Biomedical subjects

H Grosse-Wilde

Publications and source records attributed to H Grosse-Wilde.

At least 73 records · Page 4Linked to original sources

PubMed1996

Improved immune reconstitution after allotransplantation of peripheral blood stem cells instead of bone marrow.

Clinical studies are evaluating possible advantages of allogeneic peripheral blood stem cell transplantation (PBSCT) over bone marrow transplantation (BMT). We compared immune reconstitution after PBSCT (n = 20) and BMT (n = 20) in terms of lymphocyte subset counts and proliferative in vitro responses to mitogens and recall antigens (follow-up: 5 to 11 months posttransplant). Additionally, 10 PBSC harvests and 10 marrow harvests were analyzed for their composition of immunocompetent cells. Compared with BMT patients, PBSCT recipients had PB counts of naive (CD4+CD45RA+) and memory (CD4+CD45RO+) helper T cells and of B cells (CD19+) that were elevated (P < .003, P < .001, and P < .004, respectively) and proliferative responses to phytohemagglutinin (P < .0001), pokeweed mitogen (P < .02), Tetanus toxoid (P < .0005), and Candida (P < .004) that were increased. PBSCT recipients received a mean of 188 (range, 44 to 280) x 10(6) naive helper T cells and 169 (range, 18 to 296) x 10(5) memory helper T cells per kilogram; the corresponding numbers for BMT recipients were 11 (range, 4 to 24) and 10 (range, 1 to 22) x 10(5) cells per kilogram, respectively. The question of whether the documented improved in vitro immune competence after PBSCT is associated with a lower incidence of infectious complications in vivo still needs further study.

Adult↗

PubMed1996

Selective loss of human leukocyte antigen class I allele expression in advanced renal cell carcinoma.

The expression of human leukocyte antigen (HLA) class I alleles was analyzed in 65 renal cell carcinomas using one-dimensional isoelectric focusing. Normal organ tissue and peripheral blood lymphocytes were used as controls. The patients were serologically typed using the standard microcytotoxicity test. Forty-two patients were staged as pT1 or pT2, and 23 patients had advanced tumor stages (pT3/pT4). In all cases the HLA-A,B phenotypes were confirmed using one-dimensional isoelectric focusing. The expression of HLA expression was reduced in two tumors [1 x HLA-A1(pT2); 1 x HLA-A28 (pT2)]. In three carcinomas the expression of HLA-A1 was lost. One tumor showed a combined loss of HLA-A2 and HLA-B38. These selective losses occurred in tumor stage pT3 (n = 1) or pT4 (n = 3; P = 0.013, Fisher's exact test). This leads to the conclusion that the loss of HLA expression is predominantly present in advanced tumor stages.

Adult↗

PubMed1996

Assessment of peripheral blood mononuclear cell proliferation by [2-3H]adenine uptake in the woodchuck model.

The levels of incorporated exogenous [3H]thymidine of peripheral blood mononuclear cells (PBMC) of the woodchuck were low after stimulation with mitogens concanavalin A (ConA), phytohemagglutinin (PHA), and pokeweed mitogen (PWM) when compared with other cell systems. The use of EDTA as an anticoagulant for blood sampling and AIM-V medium for culturing of PBMC improved the [3H]thymidine uptake of PBMC. A pronounced uptake is observed after use of [3H]adenine instead of [3H]thymidine for PBMC proliferation measurement. One likely explanation for the difference in [3H]adenine versus [3H]thymidine uptake is that the alternative pathway for thymidine monophosphate synthesis is important: the conversion of uridine to uridine monophosphate and, thereafter, to thymidine monophosphate. The optimal conditions for mitogen-induced proliferation of PBMC of the woodchuck were 2 micrograms/ml ConA and PHA at day 4 and 0.14 micrograms of PWM/ml at day 5. No consistent differences of [3H]adenine uptake were observed between PBMC from four woodchuck hepatitis virus-infected woodchucks and five uninfected animals.

Adenine↗

PubMed1996

Bone marrow transplantation from partially HLA-matched related donors in adults with leukaemia: the experience at the University Hospital of Essen, Germany.

The Seattle group has demonstrated that bone marrow transplantation (BMT) using partially HLA-mismatched related donors is feasible in principal. However, it was unclear whether these results can also be achieved at smaller-sized BMT units. Therefore a matched pair analysis enrolling 52 BMTs from partially HLA-mismatched relatives and 52 control BMTs from HLA-identical siblings was performed at the University Hospital of Essen. Overall survival (OS) and incidence of graft-versus-host disease (GVHD) did not differ significantly after study and control BMTs (OS: 52% v 63% 1 year, 46% v 48% 5 years post transplant; acute GVHD: 37% v 35%, chronic GVHD: 67% v 55%). After study BMTs, however, therapy-related mortality (P=0.018) and incidence of graft failure (P=0.002) were increased, whereas relapse rate was reduced (11% v 27%). Two or three mismatches in HVG direction implied the same risk of graft failure as one mismatch. Therefore, (i) OS after BMT from one HLA antigen mismatched relative and from HLA-identical siblings does not differ significantly even when performed at a 'smaller' centre; (ii) two or even three HLA mismatches in host-versus graft (HVG) direction might be acceptable in family BMT for leukaemia.

Acute Disease↗

PubMed1996

Structural definition of the A*74 group: implications for matching in bone marrow transplantation with alternative donors.

We have identified two new A*74 alleles (A*7402 and 7403) in two unrelated individuals. A*7402 differs from A*7401 by a single amino acid substitution in the signal peptide and may be the result of a gene conversion event at the 3' end of exon 1. A*7403 differs from A*7401 by a single amino acid exchange in the alpha 1 domain and is most likely due to a point mutation in exon 2, since no HLA class I donor allele has been found. Since A*7402 appears to be the ancestor of the other two A*74 alleles, it is possible that A*7401 and 7403 have been created by successive point mutations. The sequences of the expressed proteins of A*7401 and 7402 are identical. The heavy chain sequence of A*7403 differs from these alleles at the crucial residue 79 which is located in the sequence stretch of the alpha 1 alpha-Helix where the Bw4/Bw6 determinants have been identified and which probably affects TCR interaction. This variation can therefore be expected to stimulate alloreactive T cells, graft rejection and graft versus host disease emphasizing the relevance for matching in bone marrow transplantation with alternative donors.

Alleles↗

PubMed1995

HLA-DP antibodies in patients awaiting renal transplantation.

Sera from 505 patients awaiting renal transplantation with known panel-reactive cytotoxic antibody (PRA) status were tested for HLA-DP antibodies of the IgG class by means of the monoclonal antibody immobilization of leukocyte antigens (MAILA) technique. The overall incidence of HLA-DP antibodies was 7.3%. A positive HLA-DP antibody status correlated only weakly (r = 0.23) with a positive cytotoxic antibody status. After retrospective analysis, patients with HLA-DP antibodies prior to retransplantation revealed a significantly (P < 0.025) higher graft function rate than HLA-DP-negative patients. One patient was found to possess IgG HLA-DP autoantibodies prior to transplantation; thus far, his graft has been functioning for more than 2 years.

Antibodies↗

PubMed1995

Progress of unrelated bone marrow donor search at the University Hospital of Essen (1991-1994).

The development of unrelated bone marrow donor search (UMDS) was analyzed in a retrospective single-center study covering the period from 1991 to 1994. The data files of the 285 enrolled UMDS patients were evaluated for the patients' underlying disease, HLA phenotype frequency, year of search start, UMDS duration/outcome, and the origin of identified donors. Within 3 months after UMDS start, a donor was found in 5%, 10%, 19%, and 49% of the searches started in 1991 (n = 57), 1992 (n = 73), 1993 (n = 77), and 1994 (n = 78, until September), respectively. The total UMDS success rate increased from 53% (1991 group) to 67% (1994 group), with 5% and 54%, respectively, of the accepted donors being supplied by German registries. Thus, the current UMDS procedure provides a suitable donor for the majority of the patients lacking a related donor, and the majority of the donors are now facilitated by national donor centers.

Bone Marrow↗

PubMed1995

Detection of panel-reactive anti-HLA class I antibodies by enzyme-linked immunosorbent assay or lymphocytotoxicity. Results of a blinded, controlled multicenter study.

A soluble HLA ELISA for the detection of anti-HLA class I IgG antibodies was developed and compared to complement-dependent microlymphocytotoxicity. ELISA plates were coated with a panel of sHLA class I antigens isolated from the culture supernatants of 46 different EBV-transformed phenotyped B-cell lines. After the incubation of the coated plates with test serum, bound antibodies were detected using a peroxidase-conjugated anti-human IgG antibody. Absorbance was read using an ELISA plate reader and assay results were analyzed by computer. Antibody specificities were determined by Fisher's exact test tail analysis. The reproducibility of ELISA assay results was evaluated in a blinded, controlled multicenter study. A total of 102 serum specimens from patients on waiting lists to receive kidney transplants were tested five times by ELISA in five different laboratories. The correlation coefficients (r) of %PRA values determined by ELISA ranged from 0.89 to 0.96, and the average agreement on qualitative assay results (antibody positive vs antibody negative) was 98%. Endpoint titration of several serum specimens demonstrated equivalent sensitivity of ELISA and microlymphocytotoxicity (using the anti-globulin antibody protocol). Most of the antibody specificities determined by ELISA were in agreement with specificities determined by microlymphocytotoxicity. To evaluate the correlation of ELISA and microlymphocytotoxicity (CDC) assay results the same 102 specimens were tested six times by CDC in five different laboratories. The interlaboratory correlation coefficient (r) of %PRA values determined by microlymphocytotoxicity ranged from 0.57 to 0.94, and the average agreement on qualitative assay results was 85%. A comparison of ELISA with microlymphocytotoxicity was performed using consensus microlymphocytotoxicity results. This showed a high correlation (r = 0.81) of %PRA values determined by ELISA and microlymphocytotoxicity. This demonstrates that the detection of anti-HLA class I antibodies by soluble HLA ELISA is a reliable alternative to microlymphocytotoxicity testing.

Antibody Specificity↗

PubMed1995

Cyclosporine-induced apoptosis in CD4+ T lymphocytes and computer-simulated analysis: modeling a treatment scenario for HIV infection.

Cyclosporine A (CsA) is a potent immunosuppressive drug which interferes in vitro and in vivo with T-cell function. CsA has been shown to arrest T-cell maturation intrathymically and to inhibit T-cell proliferation. In this study, we demonstrate that CsA induces apoptosis in the canine CD4+ CD8- T-lymphocyte cell line 401 in a dose- and time-dependent fashion. Similar results could also be obtained from human peripheral blood lymphocytes. Apoptosis is observed within 4 hours after CsA application and is not prevented by excessive addition of ConA supernatant as a source of interleukins. The induction of apoptosis in CD4+ T lymphocytes suggests a possible treatment option for human immunodeficiency virus (HIV) infections, since the major target population for the HIV would be ablated at short term. A computer-simulated analysis with the "Cybermouse" HIV model confirmed that the virus would eventually disappear and HIV-infected macrophages would also be substantially reduced if CsA were given in combination with drugs which block viral replication (3'-azido 3'-deoxythymidine or 2',3'-dideoxycytidine). This treatment scenario could be applied under controlled conditions and with supportive patient care. A further review of the literature also suggests the positive impact of CsA treatment on the progression and outcome of AIDS-related mortality.

Animals↗

PubMed1995

[Meniere disease as an autosome dominant hereditary disease].

Within one year (1993) we found a positive family history in five out of forty-eight new patients with Menière's disease, corresponding to a frequency of 10.4%. We found between two and seven cases in each of five families. The disease followed a dominant autosomal hereditary pattern over two to four generations. Audiometric and vestibulometric examination confirmed the diagnosis. Clinical emphasis was placed on trigger factors such as infections, physical and psychological stress, the autonomic system, and metabolic and endocrinological disorders. There was no evidence of a significantly high incidence of any single trigger factor. The most important result of this study, which also included human leucocyte antigen (HLA) typing, is the fact that HLA A2 was represented in 90% of patients with positive family history of Menière's disease and in 75% of patients with solitary Menière's disease in contrast to only 28.9% in the average European population. The frequency of HLA B44 was 70% in family-linked Menière's disease and 37.5% in single Menière's disease. The frequency in the average European population is 12.3%. What is even more striking is the combination HLA A2 B44, occurring with respective frequencies of 60%, 37%, and 5%. These results suggest a multi-factor etiology of Menière's disease combined with a genetic predisposition, which might be caused by mutations on the short arm of chromosome 6.

Adult↗

PubMed1995

Definition of a new HLA-B7 subtype (B*0704) by isoelectric focusing, family studies and DNA sequence analysis.

During screening of potential bone marrow donors, a previously undescribed banding position for the serologically defined HLA-B7 antigen was identified in three unrelated families using one dimensional isoelectric focusing and class I specific Western blot analysis. The isoelectric point of the new variant is more acidic than the two HLA-B7 variants that had been defined before. In each family the new B7 variant was found linked to HLA-A2 and -Cw7. Cloning and sequencing of full-length clones of complementary DNA showed that the new allele (B*0704) differs from B*0702, the common allele encoding HLA-B7, by three nucleotide substitutions within the codon for residue 156 of the mature heavy chain. As a result of these differences amino acid 156 is changed from arginine to aspartic acid, a difference consistent with the isoelectric points. The group of three nucleotide substitutions that distinguish B*0704 from B*0702 is present in other HLA-B alleles.

Alleles↗

PubMed1995

Soluble HLA class I and class II concentrations in factor VIII and PCC preparations.

Soluble HLA class I (sHLA-CI) and class II (sHLA-CII) molecules were quantitated in 11 commercially available factor VIII and prothrombin complex concentrate (PCC) preparations by enzyme-linked immunosorbent assays (ELISA). In 4 preparations, we detected traces of sHLA-CI (0.01-0.07 mg/l), and in 7 hemostatic preparations small amounts of sHLA-CII molecules (0.02-0.28 mg/l). The concentrations of these contaminant molecules are unequivocally below the mean values of sHLA in human plasma (sHLA-CI: 1.01 +/- 0.72 mg/l; sHLA-CII: 1.53 +/- 2.44 mg/l). Based on the total amount of chronically administered factor VIII or PCC, contaminating sHLA molecules may be in principle able to exert immunomodulatory effects in patients treated with such preparations.

Enzyme-Linked Immunosorbent Assay↗

PubMed1995

Biochemical definition of DLA-A and DLA-B gene products by one-dimensional isoelectric focusing and immunoblotting.

The serologically defined canine MHC gene products (DLA-A and DLA-B) were investigated by one-dimensional isoelectric focusing after detergent phase separation, and immunoblotting (1D-IEF). The animals studied (n = 36) represented three different breeds. Since DLA-A and DLA-B antigens are MHC class I and class II molecules, respectively, cross-reactive polyclonal antibodies specific for human class I or class II molecules were utilized to visualize the protein pattern. The relative positions of the bands were correlated with the DLA-A and DLA-B antigens. In most cases DLA antigens of identical serological type did not differ in their 1D-IEF pattern except for DLA-A9 and the serologically undefined DLA-B gene product DLA-B 'blank'. For DLA-A9, two subtypes (A9.1 and A9.2) were identified in Beagle and associations of A9.1 with the haplotype A9,B4 and of A9.2 with A9,B6 were seen. In contrast, the haplotype A9,B6 in Labradors was associated with A9.1. Whereas all six serologically defined DLA-A antigens possessed distinct isoelectric points (IEP), not all DLA-B antigens could be differentiated by 1D-IEF:B5 as well as B13 coband with one B 'blank' specificity (BE1). Furthermore, a second B 'blank' (BE2) could be identified by 1D-IEF, underlining the advantage of this technique in the immunogenetic analysis of DLA gene products in dogs.

Animals↗

PubMed1995

Strong association between the responder status of the FC gamma II receptor and recurrent spontaneous abortion.

Several models exist for the aetiology and therapy of recurrent spontaneous abortion (RSA). They are based in part on the assumption that an increased materno-fetal histocompatibility results in an insufficient maternal immunological recognition of the fetus, and thus renders the maternal immune system unable to sustain the pregnancy. The involvement of the FcgammaII receptor (FcgammaIIR) in RSA was suggested, since FcgammaIIR-blocking antibodies, present in normal pregnancies, could not be found in patients suffering from RSA. The FcgammaIIR is known to be functionally and structurally dimorphic, which results in a responder, respectively non-responder pattern. We used an in vitro proliferation assay to distinguish between FcgammaIIR responder and non-responder phenotypes. In 29 RSA couples we found 97% (28/29) of RSA patients and 100% (29/29) of their partners to be responders, whereas 50 fertile couples and a further 100 unrelated controls revealed distributions of responder vs. non-responder of 73% vs. 27% and 70% vs. 30%, respectively. These differences (RSA vs. controls) are highly significant (P less than or equal to 0.0001). Our results suggest further that the FcgammaIIR dimorphism might be involved in the pathogenesis of RSA.

Abortion, Habitual↗

PubMed1995

Association of increased bronchoalveolar lavage fluid albumin and serum beta 2-microglobulin with pulmonary complications after allogeneic bone marrow transplantation.

The aim of this prospective study was to identify markers in bronchoalveolar lavage fluid (BAL fluid) and serum predictive for the development of pulmonary complications in the early phase (< 50 days) post-BMT. Concentrations of BAL fluid albumin (alb) and serum beta 2-microglobulin (S-beta 2m,) were determined 10 days before BMT (BAL-B, baseline) and on day 1 post-BMT (BAL-1) in 20 patients who subsequently developed pulmonary complications (group 1) and in 66 patients who remained free of complications for a minimum of 12 months (group 2). Median BAL fluid alb concentrations were significantly (P < 0.05) higher in group 1 patients as compared to group 2 patients at BAL-B (40 vs 28 mg/l) and at BAL-1 (30 vs 15 mg/l). S-beta 2m at BAL-1 was also significantly elevated in group 1 patients (median 1.3 mg/l) compared to group 2 patients (median 1.15 mg/l). Using cut-off values for BAL fluid alb (> 23 mg/l) and S-beta 2m (> 0.8 mg/l) we identified 12 patients out of 19 who developed subsequent pulmonary complications from 12 out of 62 patients without such complications, 1 day post-BMT.

Adolescent↗

PubMed1995

Biochemical analysis of HLA-DP gene products by isoelectric focusing and comparison with cellular and molecular genetic typing results.

The polymorphism of HLA-DPA1 and HLA-DPB1 genes of 39 lymphoblastoid cell lines, representing 3 DPA1 and 14 DPB1 alleles, was studied after immunoprecipitation of DP molecules, subsequent isoelectric focusing (1D-IEF) and immunoblotting by a polyclonal DP antibody. Two 1D-IEF variants for DPA1 and 7 for DPB1 gene products were identified, including 2 new variants associated with the DPB1*1401 and DPB1*1901 alleles. The 1D-IEF polymorphism correlated clearly with molecular-genetic-defined alleles and primed lymphocyte or T cell line (PLT) specificities with exceptions observed in DUCAF (DPB1*0202) and ARBO (DPB1*0101). Both exceptional cases may reflect amino acid variations outside the hypervariable regions (HVRs) at the second exon of DPB1. The findings were supported by a comparison of HVR amino acid sequences with estimated net charges to the isoelectric points of the DPB1 molecules. 1D-IEF seems to be a valuable method for studying the expression of DPA1 and DPB1 alleles provided the molecules differ in their isoelectric points.

Amino Acid Sequence↗

PubMed1994

A research on DLA-DRB1 genotyping by PCR-RFLP. I. To select a appropriate oligonucleotide primer pair.

In order to study the DLA (Dog Leucocyte Antigen) class II region we utilized the polymerase chain reaction based restriction fragment length polymorphism (PCR-RFLP) method, which has been reported previously as an efficient and simple technique for accurate definition of the HLA class II alleles. To search for a appropriate primer pair a series of amplifications with 4 different primer pairs DLA-DR-SP/Stop, DLA-DR-SP/P3, HLA-DRB-GH46/50 and HLA-DRB-AMP-A/B were provided. Only one satisfactory amplification was obtained with the primer pair HLA-DRB-AMP-A/B. The analogous sequences of the primer pair are found in the sequence of HLA-DRB-cDNA. The amplification region of the primer pair includes also the three hypervariable regions (HVR) in the sequence of DLA-DRB cDNA. Southern blot hybridization analysis confirmed the specificity of the primer pair HLA-DRB-AMP-A/B. The results of Hae III and Hinfl digestion show high polymorphism in DLA-D region and allele specific polymorphic patterns. Therefore, it is suggested that the primer pair HLA-DRB-AMP-A/B is at present the only available and usefull primer pair in PCR-RFLP study of DLA-DRB1 gene.

Alleles↗

PubMed1994

Researches on DLA-DRB1 genotyping by PCR-RFLP. II. A study of serology and cellularly defined DLA haplotypes and their segregation.

The polymerase chain reaction based restriction fragment length polymorphism (PCR-RFLP) method was used to study DLA class II gene in dogs. Genomic DNA from 11 DLA homozygous reference dogs representing 8 different haplotypes and 2 families with a total of 16 animals were amplified by the oligonucleotide primer pair (HLA-DRB-AMP-A/B) cross-hybridizing HLA-DRB specific and fit for the amplification of DLA-DRB1 gene. The corresponding amplified DNA products were 235 base pairs. Amplified DNA was digested by 32 different restriction endonucleases, which could recognize allelic variations within DLA-DRB. After digesting only with Hae III, Hha I, Hinf I, Rsa I and Sau96 I high polymorphism was revealed respectively and 9 distinct RFLP pattern were shown, which could be correlate to the DLA haplotypes studied. The 8 cellular established DLA-D specificities present in the reference panel were defined unequivocally by PCR-RFLP and correlated with DLA-Dw5 and Dw6 two subtypes. The segregation pattern of four different DLA-DRB types could be demonstrated in two families. Based on these data we conclude that PCR-RFLP typing utilizing the above mentioned primer pair and endonucleases is a valuable tool to define DLA class II types in the dog.

Alleles↗