G. Mauff

Human C4 polymorphism: Pedigree analysis of qualitative, quantiative, and functional parameters as a basis for phenotype interpretations

SummaryTen families with 82 members were investigated for C4A- and B polymorphism in a blind trial. Phenotyping was done on neuraminidase treated sera by immunofixation and simulataneously by hemolytic overlay electrophoresis. In addition Rg, Ch, BF, C2, HLA-A, B, C, DR, and GLO were determined. After decoding the samples the reliability of blind typing was found to be 84.4% according to segregation patters. Inconsistencies occurred mostly when A 4, A 2, or A 92 were present. The detection of silent A*Q0 and B*Q0 alleles was more critical than that of “difficult” allotypes. The quantitation of the C4A/B ratio by densitometry of stained gels or by conventional immunochemical measurements of serum C4 level could not substantially improve the identification of A*Q0 or B*Q0. C4 dependent activity in radial diffusion hemolysis showed satisfactory correspondence with the number of expressed C4B alleles. At least three haplotypes with two C4A genes (duplicated A genes) were observed as ascertained from offspring analysis in accordance with the MHC segregation pattern. Individuals with the duplicated C4A gene (C4A*3. A*2. in the absence of any other expressed A allele or together with C4A*92) showed only partial inhibition of Rodgers antisera. Partial inhibition of Chido antisera was seen in individuals with C4B 2 (in the absence of other B allotypes). The findings support the hypothesis of at least two structural C4 loci. The also demonstrate the inconsistency of quantitative data in the recognition of silent alleles.

Proposal for the Nomenclature of Human Plasminogen (PLG) Polymorphism

Abstract. Since its discovery, human plasminogen (PLG) polymorphism has received widespread acceptance in population genetics and forensic haematology. Due to the large number of variant alleles described, a PLG reference typing and Plasminogen Symposium was held, at which a nomenclature proposal was inaugurated. The technology of comparing PLG variants was based on isoelectric focusing and subsequent detection by caseinolytic overlay and ‘Western’ blotting. Typing results permitted comparison of so far described variant designations and resulted in a new nomenclature proposal for PLG polymorphism. It is recommended that the two most common alleles found in all investigated races be called: PLG*A (previously also PLG*1) and PLG*B (previously also PLG*2), the known variants with acidic pI: PLG*A1 to *A3, intermediate variants: PLG*M1 to *M5, PLG*M5 being functionally inactive, and basic variants: PLG*B1 to *B3. For future classification of newly discovered variants, samples should be compared at any of the laboratories participating in the reference typing.

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 study of a French family with two duplicated C4A haplotypes

SummaryThe finding of two duplicated C4A haplotypes in a normal French family led to a detailed study of their C4 polymorphism. The father had an extremely rare A*6A*11, B* QO haplotype inherited by all of his children and the mother had the more common A*3A*2, B*QO haplotype. Two HLA identical daughters only have four C4A alleles. The fathers A11 allotype expresses Ch: 1 (Chido) rather than Rg:1 (Rodgers) and represents a new Ch phenotype Ch: 1,-2,-3,-4,-5,-6. In order to clarify the genetic background in this unusual family, DNA studies of restriction fragment length polymorphisms (RFLPs) were undertake. The fathers rare haplotype, which expresses two C4A allotypes, results from a long and a short C4 gene normally associated with the A*6, B*1 that also exhibits the BglII RFLP. As it travels in an extended MHC haplotype HLA A2, B57 (17), C2*C, BF*S, DR7 that is most frequently associated with A*6, B*1, we postulate that the short C4B has been converted in the α chain region to a C4A gene which produces a C4A protein. This report of a short C4A gene is the first example in the complex polymorphism of C4.

Campylobacter and salmonella contaminating fresh chicken meat

1853 packages of fresh chicken breast meat of German, Dutch and French origin were investigated for their contamination with Campylobacter and/or Salmonella. Swabs were taken and cultured from dripwater, meat surface, meat interior and packet bowl. Campylobacter was isolated from 619 meat samples (= 33%), Salmonella from 377 meat packages (= 20%). In 111 of these contaminated chicken samples, both Salmonella and Campylobacter were present. The contamination rate and the species spectrum observed differed depending on the origin of the packages and the time of control.

Vergleichende Untersuchungen zum Polymorphismus des Posttransferrins (Pt) und der dritten Komponente des Humankomplements (C3)

SummarySera of 541 unrelated Germans from the Cologne area were examined for their Pt and C3 types. There was correspondence between Pt A and C3 F, Pt AB and C3 FS, and Pt B and C3 S in 537 sera. C3 variants F0.65S, F0.6S, F0.5S were determined as Pt AB, C3 F0.55S was determined as Pt B. The question of possible identity and the methods used are discussed.Zusammenfassung541 Serumproben von nichtverwandten Deutschen aus dem Kölner Raum wurden auf ihre Pt- und C3-Typen untersucht. Bei 537 Seren fand sich eine Überein-stimmung der Pt A-Typen mit den C3 F-Typen, der Pt AB-Typen mit den C3 FS-Typen und der Pt B-Typen mit C3 S-Typen. Die C3-Varianten F0.65S, F0.6S, F0.5S wurden als Pt AB bestimmt, die C3-Variante F0.55S wurde als Pt B bestimmt. Die Frage der Vergleichbarkeit der Systeme sowie die Untersuchungsmethoden werden diskutiert.

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.

Humoral and cellular immunity in HIV positive and HIV negative Helicobacter pylori infected patients

The prevalence of H. pylori associated gastritis seems to be different in HIV positive and HIV negative patients. Therefore a correlation to immunodeficiency can be postulated. The histology of gastritis, status of H. pylori infection and parameters of humoral and cellular immune response were investigated in 41 HIV positive and 47 HIV negative patients, who were subjected to upper endoscopy for the evaluation of gastrointestinal symptoms. In HIV positive patients 37% had active chronic gastritis against 62% of the HIV negative patients. In 73% of HIV positive cases of active chronic gastritis H. pylori was detected by bacteriological culture and/or Warthin-Starry stain. In HIV negative patients active chronic gastritis was always associated to H. pylori infection. Production of antibodies as measured by two commercially available ELISA tests was significant in HIV positive and HIV negative patients; both tests correlated well with H. pylori detection by culture or direct microscopy. Immunoglobulin class specific immunoblots corresponded to the ELISA results in HIV negative patients but to a lesser extent in the HIV positive group which was assumed to be related to unspecific polyclonal activation in these patients. Systemic cellular immunity was investigated by proliferation assays of peripheral blood mononuclear cells (PBMC). Proliferative response to the unspecific mitogen PHA was reduced in HIV positive patients. A sonicated H. pylori antigen failed to induce lymphocyte proliferation. The antimitogenic effect was also seen in case of coincubation with PHA. This observation was independent of H. pylori and HIV infection status. We conclude that in HIV positive as in HIV negative patients active chronic gastritis is predominantly related to H. pylori infection. The prevalence of H. pylori associated gastritis in HIV positive patients is significantly reduced (p < 0.025) compared to HIV negative controls. Decreased susceptibility to H. pylori infection in HIV positive patients may not be explained by the abnormal reactivity of their humoral or cellular immune response.

Helicobacter pylori antibodies in sera of children suffering from chronic abdominal pain

107 pediatric patients aged 9 to 18 with persistent gastric complaints were examined serologically and bacteriologically for Helicobacter pylori. Helicobacter was identified in 48 (45%) of individuals. 51 (48%) of children were found to be seropositive when H. pylori antibodies were detected by the ELISA; 56 (52%) when the passive haemagglutination test was used, and 41 (38%) in the latex agglutination test. 25% of culture-negative patients were found to be seropositive. The percentage of raised H. pylori antibody titres in the control (healthy subjects) varied from 20 to 27%, depending on the method applied.

Major Histocompatibility Complex Class I to III Allotypes in Patients with AIDS-Related Complex/Walter-Reed 5, Disseminated Kaposi's Sarcoma and in Normal Controls

Abstract. In HIV‐infected patients major histocompatibility complex (MHC) class I and II (= HLA‐A, B, C, DR) association has been controversial. Of the MHC class III coded complement components C2, BF, C4A/C4B especially C4 allotypes appear of major immunogenetic relevance for their potential differences in virus neutralizing potency and immune complex formation. In the present study 29 patients with AIDS‐related complex and Walter‐Reed 5 (ARC/WR5), 35 patients with disseminated Kaposis sarcoma (KS), and 160 HIV‐negative control individuals were compared for MHC class I to III allotypes. Diagnosis of ARC and KS (WR criteria) was done by clinical and laboratory parameters, MHC testing, by standard procedures. An increase in frequency (p≤0.05) was observed between ARC/WR5 patients and controls for HLA‐B35/CW4, DRW14, a decrease for B16, CW6/DR7. However, values were not significant if corrected for the number of tested antigens. No significant differences were seen between KS and ARC patients or controls for class III allotypes, nor for previously reported associations, e.g. for B8, DR2, DR3, and especially DR5, including the DR5 splits DRW11, 12. The results indicate the lack of a strong MHC association with the investigated antigens in West German Caucasoids, and support the hypothesis of ethnic dependence of HIV‐related diseases. The HLA‐B35/CW4 increase, also associated with the duplicated C4 A*3 A*2 and the silent C4B*Q0, was more pronounced in ARC patients with progression to AIDS‐OI. The increased frequency of C4B*Q0 alleles in these patients was thought to be secondary to a hypothetical increase in ‘converted’ and dysregulated C4 genes not seen to be associated in this study.