Altered expression of cell surface membrane antigens in a common acute lymphoblastic leukemia-associated antigen-expressing neuroblastoma cell line (SJ-N-CG) with morphological differentiation.

The expression of common acute lymphoblastic leukemia antigen (CALLA) on a human neuroblastoma cell line, SJ-N-CG, was demonstrated by indirect membrane immunofluorescence, complement-dependent cytotoxicity, and quantitative absorption, using two monoclonal antibodies (J-5 and BA-3) directed against CALLA. Immunoprecipitation of solubilized 125I-labeled membrane proteins from SJ-N-CG cells with J-5 antibody revealed a protein with a molecular weight of 100,000 as determined on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Morphological differentiation of SJ-N-CG cells could be induced in the presence of 2.0 mM dibutyryl adenosine 3'-5'-cyclic monophosphoric acid for 10 days of culture. Changes in cell surface membrane antigens associated with morphological differentiation were studied by indirect immunofluorescence and complement-dependent cytotoxicity using a panel of seven monoclonal antibodies. Increases in the antigens recognized by BA-2 (detecting leukemia-associated antigen), anti-Thy-1, and antibody 390 (Thy-1 antigen) were found in "differentiated cells," while those detected by BA-1 (B-cell-associated antigen) and J-5 (CALLA) were unchanged. In contrast, the antitransferrin receptor defined by B3/25 was inhibited, and expression of B7/21-defined la-like antigen was not induced. Kinetic studies on antigenic alterations showed that the expression of BA-2-defined antigen rose on Day 2 and remained at the same level until Day 10. The expression of CALLA was not changed from Days 2 to 10. The augmentation of Thy-1 antigen was noted on Day 4 and reached the maximum on Day 10. These results show that dibutyryl adenosine 3'-5'-cyclic monophosphoric acid is capable of inducing phenotypic changes in SJ-N-CG cells. The changes of expression of some antigens on exposure of cells to dibutyryl adenosine 3'-5'-cyclic monophosphoric acid may enable us to have a greater understanding of the differentiation of neuroblastoma to a more mature ganglioneuroblastoma phenotype.

Experimental study of the maternal effects on tumor immunity of infant mice with C1300 mouse neuroblastoma.

The spontaneous regression of neuroblastoma (NB), one of the most common malignant tumors in childhood, is found to occur in 1% to 2% of patients with NB, especially in young infants. An unexpectedly favorable response to therapy is also noticed in infants suggesting the potential presence of an immune mechanism. Monoclonal C1300-S and C1300A-4 cell lines were established from polyclonal C1300 cells in our laboratory. Adult female A/J mice that had rejected 1 x 10(3) NB cells (C1300S-3) or 1 x 10(5)-10(6) NB cells (C1300A-4) were used as immunized mothers. The immunized mothers with C1300A-4 or C1300S-3 were found to have specific antibodies to C1300S-3 cells by 51Cr release assay of complement dependent cytotoxicity. Newborn mice, 24 hours after birth from immunized or nonimmunized mothers, were inoculated with 1 x 10(3) C1300S-3 NB cells. The same antibody that was assayed in the immunized mothers was detected in this offspring by the antibody-dependent cell cytotoxicity (ADCC). The tumor incidence in the offspring of the immunized mothers was found to be less than that of the offspring of the nonimmunized mothers. This study suggests that the lower tumor incidence in the offspring of immunized mothers compared with offspring of nonimmunized mothers may be attributed to their ADCC activity. Furthermore, the antibody that has the ADCC activity was proven to be immunoglobulin G by a serum absorption test using IgG absorbant. This study offers insight into the relationship between transported mother-infant immunoglobulins and on its potential control of NB.

Incidence of neuroblastoma in infancy in Japan.

Since 1974, we have been mass screening for neuroblastoma in 6-month-old infants using a VMA (vanilmandelic acid) spot test in Kyoto City. Fifteen (48.4%) of the 31 childhood neuroblastoma cases registered in this city during the recent 8-year-period, 1974-1982, were under 1 year of age. The annual number of cases per year was 3.9 cases under 15 years with 1.9 cases under 1 year of age. As the number of children under 15 years of age in this city was 292,000 according to the National Census of the Human Population in 1975, the annual incidence of this tumor in childhood was 3.9/292,000 (13.3/million). And as the number of live births was 161,153, the annual detection rate for this tumor in infancy increased to 15/161,153 (93/million) from 1974 to 1982. We found that many neuroblastomas could be detected in infancy by a VMA spot test and/or a careful physical examination.

Childhood drownings and near-drownings in Japan.

In Japan, the leading cause of death for children over 1 year old is injury, and for children aged 0-14, drowning is the second leading cause of death. The purpose of the present study was to describe the epidemiological factors of drownings and near-drowning among Japanese children and to ascertain whether there are characteristic patterns for different age groups. Epidemiologic data was obtained by questionnaire. A total of 604 cases of submersion injuries were reported from 49 hospitals located in 22 Japanese prefectures. In the present paper, victims of drowning (n = 134) and near-drowning with permanent severe brain damage (n = 51) and those of near-drowning with intact survival or mild impairment (n = 419) were investigated. Preschool-aged children, especially toddlers, are at the greatest risk of drowning and near-drowning, and for children over 2 years of age, boys have three times greater risk than girls. The bathtub is the most common place of submersion injuries in Japan, especially for children under 4 years of age. The important risk factors for the victims who died or were severely impaired were associated with duration of submersion and necessity of emergency cardiopulmonary resuscitation on arrival at hospital.

Matrix proteins induce neuroblastoma cell differentiation without altering cell growth.

The maturation of embryonal neural crest cells is thought to be regulated in part by the milieu into which these cells migrate. Neuroblastoma (NB) is a tumor of very early childhood that is thought to arise in association with the arrested differentiation of embryonal neural crest cells. In culture, neuroblastoma tumor cells differentiate in the presence of retinoic acid, which is also known to influence extracellular matrix protein synthesis. We have cultured neuroblastoma cells on laminin (LN) and fibronectin (FN) substrata to examine the role of extracellular matrix in retinoic acid (RA)-induced differentiation of these tumor cells. These proteins caused morphologic changes in NB cells indistinguishable from those caused by RA. Antiserum to each of these proteins blocked the effects induced by the corresponding protein, but neither antiserum affected the action of RA. Despite the induction of a neuronal morphologic change, matrix proteins did not alter the proliferation of NB cells. These results indicate that LN and FN modulate the differentiation of NB cells without inducing growth arrest and that RA-induced differentiation does not require these matrix proteins.

DNA contents in Wilms' tumors. A cytofluorometric study.

Cytofluorometric measurement of nuclear DNA was performed on individual tumor cells isolated from paraffin sections of Wilms' tumors. The DNA distribution of one untreated primary tumor showed the first major peak near the diploid range and the second peak in the tetraploid range. There were many cells having amounts of DNA interspersed between the diploid and tetraploid range. Polyploid cells were not observed. Most of the Wilms' tumor cells had a higher diploid DNA value than the small lymphocytes of the control cells. The primary tumor and its metastases showed similar DNA distribution patterns. After treatment, the distribution pattern showed a reduction in number of cells between the diploid and tetraploid range and in the tetraploid range, with the greater number of cells being found in the diploid range. Though the phases of the cell cycle are not always clearly detectable by cytofluorometry, the cells of the untreated Wilms' tumor were distributed into the phases of the cell cycle as follows: 65.9% in G0 and G1 phases; 31.2% in S-phase; and 2.9% in G2 and M phases.

Effects of interferon on C1300 mouse neuroblastoma.

The effects of murine interferon (MuIFN) with and without cyclophosphamide (CY) were evaluated in C1300 neuroblastoma-bearing mice. Mean survival times were prolonged by administration of 25,000 IU of MuIFN three times per week by ip injection. The combination of MuIFN and CY was more effective in the treatment of mouse neuroblastoma than either agent alone. Our results suggested that MuIFN is synergistic in combination with moderate amounts of CY in mouse neuroblastoma cells and may be given for the treatment of neuroblastoma.

Neuroblastoma. Mass screening for early detection and its prognosis.

Since July 1973, the authors began developing a mass screening system using a VMA (vanilmandelic acid) spot test on 6-to-7 month-old infants for early detection of neuroblastoma in Kyoto city, Japan. Using this method, six infants with this tumor were discovered; five of the six infants were cured, and one is under treatment. These patients showed a favorable prognosis on early diagnosis. In this article, 57 neuroblastoma patients from the Department of Pediatrics, Kyoto Prefectural University of Medicine, treated during the last 20 years, from July 1962 to June 1982, are evaluated. Since the mass screening program has run smoothly since July 1974, clinical findings are compared between 35 neuroblastoma cases before mass screening from the 12-year period from July 1962 to June 1974 and 22 cases after mass screening, during the 8-year period from July 1974 to June 1982. Before mass screening, only 20% (7/35) of the patients were discovered with neuroblastoma younger than 12 months of age and 68.6% were older than 2 years of age. After mass screening, 54.6% (12/22) of the patients were younger than 12 months of age and only 31.8% (7/22) were older than 2 years of age. Before mass screening, 17.1% (6/35) survived with five of the six surviving patients being younger than 12 months of age at the time of diagnosis; 72.7% (16/22) of the patients detected after mass screening are living now. Eleven of the 16 patients have already been cured, and the remaining 5 patients are presently undergoing treatment. A marked improvement of their prognoses is dependent on the early detection of this tumor by mass screening. To date, using the VMA spot test for early detection in infancy is convenient and effective for improvement of its prognosis.

Treatment of an infant with congenital sodium diarrhea by oral rehydration.

We have been following a male case of congenital sodium diarrhea (CNaD), who had a distended abdomen, passed watery stools with high sodium concentration, and showed metabolic acidosis in the first week of life. He also showed hyponatremia, low urine sodium, high serum aldosterone and high renin activity. Other possible causes of secretory diarrhea were ruled out. The initial effective treatment was oral supplements of water and electrolytes lost in the fecal fluid: with this he gained weight, and hyponatremia and metabolic acidosis were corrected. Loperamide hydrochloride was administered to increase intestinal absorption of sodium, as the serum prostaglandin was high. Watery diarrhea then improved, but as he passed acholic stools, we stopped and postponed the treatment for a while; the stools then became watery again. Although diarrhea has persisted, he has not shown any abnormalities in growth or psychomotor development for the first two years of life.

Intraspinal metastasis of neuroblastoma--report of a case detected at autopsy.

Metastasis to the brain or spinal cord parenchyma is extremely rare in cases of neuroblastoma. We present a 3-year-7-month-old boy with neuroblastoma, stage IV, with intraspinal metastasis. He had no neurologic manifestation except incontinentia urinae and ataxia at the terminal stage. His cranial computed tomography scan showed high density areas in both cerebellar hemispheres which seemed to be distant metastatic masses. At autopsy, the metastases in the cerebellum were found to be due to dural infiltration but in the spinal cord parenchyma of the lumbar spine metastases were detected macroscopically. There were multiple distant metastatic areas near the roots, anterior and posterior. The neuroblastoma seemed to have spread along the roots by direct invasion from the meninges. In the future, the number of patients with metastasis into the brain or spinal cord parenchyma will increase because patients with progressive disease could live for a long time as a result of intensive chemotherapy. Observation of these cases will be helpful to clarify the routes of metastasis to these areas.