Search

The Am and Bm phenotypes are characterized by weak expression of the A or B antigens, respectively, by red blood cells with a normal expression by the saliva of secretors. Deletion of the regulatory element in the first intron of the ABO gene and disruption of the GATA motif in the element were found to be responsible. In this study, we identified a novel mutation within the GATA motif (G>C substitution at position c.28 + 5830) in the regulatory element of the A allele that might diminish transcription activity causing the generation of the Am B phenotype.

Rare blood is generally defined as one that occurs at a frequency of 1:100~1000 individuals or less, and it is sometimes difficult to provide such blood types to patients because of their rarities. The Japanese Red Cross (JRC) Society lists 46 rare blood phenotypes that are divided into two categories. The rare blood types listed in Category I occur much less frequently than those listed in Category II. We screen for rare blood cells using monoclonal antibodies (MoAbs). Since 1987, our blood centre has established 93 MoAbs (32 human and 61 murine) and has provided them to the other blood centres. Many of IgG MoAbs are available on the machine by saline or bromelin method by cross-linking with anti-human or anti-mouse IgG. Thus, more than 10 000 donors with rare blood phenotypes (Category I, 748; Category II, 9314) are registered in Japan. We freeze rare blood, particularly Category I types. Since 1977, a total of 576 units of rare blood with phenotypes Di(b−), D−−, Jr(a−), Ko and Lan-, etc. have been supplied to 23 international countries. Thus, the JRC contributes to the International Panel of Donors of Rare Blood Type (IDP) which is maintained by the International Blood Group Reference Laboratory (IBGRL) in Bristol, UK. The IDP provides information on the location of rare blood donors when they cannot be found in their respective countries. We also joined the ISBT Working Party on Rare Donors which handles all matters related to rare blood. Our rare blood donor programme is successful because of international co-operation.

Radio wave emission due to rock fracture was measured at four frequency bands. Though the antenna gain is difficult to measure at the lowest frequency of 1MHz, the measuring system was successfully calibrated, and the emitted energy was estimated. The obtained results at 1MHz to 18GHz suggest the wave source to be quite narrow electrical signals, and show a possible application to the detection of an earthquake or volcanic activity.

We investigated the expression of IL-8 and the IL-8 receptor (IL-8R) in human carcinoma cells and the role of IL-8 and IL-8R in the growth of carcinoma cells. IL-8 mRNA was detected in 16 of 20 (80%) carcinoma cell lines and 20 of 24 (83.3%) cancer tissues by Northern blot analysis. IL-8R mRNA was expressed in 7 of 11 (63.6%) carcinoma cell lines by reverse transcriptase polymerase chain reaction (RT-PCR). Neutrophil chemotactic activity in the culture supernatant of carcinoma cell lines correlated with immunoreactive IL-X concentration. Growth of carcinoma cells was significantly inhibited in the presence of anti-IL-8 antibody or IL-8R antisense oligonucleotide. These results revealed that IL-8 and IL-8R are expressed in the majority of carcinoma cells and suggest that they might play a role in the growth of carcinoma cells.

Expression of low-affinity receptors for IgE (FcɛRII) on T and B cells was examined with monoclonal antibodies to cell surface antigens of FcɛRII (CD23), T cells (CD3) or B cells (CD 19) by two-dimensional analysis under laser flow cytometry system. The majority of cells bearing FcɛRII were B cells; however, a small proportion of T cells (1. 6%) bore FcɛRII in children with atopic asthma and elevated serum level of IgE. The occurrence of FcɛRII positive B cells was also increased in children with atopic asthma compared with non-atopic controls (p < 0.001). The numbers of T and B cells hearing Fc receptors for IgG (FcɛR) did not differ between atopic and non-atopic children. T cells were isolated from venous blood by rosetting with sheep erythrocytes, and then cultured in vitro with phytohemagglutinin plus phorbol-myristate-acetate. The amount of IgE-binding factors (IgE-BF), a soluble form of FcɛRII, was determined by means of enzyme-linked immunosorbent assay. The level of IgE-BF was increased in cultures from children with a atopic asthma compared with non-atopic controls (p < 0.01). The possible involvements of FcɛRII on T cells and their soluble products, IgE-BF, in the pathogenesis of atopic asthma is discussed.

During antigen presentation, CD4 functions to stabilize T cell receptor (TCR)-class II MHC interactions and coordinate Ag-induced T cell activation signals. These activation signals cause CD4 down-regulation, presumably acting to optimize T cell activation. We previously reported that oxidative stress interferes with activation-induced CD4 down-regulation in T cells. In this study, we have further investigated inhibition of CD4 down-regulation by oxidative stress and its role for T cell activation. A construct comprised of the mouse FcgammaRIIB extracellular domain and the transmembrane/cytoplasmic domains of human CD4 (FcgammaR/CD4) was expressed in a human T cell line. Oxidant actually potentiated down-regulation of the FcgammaR/CD4 chimera and induced Lck dissociation from both CD4 and FcgammaR/CD4, which is a crucial intracellular process for activation-induced CD4 down-regulation, suggesting a critical role of CD4 ectodomain in the inhibition of CD4 down-regulation by oxidative stress. Furthermore, insertion of CD4 D3-D4 membrane proximal extracellular region between FcgammaR extracellular domain and CD4 transmembrane/cytoplasmic domains in FcgammaR/CD4 chimera made this molecule behave like native CD4 molecule under oxidative stress condition. These data imply that the inhibitory effect of oxidative stress on CD4 down-regulation is executed via D3-D4 domain of CD4 ectodomain. As to its role for T cell activation, CD4 coaggregation with CD3 under the oxidative conditions enhanced activation signal induced by CD3 aggregation. Our results demonstrate that Ag-induced T cell activation which is normally concomitant with CD4 down-regulation may be disturbed through the aberrant regulation of CD4 expression by oxidative stress.

We established a T cell line, STO-5, which constitutively produced monocyte/macrophage chemotactic activity via human T cell lymphoma-leukemia-virus-induced transformation of normal human T cells.We isolated and purified a lactose-binding protein, MCF-pl5-L (MW of about 50 kD, pl of about 5) from a conditioned medium of STO-5. By using highly purified MCF-pl5-L, its biological and physicochemical properties were elucidated in comparison with C5a and MCP-1.MCF-pl5-L exhibited an evident dose-dependent monocyte chemotactic activity (MCA). MCF-pl5-L had no or little affinity for heparin unlike chemokines such as MCP-1. We further found that MCF-pl5-L exhibited potent chemotactic activity not only for monocytes but also for alveolar macrophages. In contrast, C5a and MCP-1 failed to show evident chemotactic activity for alveolar macrophages though they did show MCA. MCF-pl5-L failed to exhibit evident eosinophil and neutrophil chemotactic activities, indicating its chemotactic activity is selective for monocytes/macrophages. Regarding the biological functions of MCF-pl5-L other than MCA and chemotactic activity for alveolar macrophages, we found that MCF-pl5-L but not C5a and MCP-1 could prolong the life span of alveolar macrophages, probably by inhibiting apoptosis of macrophages, and stimulate the production of TNF-alpha from macrophages.These results suggest that MCF-pl5-L plays a role as an immune modulator for monocytes/macrophages in the site.

CD80 and CD86 were detected in high amounts on circulating T cells in the peripheral blood of some patients with systemic lupus erythematosus (SLE), using flow cytometry and monoclonal antibodies. Patients with other connective tissue diseases did not have a high percentage of T cells expressing CD80 or CD86 in their peripheral blood. CD80 was expressed mainly on CD4 T cells, whereas CD86 was expressed on CD8 T cells, and these two populations were associated with particular clinical features. These two molecules were expressed on different T-cell populations and might have different roles in the generation and regulation of immune responses. Since high expression of CD86 on T cells was detected much earlier than the appearance of clinical features and a high titer of anti-DNA antibody, it may be a useful parameter for predicting the flare of SLE.

Gastric scirrhous cancer was classified from the radiological findings into three phases (typical, latent and early phases) by cancerous involvement in the gastric wall. Radiological findings in the typical phase were divided into a giant-fold type and a no giant-fold type. We made a differential diagnosis for scirrhous cancer and other diseases. Radiological findings in the typical phase of scirrhous cancer were characteristic stiffness and rigidity of the gastric wall; in the latent phase, they were swelling and winding of mucosal folds in appropriate distended stomach; and in the early phase, interrupted folds around barium fleck. The effect of chemotherapy against a gastric scirrhous cancer is decided by distensibility of the gastric wall in radiological findings.

In order to integrate quality of life as the end point in the cancer drug therapy, a questionnaire form has been developed over the past four years by a Grant-in-Aid for Cancer Research from the Ministry of Health and Welfare, Japan. Fifty-one items generated by questionnaires were forwarded to oncologists, nurses, psychologists and 121 patients. Then questionnaires were scaled down to 36 items based upon factor analysis considering correlation to State-Trait Anxiety Inventory (STAI) and Self-rating Depression Scale (SDS). These 36 items were administered to 536 patients. Finally, a 22-item questionnaire was completed by rechecking correlations with PS, STAI, SDS and Cronbach's alpha. It includes such components as physical activities (6 items), psychological (5 items) and social variables (5 items), symptoms (5 items) and global well-being (1 item: face scale). In our treatment of G. I. cancer by chemotherapy, NC patients showed improvement (+1.56 score), especially in physical functions (+2.43 score), but with slight deterioration (-0.25 score).

Background The prevalence of bronchial asthma (BA) including exercise-induced bronchoconstriction (EIB) and airway hyperresponsivenss (AHR) is increasing not only among normal population but also among athletes and prevention of these attacks are just as life saving and crucial for the athlete. The negative impact on athletic performance and the effect of inhaled corticosteroids and ƒA2-agonists are factors to providing prevention strategies in these athletes. Objective To determine the prevalence of BA (including silent BA) and develop an effective protocol to detect BA and provide preventive measures in elite athletes. Methods Asthma questionnaires and spirometry were conducted for all regardless of their clinical history. Bronchodilator tests were administered to those who met our criteria of past history, suspicious symptoms, values of FEV1%, and concave shape of flow-volume curves. Bronchial provocation tests were held to detect EIB and AHR in some athletes with suspicious symptom. IOC-criteria were used to consider those tests to be positive. Diagnosis of BA was made by consideration of all factors. Participants 347 male and 305 female Japanese elite athletes participating at the Beijing Olympic Games 2009 and the Vancouver Olympic Games 2010. Main outcome measurements Detecting BA and silent BA is important to prevent BA attacks and begin early intervention strategies in elite athletes in view of their performance and doping violence. Results 21 athletes met IOC criteria in bronchodilator test. The provocation test was positive for asthma in four cases. The prevalence of BA among Japanese elite athletes was 12.1%. Number of Asymptomatic athletes diagnosed as BA reached to 2.9%. Conclusion The prevalence of BA among Japanese elite athletes is similar to prevalence of athletes reported and our protocol can sufficiently detect BA and silent BA as well.

Adult T cell leukemia-derived factor (ADF), originally defined as an IL-2 receptor alpha-chain (IL-2R alpha)/p55 (Tac) inducer, is a human thioredoxin homologue and has many cytokine-like activities. In this study, we examined the regulatory effect of ADF on eosinophil migration using human eosinophils and an eosinophilic subline of HL-60 human promyelocytic leukemia cells, YY-1. rADF induced migration of eosinophils from patients with hypereosinophilia, although rADF exhibited little activity on eosinophils from healthy donors. When human eosinophils were incubated with rADF (0.1-10 micrograms/ml) at 37 degrees C for 24 h, both chemotactic and chemokinetic activity of the complement anaphylatoxin peptide C5a on eosinophil migration was markedly enhanced in a dose-dependent manner. Similarly, this enhancing effect of rADF was observed in the migration assay using YY-1 cells. In contrast, rADF showed no modulation of migratory behavior of human eosinophils and YY-1 cells by IL-3, IL-5, nor granulocyte-macrophage colony-stimulating factor. Scatchard analysis of C5a receptors on YY-1 cells using 125I-C5a showed that rADF modulated neither the density nor the affinity of the cell membrane significantly. Furthermore, mutant ADF (mADF), which had no reducing activity, had no enhancing effect on C5a-induced eosinophil migration. These results indicate a possible involvement of ADF in the recruitment of eosinophils through redox regulation by a dithiol reductase activity.

An adult T cell leukemia cell line, HIL-3, constitutively secretes a factor which induces the phenotypical and functional eosinophilic differentiation of a human eosinophilic leukemia cell line, EoL-1. Biochemical characteristics of the factor, termed eosinophilic leukemia cell differentiation factor (ELDF), were examined. ELDF was precipitated by 35 to 65% saturated ammonium sulfate from the culture supernatants of HIL-3 cells (HIL-3 sup). ELDF was eluted in a peak corresponding to a molecular weight of 30 to 40 kd by gel filtration. Isoelectric focusing in the Rotofor showed that ELDF had isoelectric points of 5 to 6. ELDF was trypsin-sensitive and stable to heat treatment at 65 degrees C for 30 minutes but labile at 80 degrees C or pH lower than 3. Half of the activity adhered to lentil-lectin but not to Con-A, indicating that a part of ELDF is glycoprotein with an N-linked carbohydrate moiety, which did not seem to be essential for ELDF activity. The biochemical characteristics of ELDF and blocking experiments using cytokine-specific neutralizing antibodies suggest that ELDF is different from gamma-interferon (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-5 (IL-5) and interleukin-2 (IL-2), which may exist in HIL-3 sup, and that ELDF may be a previously unrecognized leukemia differentiation factor.

In guinea pig system, we have shown that a T cell-derived eosinophil chemotactic factor (ECF) plays a role especially in delayed-in-time tissue eosinophilia. In Kimura's disease characterized by blood and tissue eosinophilia, OKT4-positive T cells from the patients spontaneously produce an ECF (LDECF-HES; pI 6) without any additional stimulation. A monocyte-derived factor with mw of greater than 100,000 induce LDECF-HES production from T cells. On the other hand, OKT4-positive T cells from patients with parasite infection produce another ECF (LDECF-PD; pI 7-8) during mitogenic or antigenic stimulation. LDECF-PD production is selectively potentiated by ECF-potentiating factor derived from monocytes. Furthermore we have shown that the two ECF differ in their effects on eosinophil chemotaxis, eosinophil cationic protein release and Fc receptor expression. Recently we have established a T cell line, STO-2, constitutively producing 5 ECF with different pI (5, 6, 7, 8 and 9). These 5 ECF have different biological activities in eosinophil survival, Fc receptor expression and induction of specific eosinophilic granules. Comparison of chemotactic response of eosinophils from various patients with eosinophilia to respective ECF has revealed that eosinophils are heterogeneous in their chemotactic activity.

We established a T-cell line, STO-2, by human T-cell lymphoma-leukemia virus-induced transformation of normal human T cells. Partial purification with isoelectric electrophoresis revealed that STO-2 liberated several eosinophil chemotactic factors (ECF) for eosinophils from healthy individuals with different isoelectric point of PI5, PI6, PI7, PI8, and PI9. Molecular weight of all the ECF was about 30,000 to 45,000. None of the ECF except ECF-PI5 was suppressed when they were incubated with monoclonal antibodies against IL-3, IL-5, and GM-CSF together, suggesting that ECF activity of ECF-PI5 is mainly comprised of IL-3, IL-5, and GM-CSF. ECF-PI5, PI6, and PI7 also exhibited enhancing activity on ex vivo eosinophil survival whereas ECF-PI8 and PI9 failed. Expression of Fc epsilon receptor II on eosinophils was potentiated by ECF-PI6 and ECF-PI7. In contrast, expression of Fc gamma receptor III was potentiated by ECF-PI7, ECF-PI8, and ECF-PI9. ECF-PI6 could also change an eosinophilic cell line, EOL-1, to eosinophilic granule-positive cells, whereas the rest of ECF failed. The above results suggested that eosinophils attracted by an ECF exhibit their biological functions, which differ from those of eosinophils attracted by other ECF. In further experiments, the chemotactic response of eosinophils from patients with eosinophilia was compared to that of eosinophils from healthy individuals.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytokine LD78 is a member of a newly identified cytokine superfamily. We cloned the third human gene for the LD78, termed LD78 gamma and the sequence analysis showed that it is a 5'-truncated pseudogene. Exons 2 and 3 and the intron between them are highly homologous to those of the LD78 beta gene, hence, the gamma gene was probably derived from the beta gene. Southern blot analysis of human x mouse somatic hybrid cell DNAs and in situ hybridization experiments mapped all the three gene loci on human chromosome 17q21.1-q21.3. Analysis of DNAs from family members supports our previous finding that the beta and gamma genes on each of the paired chromosome 17 vary in copy number and that the LD78 alpha gene is presumably a single copy. In our analyses of cosmid clones, the LD78 beta gene and the second gene for AT744, which is also a member of the superfamily are closely linked in a head-to-head arrangement. The mechanism of generation of the three LD78 genes is discussed.

We have previously isolated two OKT4-positive T lymphocyte-derived eosinophil chemotactic factors (LDECF) with MW of about 45-60 kDa of which production is different in antigen or mitogen dependency (1-4). The production of a LDECF from patients with parasite disease (LDECF-PD) is dependent on antigen or mitogen stimulation, whereas another LDECF from patients with hypereosinophilic syndrome (HES) is independent. Further purification of these LDECF with isoelectric focusing reveals that an isoelectric point of LDECF-HES is about 6.0 and that of LDECF-PD is around 7.0 to 8.0. Little or no activity of partially purified LDECF-HES and LDECF-PD is suppressed by treatment with monoclonal antibodies against GM-CSM, IL-3, and IL-5, which activate eosinophils. LDECF-HES and LDECF-PD attract eosinophils from healthy individuals. In contrast, eosinophils from patients with HES are attracted by LDECF-HES but not LDECF-PD. LDECF-HES enhances the expression of Fc epsilon receptor II (Fc epsilon RII) and Fc gamma receptor III (Fc gamma RIII) but not that of CR1 on eosinophils, whereas LDECF-PD enhances their CR1 and Fc gamma RIII expression but not Fc epsilon RII expression. Moreover, treatment with LDECF-PD suppresses the release of eosinophilic cationic protein (ECP) from eosinophils, whereas that with LDECF-HES fails. Treatment of eosinophils with phorbol myristate acetate enhances ECP release from eosinophils but it fails to enhance the intracellular ECP level. However, the intracellular ECP level is elevated by stimulation with phorbol myristate acetate if eosinophils were previously treated with LDECF-HES but not with LDECF-PD. These results suggest that various kinds of LDECF are produced according to the nature of diseases, and that each LDECF has functional heterogeneity on eosinophils.

Adult T cell leukemia-derived factor (ADF), originally defined as an IL-2 receptor alpha-chain (IL-2R alpha)/p55 (Tac) inducer, is a human thioredoxin homologue and has many cytokine-like activities. In this study, we examined the regulatory effect of ADF on eosinophil migration using human eosinophils and an eosinophilic subline of HL-60 human promyelocytic leukemia cells, YY-1. rADF induced migration of eosinophils from patients with hypereosinophilia, although rADF exhibited little activity on eosinophils from healthy donors. When human eosinophils were incubated with rADF (0.1-10 micrograms/ml) at 37 degrees C for 24 h, both chemotactic and chemokinetic activity of the complement anaphylatoxin peptide C5a on eosinophil migration was markedly enhanced in a dose-dependent manner. Similarly, this enhancing effect of rADF was observed in the migration assay using YY-1 cells. In contrast, rADF showed no modulation of migratory behavior of human eosinophils and YY-1 cells by IL-3, IL-5, nor granulocyte-macrophage colony-stimulating factor. Scatchard analysis of C5a receptors on YY-1 cells using 125I-C5a showed that rADF modulated neither the density nor the affinity of the cell membrane significantly. Furthermore, mutant ADF (mADF), which had no reducing activity, had no enhancing effect on C5a-induced eosinophil migration. These results indicate a possible involvement of ADF in the recruitment of eosinophils through redox regulation by a dithiol reductase activity.

Splenic T lymphocytes of rats treated with complete Freund's adjuvant (CFA) spontaneously release a lymphokine that inhibits the glycosylation of IgE-binding factors during their biosynthesis and provides the factors with biologic activity: selective suppression of the IgE response. The lymphokine, which is called glycosylation-inhibiting factor, also prevents IgE-induced increases in Fc epsilon R+ cells. The glycosylation inhibiting factor is formed by stimulation of BCG-primed spleen cells with PPD and participates in the selective formation of IgE-suppressive factors. The lymphokine is derived from OX 8+ T cells in both the CFA and BCG systems. The glycosylation-inhibiting factor is a 16,000-dalton peptide, as estimated by gel filtration, and specifically binds to monoclonal antibody against lipomodulin, a phospholipase inhibitory protein. Furthermore, "lipomodulin" was detected by radioimmunoassay in the 16,000-dalton fraction that contained glycosylation inhibiting factor. The fraction did not inhibit phospholipase A2 but after alkaline phosphatase treatment, the fraction did inhibit phospholipase A2. Furthermore, purified lipomodulin obtained from glucocorticoid-treated rabbit neutrophils had the same biologic activities as glycosylation inhibiting factors; i.e., it inhibited both protein glycosylation of IgE-binding factors and IgE-induced expression of Fc epsilon R. The results collectively indicate that glycosylation-inhibiting factor is a fragment of phosphorylated lipomodulin.

Incubation with rat IgE of rat mesenteric lymph node cells obtained 8 days after infection with Nippostrongylus brasiliensis (Nb) resulted in the formation of soluble factors with affinity for IgE, i.e., IgE-binding factors(s). The factors were derived from T lymphocytes and were able to suppress an in vitro IgE response without affecting the IgG response. The minimum concentration of rat IgE for the induction of factor formation was 0.3 to 1.0 microgram/ml. Gel filtration of culture filtrates of the IgE-containing culture identified 2 components with IgE-binding activity; one component had a m.w. of between 10,000 and 20,000, and another component had a m.w. of between 25,000 and 50,000. Both factors could be purified by binding to IgE-Sepharose followed by elution at acid pH. Among the two IgE-binding factors, the lower m.w. component had the ability to suppress selectively the IgE response. In contrast to IgE-potentiating factor, which also had affinity for IgE, the IgE-suppressive factor failed to bind to lentil lectin Sepharose. Formation of IgE-specific suppressive factor was not limited to the lymphocytes obtained 8 days after Nb-infection. Incubation with IgE of lymphocytes obtained 4 wk after infection resulted in the formation of both IgE-specific suppressive factor and IgE-potentiating factor, and the activity of the suppressive factor was greater than that of the potentiating factor.

Mesenteric lymph node (MLN) cells from rats treated with complete Freund's adjuvant (CFA) spontaneously released soluble factors, which have affinity for IgE (IgE-binding factors). The factors were purified by absorption of culture filtrates of the lymphocytes with IgE-Sepharose followed by elution at acid pH. Purified IgE-binding factors selectively suppressed IgE-forming cell response of DNP-ovalbumin-primed MLN cells to homologous antigen without affecting the IgG2 response. Fractionation of purified IgE-binding factors on lentil lectin-Sepharose showed that the majority of IgE-binding factors did not have affinity for lentil lectin, whereas a minor fraction was bound to the lectin-coupled beads. It was found that IgE-binding factors lacking affinity for lentil lectin suppressed IgE response, whereas those eluted from lentil lectin-Sepharose selectively enhanced IgE response. Serum of CFA-treated animals also contained both IgE-suppressive factor and IgE-potentiating factor, and the amount of the former factor was more than the latter. The source of IgE-suppressive factor is non-B lymphocytes. Depletion of Fc, receptor(+) lymphocytes in MLN cells from CFA-treated animals did not affect the formation of IgE-suppressive factor, indicating that Fc epsilon R-bearing cells are not the source of the factor.

Previous experiments have shown that Fc epsilon receptor-bearing (Fc epsilon R(+)) T lymphocytes in mesenteric lymph nodes (MLN) of rats infected with Nippostrongylus brasiliensis (Nb) release a soluble factor that selectively potentiates the IgE response. The IgE-potentiating factor has affinity for IgE, and can be detected by the ability to inhibit rosette formation of Fc epsilon R(+) cells with IgE-coated erythrocytes. The factor was bound to IgE-coated Sepharose and was eluted from the beads at acid pH. It was also found that the IgE-potentiating factor binds to lentil lectin-Sepharose, indicating that the factor is a glycoprotein. The affinity of the factor for IgE was lost after treatment with trypsin but was maintained after treatment with neuraminidase. However, the ability of the factor to potentiate the IgE response was lost after neuraminidase treatment. The results suggested that the factor's binding site for IgE is associated with a protein (peptide) moiety but that its carbohydrate moiety is essential for its biologic activity. When MLN cells were incubated at 37 degrees C, a substantial amount of the IgE-potentiating factor was released into culture medium within 4 hr even in the presence of cycloheximide. Pretreatment of the cells with trypsin, which removed Fc epsilon R, markedly diminished the release of IgE-potentiating factor, suggesting that the factor is derived from Fc epsilon R on the cell surface.

The possible mechanism of eosinophilia was studied in rats undergoing primary infection with Nippostrongylus brasiliensis (Nb). In vivo studies showed that the kinetics of intestinal tissue eosinophilia was not directly related to those of the intestinal worm burden. Furthermore, Nb worm extract has no or only very weak in vitro eosinophil chemotactic activity, suggesting that parasite-derived eosinophil chemotactic factor (ECF) is, if at all, not a major regulator for intestinal tissue eosinophilia in this Nb rat system. On the other hand, when mesenteric lymph node (MLN) cells obtained various days after infection were cultured, potent ECF activity was detected in the cell-free supernatant from the cultures of MLN cells 15–20 days after infection, at which time marked intestinal tissue eosinophilia was observed in vivo. Production of ECF by MLN cells from Nb-infected rats seems to be spontaneous, since these cultures were performed without adding worm antigen. ECF-producing activity of day-20 MLN cells was suppressed by adding various metabolic inhibitors such as cycloheximide, mitomycin C, or puromycin. After Sephadex G-75 gel filtration, ECF activity produced by day-20 MLN cells was associated with two different molecules.

T lymphocytes in the mesenteric lymph nodes of rats infected with Nippostrongylus brasiliensis spontaneously released a soluble factor that selectively potentiated the IgE-forming cell response of antigen-primed cells to homologous antigen. The factor could enhance the IgE response of DNP-OA-primed cells to DNP-HSA and T cell-replacing factor. In contrast, the treatment of OA-primed T cells with the factor failed to enhance either the IgE or IgG response of the mixture of DNP-KLH primed cells and OA-primed T cells to DNP-OA. The results collectively suggested that the target cells of the IgE-potentiating factor are B cells. Indeed, IgE potentiating factor was absorbed by B cells rather than T cells or thymocytes. Evidence was obtained that IgE-potentiating factor could be absorbed by IgE-bearing B cells or IgE-coupled Sepharose, indicating that the factor had affinity for IgE. It appeared that the potentiating factor bound to IgE-bearing B cells and selectively enhanced the differentiation of IgE-B cells to IgE-forming cells. It was also found that the major source of the factor was Fc epsilon R-bearing T cells.

An i.p. injection of Bordetella pertussis vaccine (BP) into rats induced the formation of soluble factors that had affinity for IgE (IgE-binding factors). The factor was detected in the serum of BP-treated animals 5 to 7 days after the treatment. Their circulating lymphocytes as well as spleen cells spontaneously released IgE-binding factors in the serum of BP-treated rats and those released from their circulating lymphocytes had affinity for lentil lectin, and the ability to selectively potentiate an in vitro IgE response of DNP-OA primed cells to homologous antigen. The molecular size of IgE-potentiating factor was between 10,000 and 20,000, and was comparable to that formed by lymphocytes of rats infected with Nippostrongylus brasiliensis. Evidence was obtained that IgE-potentiating factor was derived from Fc epsilon R(+) T cells, with a T cell marker identified by monoclonal antibody W 3/25. Their production of IgE-potentiating factor may be the basis of the adjuvant effect of BP on the IgE response.

Incubation of rat lymphocytes with homologous IgE induced an increase in Fc epsilon R(+) lymphocytes and the formation of IgE-binding factors. Pretreatment of rat lymphocytes with 1 to 5 microM dexamethasone, however, prevented the IgE-induced expression of Fc epsilon R on both B and T lymphocytes. Upon incubation with IgE, T cells activated with 10 micrograms/ml Con A produced IgE-potentiating factors that had affinity for lentil lectin and selectively enhanced the IgE response. Pretreatment of the Con A-activated T cells with dexamethasone, before incubation with IgE, changed the nature of IgE-binding factors formed by the cells. The majority of IgE-binding factors formed by the dexamethasone-treated, Con A-activated cells failed to bind lentil lectin Sepharose and selectively suppressed the IgE response. An injection of 0.2 mg dexamethasone into rats infected with Nippostrongylus brasiliensis markedly diminished the proportion of Fc epsilon R(+) lymphocytes in their mesenteric lymph nodes. The lymph node cells from the infected animals spontaneously released IgE-potentiating factors in vitro. However, the majority of IgE-binding factors formed by the mesenteric lymph node cells from the dexamethasone-treated, Nb-infected animals lacked affinity for lentil lectin and selectively suppressed the IgE response. The results indicate that glucocorticoid treatment prevented the glycosylation of IgE-binding factors and thereby changed the biologic activities of the factors.

Differences between the conditions for an eosinophil chemotactic factor (ECF) and macrophage chemotactic factor (MCF) production by lymphoid cells of mesenteric lymph nodes and spleen were studied in guinea pigs. If lymphoid cells were washed less than 4 hr after concanavalin A (Con A) stimulation and were cultured for an additional 24 hr, they failed to produce ECF, whereas Con A stimulation for 1 hr before washing was sufficient to stimulate them to produce MCF. Subsequently, it was shown that heat-labile soluble factors (termed ECF-PF) with potentiating activity for ECF production are produced from macrophages by 5 micrograms/ml Con A activation. When ECF-PF were added to the cell culture with 5 micrograms/ml Con A, the lymphoid cells could produce ECF even when they were washed 2 hr after Con A stimulation and were cultured for an additional 24 hr, suggesting that ECF-PF plays a critical role in the early stage of ECF production. The lymphoid cells were also able to produce ECF even when they were cultured with ECF-PF and a suboptimal dose of Con A (1 microgram/ml) for ECF production. Protein synthesis seemed to be essential for ECF-PF production. The ECF-PF activity was associated with two separated molecular fractions with m.w. of about 50,000 to 70,000 and of 10,000 to 20,000. It is thus suggested that ECF is produced from T cells by Con A stimulation under conditions which differ, at least, from those for MCF in the requirement of ECF-PF.

Delayed tissue eosinophilia in DNP-ovalbumin-induced allergic inflammatory skin lesions of guinea pigs was markedly enhanced by previous treatment with alum hydroxy gel (Alum) or Bordetella pertussis vaccine. This enhancement seemed due to increased production of a lymphocyte-derived eosinophil chemotactic factor (ECF) at the skin site. Treatment of animals with Alum potentiated antigen-induced in vitro ECF production by lymphoid cells from spleen and mesenteric lymph node of sensitized animals. The co-culture supernatants of lymphoid cells from Alum-treated animals also potentiated concanavalin A (Con A)-induced in vitro ECF production. The potentiating effect of Alum on ECF production seemed to be ascribed to the release of soluble factors from macrophages of the Alum-treated animals. The macrophage-derived soluble factor ECF-potentiating factor (ECF-PF) selectively potentiated ECF production but not macrophage chemotactic lymphokine production by Con A-stimulated lymphoid cells from normal animals. ECF-PF activity was associated with two separate m.w. fractions: one was 50,000 to 70,000 and the other was 10,000 to 20,000. The present study provides one of the explanations for enhanced ECF production by adjuvants, such as Alum and Bordetella pertussis vaccine.

Attempts were made to induce the formation of soluble factors having affinity for IgE (IgE-binding factors) by activated T cells. Normal rat mesenteric lymph node (MLN) cells were cultured in 1 microgram/ml Con A for 48 hr or in 10 microgram/ml Con A for 72 hr, and Con A-activated lymphocytes were incubated with rat IgE. It was found that IgE induced an increase in the proportion of Fc epsilon R(+) cells in the Con A-activated cells and formation of IgE-binding factors. However, the nature of IgE-binding factors formed by the cells were different depending on the concentration of Con A for activation. Thus, IgE-binding factors formed by 10 microgram/ml Con A-activated cells had a high affinity for lentil lectin and enhanced the IgE-forming cell response of DNP-OA primed cells. In contrast, the majority of IgE-binding factors formed by 1 microgram/ml Con A-activated cells failed to bind to lentil lectin and suppressed the IgE response. Induction of Fc epsilon R by IgE on Con A-activated cells was prevented by tunicamycin, which inhibits protein glycosylation. This antibiotic did not prevent the IgE-induced formation of IgE-binding factors but changed the nature of the factors formed by 10 microgram/ml Con A-activated cells. The majority of IgE-binding factors formed in the presence of tunicamycin lacked affinity for lentil lectin and Con A, and suppressed, rather than enhanced, the IgE response. The nature and amount of IgE-binding factors formed by 1 microgram/ml Con A-activated cells was not affected by tunicamycin. The results suggest that glycosylation of IgE-binding factors during their biosynthesis is an important step in determining their biologic function.

Hypertrophic cardiomyopathy (HCM) generally shows increased systolic function of the left ventricle at rest, although patients with HCM often have decreased exercise tolerance and develop dyspnea or chest pain. The present study was to investigate of left ventricular (LV) function during exercise in 26 patients with HCM using Tc-99m equilibrium angiocardiography, and to elucidate the mechanism of impaired functional reserve during exercise. Controls consisted of 11 normal volunteers and 12 patients with chest pain syndrome who had no abnormality on coronary arteriography or left ventriculography. In patients with HCM, LV ejection fraction decreased from 65 +/- 8 (mean +/- SD)% at rest to 59 +/- 18% at peak exercise, in contrast to an increase among controls (from 56 +/- 9% to 64 +/- 9%). As compared with resting values, cardiac output increased to 168 +/- 24% at peak exercise in HCM, but the increase was significantly less than that in controls (215 +/- 47%). Stroke volume decreased gradually to 83 +/- 16% during exercise in HCM, while it increased to 114 +/- 10% at an exercise level of half intensity, and it decreased slightly to 106 +/- 16% at peak exercise. LV end-systolic volume decreased among controls to 78 +/- 27% at peak exercise, but remained unchanged in HCM (118 +/- 58%). An increase in peak ejection rate at peak exercise was less in HCM than in controls (143 +/- 26% vs 170 +/- 42%). No significant differences were observed between the two groups concerning changes in indices of LV diastolic function including LV end-diastolic volume, peak filling rate or 1/3 filling rate during exercise. In the analysis of LV function curves, pulmonary arterial diastolic pressure increased to a greater extent in HCM than in controls (19 +/- 6 mmHg vs 11 +/- 6 mmHg); whereas, an increase in the stroke work index was less in HCM (80 +/- 26 g.m/m2/beat vs 121 +/- 21 g.m/m2/beat) at peak exercise. Thus, the LV function curve shifted downward and to the right in patients with HCM. The above findings indicate that LV functional reserve during exercise is impaired, especially as to systolic function in patients with HCM, while deterioration of diastolic function may be partly compromised by elevated filling pressure.

T lymphocytes from rats infected with Nippostrongylus brasiliensis (Nb) release a soluble factor with affinity for IgE that selectively potentiates the IgE response to an unrelated antigen. The factor is derived from Fc omega receptors on T cells, binds to surface IgE on precursors of IgE-forming cells, and enhances their differentiation. The factor had a molecular weight between 10,000 to 20,000 and an affinity for lentil lectin. T cells from Nb-infected rats formed another IgE-binding factor upon incubation with rat IgE. The factor has the ability to suppress rather than enhance the IgE response. The IgE-specific suppressive factor is comparable to IgE-potentiating factor with respect to molecular weight and affinity for IgE but it fails to bind to lentil lectin. The IgE-specific suppressive factor is formed by lymphocytes complete Freund's adjuvant-treated rats. The results strongly suggest that IgE-binding factors are involved in the regulation of IgE response.

An eosinophil chemotactic factor (ECF) was produced in the cell-free culture supernatants (CFS) of peritoneal exudate cells (PEC) from guinea-pigs immunized with dinitrophenyl derivatives of ascaris extract (DNP-As), when stimulated by the antigen in vitro without activation by immune complexes. A 2 or 6 hr pulse of the antigen was sufficient for ECF production, whereas long time incubation (48 hr) was required for the production of a sufficient amount of the factor. Treatment of PEC by cycloheximide resulted in the reduction of ECF production, suggesting that protein synthesis is essential. Its generation appeared carrier-specific and the source of the factor is presumed to be lymphocytes, probably T lymphocyte. The factor with a molecular weight of 70,000 shared a common antigenicity with delayed eosinophil chemotactic factor-a (delayed ECF-a), which was isolated from the skin lesions showing delayed tissue eosinophilia in vivo.

The regulation of tissue eosinophilia induced by dinitrophenyl-ascaris extract (DNP-As) was investigated in guinea-pigs. Biphasic tissue eosinophilia peaking at 6 and 24 hr was observed in the skin lesions in Bordetella pertussis vaccine (Bp)-treated animals. In contrast, only the early phase of tissue eosinophilia was observed in Freund's complete adjuvant (FCA)-treated animals. Although less eosinophil chemotactic activity was detected in 24-hr-old skin extract of FCA-treated animals (FCA-extract), evident activity was recovered in the concanavalin A eluate (Con A-eluate) when FCA-extract was fractionated by Con A Sepharose. The chemotactic factor in Con A-eluate of FCA-extract was confirmed to be the T cell-derived eosinophil chemotactic factor, termed Delayed ECF-a, which has been isolated from allergic skin lesions by immunoadsorption. Another factor from the same skin lesions, Delayed ECF-b (which is a serum-derived one), was not detected in the FCA-extract. When eosinophils were mixed or pretreated with Con A-effluent of FCA-extract, the treated cells failed to be attracted by Delayed ECF-a, while the response to Delayed ECF-b was not affected, indicating that the inhibition was selective for Delayed ECF-a but not for Delayed ECF-b, and the eosinophil chemotactic inhibitory factor (ECIF) acts on eosinophils directly. Major ECIF activity was associated with a mol. wt. of 70,000 and minor with 12,500. Furthermore, the activity was absorbed by eosinophils but not by macrophages suggesting that eosinophils have receptor sites for ECIF. It was thus suggested that the appearance of ECIF, which is selective for the response of eosinophils to Delayed ECF-a, and decreased Delayed ECF-b production resulted in the inhibition of delayed tissue eosinophilia in FCA-treated guinea-pigs.

Glycogen storage diseases of type I, II, III, IV, V and the other muscle types, were examined electron microscopically, biochemically and physicochemically. Glycogenosomes (glycogen containing vacuoles) were found in the affected tissues of type II, type III variant of muscle glycogen storage disease, type IV and muscle type phosphorylase b kinase deficiency (disorder of the phosphorylase b kinase activation mechanism). The acid alpha-glucosidase activity was decreased only in the case of type II glycogen storage disease (Pompe's disease). The other types of glycogen storage disease showed no decrease in acid alpha-glucosidase activity. Moreover, one patient with type II disease also revealed a decrease in neutral alpha-glucosidase activity. In all cases where glycogenosomes were found, the extracted glycogen macromolecules showed some molecular abnormality or deviation when compared with normal native glycogen macromolecules.

A macrophage chemotactic factor (factor a) was extracted from DNP-ascaris extract-induced skin lesions (showing a maximal macrophage reaction) in guinea-pigs and then purified by gel filtration on Sephadex G-100 and chromatography using DEAE-Sephadex and CM-Sephadex. This material was a thermolabile protein (free of nucleic acid) with a molecular weight of about 150,000. It shared commom antigenicity with serum IgG, and its chemotactic acitivity was completely absorbed by anti-IgG and anti-light chain antibodies, but not by anti-BSA antibody. In contrast to leucoegresin, this material seemed active for macrophages but not for neutrophils. Less active factors b and c were thermostable and not absorbed by the antibodies described above. The ratio of chemotactic activity of factors a, b and c in the skin extracts was roughly 60:25:15.