Moriyama I

Detection of IL-6 in human milk and its involvement in IgA production

A large amount of interleukin-6 (IL-6) was found to be contained in human whey. The concentration of IL-6 in colostrum was significantly higher than that in serum or in milk taken 1 month after parturition. Colostrum contained many more mononuclear cells than late milk. In terms of the proportion of monocytes, T cells and B cells, however, there is no difference between colostrum and late milk. There is a significantly positive correlation between the concentration of IL-6 and the number of mononuclear cells in milk. This demonstrates that IL-6 in whey is derived in part from mononuclear cells. Stimulation of human milk mononuclear cells by Staphylococcus aureus Cowan I in the presence of anti-IL-6 antibody markedly decreased the production of IgA. This suggests that IL-6 contained in milk is closely associated with the local production of IgA in the breast.

Primary infection of Japanese infants with adult T-cell leukaemia-associated retrovirus (ATLV): Evidence for viral transmission from mothers to children

Primary infection with adult T-cell leukemia virus (ATLV) was investigated by follow-up studies on 16 ATLV-seropositive mothers and their breastfed infants in an ATLV-endemic area of Japan. Maternal antibody to ATLV decreased in all the infants, and was detectable in only three of 12 infants tested 6 months after birth. Reappearance of the antibody 9-18 months after birth was observed in only four of the 16 infants. The ATLV-bearing cells in peripheral blood were detected in all 16 mothers after delivery. None of the 16 infants showed ATLV-bearing cells in peripheral or cord blood sampled at birth, or 1, 3 or 6 months after birth. However, virus-bearing cells in the blood became detectable 9-18 months after birth in 13 of the 16 infants. Maternal antibody and virus-bearing cells were never detected in a control group of seven infants of ATLV-seronegative mothers. These findings provide evidence for the high incidence of primary ATLV infection during early infancy among infants born to ATLV-seropositive mothers and suggest maternal viral transmission. Furthermore, samples of breast milk from all 12 seropositive mothers examined contained cell-associated ATLV capable of being transmitted to peripheral leucocytes of neonates. This finding suggests that one of the possible maternal transmission routes of ATLV is via breast milk.

Bottle-feeding can prevent transmission of htlv-1 from mothers to their babies

Breast-feeding is a major factor in the vertical transmission of human T-lymphotropic virus type I (HTLV-I). We studied whether such transmission may be prevented by bottle-feeding. HTLV-I infection was detected by both HTLV-I antigen and antibody tests. Thirty bottle-fed babies were examined 24 months after birth; only one was found to be HTLV-I antigen-positive. This infection rate was lower than that for breast-fed babies in whom HTLV-I antigen was detected in 24 of the 31 24-month-old babies born to HTLV-I positive mothers in a previous study. These results suggest that most vertical transmission of HTLV-I is attributable to breast-feeding and can be prevented by bottle-feeding.

Identification of HTLV‐I Sequence in Cord Blood Mononuclear Cells of Neonates Born to HTLV‐I Antigen/Antibody‐positive Mothers by Polymerase Chain Reaction

We developed a polymerase chain reaction (PCR) method which has high sensitivity and simple technique in order to investigate the presence or absence of human T lymphotropic virus type I (HTLV‐I) provirus in cord blood mononuclear cells of neonates born to HTLV‐I carrier mothers. Out of 40, three subjects were found to contain the HTLV‐I provirus genome. These three subjects remained HTLV‐I sequence‐positive in follow‐up study. On the other hand, when examined by a conventional technique for detection of HTLV‐I‐associated antigen on peripheral mononuclear cells, all 40 neonates were HTLV‐I‐associated antigen‐negative. These results suggest that PCR is more sensitive than the conventional antigen detection method and is useful in early detection of HTLV‐I infection in neonates born to HTLV‐I carriers.

Detection of HTLV-I Genome in Seronegative Infants Born to HTLV-I Seropositive Mothers by Polymerase Chain Reaction

We applied the polymerase chain reaction (PCR) method to detect gag, env and pX sequences of human T cell leukemia virus type I (HTLV‐I) provirus in peripheral blood lymphocytes of seronega‐tive infants born to HTLV‐I seropositive mothers. Out of 22, five subjects were found to contain the HTLV‐I provirus genome. Two of the five cases were judged to be negative for not only anti‐HTLV‐I antibodies but also the viral antigens on cultivated lymphocytes by the conventional antibody/antigen detection methods. These results indicate that PCR is of great use as a simple and highly sensitive method detect HTLV‐I infection.

Characterization of Human Placental Activity for Transport of L-alanine, Using Brush Border (Microvillous) Membrane Vesicles

The uptake of taurocholate into brush border membrane vesicles prepared from human full term placenta was studied using a rapid filtration technique. The taurocholate uptake into brush border membrane vesicles was sensitive to extravesicular osmolarity, and pre-incubation of the brush border membrane vesicles with the taurocholate increased the uptake of taurocholate into the brush border membrane vesicles. These findings indicate that the uptake of taurocholate by brush border membrane vesicles represents transport into vesicles. The uptake of taurocholate into vesicles was not dependent on Na+ electrochemical gradient (extravesicular > intravesicular). But this uptake was markedly increased when the intravesicular space was rendered electrically more positive by the use of lowly permeant anions or valinomycin-induced K+ diffusion membrane potentials. These findings indicate that the taurocholate transport into brush border membrane vesicles was dependent on membrane potential. The initial rate of taurocholate transport into brush border membrane vesicles exhibited saturation kinetics with respect to the taurocholate concentration, an apparent Km of 67 microM and Vmax of 0.30 nmol/mg protein/20 sec were calculated.

Production of IL-6 (BSF-2/IFN β2) by mononuclear cells in premature and term infants

The production of interleukin 6 (IL-6) was examined in premature neonates (48 cases, including 3 miscarried fetuses) and fullterm neonates (20 cases). The IL-6 production by mononuclear cells was measured after stimulation with Staphylococcus aureus Cowan Strain I (SAC), phytohemagglutinin (PHA) or lipopolysaccharide (LPS). The production in full-term neonates was similar to that in healthy adults, whereas it was significantly lower in premature neonates without premature rupture of the membrane (PROM). However, in premature infants with PROM a normal level of IL-6 production was observed in mononuclear cells stimulated with SAC, PHA and LPS. Furthermore, there was a positive correlation between the IgM concentration in the cord serum and IL-6 production by LPS-stimulated mononuclear cells.

Characterization of the CA125 Antigen Secreted from a Newly Established Human Ovarian Cancer Cell Line (SHIN‐3)

A cell line (designated SHIN 3) was established from a 56‐year old Japanese woman diagnosed as having serous cystadenocarcinoma of the ovary. SHIN 3 produced the tumor marker, CA125. When samples were subjected to one dimensional gel electrophoresis followed by immunoblotting with anti CA125 antibody, antigen binding appeared in the low‐molecular mass fraction around 50 KDa as well as in the high molecular‐mass fraction (more than 200 KDa). Subsequent resolution of this lower molecular‐mass component by two dimensional gel electrophoresis gave rise to bands of 49 KDa near an isoelectric point of 7.3. Treatment of the CA125 antigen with 10 mM periodic acid resulted in no loss of activity. However, reduction and alkylation in 6 M guanidine HCI or treatment at 100 C for 20 min resulted in complete loss of activity. Treatment with various glycosidases did not inhibit activity, whereas digestion with proteases (except for papain) produced complete loss of activity, suggesting that the CA125 antigenic determinant is composed of con‐formationally dependent peptides.

Interleukin-2 production by human fetal lymphocytes

IL-2 production by PHA-stimulated human lymphocytes in umbilical blood was studied ontogenetically. Lymphocytes from the 16-week-old fetus produced appreciable amounts of interleukin-2 (IL-2). In 16- to 36-week-old fetuses the IL-2 production rate was found to be significantly higher than that in adults. When signal transduction, which is mediated by high affinity IL-2 receptors (IL-2R), was studied after adding exogenous IL-2 to PHA-blast lymphocytes, the response of the 24-week-old fetus was similar to that of adults. These results indicate that the attainment of IL-2 production and the function of IL-2 receptors are completed at a relatively early stage of fetal development.

IL-2 receptor expression and function on human cord blood mononuclear cells following PHA and anti-CD3 antibody stimulation

Interleukin-2 receptors (IL-2R) on mononuclear cells from human umbilical cord blood were investigated after stimulation by phytohemagglutinin (PHA) and anti-CD3 antibody. The proportion of cells expressing the Tac antigen (IL-2R alpha, p55) did not differ from that for adult mononuclear cells. However, the levels of high affinity IL-2R (H-IL-2R) and low affinity IL-2R (L-IL-2R) present on the PHA blasts of cord blood cells were shown to be twice and three times, respectively, that of adults by interleukin-2 (IL-2) binding assay. The level of low affinity IL-2R in the cord blood cells was approximately double that of adult cells after activation by anti-CD3 antibody, but no difference was observed in the levels of high affinity IL-2R. Functional investigation of IL-2R revealed that the amount of internalized IL-2 in PHA-stimulated cord blood mononuclear cells was twice that in adult mononuclear cells but there were no differences between them in the time course of internalization and degradation. Although there were significant numbers of H-IL-2R in premature (25 and 28 weeks) infants following PHA stimulation, there were markedly fewer following anti-CD3 stimulation. Prematurity of the T-cell activation system through the CD3 pathway at this stage is therefore indicated.