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Biomedical subjects

H B Jenson

Publications and source records attributed to H B Jenson.

At least 55 records · Page 3Linked to original sources

New and improved vaccines. Promising weapons against varicella, hepatitis A, and typhoid fever.

The initial motives behind development of vaccines were to protect against life-threatening infections (eg, rabies, diphtheria), to eradicate sweeping outbreaks of serious diseases (eg, paralytic poliomyelitis, smallpox), and to prevent diseases in a vulnerable population by the immunization of surrogates (eg, rubella immunization to prevent congenital rubella syndrome). Now a fourth motive emerges: prevention of less serious infections to improve quality of life. The advantages of new vaccines and immunization programs should no longer be measured exclusively in terms of the number of lives saved but should take into account direct and indirect cost savings and overall benefit to individual and societal health and well-being. Although varicella and hepatitis A infections can be life-threatening, most cases are self-limited and have no significant sequelae. Immunization is more likely to improve quality of life than to save lives. Vaccination against typhoid remains a potentially lifesaving act in developing nations, but even the newer typhoid vaccines were developed primarily to reduce the frequency and severity of adverse reactions to immunization rather than to improve the protective efficacy of the original heat-phenol inactivated vaccine. Varicella virus vaccine, hepatitis A virus vaccines, and the typhoid polysaccharide Vi capsular vaccine represent important additions to immunization agents. These vaccines are immunogenic, clinically effective, and generally safe, with infrequent and usually mild adverse reactions. Their favorable benefit-risk ratio should encourage their appropriate use. The Centers for Disease Control and Prevention, the American Academy of Pediatrics, and the American Academy of Family Physicians have already recommended varicella vaccine for universal immunization of children. Formal recommendations that hepatitis A vaccine also be routinely used for all children may be forthcoming in the next few years; in the meantime, persons at high risk should be immunized. Typhoid vaccination will likely continue to be used selectively for those who have significant contact with the organism or those who travel to typhoid-endemic countries.

Adolescent↗

Association of Epstein-Barr virus with leiomyosarcomas in young people with AIDS.

BACKGROUND: Children with the acquired immunodeficiency syndrome (AIDS) have an unusually high incidence of smooth-muscle tumors (leiomyomas and leiomyosarcomas) in addition to malignant lymphomas. We tested the hypothesis that the smooth-muscle tumors in these children are associated with the Epstein-Barr virus (EBV). METHODS: Tissue specimens of five leiomyosarcomas and two leiomyomas from six children with AIDS were studied for evidence of the human immunodeficiency virus (HIV) and EBV by in situ hybridization and quantitative polymerase chain reaction (PCR). Comparison specimens included samples of leiomyosarcoma and leiomyoma from HIV-negative children. EBV clonality of leiomyosarcomas was determined by Southern blot analysis with oligonucleotide probes for EBV terminal-repeat fragments. Tumor specimens were tested by immunoperoxidase staining for infiltration by B lymphocytes and expression of the EBV receptor. Serologic testing for EBV was performed. RESULTS: In situ hybridization showed EBV genomes in all muscle cells of the five leiomyosarcomas and the two leiomyomas from the six HIV-infected children. Quantitative PCR demonstrated strikingly high levels of EBV in tumor tissue, with as many as 4.3 genome copies per cell. Two colonic leiomyosarcomas obtained from different sites at different times from one patient contained different episomal EBV clones, signifying the presence of distinct monoclonal EBV-related tumors. We found biclonal EBV infection in the leiomyosarcoma of another patient. No EBV was detected in normal muscle or tumor specimens from HIV-negative patients. Immunostaining for the EBV receptor was strongly positive in six of the seven leiomyomas and leiomyosarcomas from the patients with AIDS. CONCLUSIONS: EBV can infect smooth-muscle cells, at least in patients with AIDS, and it may contribute to the pathogenesis of leiomyomas and leiomyosarcomas in children with AIDS. EBV seems to play no part in smooth-muscle tumors in HIV-negative children.

AIDS-Related Opportunistic Infections↗

An efficient method for routine Epstein-Barr virus immortalization of human B lymphocytes.

A variety of methods exist for the immortalization of B lymphocytes by Epstein-Barr virus due to the simplicity of such techniques to establish cell lines with stable genomic DNA. Two different methods for immortalizing lymphoblastoid cell lines were compared for differences in techniques and materials, time between initiation and immortalization, and success rate of immortalization. An incubation period in Epstein-Barr virus and the use of conditioned media improved immortalization efficiency from 86 to 98% and decreased the time (usually weeks) from culture initiation to cryopreservation. The resulting cell bank was used to produce DNA for genetic studies focusing on the genes involved in non-insulin-dependent diabetes mellitus.

B-Lymphocytes↗

Human herpesvirus 6 infection of the female genital tract.

Four of the seven human herpesviruses are recognized to replicate in the female genital tract and may be transmissible to sexual partners and newborn infants. Several of these viruses have also been implicated in the etiology of various human cancers, including tumors of epithelial cell origin. Human herpesvirus 6 (HHV-6) is a newly identified herpesvirus that causes exanthem subitum. The pathogenicity of HHV-6 within the genital tract is largely unexplored. Acellular vaginal secretions from 29 women attending a sexually transmitted diseases clinic were examined for the presence of HHV-6 DNA sequences by polymerase chain reaction. Three samples (10%) were consistently positive for HHV-6 DNA. Since HHV-6 DNA is shed in the genital tract of some women, it is possible that infectious virus is transmissible through sexual contact and to newborn infants by perinatal spread.

Adult↗

Production and characterization of murine monoclonal antibodies specific for baboon IgG heavy and light chain epitopes.

We have generated a panel of murine monoclonal antibodies (MAbs) that recognize baboon IgG epitopes. The reactivity of the MAbs with IgG from other primate species was also examined. Specificity for IgG heavy (H) or light (L) chain epitopes was determined by Western blot analysis. The H chain-specific MAbs were analyzed for IgG subclass specificity and the L chain-specific MAbs for reactivity with baboon IgM and polymeric sIgA. Finally, an ELISA was developed to demonstrate the utility of the MAbs in analysis of humoral immune responses in baboons.

Animals↗

Absence of a reservoir of Epstein-Barr virus (EBV) in normal tongue epithelium.

We examined human tongue epithelium and serum samples at autopsy for evidence of latent Epstein-Barr virus (EBV) infection. Although clinical serology revealed anti-EBV antibodies in most sera indicating past EBV infection, we found no Epstein-Barr nuclear antigen (EBNA)-coding sequences in tongue tissue by polymerase chain reaction (PCR), or Epstein-Barr-encoded RNA (EBER1) by in situ hybridization. Tongue epithelium does not appear to be a natural reservoir for latent EBV in immunocompetent hosts.

Adolescent↗

A dedicated database program for cataloging recombinant clones and other laboratory products of molecular biology technology.

A novel computer database program dedicated to storing, cataloging, and accessing information about recombinant clones and libraries has been developed for the IBM (or compatible) personal computer. This program, named CLONES, also stores information about bacterial strains and plasmid and bacteriophage vectors used in molecular biology. The advantages of this method are improved organization of data, fast and easy assimilation of new data, automatic association of new data with existing data, and rapid retrieval of desired records using search criteria specified by the user. Individual records are indexed in the database using B-trees, which automatically index new entries and expedite later access. The use of multiple windows, pull-down menus, scrolling pick-lists, and field-input techniques make the program intuitive to understand and easy to use. Daughter databases can be created to include all records of a particular type, or only those records matching user-specified search criteria. Separate databases can also be merged into a larger database. This computer program provides an easy-to-use and accurate means to organize, maintain, access, and share information about recombinant clones and other laboratory products of molecular biology technology.

Database Management Systems↗

Polymorphisms of the region of the Epstein-Barr virus genome which disrupts latency.

The nucleotide sequence of the portion of naturally occurring defective EBV (HR-1) DNA (designated het DNA) which is responsible for disruption of EBV latency has been determined and compared with the regions of the standard HR-1 viral genome from which it was derived. This rearranged 2.7-kbp DNA fragment represented an apparently nonhomologous recombination between sequences found in the BamHI W and BamHI Z fragments of the standard HR-1 genome. Only one intact open reading frame, comparable to standard HR-1 BZLF1, was present within this 2.7 kbp. No new open reading frames were created by the recombination. The BZLF1 sequence and predicted polypeptide products of standard HR-1 and het DNA were compared to B95-8 EBV. If an unspliced version of BZLF1 is used, the carboxy end of the BZLF1 polypeptides would differ considerably, principally due to an identical 28-bp insertion in both standard HR-1 and het BZLF1 relative to B95-8. However, if both virus strains use the same mRNA splicing strategy, the BZLF1 products from HR-1 and B95-8 would be similar, though distinguished by seventeen 1-bp differences which would result in nine amino acid changes. Both unspliced and spliced versions of het BZLF1 had five amino acid changes by comparison to standard HR-1 BZLF1. Differences in predicted secondary structure were found, consistent with dissimilar electrophoretic mobility of the polypeptide products. The amino acid differences between the BZLF1 polypeptide products of HR-1, het DNA, and B95-8 virus are all compatible with the creation of a protein which would function similarly to disrupt EBV latency. The differences in these polypeptides may account for some of the variation in the level of biologic activity of the BZLF1 products of different EBV strains. However, the major difference in activity between standard HR-1 and HR-1 het virus to disrupt EBV latency appears to be due to up-regulation of expression of het BZLF1 due to juxtaposition of BamHI W sequences upstream of het BZLF1.

Amino Acid Sequence↗

Two computer programs for rapid entry of DNA sequence data.

Two computer programs for the IBM personal computer are described for rapid and accurate entry of DNA sequence data. The DNA sequence files produced can be used directly by the DNA sequence manipulation programs by R. Staden (the DataBase system), the University of Wisconsin Genetics Computer Group, DNASTAR, or D.Mount. The first program, DIGISEQ, utilizes a sonic digitizer for semi-automation of sequence entry. To enter the DNA sequence each band of a gel reading is touched by the stylus of the sonic digitizer. DIGISEQ corrects for both changes in lane width and lane curvature. The algorithm is extremely efficient and rarely requires re-entering the centers of the lanes. The second program, TYPESEQ, uses only the keyboard for input. The keyboard is reconfigured to place nucleotides and ambiguity codes under the fingers of one hand, corresponding to the order of the nucleotides on the gel defined by the user. Both programs produce individual tones for each nucleotide, and certain ambiguity codes. This verifies input of the correct nucleotide or ambiguity code, and thus eliminates the need to visually check the screen display during sequence entry.

Base Sequence↗

Sequences of the Epstein-Barr Virus (EBV) large internal repeat form the center of a 16-kilobase-pair palindrome of EBV (P3HR-1) heterogeneous DNA.

We have previously characterized several genomic rearrangements of Epstein-Barr virus (EBV) DNA contained in one of the defective EBV genomes harbored by the P3HR-1 (HR-1) line (H. B. Jenson, M. S. Rabson, and G. Miller, J. Virol. 58:475-486, 1986). One recombinant clone of heterogeneous DNA (het DNA) from this defective genome is an EcoRI fragment of 16 kilobase pairs (kbp) which is a palindrome. DNA digestion fragments specific for the center of this palindrome were present in cells which contained het DNA but not in cells which lacked het DNA. Thus, the palindrome was not an artifact of DNA cloning. The organization of the center of this palindrome was studied by DNA sequencing. The comparable region of the parental HR-1 genome was also studied by DNA sequencing. The central 3,495 base pairs (bp) of the palindrome were composed of sequences derived exclusively from internal repeat 1 of EBV, represented by BamHI W fragment. At each end of the central 3,495 hp was a symmetrical recombination with sequences of BamHI-Z, located more than 50 kbp away on the standard EBV genome. The central 3,495 bp were composed of an unduplicated 341 bp flanked by two perfect palindromic repeats of 1,577 bp. The 341-bp unique region was a portion of a 387-bp region of standard HR-1 BamHI-W which was identical to the central 387 bp of the palindrome. This central 387-bp region contained numerous stretches of dyad symmetry capable of forming a large stem-and-loop structure. The palindromic rearrangement had created two novel open reading frames in het DNA derived from standard HR-1 BamHI-W sequences. These two het DNA open reading frames had different amino termini but identical carboxy termini derived from the large open reading frame in standard HR-1 BamHI-W (HR-1 BWRF1). The BamHI-W sequences found in het DNA did not include either the TATA box of standard HR-1 BamHI-W or the exons which are present in the potentially polycistronic latent mRNAs encoding EBV nuclear antigens. These marked alterations in genomic structure may relate to the unique biologic properties of virus stocks containing het DNA by creation of new polypeptides or by formation or deletion of regulatory or functional signals.

Base Sequence↗

Congenital cytomegalovirus infection with osteolytic lesions: use of DNA hybridization in diagnosis.

A case of congenital cytomegalovirus (CMV) infection with long-bone lesions is presented. The bone lesions consisted of broad regions of generalized osteopenia with irregular fragmentation and spiculization at the zone of provisional calcification. Diagnosis of CMV infection was made by DNA spot hybridization of the urine sediment to DNA from the Towne strain of CMV, demonstrating the usefulness of DNA hybridization for identification of CMV in clinical specimens. Bone lesions associated with congenital CMV infection are useful early clues to diagnosis but can be indistinguishable from those of congenital rubella syndrome.

Bone Diseases, Metabolic↗