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Biomedical subjects

J Pei

Publications and source records attributed to J Pei.

14 recordsLinked to original sources

Examination of four HLA class I pseudogenes. Common events in the evolution of HLA genes and pseudogenes.

The HLA class I gene family in lymphoblastoid cell line 721 has been studied in detail and a number of sequences in addition to the classical genes have been identified. The cloning, characterization, and nucleotide sequences of four sequences, all full length HLA class I pseudogenes, are described in this report. These pseudogenes, contained within 5.4-, 5.9-, 7.0-, and 9.2-kb HindIII fragments, each have the class I exon-intron structure as well as class I homology in their 5' and 3' flanking regions. However, all four sequences have one or more substitutions that perturb the coding region, leaving little doubt that they are in fact pseudogenes. Comparisons among these sequences and the HLA class I genes revealed that their homology with the class I genes is patchwork. Thus, although some regions have diverged, other contiguous intron-exon sequences are highly conserved. Comparisons in the 5' regions indicate that the pseudogene promoters more closely resemble the classical HLA promoters than the nonclassical promoters as none of the unique structural features found in the HLA-E, -F, or -G regulatory regions are present in any of the pseudogene promoters. Further comparisons revealed that at least two putative gene conversion events, similar to those hypothesized to have occurred in the evolution of some HLA genes, may have occurred in the evolution of some of the pseudogenes. These and other hypothetical events in the evolution of the class I gene family are discussed.

Base Sequence

Cloning and physical mapping of the HLA class I region spanning the HLA-E-to-HLA-F interval by using yeast artificial chromosomes.

The HLA class I genes are located within a 2-million-base pair (2-Mbp) region constituting the telomeric half of the human major histocompatibility complex. The large majority of the class I sequences, including the HLA-A, -E, -F, and -G genes, is found within the telomeric 1 Mbp. We report here the isolation and characterization of yeast artificial chromosome (YAC) clones that span a contiguous region of greater than 1.2 Mbp and include 14 of the 18 characterized class I sequences. Restriction enzyme mapping and the use of locus-specific probes have allowed all of the class I genes and sequences to be ordered and positioned within the region. In addition, the transcriptional orientation of the four class I genes has been determined. Using probes derived from the ends of YAC inserts and from class I pseudogenes, we describe a highly polymorphic region between the HLA-A and HLA-G genes. This region appears to be deleted in certain HLA haplotypes, shortening the distance between HLA-A and HLA-G by greater than 50 kilobase pairs (kbp). As part of the characterization of the YAC clones, unique sequence probes derived from the ends of each YAC insert were identified. When combined with probes derived from HLA genes and pseudogenes, 25 locus-specific probes spanning the 1.2-Mbp region have been identified for an average of 1 probe every 48 kbp.

Chromosomes, Fungal

Isolation and characterization of yeast artificial chromosome clones linking the HLA-B and HLA-C loci.

A 290-kilobase-pair chromosomal segment containing the genes encoding the human class I major histocompatibility complex molecules HLA-B and HLA-C as well as a class I pseudogene has been isolated on three overlapping yeast artificial chromosome (YAC) clones. One YAC clone contains both the HLA-B and HLA-C genes. These loci are located approximately 85 kilobase pairs apart, each in close association with a CpG island. Southern blotting and nucleotide sequencing showed no evidence of alteration of the structure of the cloned DNA in the YACs. End fragments from the YAC inserts have been isolated and used to confirm the overlaps between clones. These fragments can also serve as polymorphic markers for structural analysis of the major histocompatibility complex. Our data show that YAC cloning offers an attractive alternative for analysis of the structures of large gene complexes such as HLA.

Amino Acid Sequence

Association of four HLA class III region genomic markers with HLA haplotypes.

We have studied restriction fragment length polymorphism (RFLP) in the region 300 kb centromeric to the HLA-B locus. Four probes were used: one was genomic DNA derived from the tumor-necrosis factor (TNF)-beta gene, one was a cDNA for the BAT3 gene, and two single-copy genomic probes, R5A and M20A. The order of these markers from HLA-B towards the centromere is M20A, R5A, TNF and BAT3. The BAT3 and TNF-beta probes each detected two allelic bands with Taq I and Nco I digestion, respectively; the R5A and M20A probes each detected three polymorphic allelic bands with BstEII digestion. To determine if these restriction polymorphisms are preferentially associated with certain HLA-B and -DR haplotypes, a total of 153 HLA haplotypes was analyzed. The haplotypes A1, B8, DR3 and A3, B7, DR2 were each associated with a distinct combination of polymorphisms identified at these four sites, thereby demonstrating that the strong linkage disequilibrium characteristic of these haplotypes extends also to this segment of the class III region. In contrast, haplotypes that are not in positive linkage disequilibrium, such as A1,B8,DR4 and A2,B7,DR3, showed no preferential association with any of these polymorphisms. The antigens HLA-B27 and B35 were also found to be in positive linkage disequilibrium with RFLP patterns at three of these sites, and HLA-B14,B35,B44,Bw57 and Bw62 were found preferentially associated with polymorphisms at one or two of these sites, independent of the DR antigen present. These data further demonstrate that genetic linkage disequilibrium in the HLA class III region is complex and variable among different HLA haplotypes.

Chromosome Mapping

Delayed response to argon laser trabeculoplasty in monkeys. Morphological and morphometric analysis.

In eight cynomolgus monkeys we performed argon laser trabeculoplasty and evaluated morphological changes in the trabecular meshwork (TM). Four weeks after laser treatment, the treated spots were detected as regions of flattened and collapsed beams covered by a cellular sheet extending from the corneal endothelium. The inner wall of Schlemm's canal beneath these regions was flat, with no vacuoles despite fixation with intraocular pressure maintained at 15 mm Hg. Adjacent nonlasered spots had wide open intertrabecular spaces with herniations of juxtacanalicular TM and of inner wall endothelium into and across the lumen of Schlemm's canal. The herniations contained chronic inflammatory cells and large vacuoles. Statistical morphometric analysis in three monkeys established that these herniations were significantly wider, more numerous, and contained more vacuoles than did control eyes. Our findings suggest that a biologic process is involved in these reactions to argon laser trabeculoplasty. Scarring is seen to obliterate TM in spots treated by the laser, and this apparently causes the flow of the aqueous humor to be diverted through adjacent nonlasered regions that become structurally altered to compensate for this overload of flow.

Animals

Q-switched neodymium-YAG laser trabeculopuncture in monkeys.

In nine cynomolgus monkeys, trabeculopuncture was performed with a Q-switched neodymium-YAG laser, using a pulse energy of 5 to 7 millijoules and an exposure time of 14 nanoseconds. A penetration into Schlemm's canal was successfully achieved with two to four pulses; this penetration was accompanied by intraocular pressure (IOP) reduction and blood reflux into the anterior chamber. However, after eight days, IOP returned to baseline level, while white tissue was observed gonioscopically to fill in the puncture sites. Histologically, one hour after laser treatment, a blasting effect on the trabecular meshwork was observed with no signs of necrosis. Ater eight days, a hypertrophic scar formed, with the corneal endothelium extending over the scarred surface. At eight weeks and at six months after laser treatment, further shrinkage of the scar and the formation of a membrane over it was evident. Attempts to control scar formation by preventing blood reflux or injecting fluorouracil subconjunctivally for two weeks were unsuccessful. Scar formation at the trabecular puncture site severely limits the applicability of this potentially simple glaucoma treatment.

Animals

Short-term effect of argon laser trabeculoplasty in monkeys.

We performed argon laser trabeculoplasty (ALT) in four cynomolgus monkeys and evaluated the morphologic changes in the trabecular meshwork one hour and 14 hours following the laser treatment. One hour after ALT, disruption of the trabecular beams and coagulative necrosis of the tissue were evident. Fragmented cells and fibrillar tissue debris were found in the trabecular spaces, accumulating in the juxtacanalicular region. This might explain the elevation of intraocular pressure seen immediately following ALT in some glaucomatous patients. Also, many trabecular endothelial cells were rounded up, displaying different stages of leaving the beams; these cells were actively phagocytic. Such stimulation of trabecular meshwork cells with subsequent removal of tissue debris might explain the temporary nature of elevation of intraocular pressure following ALT.

Animals

HLA polymorphisms in a Han Chinese population of Sichuan.

This study reports the phenotype frequencies (PF) and gene frequencies (GF) of a homogeneous Sichuan Han Chinese population by typing of unrelated 229 individuals and 16 families for HLA-A, B, C, DR and DQ antigens. The frequencies of Bw67, Bw71, DRw12, DRw13 and DRw14 are the first to be reported in Chinese. Two two-locus haplotypes with significant linkage disequilibrium among the Sichuan have also been listed.

Asian People