Diether Schönitzer

RHD/CE typing by polymerase chain reaction using sequence-specific primers

BACKGROUND: Current DNA‐based Rh system typing strategies may detect the two RH genes and their prevalent alleles, but they are known to fail sometimes, when rare RH alleles (e.g., D category phenotypes) are encountered. It is almost impossible to find a single DNA‐based method that can accommodate the great heterogeneity within the human Rh system. STUDY DESIGN AND METHODS: An easy‐to‐perform DNA‐based method for the detection of the two RH genes and their alleles, including variant RHD alleles, was developed. By the use of one RHD/C‐, seven RHD‐, and four RHCE‐specific polymerase chain reactions, all triggered to work at identical thermocycling conditions, the DNA of 77 blood donors carrying weak D and that of 200 random donors with common D phenotype was investigated. In addition, 77 selected samples of ccDee and rare Rh system phenotypes were examined. RESULTS: Among 77 samples of weak D, one Rh33 and six DVI categories were detected, one of which showed new RHD‐specific nucleotide patterns. In DFR and CCee samples, novel variant RHD alleles were found. RHD DNA types of 200 random donors were found to be concordant with their D phenotype. For RHE and RHe genotyping, a full correlation with serologic phenotypes was found. Our method for genotyping RHC and RHc failed in some cases, because of an already published RHc allelic variation, which we have called RHc(cyt48). An estimate of the frequency of this RHc(cyt48) allele in a white population was made. CONCLUSION: The presented exon‐scanning RHD/CE polymerase chain reaction using sequence‐specific primers complements current DNA‐based Rh system typing strategies and is superior in the detection of variant RHD alleles.

PCR screening for common weak D types shows different distributions in three Central European populations

BACKGROUND: DNA sequencing showed RHD mutations for all weak D phenotypes investigated in a study from Southwestern Germany. Molecular classification of weak D offers a more reliable basis than serotyping and is relevant for optimal D transfusion strategies.

Approach to define “normal aging” in man. Immune function, serum lipids, lipoproteins and neopterin levels

In 53 healthy women with mean age 25.1 years (age range 21-34 years) and in 51 healthy women with mean age 82.1 years (age range 75-91 years), a panel of immunological and biochemical tests was performed. These tests, comprising lymphocyte mitogen responsiveness, phenotyping of lymphocytes, uptake of low density lipoprotein by T cells, serum levels of neopterin, lipids and lipoproteins, as well as routine blood chemistry, were investigated for a possible effect of age and of the classification according to the SENIEUR Protocol of admission criteria by the European Economic Communitys Concerted Action Program on Aging (EURAGE). A highly significant effect of age on serum levels of neopterin, lipids and lipoproteins was found. No clear effects, however, of SENIEUR status on these variables was detected. As expected, age had a significant impact on mitogen responsiveness of T cells. Proportional numbers of helper/inducer and cytotoxic/suppressor T cells (as well as antigen density on these cells) were not influenced by age. SENIEUR classification did not affect these immunologic variables. Thus, most of the tested variables that are not included in the SENIEUR admission criteria appear to present information not yet covered by the SENIEUR variables. Various ways for a possible revision or extension of the SENIEUR Protocol are discussed.

Prevention of transfusion of platelet components contaminated with low levels of bacteria: a comparison of bacteria culture and pathogen inactivation methods

BACKGROUND: This study compared the efficacy of bacterial detection with inactivation for reducing the risk associated with transfusion of platelet (PLT) components contaminated with low levels of bacteria.

Urinary neopterin in the diagnosis of acquired immune deficiency syndrome

Neopterin, a pyrazino-[2, 3-d]-pyrimidine derivative (Figure 1) which originates biosynthetically from guanosine triphosphate, has been found to be elevated in patients suffering from certain types of cancer or viral diseases (1). Neopterin has been demonstrated in vitro and in vivo to be a specific indicator of the solicitation of Tqymphocytes (2, 3). All diseases coupled with activation of the T4ymphocyte-macrophage axis should lead to increased neopterin levels. Results in patients with allograft rejection (unpublished observation of authors), autoimmune diseases, infections with intracellular microorganisms and malignancy support this view (4, 5, 6).

Genetic diversity of KELnull and KELel: a nationwide Austrian survey

BACKGROUND: Besides ABO and RH, the KEL blood group system, including the two antithetical antigens KEL1 and KEL2, is the most important owing to the frequent appearance of anti‐KEL alloantibodies and their considerable clinical significance. So far, only limited information was available on KEL variant alleles determining the rare silent KELnull and KELel phenotypes with absent or diminished KEL antigen expression detected only by adsorption‐elution techniques, respectively.

Novel weak D types 31 and 32: adsorption-elution–supported D antigen analysis and comparison to prevalent weak D types

BACKGROUND: Weak D types are thought to express rather quantitative than qualitative D antigen variants. Distinct type‐specific phenotypes and weak D cases with anti‐D alloimmunization, however, suggest a variable degree of D antigen alteration.

Molecular and serologic characterization of DWI, a novel “high‐grade” partial D

BACKGROUND: Accurate D antigen identification is essential for pretransfusion and prenatal evaluation to prevent anti‐D alloimmunization. Quantitative and qualitative D variants may pose typing problems and require particular consideration because of differing potential for anti‐D induction.

Prolonged iron depletion after allogeneic 2-unit RBC apheresis.

BACKGROUND: Allogeneic 2‐unit RBC apheresis is a safe procedure offering many advantages for donors and blood banks. A controlled study was performed to determine whether the recommended minimum interval of 4 months between 2‐unit RBC apheresis donations is appropriate in terms of the recovery of RBCs and the regeneration of iron stores.

Neopterin screening and acute cytomegalovirus infections in blood donors

Since October 1986 the blood bank of the University Hospital in Innsbruck, Austria, has been testing all blood donations for increased neopterin concentrations in order to further improve the safety of blood transfusions [1]. Donations with neopterin levels above 10 nmol/1 are excluded from transfusion. Increased neopterin levels indicate activation of the cellular immune system [2]. Beginning in January 1988 we tested donations with increased neopterin concentrations for acute eytomegalovirus (CMV) infection by CMV IgM ELISA. Of 93,789 donations, 1,767 (1.88%) showed raised neopterin levels. These donors were routinely informed, thoroughly examined, and asked for a control blood sample four to five weeks after the primary donation. In addition, a control group of 1,092 randomly selected donors with neopterin concentrations below the cut-off limit of 10 nmol/1 were examined following the same protocol. Neopterin was determined by radioimmunoassay (Henning, Berlin, Germany). CMV IgM/IgG was tested by ELISA (Abbott Laboratories, North Chicago, Ill., USA). CMV IgM/IgG testing was confirmed by a second ELISA (Behring, Marburg, FRG). In addition to neopterin and CMV testing, all samples were screened for hepatitis B surface antigen and human immunodeficiency virus types 1 and/or 2 antibodies (Abbott), syphilis (Fuji, Vienna, Austria), and ALT levels (Boehringer, Vienna, Austria). Samples positive for CMV IgM antibodies were also tested for rheumatoid factors, heterophil antibodies (Bio-Merieux, Carbonniere les Bains, France), and Epstein-Barr virus IgG/IgM serostatus (Epignost, Linz, Austria). Rheumatoid factor testing was performed twice by latex agglutination (Serotherapeutisches Institut, Vienna and Gamma Biologicals, Houston, Tex., USA). Of 1,767 donations with increased neopterin levels, 93 (5.26%) were found to be positive for CMV IgM antibodies. Eleven of the 93 CMV IgM seropositives also had IgG antibodies. At the control visit mean neopterin levels decreased in donors who developed CMV IgG antibodies (Table 1). In 34 samples CMV IgM antibodies without CMV IgG were detected. Ten of 1,674 donors with increased neopterin who were negative for CMV IgM/IgG antibodies at the time of donation and negative for all the other tests had CMV IgM antibodies at the time of the control; neopterin was still elevated. Three of these donors were followed up for a second time four weeks later; they presented with CMV IgM and IgG antibodies, but with decreased neopterin levels (Table 1). Acute CMV infection may explain elevated neopterin at the time of donation, the increase of neopterin preceding the appearance of CMV IgM and/or IgG antibodies by 2-4 weeks. In the group with low neopterin at donation, only three (0.3%) were CMV IgM seropositive. Thus, acute CMV infection is significantly more prevalent Table 1. Association between CMV serologic findings and neopterin concentrations in blood donors