Biomedical subjects
Publications and source records attributed to C Cermelli.
The association of a human herpesvirus 6 primary infection with a fatal case of hemophagocytic syndrome in a 3 month old baby is described. The trigger mechanism of the virus for the clinical syndrome is discussed.
HHV-7 growth on Sup-T1, an immature T-cell line, was studied using different HHV-7 isolates obtained in our laboratory. Titration of viral yields showed that all the virus isolates propagate on this cell line more efficiently than in cord blood lymphocytes, the cells usually recommended for HHV-7 growth. The permissivity of Sup-T1 to HHV-6, whose ability to replicate in these cells was still unknown, was also investigated using two virus isolates representative of variants A and B respectively. Both isolates were able to propagate on Sup-T1 and viral titres were similar to those obtained in cord blood lymphocytes. As the efficient propagation of both HHV-7 and HHV-6 isolates in Sup-T1 cultures, these cells may replace more time consuming and expensive cord blood lymphocyte preparations for the propagation of both the viruses.
Virus isolation and viral DNA detection by the polymerase chain reaction were used to investigate the presence of human herpesvirus 6 (HHV-6) and human cytomegalovirus (HCMV) in bronchoalveolar lavage from 34 human immunodeficiency virus-1 (HIV-1)-infected patients with respiratory disorders. The aim was to assess the presence of reactivated HHV-6 in lung tissues for a subsequent evaluation of the frequency of virus involvement in respiratory clinical manifestations in the course of HIV-1 infection. Bronchoalveolar lavage samples were tested for the presence of HCMV, as a routine investigation within a protocol monitoring opportunistic infections in symptomatic HIV-1 patients. Whereas HCMV DNA was detected by the polymerase chain reaction in 12 bronchoalveolar lavage specimens, 10 of which were also positive for virus isolation, all samples were negative for HHV-6 by both virological procedures. The HHV-6 DNA finding in bronchoalveolar lavage from an HIV-1-seronegative patient with renal carcinoma, investigated accidentally together with the bronchoalveolar lavage specimens from HIV-1 seropositive patients, stressed the HHV-6 polymerase chain reaction-negative results in the bronchoalveolar lavage samples under study. It is concluded that the lung may be a target organ for HCMV infection in HIV-1-seropositive patients affected by respiratory symptoms but that this does not seem to be the case for HHV-6.
HepG2 cells, a well differentiated liver cell line, were shown to be permissive for both human herpesvirus 6 (HHV-6) A and B strains by three independent methods of analysis: detection of viral antigens, viral DNA sequences and infectious virus. HepG2 cell infection with HHV-6 resulted in functional damage as shown by the increased release in the culture medium of some hepatocyte markers. Cells surviving the acute infection were serially passaged without showing cytopathic effect, but, some months later, HHV-6 DNA was still present in the cells and virus induction with a phorbol ester was successful. A possible pathogenetic role of HHV-6 in liver diseases is discussed. Experiments of HepG2 infection with human herpesvirus 7 (HHV-7) were also carried out. The lack of an efficient virus replication suggested a difficulty for HHV-7 to infect hepatic cells.
Sera from a sample of children aged 3 months to 6 years and from cord blood were tested in an indirect immunofluorescence assay (IFA) for reactivity to human herpesviruses 6 (HHV-6) and 7 (HHV-7). HHV-6 seropositivity values rose from 19% to 79.3% in the first 18 months of life, while HHV-7 seroprevalence reached a similar value (75.9%) in children aged 3-6 years. These results show that HHV-7, like HHV-6, is a prevalent virus in infancy. In cord blood sera, assayed to study infant humoral situation at birth, similar values for the two viruses (78.9% for HHV-6 and 76.3% for HHV-7) were found. HHV-6 and HHV-7 IgG antibody affinity to the corresponding antigens was assessed by the end point antibody titration in the presence and absence of urea 8M. This test distinguishes antibodies of recent (low affinity) or past (high affinity) production. Together, the data on seroprevalence and antibody affinity suggest that HHV-6 primary infection generally precedes that by HHV-7. These results are discussed in the light of a different pathogenetic role of the two viruses.
A viral isolate obtained from peripheral blood lymphocytes of an infant with a nonspecific febrile syndrome was identified as human herpesvirus 7 (HHV-7) on the basis of PCR analysis of its DNA with one set of primers specific for HHV-7. The correlation of HHV-7 with the febrile episode affecting the infant is suggested.
The antibody content to HIV-1 p24 Ag expressed as relative binding capacity to the target antigen (p24 RBC) was retrospectively quantified in serum samples from 20 HIV-1-uninfected infants born to HIV-1 seropositive mothers. p24 RBC values quantified at birth were included either in a low (0-20%) or high (80-100%) range of values, classified as group A (11 infants) and group B (9 infants), respectively. The course of maternal antibodies to HIV-1 antigens p17, p24, p31, gp41, p51, p66, gp120, and gp160 was studied in each group. A substantial difference in the amount and subsequently in the decline of maternal antibodies to gag proteins p17, p24, and p55 and to pol proteins p51 and p66 was observed in the two infant groups in contrast with a similar content and decline of the remaining antibodies. In 7 HIV-1-infected infants of whom 4 resembled infant group A and 3 infant group B for p24 RBC values, a relationship appeared between p24 antibody decline and p24 antigenemia detection.
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Papanicolaou (Pap)-stained cervical specimens from 160 squamous lesions were processed for the detection of human papillomavirus (HPV) DNA by an in situ hybridization (ISH) assay. Three biotinylated HPV DNA probes were employed, each containing HPV genotypes 6/11, HPV genotypes 16/18, or HPV genotypes 31/35/51. The HPV etiology of 86 lesions was ascertained (53.8%). In 74 out of 135 (58.8%) HPV-typed low-grade squamous intraepithelial lesions (SILs), HPV 6/11 was found in nine (6.6%), HPV 16/18 in 46 (34.2%), and HPV 31/35/51 in 19 lesions (14.1%); in 11 out of 18 HPV-typed high-grade SILs (61.1%), seven lesions (38.9%) were typed for HPV 16/18 and four (22.2%) for HPV 31/35/51. Of seven invasive carcinomas, only one (14.3%) reacted with the HPV 16/18 DNA probe. A cohort of 124 low-grade SILs were followed cytologically for a year. The results of this study are discussed in light of HPV type association and therapy.
The surface antigenic make-up of 10 HHV-6 isolates from cases of infantile disease was analyzed by immunofluorescence assay with 5 monoclonal antibodies (Mabs). Three Mabs were directed to glycoproteins (gps) expressed in cells infected with the HHV-6 variant A prototype GS, and 2 to gps expressed in cells infected with the HHV-6 variant B prototype Z29. Of the 10 viral isolates, all belonging to HHV-6 variant B, 9 showed a similar Mab reactivity, while 1, from a case of gastroenteric illness, differed widely from the others. The HHV-6 isolates studied also differed from the variant B prototype Z29 for the absence of reactivity to one of the Mabs. The choice of the HHV-6 new isolates as variant B prototypes is recommended.
546 human sera from healthy subjects, subjects with cytomegalovirus or Epstein-Barr virus infection and from cordal blood were tested for the human herpesvirus-6 (HHV-6) IgG content in an enzyme-linked immunosorbent assay (ELISA) to verify results previously obtained with an immunofluorescence assay (IFA). The HHV-6 isolate CV, obtained from a child with exanthem subitum, employed as antigen in both tests, belonged to HHV-6 variant B. The ELISA results were similar to those obtained with IFA. 445/546 (81.5%) sera had the same type of reactivity with the two serological procedures. To establish whether different HHV-6 strains may give different serological results, 117 out of the 546 sera tested with IFA against CV infected cells were also tested against cells infected with the HHV-6 isolate U1102 belonging to variant A. All the sera showed a similar reactivity.
Forty-five sera from men with bladder cancer were examined in a micro solid-phase enzyme-linked immunosorbent assay (ELISA) and in a Western-blotting (WB) assay for the presence of IgG antibodies to papillomavirus (PV) genus-antigens of bovine origin. The ELISA detected PV antibodies in 75.6% of cancer patients. This antibody frequency was significantly higher than that found in both healthy males (22.7%) and patients with urological disorders (24%). A similar correlation among the PV antibody frequencies of the three groups was found with WB assay: 60% of the neoplastic group showed PV antibodies versus 17.3% in healthy males and 32.6% in non-neoplastic patients. Within the same group, 78% to 87% sera showed the same reactivity to both assays. Of these concordant sera, PV positive sera were 55.6% in cancer patients, 13.3% in healthy adults and 19.6% in patients with urological disorders. ELISA PV antibody level in the cancer group was higher than in each of the two control groups. The meaning of the humoral response to PV genus-antigens in men with bladder cancer is discussed.
Virological studies were carried out on 3 to 36-month-old patients admitted to the Children's Hospital of the University of Modena with febrile syndrome from September 1990 to February 1991. Virological tests were carried out for human herpesvirus-6 (HHV-6), Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpes simplex virus 1 (HSV-1), adenoviruses, parainfluenza viruses 1, 2 and 3, respiratory syncytial virus (RSV) and influenza viruses A and B. Viral infections were confirmed in 60.7% patients: 39.6% were correlated with HHV-6, 5.4% with EBV, 5.4% with both HHV-6 and EBV, 5.4% with adenoviruses, 1.8% with HSV-1, 1.8% with CMV and 1.8% with an unidentified herpes-like lymphotropic virus. HHV-6 isolates were obtained from either peripheral blood lymphocytes (PBLs) or pharyngeal secretion of the infected children. HHV-6 infections included both primary infections (72%) and reactivations (28%). Among HHV-6 infected children, 40%, with exanthem subitum, had infections presenting serological evidence of primary infection and virus isolation from PBLs. The remaining cases of primary infection and the cases of reactivation were found in patients with febrile syndrome without rash (60%). HHV-6 isolates were obtained either from PBLs or pharyngeal secretions from these patients. Southern blot hybridization of the DNAs of 4 HHV-6 isolates showed that the circulating HHV-6 strains all appeared similar, but differed from the HHV-6 strain U1102 used as a positive control.
6,7-Dihydro-9H-thiazolo[3,2-a][1,2,5]thiadiazolo [3,4-d]pyrimidin-9-one, 7,8-dihydro-6H,10H-[1,2,5]thiadiazolo[3',4':4,5]pyrimido [2,1-b][1,3]thiazin-10-one and its 3-methyl derivative were prepared by reacting 6,7-diamino-2,3-dihydro-5H-thiazolo[3,2-a]pyrimidin-5-one, 7,8-diamino-3,4-dihydro-2H,6H-pyrimido[2,1-b][1,3]thiazin-6-one or its 3-methyl derivative with N-thionylaniline. A reaction mechanism is proposed. The compounds and the sodium salts of (7-amino-2,3-dihydro-5H-thiazolo[3,2-a]pyrimidin-5-on-6-yl)sulfamic acid, (8-amino-3,4-dihydro-2H,6H-pyrimido[2,1-b][1,3]thiazin-6-on-7-yl) sulfamic acid and its 3-methyl derivative were tested for antimicrobial and antimycotic activity on a number of strains, namely: E. Coli, Proteus mirabilis, P. vulgaris, Pseudomonas aeruginosa, Salmonella spp, Staphylococcus spp, Streptococcus faecalis, Bacillus subtilis, Sarcina lutea, Candida albicans, and for antiviral activity on Herpes simplex virus type 1 and Vescicular stomatitis virus. None of the compounds showed antiviral activity or exhibited biological activity against gram-negative, gram-positive bacteria or against mycetes.
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Sera from a sample of healthy Italian people were tested in an indirect immunofluorescence assay (IFA) for reactivity to a Human Herpesvirus-6 (HHV-6) strain called CV, isolated from a baby with exanthem subitum (Portolani et al., 1990). Seropositivity values of 83.78%, 92.68% and 63.64% were found in subjects aged 3 months-6 years, 6-18 years and over 18 years respectively. Sera from cordal blood, sera from subjects with evidence of active infection by Cytomegalovirus (CMV) or by Epstein-Barr virus (EBV), sera from seropositive adults to Human Immunodeficiency Virus type 1 (HIV) were also investigated for antibodies to the same HHV-6 strain. Values of antibody incidence and antibody content to HHV-6 in these groups of sera were generally higher than in the other groups. HHV-6 reinfections of both endogenous and exogenous origin and antibody cross-reactivity were considered among the reasons of this increase. HHV-6 seropositivity values in healthy Italian people and in people from different countries were also discussed in the light of the antigenic characteristics of the HHV-6 strains used and of the different test conditions.
Some derivatives of thiazolo[3,2-c]pyrimidine, pyrimido[6,1-b][1,3]thiazine, thiazolo[2,3-i]purine, [1,3]thiazino[2,3-i]purine, thiazolo[3,2-c][1,2,3]triazolo[4,5-e]pyrimidine and [1,2,3]triazolo[4',5':4,5]pyrimido[6,1-b][1,3]thiazine were prepared. The compounds were tested for antimicrobial and antimycotic activity on a number of strains, namely, Escherichia coli, Proteus vulgaris, P. mirabilis, Pseudomonas aeruginosa, Salmonella sp., Staphylococcus aureus, Streptococcus faecalis, Bacillus subtilis, Sarcina lutea, Candida albicans, Aspergillus sp., and for antiviral activity on Herpes simplex virus, Type 1 (HSV-1), Vesicular stomatitis virus (VSV) and Coxsackievirus B5 (CoxB5). The compounds proved to be devoid of activity against viruses, mycetes and gram-negative bacteria, while some of them exhibited a modest activity against gram-positive bacterial strains.
Peripheral blood cultures from children showing symptoms of generic infectious disease have been examined for human herpesvirus-6 (HHV-6). Two HHV-6-related isolates have been obtained: one from a child for whom a clinical diagnosis of exanthema subitum has been made when a typical rash appeared, and one from a child in whom the infectious symptomatology has maintained the characters of nonspecific syndrome. All the syndrome diagnosed as exanthema subitum resulted as being associated with fresh infection by HHV-6-related virus.