Nobuo Nagata

Role of Milk Whey in the Transmission of Human Cytomegalovirus Infection by Breast Milk

Breast‐fed infants are susceptible to human cytomegalovirus (HCMV) infection via breast milk. In our previous study, HCMV was isolated more frequently from breast milk at later than one month after delivery than from colostrum or early breast milk. To clarify the role of milk cells and whey in vertical infection by breast feeding, we separated breast milk into milk cells and whey and examined each fraction for the presence of HCMV. We collected breast milk from mothers who breast‐fed their infants (aged from 3 days to 2 months). The breast milk was centrifuged and separated into the middle layer (layer of milk whey) and the pellet (containing milk cells). We attempted to isolate HCMV from whey and to detect HCMV immediate early (IE) DNA in both milk whey and cells. HCMV was isolated from 7 out of 35 (20.0%) whey samples and HCMV IE DNA was detected from 15 out of 35 (42.9%) whey and/or milk cells. Detection rates of HCMV IE DNA in the whey layer and milk cells were 39.1% (25 out of 64) and 17.2% (11 out of 64), respectively. HCMV IE DNA was not detected in colostrum, but was detected in breast milk samples one month after delivery. Therefore, cell‐free HCMV shed into milk whey may have a more important role in vertical infection by breast milk than cell‐associated HCMV in the milk.

EFFECT OF GLYCYRRHIZIN, CYCLOSPORIN A, AND TUMOR NECROSIS FACTOR ALPHA ON INFECTION OF U-937 AND MRC-5 CELLS BY HUMAN CYTOMEGALOVIRUS

Reactivation of latent or persistent human cytomegalovirus (HCMV) infection of monocytes or macrophages occurs under immunosuppressive conditions. We investigated the effect of glycyrrhizin (GL), cyclosporin A (CsA), and tumor necrosis factor‐α (TNF‐α) on the viral DNA synthesis and antigen expression of HCMV in U‐937 and MRC‐5 cells. Although GL inhibited the viral antigen expression of HCMV in human monocytic cell line U‐937 and human embryonic lung cell line MRC‐5 in the study, as determined by flow cytometry and immunofluorescence assay, immediate early HCMV DNA was detected by the polymerase chain reaction. CsA or TNF had no inhibitory effect on HCMV in U‐937 or MRC‐5 cells. The HCMV infection model with U‐937 or MRC‐5 cells is of use for clarifying not only the mechanism of persistent infection but also the anti‐HCMV effect of chemical agents. J. Leukoc. Biol. 55: 24–28; 1994.

Detection of Human Cytomegalovirus DNA in Breast Milk by Means of Polymerase Chain Reaction

Three hundred and twenty‐five breast milk samples were examined for the occurrence of human cytomegalovirus (HCMV) by cell culture method. Virus was isolated from the milk in 1 of 177 samples collected within 6 days after delivery, 2 of 115 samples collected during the period of 7 days to 1 month after delivery, 10 of 33 samples collected over 1 month after delivery. Next, we tried to amplify HCMV DNA from the breast milk samples from HCMV seropositive mothers and seronegative mothers at 1 month after delivery by polymerase chain reaction. HCMV DNA was detected in 12 of 13 samples from seropositive mothers and in none of 7 samples from seronegative mothers. It was thought that all women seropositive for HCMV principally shed the virus into their breast milk at 1 month after delivery.

Congenital muscular dystrophy in Marinesco-Sjögren syndrome

The histochemical and immunocytochemical findings of biopsied muscle in a 2-year-old girl with Marinesco-Sjögren syndrome are reported. Muscle histology consisted of mild muscular dystrophy, such as that found in limb-girdle or non-Fukuyama congenital muscular dystrophy. By immunocytochemical stain using anti-dystrophin antibody, Duchenne and Becker muscular dystrophies were excluded. In addition to characteristic clinical features, including ataxia, congenital cataract, and psychomotor retardation, muscle involvement is essential to the diagnosis of Marinesco-Sjögren syndrome.

Analysis of human cytomegalovirus-infected peripheral blood mononuclear cells from infants with liver dysfunction by flow cytometry and the polymerase chain reaction

Perinatal human cytomegalovirus (HCMV) infection often involves the hepatobiliary tract, but infected individuals usually remain asymptomatic. We investigated the role of CD8+ lymphocytes in 13 infants with liver dysfunction associated with perinatal HCMV infection. In three patients more than 40% of CD8+ cells were positive for HCMV immediate early antigen (IEA) and late antigen (LA) by flow cytometry after selection of T lymphocytes subpopulations. In the other 10 infants, 20% to 30% of CD8+ cells were positive for HCMV IE A and LA. HCMV IE DNA was detected in CD8+ cells from one infant, in CD4+ cells from one infant, and in both CD4+ and CD8+ cells from three infants. HCMV infection of CD8+ cells may play an important role in the process of perinatal primary infection. J. Leukoc. Biol. 56: 187–191; 1994.

Replication of human cytomegalovirus in the cells of the U937 monocytoid cell line

Human cytomegalovirus (HCMV) infection in immunocopromized hosts sometimes occurs as a result of reactivation. Cells of the monocytemacrophage linkage are suggested to be a site of latency and persistence for HCMV. The human monocytic cell line U937 was infected with the AD169 strain and a clinical isolate of HCMV. The expression of surface antigens on the cells was assessed by flow cytometry. The polymerase chain reaction (PCR) was used to detect viral DNA from infected cells. CMV immediate early antigen, early antigen, and late antigen (LA) were detected from both clinical isolate- and AD169-inoculated U937 cells by flow cytometry. CMV DNA which code major immediate early gene (US3) and LA gene (US14) were detected from the clinical isolateinoculated U937 over a period of 31 days as tested by PCR. These U937 cells proliferated as well as uninfected U937 cell, but only a small number of AD169-inoculated U937 cells survived after 14 days of inoculation. Interleukin-2 activities were detected in the media on days 24–40 after inoculation with AD169. This chronic CMV infection model of U937 might be utilized to study the mechanisms of persistence and reactivation.

Minocycline‐induced hemolytic anemia

A case of drug‐induced immune hemolytic anemia is described. A 2 year old boy exhibited sudden anemia and hemoglobinuria after administration of minocycline (MINO). The specific immunoglobulin G antibody against MINO was demonstrated in the patients serum by western blotting. This is a rare example where anti‐minocycline immune complex‐mediated hemolysis was responsible for an intravascular hemolytic process.

Dipyridamole‐provoked chest pain implies severe coronary artery disease in children

The diagnostic significance of dipyridamole‐provoked chest pain was studied in 17 children with severe coronary arterial stenotic lesions (CAL) complicated with Kawasaki disease. Although dipyridamole induced chest pain in seven patients (symptomatic group), 10 reported no pain (asymptomatic group). In the asymptomatic group, seven children had one vessel disease (1VD) of right coronary artery (RCA) and the other three had two vessel disease (2VD) involving the RCA and left anterior descending artery (LAD). Four multivessel disease patients, one three vessel disease (3VD) and three 2VD of LAD and RCA, and three 1VD of LAD, were symptomatic. In the thallium scans, all patients, except two of the asymptomatic group, showed perfusion abnormalities. In addition, the extent score of the symptomatic group was significantly worse than that of the asymptomatic group (P = 0.01). While only one in six of the asymptomatic group showed abnormal ST depression on treadmill exercise electrocardiography, all patients in the symptomatic group (P = 0.02) showed ischemic ST depression. These findings suggest that the occurrence of chest pain after medication with dipyridamole closely correlates with the severity of CAL in children.

Dystrophin analysis in the differential diagnosis of autosomal recessive muscular dystrophy of childhood and Duchenne muscular dystrophy

We report 2 patients with childhood autosomal recessive muscular dystrophy. Both patients had slight muscle weakness without enlargement of the calf muscles or involvement of the facial muscles. Their clinical courses are static. Muscle histology revealed characteristic features of muscular dystrophy. Dystrophin was identifiable in the sarcolemma of both patients by immunocytochemical staining with an antidystrophin antibody. At an early age, immunocytochemical analysis with antidystrophin antibody was useful in distinguishing between childhood autosomal recessive and Duchenne muscular dystrophies.

Reactogenicity and immunogenicity of measles–rubella combined vaccine in school-entry-aged subjects with naturally acquired measles immunity

The reintroduction of measles–rubella combined (MR) vaccination to Japan raised concerns about adverse events as well as immunogenicity related to booster immunization in subjects with naturally acquired immunity to measles or rubella.