Quantitation of cytomegalovirus DNA in lung tissue of bone marrow transplant recipients

Shibata, Mitsue; Terashima, Makoto; Kimura, Hiroshi; Kuzushima, Kiyotaka; Yoshida, Jun; Horibe, Keizo; Morishima, Tsuneo

doi:10.1016/0046-8177(92)90404-q

Cited by 23 publications

(9 citation statements)

References 25 publications

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“…For patient nos. 1, 2, 4, 5, and 6, congenital infection was confirmed by isolation of CMV or detection of virus DNA by polymerase chain reaction (PCR) [Shibata et al, 1992;Tanaka et al, 2000] from urine sampled within the first week of life. Urinary CMV DNA was detected by PCR in patient no.…”

Section: Blood Donorsmentioning

confidence: 99%

Flow cytometric analysis of cytomegalovirus‐specific cell‐mediated immunity in the congenital infection

Hayashi

Kimura

Morishima

et al. 2003

Journal of Medical Virology

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Flow cytometric analysis of CD4(+) and CD8(+) T cells specific to human cytomegalovirus (CMV) was undertaken in seven patients with congenital CMV infection, six healthy infants who had acquired infection, and six CMV-seropositive adults. Intracellular cytokine assays showed that 0.03-2.23% of CD4(+) T cells in the healthy infants and adults produced interferon-gamma (IFN-gamma) in response to CMV antigens. In contrast, such CD4(+) T cells were almost undetectable in patients with congenital CMV infection who were younger than 2 years of age. Tetrameric major histocompatibility complex/peptide complex analysis demonstrated HLA*A2402-restricted phosphoprotein 65-specific CD8(+) T cells to be present in most healthy infants and adults tested, but almost absent in the patients. Interestingly, CMV-specific CD4(+) T-cell responses were observed in two patients with congenital infection beyond the age of 5 years. The present study points to impairment of CMV-specific cellular immunity in patients in single-cell levels with congenital CMV infection during the infant period and possible restoration in later childhood.

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Section: Blood Donorsmentioning

confidence: 99%

Flow cytometric analysis of cytomegalovirus‐specific cell‐mediated immunity in the congenital infection

Hayashi

Kimura

Morishima

et al. 2003

Journal of Medical Virology

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“…Determination of EBV DNA by the polymerase chain reaction EBV DNA was detected and quantified by polymerase chain reaction (PCR), as previously described [32][33][34]. Briefly, DNA extracted from the patient's plasma (0 .…”

Section: Cytokine Assaysmentioning

confidence: 99%

Establishment of anti-Epstein–Barr virus (EBV) cellular immunity by adoptive transfer of virus-specific cytotoxic T lymphocytes from an HLA-matched sibling to a patient with severe chronic active EBV infection

Kuzushima

Yamamoto

Kimura

et al. 1996

Clinical and Experimental Immunology

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SUMMARYWe describe an experience of a specific immune transfer treatment in a patient with chronic active EBV infection. The patient had low anti-EBV T cell-mediated cytotoxic activity in his peripheral blood mononuclear cells (PBMC), which may have been the primary cause of the disease. An EBV-specific cytotoxic T lymphocyte (CTL) line was established from PBMC obtained from the patient's sister whose human leucocyte antigens (HLA) are identical to patient's. The patient received three courses of intravenously administered CTL at 3-week intervals. The number of the cells was increased with each course of treatment. After infusion of the T cell line, anti-EBV CTL activity was detected in the patient's PBMC. CTL activity increased markedly after the second course of immune transfer therapy. The amount of EBV DNA in the patient's plasma showed transient but repeated decreases. Serum levels of tumour necrosis factor-alpha (TNF-), which had elevated before treatment, began to decrease after initiation of treatment. No adverse effects were directly associated with CTL infusions. Despite having previously received a pneumococcal vaccine and prophylactic antibodies, the patient died of infection caused by Streptococcus pneumoniae bacteraemia 27 days after the third infusion. Although the longterm efficacy and safety of this therapy remains to be established, our findings suggest that adoptive transfer of CTL specific for EBV obtained from an HLA-matched donor might be a promising treatment for patients with chronic active EBV infection.

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“…Although quantitative analysis is important to diagnose or monitor virus-related disease [Shibata et al, 1992;Ando et al, 1993;Shinkai and Spector, 1995;Yamamoto et al, 1995;Boeckh and Boivin, 1998], methods such as competitive-quantitative PCR assay or quantitative viral isolations take a long time and are labor intensive. They are not convenient for rapid-decision making in clinical situations.…”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis of cytomegalovirus load using a real-time PCR assay

et al. 2000

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A novel real-time PCR assay system was developed to quantify the cytomegalovirus (CMV) genome load. The real-time PCR assay could detect from 6 to over 10(6) copies of CMV-DNA with a wide linear range. The virus load of immunocompromised patients with symptomatic CMV infections was quantified and compared to that of asymptomatic ones. In symptomatic patients, all 17 peripheral blood leukocytes were positive for CMV DNA, and its mean value was 10(3.3) copies/10(6) cells. On the other hand, only 9 of 38 samples (24%) were positive in the asymptomatic patients, and its mean titer was lower (10(2.0) copies/10(6) cells) than that of the symptomatic group (P = 0.002). In plasma, the virus genome was detected in 13 out of 17 samples from symptomatic patients (76%), and its mean value was 10(4.0) copies/ml. In contrast, for the asymptomatic group, only one out of 36 samples were positive (3%). Finally, this system was used to monitor two patients with CMV infections serially. The CMV DNA copy number changed with their clinical symptoms and anti-CMV therapy, and virtually paralleled the result of the pp65 antigenemia assay in both cases. In one patient with the cord blood transplantation, however, the CMV DNA became positive faster than the antigenemia assay. These results indicate that this assay is sensitive and useful for estimating the CMV genome load not only in peripheral blood leukocytes but also in plasma. It can be very helpful for diagnosing CMV-related diseases and monitoring the virus load in patients with CMV infections.

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Quantitation of cytomegalovirus DNA in lung tissue of bone marrow transplant recipients

Cited by 23 publications

References 25 publications

Flow cytometric analysis of cytomegalovirus‐specific cell‐mediated immunity in the congenital infection

Flow cytometric analysis of cytomegalovirus‐specific cell‐mediated immunity in the congenital infection

Establishment of anti-Epstein–Barr virus (EBV) cellular immunity by adoptive transfer of virus-specific cytotoxic T lymphocytes from an HLA-matched sibling to a patient with severe chronic active EBV infection

Quantitative analysis of cytomegalovirus load using a real-time PCR assay

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