Simone Mayer

Restriction fragment length polymorphism of DP genes defines three new DP alleles

The class II major histocompatibility complex, coded on chromosome 6 in man, is divided into the DR, DQ, and DP regions, with each region containing one or several c~ and/or [3 genes. The DP region is subdivided into SX and DP (formerly known as SB) subregions, each composed of one o~ and/3 gene, the gene order being SX¢, SX~, DP~, DP~ (Korman et al. 1985). SX~ and SX~ showed on some haplotypes a deletion in their exon sequence, thus suggesting that they are pseudogenes (Boss et al. 1985, Kappes and Strominger 1986). DP~ and DP, code, respectively, for a M r 28 000 DP¢ and a M r 34 000 DP~ glycoprotein chain. The chains are noncovalently associated and are expressed as DP antigens on the cell surface (Giles and Capra 1985). DP antigens have been mainly studied by using the primed lymphocyte typing (PLT) assay (Bourgue et al. 1982), which allowed determination of six DP polymorphic specificities at the Ninth Histocompatibility Workshop, whereas approximately 30% of the alleles corresponded to blank alleles (Wank and Schendel 1984). Restriction fragment length polymorphism (RFLP) studies have been performed in our laboratory by testing the genomic DNA of eight HLA genotyped families selected because they showed a clear-cut segregation from DP1 to DP5 antigens. Digested samples of DNA were electrophoresed through 0.7% agarose gels at 30 V for 40 h in Tris-EDTA-acetate buffer and were then transferred onto BIOTRACE membranes (Gelman Science Inc.) overnight with 0.4 N NaOH. After prehybridization, the transferred membranes were hybridized with a hybridization mix containing nick-translated 32p-labeled probes whose specific activity was 1-2X 108 cpm/gg. The hybridized membranes were washed twice in 2 x standard sodium citrate (SSC) at laboratory temperature for 5 rain, twice in 2 x SSC plus 0.5 % sodium dodecyl sulfate (SDS) at 65 °C for 15 rain, and once in 0.1 x SSC plus 0.5% SDS at 65 °C for 15 rain, and then autoradiographed (Kodak XAR-5). DP typing of the eight families is shown in Table 1. No DP6 family was studied but DP blank antigens were carried by some individuals. As an example of the results obtained using Southern blot analysis with DP~ (Auffray et al. 1984) and DP~ probes (Roux-Dosseto et al. 1983), autoradiograms obtained in five of these families with the

Treatment of Acquired C1-Inhibitor Deficiency with Danazol

Excerpt Hereditary angioedema is characterized by a marked deficiency of the inhibitor of activated first component of complement (C1 inhibitor). Some cases of acquired C1-inhibitor deficiencies an...

MIXED-LYMPHOCYTE REACTION AS ASSAY FOR IMMUNOLOGICAL COMPETENCE OF LYMPHOCYTES FROM PATIENTS WITH HODGKIN'S DISEASE

Abstract Peripheral-blood lymphocytes from Summary eleven untreated patients with Hodgkins disease have been tested for transformation under stimulation by allogeneic cells and phytohaemagglutinin (P.H.A.) in vitro. Mixed-lymphocyte reaction (M.L.C.) was clearly positive in nine patients whatever their clinical stage or histological class. It is suggested that M.L.C. is a more useful tool than P.H.A. stimulation for the study of lymphocyte immunity capacity in vitro. Lymphocyte immunological competence in Hodgkins disease needs to be re-evaluated.

Linkage between C2 deficiency and theHLA-A10,B18,Dw2/BfS haplotype in a French family

A linkage between C2 deficiency and the HLA-A10,B18/BfS antigens has been found in a French family from the Strasbourg area. The propositus, suffering from a chronic glomerulonephritis, is homozygous forHLA-A10,B18/BfS and totally C2-deficient. The parents and the brother are heterozygous for C2 deficiency and share theHLA-A10,B18/BfS haplotype. MLC tests and HLA-D typing revealed that the homozygous C2-deficient patient is also homozygous at theHLA-D locus for the w2 specificity. Evidence was obtained for a heterogeneity of the HLA-Dw2 specificity. This observation confirms the remarkable association between C2 deficiency and theHLA-A10,B18,Dw2 haplotype.

Stage-related decrease in natural killer cell activity in untreated patients with mycosis fungoides

SummaryNatural killer activity of peripheral blood mononuclear cells against the human cell line K 562 was evaluated in 11 patients with mycosis fungoides and simultaneously in 10 age-and sex-matched controls. In the patient group, nine had no previous treatment and in two topical therapy had been discontinued more than 3 months before. None had any associated disease or concurrent therapy that could interfere with the immune system. Patients with early disease showed a mean specific lysis and a range of individual data similar to the controls whereas patients with advanced disease had a significant defect of natural killer activity at effector: target ratios of 100:1, 50:1, and 25:1, as shown by the Mann-Whitney test. Preincubation of effector cells with α-interferon for 1 h in a single patient with low natural killing capacity led to a clear increase of the specific lysis, suggesting reduced functional activity rather than depletion of effector cells.

DP RFLP Studies of the 72 Core Cell Lines Selected for the Southern Blot Protocol

DNA was digested with the requested 12 enzymes and hybridized with the DPα and DPβ probes given to the participating laboratories. Only clusters correlating with the PLT-defined DP specificities were retained from the Workshop computer analysis.

Comparative study of autologous, total E and ‘active’ E-rosette-forming T lymphocytes in untreated Hodgkin's disease

SummaryIn order to evaluate the value of autologous rosettes, a marker for a T lymphocyte subpopulation, in the diagnosis of T lymphocyte defects, we compared the ability of T cells from untreated patients with Hodgkins disease to form autologous, total E and ‘active’ E rosettes. While the mean percentage of total E and ‘active’ E rosettes was significantly decreased in the patients compared to normal controls (0.01<P<0.02), no difference was observed for autologous rosettes (0.6<P<0.7). Only two patients showed decreased ‘active’ E rosettes whilst eight had decreased total E rosettes. Individual data in the patients showed a wide dispersion of autologous rosettes with both high and low percentages whatever the relative number of the two other T cell markers. No correlation was observed between any of the three types of rosettes in the patients whereas autologous rosettes were clearly correlated with total E (0.01<P<0.02) but not with ‘active’ rosettes in normal controls. Neither autologous nor ‘active’ rosettes could be correlated with skin test results.

Membrane markers, karyotypic abnormalities, ultrastructure and functional properties of lymphocytes in a case of ‘D-cell’ chronic lymphatic leukemia

D cells are lymphocytes bearing both receptors for the third complement component and the ability to form spontaneous rosettes with SRBC. We report the case of a patient with a D-cell chronic lymphatic leukemia who presented a long evolution without treatment and whose leukemic cell characteristics have been extensively studied. Cytogenetic analysis showed numerous karyotypic abnormalities among leukemic cells; all metaphases were hypodiploid and arranged in four different clones; seven marker chromosomes were present. The cells were found to bear human T-cell specific antigen, the T helper/inducer phenotype, HLA-A and HLA-B determinants, but no HLA-DR antigens. They displayed a high proliferative response to PHA and Con A, no response to PWM stimulation, and possibly the capacity of allogeneic stimulation in the mixed lymphocyte culture system. Assays for cell-mediated cytotoxicity in the CML system, and for K and NK activities were negative.

Selective IgA Deficiency and Idiopathic Thrombocytopenia

Excerpt To the editor: Selective IgA deficiency has been described in apparently healthy people and in association with significant disorders (1). Only one case associated with idiopathic thrombocy...