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21.
Development of a PCR-SSOP approach capable of defining the natural killer cell inhibitory receptor (KIR) gene sequence repertoires 总被引:10,自引:0,他引:10
A molecular typing method based on polymerase chain reaction (PCR) amplification of three different target domains (immunoglobulin domains 1 and 3, and the transmembrane-cytoplasmic domain), followed by hybridisation with 26 digoxigenin-labelled sequence-specific oligonucleotide probes (SSOP) has been established for the polymorphic killer inhibitory receptor (KIR) genes. In addition to identifying the 12 KIR subfamilies, our PCR-SSOP typing approach could also distinguish the putative alleles, NKB1 and NKAT3, that comprise the KIR3DL1 subfamily. Ninety unrelated blood donors and 13 families (52 individuals), including both parents, were subjected to our KIR PCR-SSOP typing approach. All 12 KIR subfamilies, including a 2DS5 variant sequence, were present in the 90 individuals and displayed varied phenotype frequencies: 2DL1 (0.96), 2DL2 (0.31), 2DL3 (0.95), 2DS1 (0.56) 2DS2 (0.51), 2DS3 (0.27), 2DS4 (0.96), 2DS5v (0.35), 3DS1 (0.47), 3DL1 (0.96), 3DL2 (1.0) and 2DL4 (1.0). A total of 23 different KIR phenotypes were defined in this study, and 10 of these were only found on one occasion in one individual, indicating considerable diversity in the KIR phenotype profiles within the Irish population. Most individuals (93%) possessed the complement of inhibitory KIR specificities for the three well-defined HLA-B and -C ligands. An unusual probe pattern for 3DS1 was observed in 3 individuals indicating a variant 3DS1 gene sequence with changes at nucleotide positions 1185-1186, within the cytoplasmic domain. Sequencing analysis revealed a new single nucleotide polymorphism in exon 3 of 3DL1 NKB1(195, G-A) and a 22-bp deletion polymorphism in exon 5 of 2DS4 (nucleotides 777-798 deleted). A number of strong KIR associations were observed, namely 2DL1 with 2DL3, 2DS4 with 3DL1, 2DL2 with 2DS1/2DS2/2DS3, 2DS1 with 2DS3/2DS5v/3DS1, 2DS2 with 2DS3 and 2DS5v with 3DS1. Analysis of the KIR segregation observed in the 13 families confirmed these strong associations and permitted the definition of a number of partial KIR haplotypes, e.g. 2DL2-2DS1-2DS2-2DS3-3DL1. The segregation analysis concluded that at least 3 distinct gene loci encode 2DL1-4 and at least 4 gene loci encode the non-inhibitory KIR2DS1-2DS5. In the case of 3DL1-2 and 3DS1, our data suggests 3 gene loci, one for each subfamily. 相似文献
22.
Michele C. Hindi-Alexander Ph.D. Maria A. Zielezny Ph.D. Naris Montes R.N. M.S. Bonnie Bullough Ph.D. Elliott Middleton Jr. M.D. Dutzu H. Rosner M.D. William M. London Ed.M. 《The Journal of allergy and clinical immunology》1985,75(6):709-715
After literature reports linking fibrocystic breast disease (FBD) to methylxanthine ingestion, a pilot study was undertaken to investigate the possible contribution of theophylline to this effect. The major goal of this project was to measure the effect of theophylline therapy on FBD in asthmatic women. All women attending an allergy clinic or an obstetrics/gynecology clinic over a 9-month period were examined to clinically assess FBD and were asked to complete a detailed questionnaire covering health history, other risk factors, and drug and dietary methylxanthines. The sample included 62 asthmatic women, 66 allergic but not asthmatic women, and 72 nonallergic and nonasthmatic women. By use of the FBD clinical taxonomy with its 19-point scale going from 0 to 18 that was developed for this study, the three groups did not differ significantly in terms of mean severity of FBD. On analyzing the effect of each of the methylxanthines on FBD severity, there is clear evidence that total methylxanthines was a contributing factor in FBD severity with or without adjustment for relevant variables, such as age, menopause, pregnancies, and groups. Theophylline was significant only when adjustments were made for age, pregnancy, and menopause in contrast to caffeine that was only significant with no adjustments. 相似文献
23.
Frequency of cytokine polymorphisms in populations from western Europe,Africa, Asia,the Middle East and South America 总被引:15,自引:0,他引:15
Meenagh A Williams F Ross OA Patterson C Gorodezky C Hammond M Leheny WA Middleton D 《Human immunology》2002,63(11):1055-1061
PCR-SSOP identification procedures for IL-2, IL-6, IL-10, TNF-alpha and TNF-beta cytokine polymorphisms have been developed. Application of the procedures to a range of diverse geographically distributed populations has identified ethnic differences within the groups studied. Five populations were investigated, Northern Ireland, South African Zulu, Omani, Singapore Chinese and Mexican Mestizos. 相似文献
24.
High incidence of the Cys 282 Tyr mutation in the HFE gene in the Irish population -implications for haemochromatosis 总被引:3,自引:0,他引:3
S. Murphy M.D. Curran N. McDougall M.E. Callender C.J. O'Brien D. Middleton 《Tissue antigens》1998,52(5):484-488
Abstract: A simple PCR-SSOP approach based on a single PCR product has been developed to screen the HFE gene for the haemochromatosis-associated mutations Cys 282 Tyr and His 63 Asp. Using this approach the prevalence of these mutations in a cohort (30) of haemochromatosis patients and normal controls (404) was determined. Ninety percent of the haemochromatosis patients were homozygous for the Cys 282 Tyr mutation. In the normal population we found an increased incidence of the Cys 282 Tyr mutation (17.3%; 95% confidence limits 0.136–0.209) which was also reflected in the higher frequency of Cys 282 Tyr homozygotes (1.24%; 95% confidence limits 0.0015–0.0232). Linkage disequilbrium analysis confirmed the association between A*03 and Cys 282 Tyr. However, strong linkage disequilibrium occurred with the HLA-A*03-associated allele HLA-B*14 but not the HLA-A*03-associated allele HLA-B*07. The His 63 Asp was found to be in linkage disquilibrium with HLA-A*29. 相似文献
25.
High resolution PCR-SSOP typing methods for HLA-B identification have been established and applied to a Northern Ireland population, using large enough numbers to give dependable allele frequencies. The six systems, which operate independently of each other, are intended for use as secondary typing systems following HLA-B identification with a medium resolution PCR-SSOP technique. 相似文献
26.
Diagnosis of fastidious enteric adenoviruses 40 and 41 in stool specimens 总被引:3,自引:12,他引:3 下载免费PDF全文
Thirty-five stool specimens, collected over a 14-week period from pediatric gastroenteritis patients and shown to contain adenovirus by electron microscopy, were inoculated onto 293 and HeLa cells. Virus isolates were characterized by serum neutralization and restriction endonuclease cleavage analysis of viral DNA from infected cells. Adenovirus was isolated upon primary inoculation of 293 cells from all 35 specimens shown to contain adenovirus by electron microscopy. Fastidious adenoviruses 40 and 41 (Ad40 and Ad41) were found in 17 (49%) of the stool specimens, and 4 of these specimens contained a conventional species (Ad1, Ad1, Ad18, Ad31) as well as Ad40. This was first manifest by the observation that four of the isolates which initially grew only in 293 cells acquired the capacity to grow in HeLa cells upon subsequent passage. In each case, the conventional species was undetectable by DNA analysis in the original inoculum but was selected in 293 cells and became the only one detectable by the second passage. Four other specimens, containing Ad1 or Ad31 alone, failed to grow initially in HeLa cells but did grow in 293 cells. The results of this study demonstrate therefore that (i) 293 cells are more sensitive than HeLa cells for the isolation of conventional as well as fastidious enteric adenovirus species and (ii) identification of viruses from patient specimens should involve minimal passage of the virus in cell culture, as a single passage can result in misdiagnosis of the virus associated with the infection. 相似文献
27.
C Arbesman I L Bernstein C W Bierman J S Bocles R Katz P L Lieberman K Mattucci E O Meltzer E Middleton J Noyes D S Pearlman H L Pence R G Slavin S L Spector 《The Journal of allergy and clinical immunology》1983,71(6):597-603
Fluocortin butyl (FCB) is a newly synthesized corticosteroid with a high ratio of topical to systemic activity. FCB was studied in a multi-center, double-blind, placebo-controlled trial of therapy of perennial rhinitis. The study was conducted between January and May 1981. Patients evaluated suffered from either chronic allergic or chronic nonallergic rhinitis or both. A total of 306 patients from 16 investigative centers were evaluated by comparing FCB to placebo. Three separate dosage regimens were employed. Patients received a total daily dose of 2, 4, or 8 mg. FCB was found to be an effective therapeutic agent. It reduced symptoms of nasal congestion, rhinorrhea, postnasal drainage, and sneezing. It also markedly reduced the use of concomitant medications (chlorpheniramine maleate and/or pseudoephedrine). Relief of symptoms was noted as early as the first week of therapy, and the degree of improvement increased progressively during the study. There was little difference between the relief produced by the 4 mg and 8 mg regimens. Both of these were superior to the 2 mg regimen. The drug was well tolerated; no significant side effects were noted. 相似文献
28.
Effect of adrenergic compounds, aminophylline and hydrocortisone, on in vitro immunoglobulin synthesis by normal human peripheral lymphocytes 总被引:2,自引:0,他引:2
Suspensions of normal human peripheral lymphocytes were exposed briefly to various concentrations of propranolol, isoproterenol, epinephrine, or aminophylline (both without and with added hydrocortisone) and then incubated for 72 hours at 37 ° C. After this incubation the amount of immunoglobulin (Ig) synthesized and secreted into the cultured supernatants was quantitated by radiaimmunoassay. Significantly increased Ig formation compared to controls was observed with the following concentrations of the experimental compounds (without hydrocortisone): 10?9 to 10?7M propranolol, 10?10 to 10?7M isoproterenol, 10?8M epinephrine, and 10?7 to 10?5M aminophylline. When cell suspensions were exposed briefly to these same compounds but with hydrocortisone added, the following concentrations of compounds significantly increased Ig synthesis: 10?10 to 10?8M propranolol, 10?10 to 10?6M isoproterenol, 10?9 to 10?7M epinephrine, and 10?8 to 10?5M aminophylline. Of the four compounds studied, only the epinephrine data suggest that hydrocortisone enhanced the stimulation of Ig synthesis by this catecholamine. 相似文献
29.
Counter-immunoelectro-osmophoresis for the detection of infantile gastroenteritis virus (orbi-group) antigen and antibody. 总被引:7,自引:2,他引:7 下载免费PDF全文
P J Middleton M Petric C M Hewitt M T Szymanski J S Tam 《Journal of clinical pathology》1976,29(3):191-197
A moderatley sensitive, rapid, and economical test scheme for the detection of infantile gastroenteritis virus (IGV) in stool or antibody in serum has been developed and evaluated. The test scheme with minor modifications was an adaptation of a counter-immunoelectro-osmophoresis system we once used for the detection of hepatitis B antigen. Large numbers of stool samples may be screened during half a working day for the presence of IGV using reference antiserum to IGV prepared in guinea-pigs. Serological studies of a diagnostic but not epidemiological nature may also be performed with equal facility by this same test scheme using highly purified IGV antigen derived from stool. 相似文献
30.
The development of respiratory syncytial virus-specific IgE and the release of histamine in nasopharyngeal secretions after infection 总被引:16,自引:0,他引:16
R C Welliver D T Wong M Sun E Middleton R S Vaughan P L Ogra 《The New England journal of medicine》1981,305(15):841-846
We studied the development of respiratory syncytial virus (RSV)-specific IgE and the release of histamine in nasopharyngeal secretions from 79 infants with various forms of respiratory illness due to RSV. RSV-IgE was measured by an enzyme-linked immunosorbent assay; specificity was confirmed by appropriate blocking experiments. Histamine content in the secretions was determined by fluorimetric methods. RSV-IgE was detectable in only one of 19 patients with RSV infection without wheezing, but was detectable in the majority of 60 patients with wheezing (P less than 0.01). Titers of RSV-IgE were significantly higher in patients with wheezing (P less than 0.05). Histamine was detectable in secretions of some patients with all forms of illness but was detected significantly more often (P = 0.05) and in higher concentrations in patients with wheezing. Peak titers of RSV-IgE and concentrations of histamine correlated significantly with the degree of hypoxia (P less than 0.001). Formation of RSV-specific IgE and release of histamine may adversely affect the outcome of RSV infection. 相似文献