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J Parlevliet

Publications and source records attributed to J Parlevliet.

16 recordsLinked to original sources

Influence of thawing method on motility, plasma membrane integrity and morphology of frozen-thawed stallion spermatozoa.

Different thawing methods are used for stallion semen, however, it is unclear which method is the optimal one. To determine if the thawing temperature has an effect on semen quality, we compared 2 thawing temperatures, 75 degrees C and 37 degrees C. The following parameters were used to measure sperm quality: sperm motility, sperm viability, plasma membrane integrity and sperm morphology. Twenty-three ejaculates from 10 Dutch Warmblood stallions were thawed either at 37 degrees C for 30 sec or at 75 degrees C for 7 sec. Sperm motility was evaluated by a Hamilton Thorn Motility Analyser. Plasma membrane integrity and sperm viability were evaluated by using a live/dead fluorescein stain containing a calcein AM probe and ethidium homodimer-1 probe. The eosinaniline blue staining method was used to evaluate the percentage of live and dead cells, as well as sperm morphology. There was no significant difference (P = 0.84) between sperm motility after thawing at 37 degrees C and 75 degrees C. There was also no significant difference (P = 0.053) between the percentage of live spermatozoa using the calcein AM/ethidium homodimer stain after thawing at 37 degrees C and 75 degrees C. There was, however, a significant difference (P = 0.032) between the percentage of live spermatozoa using the eosin-aniline blue stain after thawing at 37 degrees C compared with that at 75 degrees C. In conclusion, our laboratory results indicated that stud farms using frozen semen should thaw the straws at 37 degrees C instead of 75 degrees C. The lower temperature is easier to work with, as thawing at the higher temperature requires special equipment and has to be timed very carefully to avoid damage to the spermatozoa.

Journal Article↗

Six newly identified HLA-DRB alleles: DRB1*1121, *1419, *1420, *1421, DRB3*0203 and DRB5*0103.

Seven samples with irregular PCR-SSO hybridization patterns, observed during routine HLA-DRB typing, were studied in more detail. Group-specific amplification, followed by hybridization with relevant SSOs strengthened the suggestion that these samples contained new DRB alleles. DRB exon 2 segments were amplified, cloned and sequenced and revealed: DRB1*1121 [MUL] is similar to DRB1*1102 in which codon 85 changed from GTT(V) into GTC(A); DRB1*1419 [AKKAL] is similar to DRB1*1402 with codon 71 changed from AGG(R) into AAG(K); DRB1*1420 [OND-52971] is a DRB1*1406 with codon 37 changed from AAC(N) into TTC(F); DRB1*1421 [TGI] is similar to DRB1*1417 with codon 71 changed from AGG(R) into AAG(K); DRB3*0203 [POS] is similar to DRB3*0202 in which codons 37-38 are changed from TAC GCG(YA) into TCC GTC(SV); DRB5*0103 was found in two unrelated individuals of Oriental origin [IND-24 and IND-59] and is similar to DRB5*0102 in which codon 71 AGG(R) changed into ACG(T). This particular sequence variation at position 71 has not yet been described. The new DRB sequences were confirmed using the sequencing based typing technique. Low resolution PCR-SSP typing failed to amplify two of the DRB1*14 variants, whereas high resolution PCR-SSP resulted in aberrant patterns. Class II alloantisera identify the codon 71 changes in DRB1*1419 and *1421 with respect to the MC1 ('DR1+DR4') epitope.

Alleles↗

Relation between stallion sperm binding to homologous hemizonae and fertility.

The hemizona assay (HZA) has been developed as a diagnostic test to predict the fertilisation potential of human spermatozoa. The aim of this study was to develop an HZA for stallion spermatozoa and to investigate a possible relationship between fertility and the outcome of the HZA in this species. Equine oocytes were obtained from ovaries collected at a slaughterhouse and by transvaginal, ultrasound-guided follicle aspiration. They were then denuded from cumulus cells and stored in salt solution at 4 degrees C until use. On the day of the experiments the oocytes were bisected, thus providing 2 equal matching hemizonae from each oocyte. Semen samples from Dutch Warmblood stallions with known fertility data were used to assess the number of spermatozoa bound to the outer side of the hemizona after incubation in vitro. Sperm binding to matching hemizonae of a particular stallion was similar and confirmed the feasibility of using the HZA for the horse. Sperm hemizona binding capacity of 10 pairs of stallions was compared by incubating 1 hemizona with the semen of a stallion and the matching hemizona with the semen of another stallion from the same stud farm. Five matching pairs of hemizonae were used for each pair of stallions. There was a significant relationship between the mean number of spermatozoa bound to matching hemizonae and the fertility indices of stallions from each stud farm (P < 0.0001). It is concluded that HZA can be used as a valuable parameter in stallion semen analysis.

Journal Article↗

Increasing complexity of HLA-DR2 as detected by serology and oligonucleotide typing.

Serological and oligonucleotide typing was performed on a number of HLA-DR2-positive cells from different ethnic origin, including DR2 haplotypes with various DQ associations. Exons 2 of DRB1 and DRB5 of DR2-positive individuals were locus-specific amplified and hybridized with a number of different oligonucleotides capable of discriminating between the various Dw2, Dw12, Dw21, and Dw22 associated sequences. The linkage of DRB with DQA1 and DQB1 in these haplotypes was analyzed. Among the DR2- positive cells we could define 10 different DR DQ haplotypes by serology and 13 by oligonucleotide typing. The DR2.ES specificity is a serological DRw15 variant which could not be discriminated by oligonucleotide typing from a DRw15 DQw5 haplotype. The DR2.JA variant represents a unique DRB1*1602 DRB5*0101 haplotype. The DR1+2s haplotype consists of a DRB1 DQ region from a Dw1 and a DRB5 gene from a Dw2 haplotype. Its short DR2 serum pattern can be explained by the absence of a DR2 DRB1 gene product. DRB5*0101 sequences were found in association with DRB1*1501, *1502, *1602, and *0101 alleles. Since the DRB5 gene is capable of such different associations it is comparable to the DRB3 and DRB4 genes. This may have implications for the definition of the broad DR2 specificity which is predominantly encoded by the DRB5 gene product. New DR2 haplotypes included the following DQ combinations: DQw2-positive DQA1/B1*0301/0201 and DQw6-positive DQA1/B1*0102/0601 and *0102/0603 haplotypes.

Amino Acid Sequence↗

Polymorphic and monomorphic HLA-DR determinants on human hematopoietic progenitor cells.

The expression of monomorphic Ia-like antigens and polymorphic (allotypic) HLA-DR determinants on CFU-GM, BFU-E, CFU-E, and CFU-GEMM was studied in bone marrow and peripheral blood cells from normal healthy individuals. Using various polyclonal and monoclonal anti-Ia-like antibodies, the presence of HLA-DR backbone antigens was shown on all hematopoietic progenitor cells (HPC) studied, both in complement-dependent cytotoxicity assays and in fluorescence-activated cell sorting (FACS). The expression of allotypic determinants was demonstrated on all HPCs, using the HLA-DR typing sera anti-HLA-DR1, 2, 3, 4, 5, and 7. The Class II antigen MT-2 was also shown on all HPCs, using both monoclonal and alloantisera, whereas the MB-1 (DC-1) determinant could not be demonstrated on HPCs. This might open the possibility of removing MB-1-positive malignant cells from the graft in autologous bone marrow transplantation.

Antilymphocyte Serum↗

Complement-dependent cytotoxicity in the analysis of antigenic determinants on human hematopoietic progenitor cells with HLA-DR as a model.

Complement-dependent cytotoxicity (CDC) assays using anti-Ia antisera have resulted in controversial conclusions about the expression of HLA-DR determinants on human hematopoietic progenitor cells (HPC). The expression of these antigens on CFU-E in particular could often not be demonstrated. Since this is contradictory to our own results, we decided to study the influence of both the antibody and complement concentrations in CDC assays using murine monoclonal and polyclonal anti-Ia (HLA-DR "backbone") antibodies and human anti-HLA-DR typing sera. We found that with use of the same anti-Ia antibody concentrations, elimination of CFU-E required significantly more complement than CFU-GM and BFU-E. In CDC assays with antipolymorphic HLA-DR antisera, complete kill of both CFU-E and BFU-E required significantly more complement than of CFU-GM. Insufficient complement concentration could be partly overcome by increasing the antibody concentration. Intrinsic insensitivity of CFU-E to low concentrations of complement could be excluded by experiments using monoclonal anti-HLA-A/B/C "backbone" antibodies. Furthermore, FACS experiments demonstrated that the density of HLA-DR determinants on CFU-E is lower than on BFU-E and CFU-GM. These data show that in CDC assays, antigens with low expression on HPC can easily be overlooked.

Colony-Forming Units Assay↗

Reanalysis of the HLA-DRw6 complex.

The serological definition of the HLA-DRw6 specificity has been complicated by the lack of monospecific reagents. During the 8th International Histocompatibility Workshop several subspecificities of DRw6 were proposed. In the present study we have reanalysed the reactions of 8th Workshop and local sera in families and homozygous typing cells. These studies have revealed four DRw6 related reaction patterns. The patterns are described and compared with those reported by others. Two of these specificities are commonly observed in Dutch Caucasoids, one related with Dw6 and one with Dw9. The other DRw6 related reaction patterns are very infrequent in this population. In addition, the relation between a locally defined specificity LB5x8 and two new specificities 8wDRw13 and 8wDRw17 is discussed.

Adult↗

Improved detection of HLA-Cw5 and Cw7 by two colour fluorescence technique.

Reactivity of two Cw5 and three Cw7 antisera was compared in the standard NIH and the Two Colour Fluorescence (TCF) techniques. Contrary to the NIH, the TCF method yielded strong positive reactions which enabled clear assignment of the Cw5 and Cw7 specificities. The TCF technique combines three factors that increase the sensitivity of serum reactions: 1. typing on B lymphocytes, 2. prolonged incubation and 3. staining of the lysed cells with ethidium bromide.

Cross Reactions↗

Cell membrane polymorphisms coded for in the HLA-D/DR region. I. Relation between D and DR.

The relation of HLA-D and -DR determinants was studied in Dutch Caucasoids. The recognition of subgroups of DR4, DR5, and DR7, and the specificities LB12 and LB13 are described. Phenotype and gene frequencies and a Hardy--Weinberg analysis of DR and local (LB) B-cell groups are given. Excellent correlation between D and DR typing was obtained when HTCs were studied by selected B-cell antisera. When the same sera were used to type a panel of D typed cells, the correlation was decreased (with the exception of DR3 and Dw3). In the case of discrepancies the DR specificity, but not the corresponding D specificity, always could be found and not the other way around. The data fit best the assumption that HLA-D and -DR are carried by the same molecule, although they might be different determinants on this molecule. A number of possible explanations for the observed discrepancies has been given.

Cell Membrane↗

Influence of previous immunization on skin graft survival.

The results of skin grafts transplanted in immunized and nonimmunized recipients was analysed. Specific sensitization for HLA-A or B determinants shortens graft survival if the recipients were immunized by s.c. injections of leukocytes. When the recipients had been pregnant, no such influence of specific HLA-A or B sensitization could be demonstrated. The variance in mean survival times of grafts exchanged between mixed lymphocyte culture (MLC)-positive donor-recipient combinations was significantly smaller than the variance in mean survival time (MST) of grafts exchanged between MLC-negative combinations. This difference could be the result of the influence of allograft immune-activating determinants of different strength in the MLC-negative donor-recipient combinations. Also the variance in MST of grafts in immunized recipients was significantly larger than the variance in MST of grafts in nonimmunized recipients. Apart from the obvious effect of HLA-A and B sensitization, other less well documented factors must have influenced graft survival. We did not find evidence for a graft enhancing effect of B cell-specific antibodies.

Antibodies↗

The lymphocyte activating influence of the HLA-A region.

Six individuals with an established recombination between the HLA-A and the HLA-B loci of the major histocompatibility complex have been tested in unilateral mixed lymphocyte cultures with other members of their families. A weak, but statistically significant lymphocyte activating influence of the HLA-A chromosomal region has been observed.

Adult↗