Robert W. Atchison

Adenovirus-Associated Defective Virus Particles

Small, DNA-containing particles were separated from preparations of a simian adenovirus. These particles differed antigenically from the adenovirus. Replication of the particles in cell cultures was obtained only when theywere inoculated simultaneously with adenoviruses. This suggests that these adenovirus-associated particles behave as defective viruses.

Studies on the relationship between adeno-associated virus type I (AAV-1) and adenoviruses. I. Replication of AAV-1 in certain cell cultures and its effect on helper adenovirus.

Abstract AAV type 1 has been shown to replicate in several species of cell cultures with the aid of certain helper adenoviruses from man, monkey, mouse, and dog. However, during the course of AAV replication the yield of the helper adenovirus was decreased. The reduction in yield of adenovirus was directly related to the quantity of AAV present and inversely- to the dose of the adenovirus challenge. The decrease in adenovirus yield was shown to be due to a reduction in the number of cells yielding virus.

Studies on the relationship between adeno-associated virus type 1 (AAV-1) and adenoviruses. II. Inhibition of adenovirus plaques by AAV; its nature and specificity.

Abstract Plaque formation by simian adenoviruses, SV15 and SV34, and a simian papovavirus, SV40, was prevented in cell cultures treated with AAV. Specific antiserum to AAV inhibited this effect and thus provides a quantitative means for measuring AAV neutralizing antibody. The nature of the adenovirus inhibition does not suggest an interferon-like mechanism, but suggests an interference mediated directly by the AAV particle. This phenomenon, of adenovirus plaque inhibition, has been utilized in an attempt to determine the minimum number of interfering AAV particles present in a sample.

The role of herpesviruses in adenovirus-associated virus replication in vitro

Abstract Herpes simplex virus (HSV) and infectious bovine rhinotracheitis virus (IBR) were shown to be helper viruses for partial replication of defective adenovirus-associated viruses (AAVs) in cell cultures. The specificity of the helper activity was established using antiserum and/or ether treated HSV in helper experiments with AAV-1. Newly synthesized AAV antigen(s), presumed to be virus coat protein(s), was observed by fluorescent antibody staining, but no production of infectious AAV progeny particles could be shown. Likewise, no AAV particles were visible in cell culture harvests examined by electron microscopy. The evidence suggested a lack of AAV particle production under these circumstances, and a different or more limited type of helper function(s) from herpesviruses as compared to adenoviruses. Other DNA- and RNA-containing animal viruses that were tested were found to be lacking any helper activity.

Evaluation of Chlamydia pneumoniae and Mycoplasma pneumoniae as Etiologic Agents of Persistent Cough in Adolescents and Adults

ABSTRACT Chlamydia pneumoniae and Mycoplasma pneumoniae were evaluated as agents of persistent cough in adolescents and adults (n = 491). Tests of 473 respiratory specimens by culture or PCR or both identified four episodes (0.8%) of M. pneumoniae-associated illness and no episodes of C. pneumoniae illness, suggesting that these bacteria do not frequently cause persistent cough.

The development of type 1 (insulin-dependent) diabetes mellitus: Two contrasting presentations

SummaryGenetic, immunological and viral factors have been implicated in pathogenesis of Type 1 diabetes mellitus. The development of Type 1 diabetes in two siblings of patients with Type 1 diabetes studied as part of a large epidemiological study, is described. One case, a 13-year-old male not sharing either HLA haplotype with his diabetic sister, had virtually normal glucose tolerance 80 days before symptomatic presentation. He showed serological evidence of infection by Coxsackie CB4 (at diagnosis) and influenza A virus (soon after diagnosis). The other case, a 15-year-old male, had impaired glucose tolerance for over 500 days (i. e., since the diagnosis of diabetes in his HLA-identical brother) before symptomatic presentation which was not associated with serological evidence of acute viral infection. The former case was negative for islet cell antibody (cytoplasmic) when first seen though positive at diagnosis, while the latter was positive throughout. These two cases suggest contrasting interactions of the main pathogenetic factors associated with Type 1 diabetes.

Attempted Type Specific Diagnosis of Dengue Virus Infection by the Indirect Fluorescent Antibody Method Directed at Differentiating IgM and IgG Responses

The need for paired acute and convalescent phase sera for diagnostic purposes in virology has long been recognized. Along these lines, some important problems are: (a) getting an early enough acute phase serum, (b) obtaining only a convalescent phase serum perhaps none at all, (c) choice of most suitable test method (s) to use with likely small quantities of specimen(s) obtained and (d) perhaps most important of all, furnishing a diagnostically useful result to the attending physician at the time he needs it. This latter requirement is incompatable with awaiting a convalescent phase serum. Furthermore, unless the physicians early needs are met, his subsequent lack of cooperation further complicates obtaining the needed sera from the next patient. The solution in so far as serology is concerned apparently lies in the detection of early specific antibody in the IgM globulin fraction and distinguishing this from that of any preexisting antibody in the IgG globulin which may also react with the antigen used. Since fractionation procedures to separate IgG or selectively inactivate IgM are time consuming or impractical in large scale routine diagnostic work, the indirect fluorescent antibody technique employing highly specific conjugates against human IgM and human IgG appears to be most desirable. Diagnostic use of such a procedure in virology was introduced by Brown and his associates in 1968 (1, 2) for mumps, but to our knowledge this has not been applied in diagnostic arbovirology prior to the studies reported here. The major interest in this laboratory is in the area of the arboviruses and currently in the dengue (DEN) subgroup of group B most specifically, therefore these viruses were employed. Another reason for selecting group B and the DEN subgroup relates to the lack of type specificity, using current serological tests, of sera of persons having a second or subsequent infection with a group B virus.

VIROLOGIC, IMMUNOLOGIC, AND EPIDEMIOLOGIC ASSOCIATIONS WITH AIDS AMONG GAY MALES IN A LOW INCIDENCE AREA a

The Pittsburgh tri-state area is presently a low incidence area for AIDS and AIDSrelated complex (ARC). There are only six confirmed cases of full-blown AIDS using the Centers for Disease Control criteria. The first case occurred in 1980, the second appeared in 1982, and four were discovered in 1983, which is two cases per million in the Pittsburgh area. This is lower than the 5 to over 30 cases per million noted in such cities as Los Angeles, San Francisco, Miami, Houston, and New York. We estimate that there are only about 20 known cases of ARC in Pittsburgh. We have confirmed 10 of these in our study using the definition of noncontiguous, extrainguinal lymphadenopathy persistent for more than three months, without any association with known, non-AIDS-related causes. All cases of AIDS and ARC have been in non-Haitian gay males with no history of intravenous drug abuse. We began our pilot study earlier this year to examine the basis for this low incidence of AIDS. There are two major hypotheses that could explain the low incidence. First, the putative infectious agent or agents causing AIDS and ARC may not be widely introduced into our gay population. Second, and not necessarily exclusive from the first, is that various lifestyle, virologic, and immunologic characteristics that have been associated with AIDS differ between gay males in Pittsburgh and those in high incidence areas. Our study population consisted of 57 gay male volunteers who were healthy by physical exam and history. These subjects were all white and were predominantly in their 20s and 30s. The healthy subjects began having gay sex in their late teens. They were relatively sexually active as evidenced by an average of over 30 different sex partners in the year preceding our study. About one-third of our subjects participated in sex at gay baths. Those studied were also sexually active as shown by a relatively high level of self-reported episodes of sexually transmitted diseases (mean of 3) and history of hepatitis (26%). Over 60% participated as the passive recipient in anal intercourse.

The Changes in Fluorescence of Acridine Orange-Stained Hela Cells During Ultraviolet Illumination

HeLa cells were stained with a 1/12,000 concentration of acridine orange at pH 7.2 for 3 min and the fluorescence emission was measured quantitatively for effects of ultraviolet illumination with durations including intervals between 5 and 210 min. The total photometric fluorescence intensity increased for the first 30 min, then decreased with illumination time. The initial maximum fluorescence intensity occurred at 525 nm and shifted progressively to shorter wavelengths. Fluorescence intensity above 580 nm decreased with increasing duration of illumination time while that below 580 nm showed an initial increase in intensity followed by a gradual fading.

Small cell lung cancer antigen expression differs on ‘classic’ and ‘variant’ SCLC and carcinoid cells

Abstract The panel of 98 monoclonal antibodies (MOABS), (mouse or rat) provided by the Second International Workshop on Small Cell Lung Cancer Antigens was tested by indirect immunofluorescence and flow cytometry using prototype cell lines [NCI-H69 (SCLC ‘classic’), NCI-N417 (SCLC ‘variant’), NCI-H727 (carcinoid) [A.F. Gazdar et al. (1985, 1988)] peripheral blood lymphocytes (PBL), and sea urchin coelomocytes [Koros, A.M.C. et al. (1987, 1988, 1989).] All cells had some reactivity with some of the MOABS. There is however, differential expression of antigens amongst the prototype cell lines, which may provide a useful method for phenotyping human lung cancers. More MOABS react with the SCLC ‘classic’ NCI-H69 than with any of the other cell types (MOABS, 5, 12, 16, 18, 21, 28, 31, 34, 36, 41, 45, 47, 48, 53, 56, 58, 69, 70, 71, 79, 87, 91). MOABS 45-49 seem most useful in distinguishing between SCLC and carcinoid cultured cell lines. Those MOABS which do not react with normal PBL (MOABS 3, 5 and 12 especially) should be tested further for possible utility as therapeutic agents in patients whose tumours have escaped conventional therapy. In contrast to our earlier observations of neuroendocrine markers (CD56, CD57) on SCLC, neuroendocrine cells, human natural killer cells, and sea urchin coelomocytes, few (14/50) antigens identified by the present panel appear to be evolutionarily conserved.