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Background Adenovirus (ADV) infections can have a high mortality in immunocompromised patients and are difficult to treat. Objectives and study design We retrospectively analyzed occurrence and risk factors of ADV infection in 399 adults with hematological disorders undergoing hematopoietic stem cell transplantation (allo-HSCT), focusing on alternative donor transplantation (ADT) and disseminated disease. Results ADV infection occurred in 42 patients (10.5%). Disease was localized in 18 and disseminated in 6 patients. ADV infection was observed in 15% after ADT, performed in 29% of all recipients, and was less frequent (6%) in T-cell-replete (TCR) haploidentical transplantation using post-transplantation cyclophosphamide (PTCY) than in other ADT protocols. Lower age, the use of alternative donor grafts and acute graft-versus-host disease (GvHD) ≥ grade II were risk factors for ADV infection. After failure of standard antiviral treatment, three patients with disseminated ADV disease received one dose of ADV-specific T cells, resulting in virological response in 2/3 patients, clearance of ADV viremia in 2/2 patients, and survival of 1/3 patients; both patients with pneumonia died. Conclusions ADV infection was of moderate occurrence in our adult recipients of allo-HSCT despite a high proportion of potential high-risk patients receiving ADT. TCR strategies using PTCY might limit ADV complications in haploidentical transplantation. Despite feasible adoptive therapy strategies, outcome of disseminated disease remains dismal.

Several enzyme immunoassays (EIA) for the detection of parvovirus B19 IgM (anti-B19 IgM) are now commercially available. In this study, seven commercial EIAs (Biotrin, DAKO, Viramed, Viratech, R-Biopharm, Mast) were compared with an in-house EIA (MvP-EIA) using native viral B19 particles and the reference IgM radioimmunoassay (MACRIA). A total of 88 sera were tested. Results agreed in 39/88 (44.3%) sera, whereas 47/88 (53.4%) were discrepant and 2/88 (2.3%) gave an equivocal result. Assay sensitivity ranged from 70.3 to 100% and specificity, from 75.9 to 100%. The best results were obtained with two EIAs (Biotrin, DAKO) using baculovirus-expressed B19 proteins as antigen. This study has shown that baculovirus-expressed B19 antibody tests are suitable tools for detecting anti-B19 IgM.

Summary A single tube, reverse transcriptase/polymerase chain reaction (RT-PCR) was developed and evaluated for detecting a 400-bp product of the small RIMA of sandfly fever virus, serotype Toscana (TOS). For more sensitive detection of genomic TOS RNA, a nested PCR amplifying a 243-bp cDNA within the RT-PCR product was established. Nucleotide sequence analysis of first- and second-round PCR products using the dideoxy cycle sequencing technique confirmed a previously published sequence of the TOS reference strain (ISS. Phi. 3). By nested PCR, genomic TOS RNA was amplified from two consecutive sera taken 3 and 7 weeks after the onset of illness in one patient, and from CSF of a second patient obtained at the onset of meningitis. Authenticity of amplified PCR products was confirmed by nucleotide sequence analysis, revealing a sequence identical to the TOS reference strain. RT-PCR and nested PCR are useful for laboratory diagnosis and studies of the molecular epidemiology of TOS infection.

cALL positive blast cells from peripheral blood of four children as well as cells from the Reh line were cultivated in DC in order to promote cell differentiation. DC were implanted into the peritoneal cavity of pre-irradiated CBA mice. At different intervals during the culture period changes in the membrane marker profile were determined by direct immunofluorescence after labeling the cells with AcALLG, ATCG and polyvalent AIg. In addition, the ability of cells to form rosettes with SRBC, AET-treated sheep erythrocytes and mouse red blood cells was tested. In the course of the culture the majority of cALL positive cells from three patients (T.H., V.M., I.G.) simultaneously developed a positive reaction with polyvalent AIg. Furthermore, a remarkable portion of cells in V.M. turned out to become positive with only AIg. Comparable results together with an increasing percentage of M-RFC were obtained in the fourth patient, C.M., using AIg(Fab) 2 . Besides this development, quite a few cells expressing the T-cell antigen occurred in the latter two patients. In the case of the Reh line, the cells developed T-cell antigen, and in one of two experiments they further acquired an E-receptor. Conversely, in the second experiment a receptor for mouse red blood cells was detected. Our data suggest that malignant cells carrying the cALLA are representative of an early developmental stage of lymphatic ontogeny.