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Close association between shape alteration and loss of immunity to superinfection in a wild-type Klebsiella pneumoniae stable lysogen which can be both immune and nonimmune to superinfection.

Klebsiella pneumoniae MirM7 is a wild-type strain which grows as cocci at pH 7 and above and as rods at pH 6.5 and below. Cultures of this strain and an auxotrophic derivative, MirM7b, have been found to undergo spontaneous lysis after purification from possible contaminating viruses. Lysates always contained two phages, FR2 and AP3, most often at high titers. FR2 and AP3 plated with the same efficiency on both MirM7b and K59 (another K. pneumoniae strain sensitive to FR2 and AP3) and lysogenized 45 and 54% of the K59-infected cells, respectively. These findings raise the possibility that MirM7b is lysogenic for FR2 and AP3, although nonimmune to their superinfection. The fact that mitomycin C and N-methyl-N'-nitro-N-nitrosoguanidine can induce phages FR2 and AP3 from MirM7b confirmed this possibility. When MirM7b was infected with FR2 several strains immune to FR2 and AP3, which were all rod shaped, were obtained. Furthermore, 19 derivatives, rod shaped at all pH's have been isolated from MirM7b. They were all immune to both FR2 and AP3. From mating experiments between the MirM7b donor derivative, strain M720, and either K59 or MirCV5, a rod-shaped MirM7b derivative cured from the prophages, cysteine recombinants were obtained which were most often (80%) immune to FR2 and AP3. Nonimmune and still lysogenic recombinants were obtained by mating M720 with a rod-shaped immune MirM7b derivative; the majority of the non-immune strains maintained the rod shape. Five coccus-shaped recombinants were also isolated; they were nonimmune to superinfection. Several physiological properties of strain MirM7b and the other nonimmune coccal recombinants have been studied in comparison with those of the rod-shaped immune derivatives. All of the coccal strains have shown several alterations with respect to the rods. The role of possible derepressed prophage genes in the various physiological alterations of MirM7 is discussed, and the analogies between this system and those of vertebrate cells transformed by proviruses are stressed.

Bacteriophages

Superinfection exclusion and changes in cellular transport processes in phage infected Salmonella typhimurium.

The changes induced by bacteriophage P22 in the cellular transport process(es) of the host Salmonella typhimurium (Taneja et al., 1975; Khandekar et al., 1975; Bandyopadhyay and Chakravorty, 1976) involve interactions between the superinfection exclusion system of the resident prophage and the C immunity region of the superinfecting phage. The sie A gene of the prophage interferes with the changes in the cellular transport process induced by the superinfecting phage. However, if the superinfecting phage carries active C1 and C2 genes there is no such interference. Thus the C1 and C2 genes of the superinfecting phage seem to be expressed in the sieA+ lysogen.

Biological Transport

Bacteremic superinfections of patients with bacteremia: occurrence, bacteriology, mortality, and duration of hospitalization at Boston City Hospital during 12 selected years between 1935 and 1972.

During the course of hospitalization of 6,414 patients with bacteremic infections identified at Boston City Hospital (Boston, Massachusetts) during 12 selected years between 1935 and 1972, 6.0% developed verified bacteremic superinfections with organisms not isolated or identified in the primary bacteremic infection. Analysis of these cases reemphasizes the increasing occurrence of serious hospital-acquired infections despite the successive introduction and intensive use of a large number of effective antibacterial agents. The bacteremic superinfections, like the primary hospital-acquired bacteremias, increased in incidence over the years, particularly since 1961; they were more frequent and were associated with a higher mortality rate and longer duration of hospital stay in the primary hospital-acquired cases than in the patients in whom bacteremia was considered to be community-acquired. The organisms in superinfections were similar to those in primary hospital-acquired bacteremias. Superinfection was more frequent among the patients who died than among the survivors.

Bacterial Infections

Mechanisms of infection with Epstein-Barr virus. I. Viral DNA replication and formation of noninfectious virus particles in superinfected Raji cells.

Human lymphoblastoid Raji cells, which do not produce virus, supported replication of Epstein-Barr virus (EBV) upon superinfection. Early antigen, viral capsid antigen, and virions were produced in Raji cells superinfected with EBV. Viral DNA replicated under complete inhibition of host cell DNA synthesis to the extent that a few micrograms of EBV DNA were recovered from 107 superinfected Raji cells, corresponding to 5,000 viral genomes/cell. Homology of the synthesized viral DNA to parental EBV DNA was more than 90%. Virions produced by the Raji cells contained a 55S DNA but failed to induce early antigen, viral capsid antigen, and viral DNA synthesis after a second superinfection of Raji cells.

Antigens, Viral

Mouse natural killer (NK) cell activity against human cell lines is not influenced by superinfection of the target cell with xenotropic murine C-type virus.

Mouse natural killer (NK) cells can lyse a variety of syngeneic, allogeneic and xenogeneic target cells in short-term 51Cr release assays. The target specificity of NK cells is not known, but endogenous C-type viral antigens have been suggested as possible target structures. To test this hypothesis, human lymphoid lines were superinfected with xenotropic mouse C-type virus either by repeated dosage through nude mice or by in vitro superinfection with the supernatants of nude-mouse-passaged lines. The appearance of surface-associated MuLV antigens after superinfection was confirmed in a complement-dependent cytotoxicity test. Subsequently, the NK sensitivity of each infected line was compared with its non-infected counterpart in direct cytolytic and competition assays. None of these two assay systems showed a consistent difference in NK sensitivity of infected and non-infected cell lines. These findings do not lend support to the concept tht murine C-type viral antigens are responsible for NK sensitivity.

Animals

Recovery of transforming EBV from non-producer cells after superinfection with non-transforming P3HR-1 EBV.

Cells of the Raji and NC37 lines can be induced by chemical inducers, such as BrdUrd and IdUrd, or the tumor-promoter TPA to EA-expression only, but do not reveal any VCA synthesis. After superinfection by nontransforming P3HR-1 EBV, however, a varying percentage of the cell population shows VCA synthesis and releases infectious viral particles. The recovered virus differs biologically from P3HR-1 EBV since it transforms human umbilical cord blood lymphocytes into EBNA-positive lymphoblastoid cell lines. Cells of these established lines are susceptible to renewed infection by P3HR-1 EBV which results in EA induction and VCA synthesis. Only cells of one line, NC37-R1, spontaneously produce VCA and EBV particles, which reveal transforming properties and do not induce EA upon superinfection of Raji cells. Infection of P3HR-1 EBV-converted BJA-B cells also leads to EA and VCA induction and the release of viral particles. In contrast to particles recovered from Raji and NC37 cells, no transforming activity was detectable in these virus preparations. According to these data, we propose that viral genomes persisting within Raji and NC37 cells are defective and become complemented by the superinfecting P3HR-1 virus.

Antigens, Viral

Transforming activity of Epstein-Barr virus obtained by superinfection of Raji cells.

Epstein-Barr virus obtained by superinfection of Raji cells with Epstein-Barr virus recovered from P3HR1 cells (HRI virus) transformed human lymphocytes, but it did not superinfect Raji cells. A human lymphoblastoid cell line, HLB, established by such transformation contained 22 Epstein-Barr virus genomes per cell and Epstein-Barr virus-associated nuclear antigen, and a few cells contained early or viral capsid antigen complexes. Chromosomal analysis revealed that HLB-cells were diploid with normal female karyotypes. Replication of Epstein-Barr virus DNA and inhibition of host cell DNA synthesis were observed in HLB cells after superinfection with HR1 virus.

Antigens, Viral

Selective resistance to togaviral superinfection in mice with tolerant lymphocytic choriomeningitis virus infection.

Mice infected neonatally with lymphocytic choriomeningitis virus (LCMV) developed partial and complete resitance to cerebral superinfection with tick-borne encephalitis virus (TEV) in 10 and 20 days after birth, respectively. This resistance lasted at least till the age of 40 days. LCMV tolerant mice neither succumbed to TEV infection, nor circulated TEV in their blood. Moderate, gradually decreasing TEV titres were detected in the brains and TEV-induced brain interferon was lower than in control mice of the same age. TEV superinfection caused a significant depression of the blood titre of tolerated LCMV while the titres in the brains remained equal to those in tolerant but not superinfected mice. LCMV tolerant mice showed a similar resistance to another togavirus (chikungunya) but not to encephalitogenic picorna-, herpes- and rhabdoviruses.

Animals

Somatic cell hybrids between human lymphoma cell lines. V. IdUrd inducibility and P3HR-1 superinfectability of Daudi/HeLa (DAD) and Daudi/P3HR-1 (DIP-1) cell lines.

We have studied two types of somatic cell hybrid with regard to expression of the Epstein-Barr virus (EBV) cycle and its regulation. The first, DIP-1, a hybrid formed between two human lymphoma EBV producers (Daudi and P3HR-1), contained EBV DNA, expressed the virus-determined nuclear antigen (EBNA), andwas a producer of the EBV-associated antigens EA (early antigen) and VCA (viral capsid antigen). The second, DAD, a hybrid series of clones formed between Daudi and a HeLa cell derivative (D98), differed with regard to the expression of EBNA, EA, VCA and the content of EBV DNA. EA was regularly induced in the EBV DNA-containing hybrids following treatment with iododeoxyuridine (IdUrd). This induction was greater in lines spontaneously expressing EA. In two hybrids, DIP-1 and DAD10, VCA and virus DNA synthesis were also induced in the presence of IdUrd, the latter being detected by in situ hybridization with P3HR-1 EBV complementary RNA. Finally, while DIP-1 was superinfectable by the P3HR-1 EBV strain, the DAD series of hybrids were refractory to P3HR-1 superinfection and lacked EBV receptors.

Antigens, Viral

A virus-specified mechanism for the prevention of multiple infection--T7- and T3-mutual and superinfection exclusion.

Co- and superinfection of cells with T3/T7 result in exclusion (mutual or superinfection exclusion). The exclusion mechanism is also directed against homologous (or identical) virus. Exclusion is established after the adsorption but before the genome becomes available for gene expression or replication, that is only one virus per cell develops. The exclusion is triggered by a constituant of the viral particle. An early T7 gene (M gene) (Schweiger et al., 1975) is essential for the formation of exclusion competent virions.

Bacterial Proteins

Superinfection epithelial nasopharyngeal carcinoma cells with Epstein-Barr virus.

Attempts were made to superinfect epithelial explant cell cultures prepared from nasopharyngeal carcinomas with Epstein-Barr virus. Virus-specific markers were observed in such cultures 3 days after superinfection. In addition, expression of Epstein-Barr virus early antigens was observed in epithelial cell explant cultures treated with iododeoxyuridine. The results suggest that epithelial cells of at least certain nasopharyngeal carcinomas possess the receptor for the Epstein-Barr virus and that the latent virus genome can be induced from epithelial cells prepared from such tumors.

Antigens, Viral

Protective effect of murine sarcoma virus-superinfected mouse tumor cells against outgrowth of corresponding noninfected tumor.

Cultured lines of three newly established methylcholanthrene-induced tumors, MBK and MBL in CBA and MC57M in C57BL mice, and two mammary tumors, SBfnHC in CBA and S3W in ASW mice, were superinfected in vitro with Moloney sarcoma or leukemia virus (MSV, MLV). After superinfection, they expressed the Moloney virus-determined cell surface antigen (MCSA) and murine C-type viral p30 antigen, and produced NB-tropic C-type virus. The virus-infected tumors became more rejectable in normally susceptible syngeneic mice compared with the original noninfected line. There was no difference in 400-rad irradiated hosts. Mice that have rejected the virus-infected tumors showed an increased resistance to the corresponding noninfected tumor. The protective effect was comparable, with only one exception, to the immunizing effect of irradiated, noninfected cells. In vitro tests showed that small numbers of viable MSV-infected MBL generated cytotoxic spleen cells against both uninfected and MSV-infected MBL in syngeneic mice, while the same numbers of viable noninfected MBL did not induce cytotoxic cells. Relatively large numbers of irradiated MBL and MSV-MBL had a similar activity in inducing cytotoxic spleen cells against MBL in syngeneic hosts.

Animals

Suitability of Epstein-Barr virus DNA obtained from superinfected Raji cells for complementary RNA hybridization studies.

A method was established to obtain a high yield of Epstein-Barr virus (EBV) DNA for nucleic acid hybridization studies on latent virus DNA in transformed cells. Superinfection of Raji cells with EBV concentrated from HRI cell cultures produced a 600-fold higher yield of EBV DNA than direct isolation of EBV from HRI cell cultures. The virus DNA thus prepared from Raji cells superinfected with EBV was radioisotopically and spectrophotometrically pure and served as a satisfactory template for the preparation of cRNA specific to EBV DNA.

Burkitt Lymphoma

Appearance of early and late components of Epstein-Barr virus-associated membrane antigen in Daudi cells superinfected with P3HR-1 virus.

This study investigated the synthesis of membrane antigen (MA) as well as virus capsid antigen (VCA) and early antigen (EA) in Daudi cells which had been superinfected with the P3HR-1 strain of Epstein-Barr virus (EBV) and then treated with trypsin to remove initially absorbed MA-positive material from the cell surface. Synthesis of MA, VCA and EA was completely inhibited by puromycin. A marked reduction in the frequency of MA positive cells was observed in superinfected cells cultured in the presence of either cytosine arabinoside (Ara-C) or phosphonoacetate (PA); however, a small fraction of MA synthesis occurred, suggesting an inhibitor insensitive component in MA, A differential absorption of EBV antibody-positive human serum with the Ara-C treated or untreated infected cells detected two antigenically different components in MA: early (Ara-C insensitive) and late (Ara-C sensitive) MA.

Antigens, Viral

Prophage substitution and prophage loss from superinfected Escherichia coli recA(P1) lysogens.

It is shown that the plasmid prophage P1 can be displaced by a superinfecting P1 phage in Escherichia coli recA(P1) lysogens. Six widely separated phage markers were used to distinguish between residual recombination and total substitution. It is further shown that superinfection of recA lysogens can lead to loss of both phage (curing). These two phenomena, previously reported in Rec+ strains, are thus independent of host recombination and may result from perturbations of some function involved in plasmid maintenance.

Chloramphenicol

Differential expression of D and R components of Epstein-Barr virus early antigen after superinfection and after induction with 5-iododeoxyuridine.

Three non-productive human lymphoblastoid cell lines, NC-37, RAJI and L-1, either superinfected with Epstein-Barr (EB) virus from P3HR-1 cells or treated with 5-iododexyuridine (IUDR), were studied for the development of early antigen (EA) of EB virus. In all three superinfected cell lines, both the restricted (R) and diffuse (D) componets of EA were synthesized, with R-component formation preceding D-component synthesis. After IUDR treatment both components of EA were only formed in RAJI cells, while in NC37 and L-1 cells only the R-component was synthesized.

Antigens, Viral

Cell surface markers on epithelial-Burkitt hybrid cells superinfected with Epstein-Barr virus.

Attempts were made to superinfect two epithelial-Burkitt hybrid cell lines, designated D98/HR-1 and D98/Raji, with Epstein-Barr virus (EBV) and to investigate the expression of some cell surfacr markers including histocompatibility antigens, and the presence of B-cell markers, such as receptors for the third complement component and for monkey red blood cells. Successful superinfection of D98/HR-1 cells with EBV was made evident by the expression of early antigen and, to a lesser extent, virus capsid antigen. Only a rare D98/Raji cell was found to be positive for early antigen. The histocompatibility antigens of the parental cell lines D98, HR-1, and Raji were expressed on the surfaces of the hybrid cells. Receptors for third complement components b and d were not detected on the hybrid cells or on the D98P OR HR-1 cell lines; they were found, however, on the Raji cells, indicating that EBV receptors and complement receptors can be separated. The significance of the infection of the hybrid cells with EBV and the expression of cell surface markers is described.

Antigens, Viral

[Echinococcus multilocularis: superinfections in experimentally infected mice (author's transl)].

The influence of subcutaneous superinfections following an intraperitoneal primary infection after 4 and 14 weeks respectively has been examined in Echinococcus multiolcularis infected NMRI-mice. It could be shown that the growth of subcutaneous echinococci is inhibited in superinfected mice (372 mg) if compared with control mice (903 mg parasite weight). Other differences (composition of serum proteins and white blood cells, hemoglobin content) are exclusively due to the primary intraperitoneal infection.

Animals