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C Ling

Publications and source records attributed to C Ling.

54 records · Page 3Linked to original sources
PubMed1991

Vaccine-induced CD4+ T cells against the simian immunodeficiency virus gag protein. Epitope specificity and relevance to protective immunity.

We have examined the induction and epitope specificity of T cells for the simian immunodeficiency virus (SIV) gag p27 protein in macaques immunized with either a recombinant SIV gag protein or an inactivated SIV vaccine. CD4+ MHC class II-restricted T cell lines and clones derived from five immunized macaques recognized a total of seven peptides in three immunodominant regions of p27. Two T cell clones generated from one of the lines, recognized a single 20 amino acid peptide that overlapped with a region previously shown to include a CTL epitope from SIV-infected macaques. Although this epitope is in a conserved region of the gag protein of SIV, its recognition by a CD4+ T cell clone was abrogated by sequence variation in the equivalent HIV protein. The specificity of the T cell lines for synthetic peptides demonstrated considerable overlap between T cells generated by immunization with the recombinant gag protein and inactivated SIV. However, in contrast to the protective efficacy of the whole virus vaccine in the syntex adjuvant formulation, immunization with the p27 protein with alum failed to generate a protective immune response. Furthermore, despite the consistent gag-specific T cell responses induced by the recombinant protein, there was no evidence of an enhanced antibody response to envelope (env) after live SIV challenge.

Amino Acid Sequence↗
PubMed1991

Use of a fluorescent DNA analog for fluorometric detection of DNase activity.

The enhancement of fluorescence of the DNA analog poly(d epsilon A) following nucleolytic degradation to mononucleotides was found to be a convenient signal for studying nuclease, especially exonuclease, activity. This measurement, which is simple to obtain and extremely sensitive, detects various kinds of DNases and can be applied to the detection of nucleases in the course of protein purification. The signal change can be observed continuously during the reaction and easily converted to the amount of liberated mononucleotide. The method is thus suitable for quantitative and kinetic studies of exonuclease activity.

DNA Probes↗
PubMed1991

Studies on the immunogenicity of Chinese hamster ovary cell-derived recombinant gp120 (HIV-1IIIB).

Recombinant DNA-derived gp120 (HIV-1IIIB) expressed in chinese hamster ovary cells elicited specific humoral and cell-mediated immune responses in a variety of mammals. Antisera from immunized rabbits, sheep and goats recognized virus-derived gp120 and its precursor (gp160). Neutralizing antibodies were also elicited, but only in a few animals, and this may be related to the protein's susceptibility to cleavage through the neutralizing domain. However, in rabbits the degree of cleavage of gp120 had little or no effect on its antigenicity or immunogenicity. All antisera had limited cross-reactivity to envelope glycoproteins from a panel of HIV-1 isolates suggesting that immunodominant antibody epitopes are in variable regions of the recombinant gp120. Antigen-specific T-cell responses were detected in immunized macaques and were found to be stronger and more prolonged when gp120 was administered in Freund's adjuvant rather than alum.

Animals↗
PubMed1990

The production and purification of PCR-derived recombinant simian immunodeficiency virus p27 gag protein; its use in detecting serological and T-cell responses in macaques.

The polymerase chain reaction (PCR) was used to amplify a region of the gag gene, encompassing the core protein p27, from genomic DNA of cells infected with SIVmac251 (32H isolate). The 767 base pair PCR product was cloned into the bacteriophage M13 and fully sequenced before sub-cloning into the expression vector pUC19. The 30 kilodalton (kDa) fusion protein of lacZ-p27 was expressed as a soluble protein in E. coli JM101 cells and purified to greater than 90% purity by affinity chromatography. The affinity purified product was used in serological and T-cell assays to assess immune function in cynomolgus macaques immunised or challenged with immunodeficiency virus derived material. This reagent and accompanying methods provide valuable assays for monitoring the efficacy of vaccines for SIV as a model for human AIDS.

Animals↗
PubMed1990

Polyadenylated messenger RNA in paired helical filament-immunoreactive neurons in Alzheimer disease.

An antibody raised against isolated paired helical filaments (PHF) was used to identify tangle-bearing (PHF+) neurons in autopsy brain tissue from six Alzheimer disease (AD) patients and six age-matched controls (AMC). A comparison of the levels of polyadenylated messenger RNA [poly(A)+ mRNA] in PHF+ and PHF- neurons of similar cross-sectional area in temporal and parietal lobe and cerebellum from four AD and four AMC brains was made by analysis of in situ hybridization of [3H] polyuridylate [poly(U)] to intracellular poly(A)+ mRNA. In PHF+ neurons, the level of poly(A)+ mRNA was approximately two-thirds that in similar-sized PHF- neurons in either AD or AMC. The level of poly(A)+ mRNA in PHF- neurons in regions of the brain that have more of the histopathologically defined effect in AD was similar to that in regions with less effects.

Aged↗
PubMed1989

Auger electron contribution to bromodeoxyuridine cellular radiosensitization.

Halogenated thymidine analogs become incorporated into the DNA of proliferating cells during S-phase and may be used clinically to radiosensitize tumors that are otherwise poorly responsive to radiation. Although radiosensitization has been studied for years, mechanisms of radiosensitization are poorly understood. One possible mechanism involves the release of short range, high-LET, Auger electrons following photoelectric absorption of an X ray by the K-shell of the incorporated halogen. Such absorption occurs only with X ray energies slightly greater than the K-shell binding energy. We report the results of an experiment designed to measure this effect, in which cultured monolayers of Chinese hamster V79 cells, with 32% replacement of thymidine by bromodeoxyuridine (BUdR), were exposed to monoenergetic X rays just below (13.450 KeV) or above (13.490 KeV) the K-edge (13.475 KeV) of bromine. Enhancement ratios calculated in five different ways were slightly increased (3-12%) above the K-edge compared to below. However, only a calculation using a linear-quadratic fit to the data and a surviving fraction of 0.01 demonstrated a statistically significant increased enhancement ratio (12%) above the K-edge. We conclude that Auger electrons produced following photoelectric absorption of X rays by the K-shell of bromine contribute minimally to observed BUdR cellular radiosensitization.

Animals↗
PubMed1989

Brain interleukin 1 and S-100 immunoreactivity are elevated in Down syndrome and Alzheimer disease.

Interleukin 1, an immune response-generated cytokine that stimulates astrocyte proliferation and reactivity (astrogliosis), was present in up to 30 times as many glial cells in tissue sections of brain from patients with Down syndrome and Alzheimer disease compared with age-matched control subjects. Most interleukin 1-immunoreactive glia in Down syndrome and Alzheimer disease were classified as microglia. The number of interleukin 1 immunoreactive neurons did not appear to differ in Down syndrome and Alzheimer disease compared with control brain. Numerous temporal lobe astrocytes in Alzheimer disease and postnatal Down syndrome were intensely interleukin 1-, S-100-, and glial fibrillary acidic protein-immunoreactive and had reactive structure. Interleukin 1 levels in Alzheimer disease temporal lobe homogenates were elevated, as were the levels of S-100 and glial fibrillary acidic protein, two proteins reportedly elevated in reactive astrocytes. These data suggest that increased expression of S-100 in Down syndrome, resulting from duplication of the gene on chromosome 21 that encodes the beta subunit of S-100, may be augmented by elevation of interleukin 1. As a corollary, the astrogliosis in Alzheimer disease may be promoted by elevation of interleukin 1.

Adult↗
PubMed1987

Fast abortive initiation of uvrA promoter in a supercoiled plasmid studied by stopped-flow techniques.

In order to follow the fast kinetics of abortive initiation (lag time from 1 ms to 10 s), we have built a stopped-flow apparatus equipped for fluorescence detection. The small volume used for each assay (35 microliters), and the short dead time (approximately 0.5 ms) are the essential advantages of this apparatus. Supercoiling of DNA affects considerably the initiation of transcription from the uvrA promoter. It decreases the lag time due to the isomerisation process 3-fold. Nevertheless, it does not change significantly the product KBk2, which is indicative of promoter strength and shows that uvrA is an 'association-limited' promoter. The presence of the LexA repressor increases the lag time considerably. At least for small RNA polymerase concentrations this increase is stronger for supercoiled than for linearized DNA.

Bacterial Proteins↗
PubMed1980

Non-A/non-B hepatitis in experimentally infected chimpanzees: cross-challenge and electron microscopic studies.

Inoculation of eight chimpanzees with factor VIII, factor IX, or "H" strain plasma resulted in enzymatic and histopathologic evidence of non-A/non-B hepatitis in all eight animals. Challenge of two chimpanzees convalescent from factor VIII-induced disease with either factor IX or "H" strain plasma resulted in non-A/non-B hepatitis only in the animal inoculated with factor IX materials. Reciprocal cross-challenge of a chimpanzee convalescent from factor IX-induced disease with factor VIII also produced unequivocal enzymatic and histopathologic evidence of non-A/non-B hepatitis. Cross-challenge of a chimpanzee convalescent from "H" strain-induced non-A/non-B hepatitis with factor VII did not cause a second bout of non-A/non-B hepatitis. These findings suggest the factor VIII materials and "H" strain plasma used in these studies share a common etiologic agent (or agents), but that factor VIII and factor IX may contain two distinct agents. Electron microscopic (EM) examination of thin-sectioned, acute-phase liver biopsies from all but one of the chimpanzees receiving the primary inocula revealed the presence of abnormal hepatocyte cytoplasmic structures previously shown to be associated with non-A/non-B hepatitis. Crystalline structure containing 25 to 30 nm particles were visualized by EM in the cytoplasm of endothelial or Kupffer cells in acute-phase liver biopsies obtained from three chimpanzees inoculated with either factor VIII materials or "H" strain plasma.

Animals↗
PubMed1977

Incidence of HBsAg, anti-HBs and subtypes ad and ay in a general hospital: correlation with clinical data and its significance as a public health problem.

During a period of four months, 9,418 individuals, comprising new hospital admissions, new employees and blood donors, were tested for HbsAg and 1,212 for anti-HBs. A high incidence of positivity for both was found in the different groups under investigation. Subtyping was carried out on acutely ill patients and carriers. Sbtype ay was common in acutely ill patients and among addicts and subtype ad in nonhepatitis patients and healthy carriers. The significance of these findings and possible measures for prevention of spread of the infection are discussed.

Antibodies↗
PubMedDate not supplied

Target-specific morphology of retinal axon arbors in the adult hamster.

The B fragment of cholera toxin (CT-B) provides a highly sensitive anterograde tracer for labeling retinofugal axons, revealing dense projections to known central retinorecipient nuclei, and sparse but distinct inputs to regions that have not been traditionally recognized as targets of direct retinal projections. In hamsters, we can identify CT-B labeled retinal axons in more than 25 cell groups in the mesencephalon, diencephalon, and basal telencephalon. CT-B labeling additionally delineates the complete arbor morphology, especially in regions that receive a sparse input, offering hitherto unknown views of retinal axon ramifications. We present here the terminal morphology of retinal axons in the lateral geniculate body and superior colliculus, verifying earlier studies, and also document novel findings on the configuration of retinal axon endings in the ventral nucleus of the lateral geniculate body, intergeniculate leaflet, suprachiasmatic nucleus, and in the nuclei of the accessory optic tract. Additionally, the trajectory and terminal morphology of retinal afferents to the hypothalamus, preoptic area, and basal telencephalon are detailed. The results are discussed in the context of possible functional roles for some of these projections.

Animals↗
PubMedDate not supplied

Dose parameters of 125I and 192Ir seed sources.

As mandated by an NCI brachytherapy contract, we measured dosimetric parameters for 192Ir seeds and two models of 125I seeds. Measurements were with LiF powder in a water-equivalent phantom. Data were corrected for background, sample mass, and finite detector volume. Selected parameters were also investigated through Monte Carlo calculations. Results are presented in terms of a dose parametrization that is described in detail, and are compared to published data. Our results agreed well with published data for relative quantities such as radial and angular dose dependence. Our measured value for the 192Ir dose factor was 4.55 cGy(H2O) cm2 mCi-1 h-1, also in good agreement with commonly used values. However, the measured dose factors for 125I seed models 6702 and 6711 were 1.18 and 1.06 cGy(H2O) cm2 mCi-1 h-1, values well below those in general use.

Brachytherapy↗
PubMedDate not supplied

Hypopituitarism and chorioretinopathy in two siblings.

Two male siblings with chorioretinopathy and pituitary dysfunction (CPD) were found to have empty sellas on CT scan. Extensive ophthalmological and endocrinological investigation revealed the absence of gonadotrophins in both brothers and growth hormone deficiency in one. ACTH, TSH, and posterior pituitary function were normal. Karyotyping in one brother revealed a 46XY complement. Ophthalmological evaluation of three other siblings and both parents revealed normal vision and no evidence of retinopathy. The brothers represent two additional examples of CPD syndrome, suggesting a genetic etiology for this syndrome complex. The empty sellas present add another facet to the central nervous system disorders found in CPD syndrome.

Adolescent↗
PubMedDate not supplied

Protection against SIV infection in macaques by immunization with inactivated virus from the BK28 molecular clone, but not with BK28-derived recombinant env and gag proteins.

Vaccination of cynomolgus macaques with beta-propiolactone inactivated SIVmacBK28 in Freund's adjuvant induced low but detectable levels of anti-SIV envelope (env) antibodies and T-cell responses and protected against challenge with the 32H isolate of SIVmac251 grown in C8166 cells. In contrast, purified recombinant SIV env and gag proteins derived from BK28 formulated in Syntex adjuvant generated consistent and long-lived cellular and humoral immune responses to SIV env, but failed to protect against infection with the 32H virus. Thus, protection against a heterogeneous challenge stock is possible by immunization with a molecularly-cloned virus, but not with recombinant proteins from the same molecular origin. High levels of anti-cell antibodies induced by the whole virus vaccine, but not by recombinant proteins, may have contributed to the protection observed.

Animals↗
PubMedDate not supplied

Immunisation of macaques with SIV env recombinants: specificity of T cell and antibody responses and evaluation of protective efficacy.

Macaques were immunised with lentil lectin purified recombinant SIVmac (BK28) derived gp160 (rgp160) with or without live vaccinia (vac)-env (BK28) priming, followed by a final boost with solid matrix antibody antigen (SMAA)-gp160 (J5) complexes and challenged with the SIVmac molecularly cloned virus J5M. Rgp160 and vac-env plus gp160 induced strong Ab responses against the homologous virus. Live vac-env did not enhance or prolong the antibody response, however, T cell responses were stronger. Analysis of the specificity of the immune response demonstrated that sequence variation within SIVmac viruses can affect antibody and T cell recognition. A single booster immunisation with the heterologous SIVmac J5 env recombinant protein was not sufficient to protect against the molecularly cloned virus J5M. These findings further illustrate the difficulty of generating protective immunity with immunogens based on single sequence recombinants.

Amino Acid Sequence↗