Pathogenesis and treatment of human genital papillomavirus infections: a review.

HPV infection of the genital tract is common and anogenital warts or condyloma acuminatum is increasing rapidly in incidence. In addition, certain HPV types are closely associated with genital tract malignancies. Although recent advances in molecular biology have led to an increased understanding of the organization and functions of the papillomavirus genome, the pathogenesis of HPV infections and host responses to these diseases remain poorly understood. Treatment of anogenital warts is difficult and no completely satisfactory treatment modality is currently available. Comparatively few therapeutic modalities have been thoroughly evaluated, although recent studies of intralesionally and parenterally administered interferons have demonstrated beneficial effects of interferon compared to placebo. Additional studies of treatment for condyloma acuminatum are needed and should include the use of biologic response modifiers such as interferons, as well as antiviral drugs, with or without conventional methods of local therapy.

Vertebral osteomyelitis due to Staphylococcus warneri attributed to a Hickman catheter.

A patient with a long-term right atrial (Hickman) catheter developed vertebral osteomyelitis due to Staphylococcus warneri. Documentation of this event--to our knowledge previously unreported--was made possible by use of special studies including plasmid profiles of the coagulase-negative staphylococcal isolates.

Identification of human papillomavirus type 11 E4 gene products in human tissue implants from athymic mice.

A new method has recently been described for the growth of human papillomavirus type 11 (HPV11), an agent associated with genital warts, in human tissue xenografts implanted under the renal capsules of athymic nude mice (J. W. Kreider et al., 1986, J. Virol. 59, 369-376; 1987, J. Virol. 61, 590-593). With this model it is now possible to study productive HPV11 infection under controlled laboratory conditions. To identify proteins encoded by the HPV11 E4 open reading frame in infected implants, we have cloned an HPV11 E4 genomic DNA fragment representing all of the E4 region thought to be expressed in vivo, as evidenced by cDNA cloning and R loop mapping. The cloned HPV11 fragment was expressed in Escherichia coli as a cro-beta-galactosidase E4 fusion protein (Gal-E4 fusion). Rabbit antibodies raised against the Gal-E4 fusion protein were affinity purified using an HPV11 E1 E4 fusion protein. The E1--E4 protein was synthesized independently by expressing an HPV11 E1--E4 cDNA in E. coli using a second expression vector. Affinity-purified anti-E4 antibodies identified putative E4 proteins of 10 and 11 kDa in both the condylomatous cyst walls and in the desquamated cells in the cavities of HPV11-infected human skin implants from athymic mice. Similar proteins were not detected in uninfected controls. Implications for use of the athymic mouse system are discussed.

Detection of human papillomavirus types 6 and 11 E4 gene products in condylomata acuminatum.

Polyclonal antiserum to an Escherichia coli-produced beta-galactosidase/E4 fusion protein of human papillomavirus type 6b (antiserum 256), and affinity purified HPV 11 anti-E4 antibodies were tested for reactivity in Western blots with bacterially expressed trpE/E4 fusion proteins of HPV types 6b, 11, 16, and 18. To further characterize the affinity purified anti-E4 antibodies, a dot-immunobinding assay was performed using overlapping synthetic HPV 11 E1E4 peptides as antigens. Protein extracts of condylomata acuminatum from 18 patients containing HPV type 6 or 11 DNA sequences were tested in Western blots using antiserum 256 or affinity purified HPV 11 anti-E4 antibodies. In the Western blots of the trpE proteins, antiserum 256 identified the HPV types 6b and 11 fusion proteins; the affinity purified HPV 11 anti-E4 antibodies identified only the HPV 11 fusion protein. In the dot-immunobinding assay, three HPV 11 peptides were recognized, each containing a shared 8 amino acid sequence that differs significantly from the corresponding sequences of HPV types 6b, 16, or 18. In the Western blots of protein extracts from 18 condylomata acuminatum samples shown to contain HPV types 6 or 11 DNA, putative E4 gene products were identified in six samples by antiserum 256. The affinity purified HPV 11 anti-E4 antibodies identified putative E4 gene products in one of these same six lesions, which was shown to contain HPV 11 sequences by the Southern blot method. All six samples containing E4 gene products were from women. Three of these women were pregnant, one had serum antibodies to the human immunodeficiency virus, and one was a renal transplant recipient receiving glucocorticoids.(ABSTRACT TRUNCATED AT 400 WORDS)

Expression of the full-length products of the human papillomavirus type 6b (HPV-6b) and HPV-11 L2 open reading frames by recombinant baculovirus, and antigenic comparisons with HPV-11 whole virus particles.

The L2 open reading frames (ORFs) of human papillomavirus (HPV) types 6b and 11 were expressed as full-length non-fusion proteins in Spodoptera frugiperda (Sf-9) cells using recombinant baculovirus. Both proteins were detected on Western blots as immunoreactive bands which migrated with apparent Mrs of 76K and 78K, respectively, and contained both cross-reactive and type-specific epitopes, as determined by polyclonal antisera directed against defined subregions of the HPV-6b and HPV-11 L2 ORFs. In addition, the minor capsid protein of HPV-11 particles co-migrates with the HPV-11 L2 ORF product and is immunoreactive with HPV-11 L2-specific antisera. These observations indicate that the anomalous electrophoretic mobilities of papillomavirus L2 ORF proteins can be explained without invoking post-transcriptional processing events and that the minor capsid protein of HPV-11 is antigenically and biophysically related to the HPV-11 L2 ORF product.

Expression in Escherichia coli of seven DNA fragments comprising the complete L1 and L2 open reading frames of human papillomavirus type 6b and localization of the 'common antigen' region.

Molecular cloning was used to express human papillomavirus type 6b (HPV-6b) antigens in Escherichia coli. Seven genomic DNA fragments of HPV-6b which together comprise the complete L1 and L2 open reading frames, known to code for capsid proteins, were cloned and expressed in E. coli as both beta-galactosidase and TrpE fusion proteins. Western blots of HPV-6b beta-galactosidase fusion proteins using 'genus-specific' antisera produced by immunization of rabbits with disrupted bovine papillomavirus type 1 (BPV-1) showed that polypeptides encoded by two DNA fragments from the mid portion of L1 of HPV-6b were cross-reactive. Only one of these two polypeptides reacted with antisera raised against disrupted HPV-1, directly demonstrating that this polypeptide contains the papillomavirus 'common antigen'. The cross-reactive region was confirmed by reversing antigen and antibody. Polyclonal antisera were raised against the seven HPV-6b beta-galactosidase fusion proteins and tested against BPV-1 virion proteins on Western blots. Only antiserum against the mid portion of L1 of HPV-6b reacted with the BPV-1 major capsid protein. HPV-6b fusion proteins were also used to test human sera for antibodies reactive in Western blots. Serum samples from 38 patients with documented HPV-6 infections and from 22 presumably uninfected controls were tested. Antibodies were not detected in any of the sera to any of the seven fusion proteins. HPV-6b beta-galactosidase fusion proteins are antigenic and can be used on Western blots to localize immunologically reactive sub-regions of proteins by reacting protein fragments with antisera from immunized animals. However, alternative methods will be required to detect anti-HPV antibodies in human sera.

Randomised comparison of ceftriaxone and cefamandole therapy in lower respiratory tract infections in an elderly population.

Patients with pneumonia or bronchitis were randomized to receive ceftriaxone or cefamandole. A total of 30 of 38 patients were evaluable, 16 in the ceftriaxone group (average age 66.3 years) and 14 in the cefamandole group (average age 69.4 years). All but one had underlying diseases. Patients usually received 1 g of ceftriaxone intravenously every 12 h (mean duration 8.7 days) or 1.5 g of cefamandole intravenously every 6 h (mean duration 8.2 days). Adverse experiences attributable to the drugs were confined to one episode of discomfort at the infusion site in each group. Bacteriological results with ceftriaxone were 83% cured, 11% superinfected after eradication of pretherapy isolate, and 6% failed. Bacteriological results with cefamandole were 76% cured, 24% failed. Clinical results with ceftriaxone were 38% cured, 56% improved, 6% failed. Clinical results with cefamandole were 57% cured, 21% improved, 21% failed. Emergence of a resistant Serratia marcescens was seen in a ceftriaxone-treated patient. Disc diffusion susceptibility testing identified six of the seven pretherapy nonfastidious Gram-negative isolates as susceptible; however, two of the six could not be eradicated with the assigned drug and another two were eradicated with ensuing super-infection with susceptible isolates of Pseudomonas aeruginosa. In contrast, MBCs were an accurate guide to clinical outcome with nonfastidious Gram-negative bacilli.

Resistance to cefamandole: a collaborative study of emerging clinical problems.

Cefamandole resistance in five patients was studied. Microorganisms emerged resistant to cefamandole during therapy with the drug in three patients with complicated infections. This resistance was associated with an enhanced production of beta-lactamase and/or with a change in the substrates and the isoelectric focusing patterns of the enzymes. Cross-resistance to other beta-lactam antibiotics developed concurrently in isolates from these patients. Disk diffusion tests did not detect resistance to cefamandole in the pretreatment isolate from the fourth patient; this isolate produced inactivating enzymes, and resistance was detected only in broth dilution tests. In the fifth patient, infection with a cefamandole-resistant Enterobacter developed during postoperative therapy with the drug. Resistance to cefamandole in the isolate from this patient was unstable and was associated with inducible beta-lactamase activity. These examples emphasize the need for close monitoring of patients who are given cefamandole and for thorough in vitro evaluation of isolates from the patients both before and after treatment.