

Sharing of major histocompatibility antigens between spouses has been claimed to play an aetiological role in couples with unexplained, habitual spontaneous abortions. On the other hand there is evidence that a subset of habitually aborting women exists, in whom autoimmune aberrations seem to be of importance. In the present study, 56 couples with unexplained habitual abortions were histocompatibility typed for HLA-A, -B and -DR specificities, as were a group of 33 fertile control couples with two or more children and no spontaneous abortions. The DR-typing was done partly by use of the restriction fragment length polymorphism (RFLP). There was no significant difference in the distribution of histocompatibility antigens between the habitually aborting women and the control women. No increased HLA-antigen sharing could be demonstrated between spouses in aborting couples compared with controls, neither in each single locus nor in the combination of all loci (all P-values greater than 0.2). Neither could any increased HLA-antigen sharing in sub-groups of aborting couples with and without autoimmunity be demonstrated. It can be concluded from these data that compatibility between spouses for HLA-A, -B and DR-antigens per se only plays a minor role in the aetiology of habitual abortion.

HLA-DR typing using restriction fragment length polymorphism (RFLP) with one enzyme and two probes.

A panel of 43 homozygous and 36 heterozygous highly selected cells, representing the most common DR-specificities, were investigated with the DNA hybridization technique. By using a single restriction enzyme, TaqI, and two probes, DR beta and DQ alpha, it was possible to construct assignment criteria giving a reasonable definition of DR1, 3, 4, 5, 7 + w9, w8, w10 and w11. The criteria sometimes require that certain bands must not be present. Therefore, in certain genotypic combinations, the presence or absence of the particular specificity on one haplotype cannot be decided. This is a problem only for DR2 and DRw6, which for this reason cannot be assigned in about 1/3 to 1/4 of the cases. The association between RFLP assignment and serological assignment is not absolute, the correlation coefficients ranking from 0.62 to 1.0. In the case of false negative RFLP assignment, this may be due to genetic heterogeneity, as in the case of a DR2 individual who proved to be Dw12 and not Dw2 associated. It is often stated that interpretation of the RFLP pattern is particularly difficult in random or heterozygous individuals compared to proven homozygotes. This is not the case in the present study, where in fact correlation coefficients between RFLP and serologically determined DR specificities were higher in the heterozygotes (range 0.79-1.00).

Post mortem HLA-DR typing of donor corneas using DNA-hybridization technique.

With donor material kept under banking conditions, corneal grafting is today an elective routine surgical procedure. Increasing evidence suggests that the clinical results may be improved by HLA matching. However, serological tests require that a blood sample be taken while the donor is still alive or shortly post mortem. We report HLA-DR donor typing with a DNA hybridization technique using DNA isolated from the donor eye more than 24 h post mortem. The practical implications of this procedure are briefly discussed.

Antibodies to isolated hepatocytes in chronic active liver disease. Identification and diagnostic significance of bile canalicular antibodies.

A branched or net-like immunofluorescence pattern was demonstrated across the surface of isolated rabbit hepatocytes. We presume that this staining pattern is related to bile canaliculi on the cell surface, since it was closely correlated with the presence of bile canalicular antibodies, as detected by immunofluorescence on liver sections. Reaction with the putative bile canaliculi on rabbit hepatocytes was produced by 20 of 26 sera from patients with chronic active liver disease, 13 of 120 sera from patients with various liver diseases, and 1 of 40 normal blood donor sera.

Demonstration of organ-nonspecific antigens in liver-specific protein.

Liver-specific protein was isolated from bovine, rabbit, and human liver. A corresponding kidney protein was isolated from rabbit kidney. The liver-specific protein and kidney-specific protein were coupled to activated CH-Sepharose 4B. Affinity chromatography was performed on columns of liver-specific protein and kidney-specific protein CH-Sepharose 4B, with sera containing antimitochondrial antibodies, smooth muscle antibodies, bile canalicular antibodies, reticular tissue antibodies, and antinuclear antibodies. The material eluted from the affinity chromatography columns was studied for immunofluorescence reaction against sections of liver, kidney, and stomach. Reactions were found of the liver-specific protein and kidney-specific protein column eluates against mitochondria, smooth muscle, double-stained bile canaliculi, reticular tissue, and nucleoli. Thus, the antigens belonging to these cell and organ constituents must have been present in the liver-specific protein (and kidney-specific protein) preparations, indicating that liver-specific protein is not a pure liver cell membrane-specific protein.

Anti-actin antibodies revealed by counter-immunoelectrophoresis. Relation to smooth muscle antibodies and bile canalicular antibodies.

In investigations by counter-immunoelectrophoresis, anti-actin antibodies were found in 59% of patients with chronic hepatitis and in 8% of patients with non-hepatic diseases and normal blood donors. Anti-actin antibodies were found more frequently in patients with hepatitis and IgG smooth muscle antibodies than in other groups of diseases and normal subjects with IgG smooth muscle antibodies. Anti-actin antibodies showed no correlation with bile canalicular antibodies.

Peripheral immunofluorescence of hepatocytes. Relation to smooth muscle antibodies and bile canaliculi.

By means of indirect immunofluorescence microscopy, a peripheral fluorescence reaction with hepatocytes was found, surrounding the entire cell, with sera from patients with cancer and acute hepatitis and from normal blood donors. It was proved that this reaction was not related to bile canaliculi, contrary to bile canaliculi antibody demonstrated previously from patients with chronic active liver disease. By using fluorescein-conjugated anti-human IgG, the reaction with the periphery of the hepatocytes was produced to a more or less pronounced degree with almost all sera studied. The reaction seems not to be directed against contractile proteins on the liver cell membrane as only a minor part of these sera had also IgG smooth-muscle antibody, and anti actomyosin antibody obtained by affinity chromatografy did not react with the hepatocyte. When applying fluorescein-conjugated anti-human IgM the reaction with the periphery of the hepatocytes was observed in one fourth of the patients with acute hepatitis and only sera which contained also IgM smooth-muscle antibody.