Clinical characteristics of monochorionic twins with large hemoglobin level discordance at birth.

OBJECTIVES: To evaluate neonatal outcomes and clinical characteristics of monochorionic diamniotic (MCDA) twins with a large intertwin hemoglobin (Hb) difference at birth. METHODS: This was a retrospective cohort study of MCDA twin gestations delivered at Osaka Medical Center and Research Institute for Maternal and Child Health between 2003 and 2012. Cases of pregnancy termination, acardiac twins or intrauterine death were excluded. A large intertwin Hb difference at birth was defined as > 8.0 g/dL according to the postnatal criteria for twin anemia-polycythemia sequence (TAPS). The intertwin reticulocyte count ratio (RCR) was calculated by dividing the reticulocyte count of the anemic twin by that of the polycythemic twin. Cases with Hb differences were divided into two groups according to the RCR, TAPS when the RCR was > 1.7 and acute fetofetal hemorrhage (AFFH) when the RCR was ≤ 1.7. Neonatal outcomes were compared between the TAPS and AFFH groups. RESULTS: During the study period, 432 MCDA twin pregnancies of a total of 532 born at our hospital were analyzed. There were 12 (2.8%) cases of a large intertwin Hb difference. The median gestational age at birth of these cases was 34 (range, 23-38) weeks, and all were delivered by Cesarean section. There were seven (1.6%) cases of TAPS and five (1.2%) of AFFH. The neonatal survival rate was 91.7%; in one pair of twins with TAPS neonatal death occurred. All (100%) cases with TAPS and two (40%) with AFFH required blood transfusion or partial-exchange transfusion for at least one infant. CONCLUSIONS: Although the incidence of TAPS and AFFH may be low in MCDA twins, many affected neonates required treatment for hematological abnormalities. Delivery of MCDA twins via Cesarean section does not appear to prevent AFFH, despite the absence of labor.

Prevalence of radiographic knee osteoarthritis and its association with knee pain in the elderly of Japanese population-based cohorts: the ROAD study.

OBJECTIVE: We investigated the prevalence of radiographic knee osteoarthritis (OA) and knee pain in the Japanese elderly using a large-scale population of a nationwide cohort study, Research on Osteoarthritis Against Disability (ROAD), and examined their association. METHODS: From the baseline survey of the ROAD study, 2,282 participants > or =60 years (817 men and 1,465 women) living in urban, mountainous and seacoast communities were analyzed. The radiographic severity at both knees was determined by the Kellgren/Lawrence (KL) grading system. KL> or =2 and KL> or =3 knee OA were examined separately to assess osteophytosis and joint space narrowing (JSN). RESULTS: The prevalence of KL> or =2 OA (47.0% and 70.2% in men and women, respectively) was much higher than that of previous studies in Caucasians, while that of KL> or =3 OA was not much different in men. Age, BMI, female sex and rural residency were risk factors for radiographic knee OA, knee pain and their combination. The prevalence of knee pain was age-dependent in women, but not in men. Knee pain was more strongly associated with KL> or =3 OA than with KL=2, and the association was higher in men than in women. Female sex was a strong risk factor even in the subgroup without radiographic knee OA (KL=0/1). CONCLUSION: The present cross-sectional study revealed a high prevalence of radiographic knee OA in the Japanese elderly. Knee pain was strongly associated with JSN especially in men, while women tended to have knee pain even without radiographic OA.

Epidemiology of lumbar osteoporosis and osteoarthritis and their causal relationship--is osteoarthritis a predictor for osteoporosis or vice versa?: the Miyama study.

SUMMARY: In a 10-year follow-up of a population-based cohort of Japanese subjects, incidences of and causal relationships between osteoporosis (OP) and osteoarthritis (OA) at the lumbar spine were clarified. OP might reduce the risk of subsequent OA at the spine in women, but not in men. INTRODUCTION: The aim of this study is to clarify the contribution of osteoarthritis (OA) to osteoporosis (OP) and vice versa. METHODS: A population-based, epidemiological study was conducted in a Japanese rural community. From 1,543 participants aged 40-79 years, 200 men and 200 women were selected and followed up for 10 years. Bone mineral density measurements were repeated after 3, 7, and 10 years, and X-rays were repeated after 10 years. RESULTS: The incidence of lumbar OP per 10,000 person-years for persons in their 40s, 50s, 60s, and 70s was 0, 0, 109.5, and 151.1 for men and 124.2, 384.0, 227.3, and 239.5 for women, respectively. The cumulative incidence of lumbar OA over 10 years aged 40-79 years was 25.8% in men and 45.2% in women. Cox's proportional hazards model showed no significant relationship between the presence of lumbar OA at the baseline and incidence of lumbar and femoral neck OP in both genders. A significant relationship was demonstrated between the presence of lumbar OP, not femoral neck OP, at the baseline and cumulative incidence of lumbar OA in women (odds ratio, 0.20; 95% confidence interval, 0.05-0.80; P = 0.02). CONCLUSION: OP in women appears to reduce the future incidence of OA at the lumbar spine.

Prevalence of radiographic lumbar spondylosis and its association with low back pain in elderly subjects of population-based cohorts: the ROAD study.

OBJECTIVES: Although lumbar spondylosis is a major cause of low back pain and disability in elderly people, few epidemiological studies have been performed. The prevalence of radiographic lumbar spondylosis was investigated in a large-scale population study and the association with low back pain was examined. METHODS: From a nationwide cohort study (Research on Osteoarthritis Against Disability; ROAD), 2288 participants aged > or =60 years (818 men and 1470 women) living in urban, mountainous and coastal communities were analysed. The radiographic severity at lumbar intervertebral levels from L1/2 to L5/S was determined by Kellgren/Lawrence (KL) grading. RESULTS: In the overall population the prevalence of radiographic spondylosis with KL> or =2 and > or =3 at the severest intervertebral level was 75.8% and 50.4%, respectively, and that of low back pain was 28.8%. Although KL> or =2 spondylosis was more prevalent in men, KL> or =3 spondylosis and low back pain were more prevalent in women. Age and body mass index were risk factors for both KL > or =2 and KL> or =3 spondylosis. Although KL = 2 spondylosis was not significantly associated with low back pain compared with KL = 0 or 1, KL> or =3 spondylosis was related to the pain only in women. CONCLUSIONS: This cross-sectional study in a large population revealed a high prevalence of radiographic lumbar spondylosis in elderly subjects. Gender seems to be distinctly associated with KL> or =2 and KL> or =3 lumbar spondylosis, and disc space narrowing with or without osteophytosis in women may be a risk factor for low back pain.

Fully automatic quantification of knee osteoarthritis severity on plain radiographs.

OBJECTIVE: Although knee osteoarthritis (OA) is a major public health issue causing chronic disability, there is no objective or accurate method for measurement of the structural severity in general clinical practice. Here we have established a fully automatic program KOACAD (knee OA computer-aided diagnosis) to quantify the major OA parameters on plain knee radiographs, validated the reproducibility and reliability, and investigated the association of the parameters with knee pain. METHODS: KOACAD was programmed to measure joint space narrowing at medial and lateral sides, osteophyte formation, and joint angulation. Anteroposterior radiographs of 1979 knees of a large-scale cohort population were analyzed by KOACAD and conventional categorical grading systems. RESULTS: KOACAD automatically measured all parameters in less than 1s without intra- or interobserver variability. All parameters, especially medial joint space narrowing, were significantly correlated with the conventional gradings. In the parameters, osteophyte formation was associated with none of the joint space parameters, suggesting different etiologic mechanisms between them. Multivariate logistic regression analysis after adjustment for age and confounding factors revealed that medial joint space narrowing and varus angulation of knee joints were risk factors for the presence of pain (594/1979 knees), while neither lateral joint space nor osteophyte area was. CONCLUSION: KOACAD was shown to be useful for objective, accurate, simple and easy evaluation of the radiographic knee OA severity in daily clinical practice. This system may also serve as a surrogate measure for the development of disease-modifying drugs for OA, just as bone mineral density does in osteoporosis.

Association between height loss and bone loss, cumulative incidence of vertebral fractures and future quality of life: the Miyama study.

INTRODUCTION: The study aimed to clarify associations between height loss, bone loss and the quality of life (QOL) score among general inhabitants of Miyama, a rural Japanese community. This population-based epidemiological study was conducted in Miyama, a village located in a mountain area in Wakayama Prefecture, Japan. METHODS: A list of all inhabitants comprising 1,543 inhabitants (716 men, 827 women) born in this village between 1910-1949 was compiled. From the above whole cohort, a subcohort to measure bone mineral density (BMD) was recruited, consisting of 400 participants, divided into four groups of 50 men and 50 women each, and stratified into age decades by decade of birth-year (1910-1919, 1920-1929, 1930-1939 or 1940-1949). BMD measurement, physical measurements of height (cm) and body weight (kg) were taken, and body mass index (BMI; kg/m(2)) were calculated. BMD and anthropometric measurements were repeated on the same participants at 3, 7 and 10 years after baseline measurement (1993, 1997 and 2000). RESULTS AND DISCUSSION: Among 299 of 400 participants, changes in height over 10 years for men in their 40s, 50s, 60s and 70s were -0.7 cm, -0.5 cm, -1.2 cm and -1.5 cm, respectively, compared with -0.7 cm, -1.4 cm, -2.1 cm and -3.7 cm in women, respectively. No significant relationships between change in height and rate of change in BMD at the lumbar spine and femoral neck after adjustment for age in men (lumbar spine, beta = 0.058, standard error of the mean (SE) = 0.031, P = 0.501, R(2) = 0.038; femoral neck, beta = 0.100, SE = 0.038, P = 0.228, R(2) = 0.121) were identified. By contrast, among women, a significant positive association was identified between height change and change rate of BMD at the lumbar spine after adjusting for age (beta = 0.221, SE = 0.039, P = 0.012, R(2) = 0.069), while no significant relationship was found between height change and change rate at the femoral neck (beta = 0.107, SE = 0.039, P = 0.229, R(2) = 0.048). No significant relationship was noted between vertebral fractures (VFx) and height at baseline in men and women (men: odds ratio (OR) 0.93, 95% confidence interval (CI) 0.81-1.05, P = 0.24; women: OR 0.97, 95% CI 0.87-1.08, P = 0.58) or between VFx and height loss (men: OR 1.31, 95% CI 1.00-1.71, P = 0.051; women: OR 1.20, 95% CI 0.94-1.53, P = 0.14). In both men and women, no significant relationship was identified between utility of the EuroQol EQ5D questionnaire and height at baseline (men: beta = -0.148, SE = 0.003, P = 0.202, R(2) = 0.076; women: beta = 0.127, SE = 0.004, P = 0.235, R(2) = 0.048), and height change (men: beta = -0.078, SE = 0.008, P = 0.452, R(2) = 0.065; women: beta = 0.053, SE = 0.010, P = 0.608, R(2) = 0.038).

Novel mutation in exon 18 of the cartilage oligomeric matrix protein gene causes a severe pseudoachondroplasia.

Pseudoachondroplasia (PSACH) is a common skeletal dysplasia characterized by disproportionate short stature, early-onset osteoarthrosis, and dysplasia of the spine, epiphysis, and metaphysis. Multiple epiphyseal dysplasia (MED) is a similar but less severe disorder characterized by dysplasia of the epiphysis. Both disorders are caused by mutations in the cartilage oligomeric matrix protein (COMP) gene. COMP mutations cluster in a region of the gene that encodes calmodulin-like repeats (CLRs) and correlate closely with disease severity. Typically, mutations in exon 13 that composes the seventh CLR produce severe PSACH phenotypes, whereas mutations found elsewhere in the gene produce mild PSACH or MED phenotypes. We have identified a PSACH patient carrying a novel mutation in exon 18 of COMP that composes the C-terminal globular domain. This mutation produced a severe PSACH phenotype with marked short stature and deformities of the spine and extremities. Our results extend the range of disease-causing mutations within the COMP gene and demonstrate the importance of the additional domain of COMP protein in its in vivo function.

Identification of sequence polymorphisms in two sulfation-related genes, PAPSS2 and SLC26A2, and an association analysis with knee osteoarthritis.

Osteoarthritis (OA) is one of the most common musculoskeletal disorders and is characterized by degeneration of articular cartilage. Sulfation of extracellular matrix proteins in articular cartilage is an important step in maintaining normal cartilage metabolism. Two sulfation-related genes have been reported as the causal genes of severe chondrodysplasias: mutations in PAPSS2 (3'-phosphoadenosine 5'-phosphosulfate synthase 2) cause spondylo-epimetaphyseal dysplasia (SEMD), and mutations in SLC26A2 (solute carrier family 26, member 2) cause diastrophic dysplasia. Given their critical roles in cartilage metabolism and the severe phenotypes that result from mutations in these genes, we examined PAPSS2 and SLC26A2 as candidate susceptibility loci for OA. We identified sequence polymorphisms in the coding and core promoter regions of these genes and analyzed their potential association with knee OA within the Japanese population. Ten sequence polymorphisms were detected in PAPSS2 and five in SLC26A2. An association analysis showed suggestive association of one minor polymorphism in the promoter region of SLC26A2. This 4-bp adenine deletion allele, del4A, was over-represented in knee OA (P = 0.043, odds ratio = 3.43) and is thought to confer a minor susceptibility to knee OA within the Japanese population. Haplotype analysis showed no evidence of association with the two genes, however, excluding them as major susceptibility loci for knee OA.

Identification of sequence polymorphisms of the COMP (cartilage oligomeric matrix protein) gene and association study in osteoarthrosis of the knee and hip joints.

Osteoarthrosis (OA) is a common cause of musculoskeletal disability characterized by late-onset degeneration of articular cartilage. Although several candidate genes have been reported, susceptibility genes for OA remain to be determined. Hereditary osteochondral dysplasias produce severe, early-onset OA and hence are models for common idiopathic OA. Among them are pseudoachondroplasia and multiple epiphyseal dysplasia, both of which are caused by mutations in the cartilage oligomeric matrix protein (COMP) gene. Therefore, COMP may be a susceptibility gene for OA. We screened for polymorphisms by direct sequencing of all exons of the COMP gene with their flanking intron sequences and the promoter region. We identified 16 polymorphisms, of which 12 were novel. Using six polymorphisms spanning the entire COMP gene, we examined the association of COMP in Japanese patients with OA of the knee and hip joints. Genotype and allele frequencies of the polymorphisms were not significantly different between OA and control groups, and there was no significant difference in haplotypes. These results do not support an association between COMP and OA in the Japanese population.

Long-term results of rotational acetabular osteotomy in patients with slight narrowing of the joint space on preoperative radiographic findings.

Between 1975 and 1984, we performed rotational acetabular osteotomy in 22 female patients with painful hip dysplasia. At the time of surgery, the patients were in their twenties, and radiographs showed slight narrowing of the joint space. Of these patients, 15 were followed-up for 15 to 22 years (average, 19.8 years) after surgery. The preoperative severity of coxarthrosis in all 15 hips was graded as stage II, according to the classification of coxarthrosis advocated by the Japanese Orthopaedic Association. All 15 patients available for follow-up had had no additional operations on the operated side during the follow-up period. At the time of follow-up, the patients were aged 41 to 48 years (average, 44.3 years). Of the 15 patients, 12 had little or no pain and 14 could walk for more than 30 min without a cane; the severity of coxarthrosis was graded stage I in 3 hips, stage II in 4 hips, stage III in 5 hips, and stage IV in 3 hips. We conclude that rotational acetabular osteotomy is efficacious for patients who have preoperative radiographic findings of slight narrowing of the joint space.

Unresponsiveness of intrahepatic lymphocytes to bacterial superantigen: rapid development of suppressive Mac-1(high) cells in the mouse liver.

We previously found that a small dose (2 microg per mouse) of staphylococcal enterotoxin B (SEB) induced early emerging unresponsiveness in intrahepatic-lymphocyte populations (IHLs). The purpose of this study was to reveal the inducing role of accessory cells involved in IHLs in this phenomenon. IHLs prepared at 3 to 24 hours after SEB injection failed to proliferate in response not only to SEB but also to SEA, representing ligand-nonspecific unresponsiveness, whereas spleen cells (SPCs) and mesenteric lymph-node cells showed transient proliferation. Unresponsiveness in IHLs was related to a deficit of their accessory cell function as measured by coculture of irradiated IHLs and antigen-specific, type 1 T-helper (Th1) clone cells. High levels of nitrite were detected in the culture supernatant. Supplement of N(G)-monomethyl-L-arginine lowered nitrite levels and concurrently restored the proliferative response of Th1 cells, indicating the involvement of nitric oxide in suppression. Adherent cells prepared from IHLs well reproduced these results. As shown by flow cytometry, Mac-1(high) Ia(+) cells, which mainly included F4/80(+) cells (macrophages) and a minor population of CD11c(+) cells (dendritic cells), increased in proportion in IHLs but not in SPCs at 6 to 24 hours. Depletion of Mac-1(high) cells from IHLs with antibody-coated magnetic beads recovered the proliferative response. Depleted Mac-1(high) cells had a monocytoid appearance. In immunostained sections, Kupffer cells came to highly express both Mac-1 and Ia at 12 hours. These results indicate that Mac-1(high)Ia(+) adherent cells, largely Kupffer cells activated by SEB, nonspecifically suppress the proliferation of Th1 cells via nitric oxide production before manifestation of ligand-specific unresponsiveness.

Cultured murine parenchymal liver cells induce differentiation of bone marrow cells to macrophage-like cells which present antigen to Th1 clones but inhibit their proliferation by nitric oxide and prostaglandins.

Suppressor cells were developed from nylon wool nonadherent CD4(-)8(-)TCRbeta(-) bone marrow cells cocultured with parenchymal liver cells for 2.5 days. The major suppressor cell population consisted of nylon wool/plastic dish-adherent, phagocytic Mac-1(+) CD3(-)4(-)8(-) cells (Ad cells), with 34% of the Ad cells being F4/80(+). These Ad cells suppressed the antigen-specific proliferation of Th1 clones in an MHC-nonrestricted manner. They showed a dose-dependent increase in suppressive activity, with both NO and PGE(2) levels in the culture supernatant rising with Ad cell concentration. OVA-pulsed Ad cells (OVA-Ad cells) were found to stimulate IFN-gamma production, resulting in an elevation of the NO and PGE(2) levels in wells containing OVA-specific Th1 clones. No DNA synthesis by these clones was detected in the absence of N(G)-monomethyl-l-arginine and indomethacin, yet the proliferation of the clone was induced in the presence of these chemicals. As proliferation is inhibited by NO and PGE(2) the Ad cells give the impression that they have no antigen-presenting function. This function is MHC-class-II-restricted. If cells such as Ad cells did actually exist in the hepatic sinusoid, they could by their nature play a major role in inducing the early emerging unresponsiveness of T cells in the liver which we reported in a previous paper.

Role of the liver in T cell differentiation--generation of CD3-CD4+/CD8+TCRbeta- cells and CD3-4-8-TCRbeta+ cells from CD4-8-TCRbeta- athymic nude bone marrow cells by culture with parenchymal liver cells.

To investigate the influence of the liver on differentiation of hematopoietic stem cells/pro-T cells, TN-NWP-BMC (athymic nude bone marrow cells that were treated with anti-TCRbeta, anti-CD4, and anti-CD8 Abs plus complement and then passed through a nylon wool column) were cultured on parenchymal liver cells. After culture for 2.5 days, CD3-4-8-TCRbeta+ cells and CD3-CD4+/CD8+TCRbeta- cells were developed from TN-NWP-BMC. TCRVbeta8+ cells comprised 19.9% of CD3-4-8-TCRbeta+ cells, and Vbeta8 mRNA was detected in the CD3-4-8-TCRbeta+ cells by reverse transcriptase-polymerase chain reaction. The CD3-CD4+/CD8+TCRbeta- cells contained not only single-positive cells but also CD4+8+ double-positive cells. The CD8 protein consisted of 88.9% CD8alpha+beta-, 10.1% CD8alpha+beta+, and 1% CD8alpha-beta+ molecules. From these results and the finding of co-expressed antigens, CD3-4-8-TCRbeta+ cells and CD3-CD4+/CD8+TCRbeta- cells appear to be immature cells not committed to a certain cell lineage.

A unique response to staphylococcal enterotoxin B by intrahepatic lymphocytes and its relevance to the induction of tolerance in the liver.

It has been reported that the intrahepatic lymphocyte (IHL) population is somewhat differently constituted from lymphocytes in other lymphoid tissues. Staphylococcal enterotoxin B (SEB) is a superantigen which can induce T-cell tolerance in mice. The authors investigated the in vitro and in vivo responses of mouse IHL to SEB. An intravenous injection of SEB did not result in the augmentation of the proliferative response of IHL, while mesenteric and splenic lymphocytes (mLNC and SPC, respectively) had augmented responses. Interleukin-2 (IL-2) mRNA was clearly detected in mLNC and SPC by reverse transcriptase-polymerase chain reaction (RT-PCR) shortly after the administration of SEB, but it was scarcely expressed in IHL. The expression of CD25 (IL-2 receptor) was also augmented in mLNC and SPC in the early period, while it was not changed in IHL. These findings suggested that the time required for tolerance induction is different locally and that the loss of augmentation of IL-2 and IL-2 receptor production by IHL may be relevant to the rapid induction of T-cell tolerance in the liver.

Production of B cell differentiation factors by mouse parenchymal liver cells.

Previously we reported that most antibody secreting cells secreted IgA in the liver. Here we assessed the possibility that parenchymal liver cells (PLC) produced factors, transforming growth factor (TGF)-beta and IL-5, which participate in the differentiation of B cells to IgA-secreting cells. We showed that TGF-beta activity was present in the culture supernatant of PLC, and IL-5 activity was in the lysate of PLC. Moreover, it was confirmed that IL-5 protein produced by PLC was mainly localized in the cell membrane by histochemical staining. The findings that both TGF-beta and IL-5 were produced by PLC should provide useful information concerning the fact that IgA-secreting cells were dominant in the liver.

Interleukin 7 receptor-deficient mice lack gammadelta T cells.

The interleukin 7 receptor (IL-7R) plays a crucial role in early B- and T-cell development. It consists of a unique a chain and a common gamma chain [IL-2 receptor gamma chain (IL-2Rgamma)]. Gene inactivation of IL-7, IL-7R, and IL-2Rgamma resulted in severe impairment of B and T lymphopoiesis in mice. In addition, IL-2Rgamma-deficient mice lack gammadelta T cells in the skin and have the impaired development of natural killer (NK) cells and intraepithelial lymphocytes. To explore the role of IL-7/IL-7R system in gammadelta T- and NK-cell development, we have generated and analyzed IL-7R-deficient mice. gammadelta T cells were absent from skin, gut, liver, and spleen in the deficient mice. In contrast, alphabeta T and B cells were detected in reduced, but certain, numbers, and NK cells developed normally. The gammadelta T-cell development in fetal and adult thymus was also completely blocked. These results clearly demonstrate that the signal from IL-7R is indispensable for gammadelta T-cell development in both thymic and extrathymic pathways. On the contrary, it is suggested that NK-cell development requires cytokine(s) other than IL-7.

Characteristic immunological tolerance in intrahepatic lymphocytes induced by bacterial superantigen SEB.

We have reported that the liver is closely associated with the hematolymphoid system and that the liver, as a lymphoid organ, may play an important role in the differentiation of lymphocytes and oral tolerance. To investigate the relevance of the liver to immunological tolerance, we compared the response pattern to Staphylococcus enterotoxin B (SEB) in intrahepatic lymphocytes (IHL) with those of spleen cells (SPC) and mesenteric lymph node cells (mLNC) during tolerance induction. Tolerance was induced by i.v. injections of SEB in all three organs. IHL had a distinct pattern from the other cells in the induction of unresponsiveness to SEB. While SPC and mLNC have a proliferation phase before the reduction of DNA synthesis, IHL become tolerant immediately without this phase. When IHL became tolerant in the early period after SEB injection, their nylon-wool column non-adherent (NW-NA) IHL were not recovered from unresponsiveness in vitro in spite of the addition of irradiated normal SPC as accessory cells. NW-NA IHL did not secrete IL-2 during this process, indicating that NW-NA IHL were in a state of functional anergy soon after the i.v. administration of SEB. Phenotypic analysis revealed that the characteristic changes of IHL during the induction of unresponsiveness might be associated with the kinetic modulation of CD4+ V beta 8+ T cells. These results suggest that the liver may hold a different mechanism for the induction of tolerance to SEB from that of SPC and mLNC.

[Modification of surface phenotype and function in intrahepatic lymphocytes during liver regeneration after partial hepatectomy].

In the intrahepatic lymphocyte fraction (IHL) of mice whose livers have been perfused by Ca+2 and Mg+2 free Hank's balance salt solution, flow cytometric analysis reveals various cells similar to those found in the spleen (CD3+ cell, CD4+ cell, CD8+ cell, alpha beta TCR+ cell, gamma delta TCR+ cell, Thy1.2+ cell, B220+ cell and asialo-Gm1+ cell). The cytotoxic activity against YAC-1 cells in IHL is significantly higher than that in spleen cells. Furthermore, IHL has cytolytic activity against syngeneic thymoma BW5147 cells which is not found in spleen cells. Both cytotoxic activities of IHL are greatly weakened by the pretreatment of IHL with anti asialo-Gm1 Ab and rabbit complement. During liver regeneration after a partial hepatectomy, the percentages of lymphocyte subsets of IHL such as alpha beta TCR+ cells, gamma delta TCR+ cells and asialo-Gm1+ cells transiently increase 2 days after the partial hepatectomy and their cytotoxic activities against YAC-1 and BW5147 cells also reach a peak at that time. It is well known that the peak of mitotic index in regenerating parenchymal liver cells peaks 2 days after a partial hepatectomy and that the number of Pit cells peaks 10 to 14 days after. Compared with the time for reaching the highest mitotic index in regenerating parenchymal liver cells, the proliferation of Pit cells comes very late. It has not been clear which proliferation process in parenchymal liver cells is suppressed by activated Pit cells. Our findings in this paper reveal that the percentages and the cytotoxic activities of asialo-Gm1+ cells in IHL reach their peak 2 days after an operation.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth control of primary culture hepatocytes by nonparenchymal liver cells. Role of interferon produced by liver sinusoidal cells.

Using the primary culture of liver cells, we showed that interferon produced by nonparenchymal liver cells inhibits the proliferation of cultured parenchymal liver cells. DNA synthesis of parenchymal liver cells was suppressed not only by their coculture with nonparenchymal liver cells but also by the addition of the culture supernatant of nonparenchymal liver cells. The suppressive activity of the supernatant correlated closely with the interferon (alpha + beta) level in the supernatant and was reduced by anti interferon (alpha + beta) serum. Furthermore, purified interferon (alpha + beta) also suppressed parenchymal liver cell proliferation in a dose-dependent manner and the suppression was released by anti interferon (alpha + beta) serum. The interferon level of the supernatant necessary for suppressing parenchymal liver cell proliferation, however, was extraordinarily low compared with purified interferon. The possibility exists that IFN in the culture supernatant of nonparenchymal liver cells works synergistically with other factors in the supernatant to suppress the cell proliferation.