Multiple sclerosis: immunogenetic analyses of sib-pair double case families. II. Studies on the association of multiple sclerosis with C2, C4, BF, C3, C6, and GLO polymorphisms.

The complement component polymorphisms of C2, C4, BF, C3, C6, and the enzyme polymorphism GLO were studied in 13 sib-pair double case families with multiple sclerosis. A significant association was seen between MS patients and the C4 haplotype A4,B2 as compared with their healthy siblings. This finding seems to parallel reports on C2 hypocomplementemia in MS patients since C4 A4,B2 in normal individuals was also seen to be in linkage disequilibrium with the C2 deficiency allele (C2QO) by other investigators.

The BF F subtypes are detectable in the Ba fragment of factor B.

The unanimous recognition of the two subtypes FA and FB of the BF*F allele has repeatedly been challenged. In the present investigation we are reporting about the unequivocal and simple detection of the subtypes on the Ba fragment of factor B by immunofixation isoelectric focusing after conversion with inulin. The common BF phenotypes F, S, and FS could be diagnosed in addition to the subtypes of BF*F which were observed in two regions acidic of the F major band. By comparison of standard phenotypes the subtypes in the Ba fragment corresponded to those of native factor B. All BF bands could be attributed to the Ba fragment by developing Western Blots with monoclonal antibodies directed against Ba. The distribution of the major BF phenotypes and alleles and the BF F subtypes in a population sample of 527 unrelated individuals from F.R.G. was in Hardy-Weinberg equilibrium. The allele frequency was determined to be 0.0731 for BF*FA, and 0.1053 for BF*FB. The advantages of determining the subtypes on the Ba fragment are: broadening of the FA/FB corridor, a more reliable diagnosis of phenotypes, improved distinction between homozygous FA and heterozygous FAFB types, and recognition of common BF phenotypes as well as subtypes in aged sera. It is suggested that the problem in the designation of BF F subtypes by different groups should be resolved by an international reference typing.

[Serological diagnosis of Campylobacter jejuni infections]. / Serologiczna diagnostyka w zakazeniach Campylobacter jejuni.

Antibody level to Campylobacter in 28 sera of patients of whom Campylobacter infection was confirmed by germ isolation from feces was tested. The investigation was performed using passive haemagglutination technique and as antigens heated and acid glycine extraction prepared from homologous and reference strains. For the method used the heated antigen proved to be superior. Out of 28 tested patients of whom 92.8% were children, 8 sera were positive, 9 doubtful and 9, derived mainly from neonates (0-14 month of age), were negative.

Another family with a silent allele of properdin factor B polymorphism (BF QO).

In five of eight members of a three generation family the existence of a silent allele of the properdin factor B polymorphism (BF QO) was indicated by immunofixation of BF electrophoretic variants and by the hemolytic overlay after isoelectric focusing of BF allotypes. This was further supported by the results of HLA-A, B, C, DR, C2, C4A, C4B, GLO-typing. BF protein was decreased in all heterozygous BF deficient family members. The absolute hemolytic activity, however, was obviously compensated for by an increased relative functional activity of the normal S or F alleles on the other chromosome.

Genetic polymorphism of the fourth component of human complement.

A modified electrophoretic system for the determination of C4 polymorphism has been found with which the new allotype F1 could be detected. The system has been applied to the population distribution of C4 in 266 unrelated Germans, further to association and linkage studies. Gene frequency was 0.3985 for C4F, 0.5526 for C4S and 0.0489 for the rare C4 genes. In the population sample, significant association between Bf and C4 but no indication for close association between C4 and GLO I has been found. In the families HLA, Bf and C4 segregated together.

Properdin factor B (glycine-rich beta-glycoprotein or C3 proactivator)-polymorphism: genetic and biochemical aspects. First application to paternity cases.

Determination of genetic properdin factor B(Bf) polymorphism was carried out in immunofixation electrophoresis. Genetics of factor B were also studied after ageing, conversion with cobra venom and neuraminidase. In population studies the distribution of factor B in a West German population of 1245 non-related individuals was found to be: Bf F 2.73%, Bf FS 28.43%, Bf S 65.38%. Rare phenotypes (F 1F,F 1S, FS 1, SS 1) were seen in 3.46%. In addition a new variant, designated F1.6S, was observed. The application of factor B polymorphism to 68 paternity cases is discussed.

[Transferrin variants: significance and identification in paternity cases (author's transl)]. / Die Bedeutung und Zuordnung von Transferrin-Varianten in der Abstammungsbegutachtung

Transferrin phenotypes were determined in 3380 sera of unrelated persons of the western region of Germany with 97.60 percent for TfC and 2.40 percent for Tf variants. Identification was achieved by immunochemical means or through autoradiography. Relative mobilities in some variants were measured using Tf B2C (0.7) as reference. Application of Tf variants is demonstrated in paternity cases.

[Studies on the antigenicity of synthetic ACTH (beta1-24-corticotropin) in normal subjects (author's transl)]. / Untersuchungen zur Antigenität von synthetischem ACTH (beta1-24-Corticotropin) bei freiwilligen Versuchspersonen

9 healthy volunteers were subjected to a 3-week treatment with synthetic ACTH. Antibody response against beta1-24-corticotropin (Synacthen) was tested by passive transfer in the Prausnitz-Küstner reaction, complement fixation, passive hemagglutination and agar-gel diffusion. Failure of the healthy persons to produce reaginic or non-reaginic antibodies is compared titerature.

C3 polymorphism in polyacrylamide-gel electrophoresis.

C3 polymorphism in polyacrylamide-gel electrophoresis was identified by crossed immunoelectrophoresis using anti-C3/C3c-serum. Through ageing, treatment of sera with cobra venom factor, endotoxin or with neuramindase, polymorphic bands were seen also in a conversion product and antigenically attributed to C3c. Rare phenotypes were observable in native C3 and in C3c.

Esterase D polymorphism: high-voltage agarose-gel electrophoresis and distribution of phenotypes in different European populations.

Esterase D phenotypes were determined in 1082 non-related individuals from the western region of Germany by agarose-gel electrophoresis. Gene frequencies were compared with previous data and all European populations studied so fare agreed with the Hardy-Weinberg equilibrium. Mean gene frequencies for Europeans are: EsD1 0.8888, EsD2 0.1112.

Yersinia enterocolitica serodiagnosis: a dual role of specific IgA. Evaluation of microagglutination and ELISA.

The microagglutination technique for the detection of antibodies against Y. enterocolitica, serovars 3 and 9 (corresponding to O-groups I and V), was compared with the conventional tube agglutination. An immunoglobulin class specific, indirect ELISA (polyvalent immunoglobulin, IgG, IgM, and IgA) was established employing as antigens formalinized whole bacteria ("OH"-antigens) and LPS preparations (hot phenol-water extraction). ELISA titers and net absorbancy (ELISA-"units") of single serum dilutions were in good agreement; the same was true for ELISA and agglutination results. Specificity (against healthy controls) and sensitivity of both serologic techniques were comparable. Cross-reacting antibodies against serovars 3 and 9 could be identified in the ELISA. Correct serovar-specific diagnosis was possible in 95% with a single assay (polyvalent Ig assay with LPS-antigen). The sensitivity of the LPS-ELISA was superior to the "OH" antigen assay after infections by serovar 3 strains, and antibodies were detected with LPS preparations for a longer period following reconvalescence. Specific IgA, due to its rapid decrease during reconvalescence, on one hand impresses as a valuable marker for the differentiation of recent disease from uncomplicated past infections, while persistence of IgA appears to be associated with Yersinia-induced arthritis. Persisting IgM but rarely IgA titers were characteristically found in patients with prolonged enteric yersiniosis.

[Menstrual toxic shock syndrome in a 17-year-old girl]. / Menstruales Toxisches-Schock-Syndrom bei einem 17jährigen Mädchen.

In a seventeen-year old female patient fever, vomiting, conjunctivitis, pharyngitis, hypotension, exanthema, disorientation and severe myalgia were observed on the second day of menstruation. The typical symptoms suggested the clinical diagnosis of toxic shock syndrome (TSS). During the period of reconvalescence desquamations on hands and feet occurred. From vaginal swabs and the tampons Staphylococcus aureus was recovered. In supernatants from cultures the strain was found to produce toxic shock toxin (TST). Antibodies against TST in the patients serum were not detectable for a period of 70 days after onset of the disease. The patient recovered within three weeks, relapses were not observed.

Early onset sensorineural hearing loss: association studies with major histocompatibility class III (complement) markers.

In 39 families with at least one child suffering from moderate or severe bilateral sensorineural hearing loss (SNHL), major histocompatibility complex (MHC) class III complement phenotypes were retrospectively determined by standard methods; MHC class I segregation data was also available. The families were treated in the Hospital for Communication Disorders and selected for the HLA-B16 and B18 specificities, respectively. Haplotype and allele frequencies were derived from segregation analysis in the families. From 31 unrelated children with random and familiar forms of SNHL significant deviations in the distribution were seen for the following MHC class III alleles using as a control population 60 German healthy individuals: duplicated C4A alleles (C4"DA") p = 0.009, silent C4A alleles (C4A*Q0) p = 0.006, duplicated heavy C4 beta-chain alleles (C4 beta"DHH") p = 0.0003, and silent C4 beta-chain alleles (C4 beta*Q0) p = 0.0075. In serum samples from patients with an assumed genetic disposition according to clinical criteria indications for an association were found for C4"DA" (p = 0.03), C4A*Q0 (p = 0.003), C4B*3 (p = 0.046), C4 beta"DHH" (p = 0.004), and C4 beta*Q0 (p = 0.02). The underrepresentation of C4A*Q0 may be an indicator for aberrant or duplicated C4 alleles on the same haplotype or exhibit a protection mechanism for acquiring the inheritable forms of early onset SNHL.