Shigenori Higuchi

The ratio of neutrophils to lymphocytes and the phenotypes of neutrophils in patients with early gastric cancer

Abstract We examined 55 patients (40 male, 15 female) who were diagnosed from 1987 to 1991 as having early gastric cancer (EGC) stage I according to the general rules of classification of the Japanese Research Society for Gastric Cancer. Of the 55 patients, 42 (30 male, 12 female) were alive in April 1992. The prognosis correlated well with the ratio of neutrophils to lymphocytes (N/L ratio) but not with the total number of white blood cells in the peripheral blood. The patients were divided into two groups according to their N/L ratio. Of the 29 patients with an N/L ratio less than 2, 27 were alive in 1992, whereas only 15 of the 26 patients with an N/L ratio of 2 or more were alive (χ2 analysis, P = 0.0022). We further examined the phenotypes of neutrophils from 29 other patients with EGC at the time of diagnosis before surgical operation. These patients were divided into two groups: 17 patients with a low N/L ratio (less than 2) and 12 patients with a high N/L ratio (2 or more). CD10 and CD35 expressions on neutrophils from the patients with a low N/L ratio were lower than those from the patients with a high N/L ratio. The N/L ratio correlated well with both CD10 and CD35 expression, whereas no correlation was observed between the numbers of neutrophils and the expression of these phenotypes. The respiratory burst of neutrophils from the patients with a high N/L ratio was higher than that of neutrophils from the patients with a low N/L ratio, though there was no correlation in the phagocytic activity between both groups. It was thus suggested that the heterogeneity of neutrophils is, at least partly, related to the prognosis of patients with EGC.

Characterization of the human IL-5 receptors on eosinophils

Interleukin 5 (IL-5) receptors on the cell surface of human eosinophils and other hematopoietic cells were characterized using radiolabeled recombinant IL-5. The binding of 35S-labeled murine IL-5 to eosinophils from normal human peripheral blood was rapid and saturable within a 30-min incubation at both 4 and 37 degrees C. The binding of 35S-labeled murine IL-5 to eosinophils was inhibited by an excess of unlabeled murine and human IL-5 or by an anti-murine IL-5 monoclonal antibody (NC17) but not by other human cytokines. Scatchard plot analysis revealed that human eosinophils have a single class of high affinity receptor (Kd 170-330 pM; number of binding sites: 260-380/cell). IL-5 receptors on eosinophils from four patients with eosinophilia displayed similar characteristics. Affinity cross-linking experiments resulted in the identification of human IL-5 receptor on eosinophils with a molecular mass of 55-60 kDa. Among the various cells besides eosinophils and cell lines that we could test, a subline of HL-60 (YY-1 cells) was found to display a significant number of IL-5 receptor. These results suggest that IL-5 may act on limited types of cells in the human system.

Nutritional Copper Deficiency in Severely Handicapped Patients on a Low Copper Enteral Diet for a Prolonged Period: Estimation of the Required Dose of Dietary Copper

Six cases of nutritional copper deficiency were identified in a nursery institute for severely handicapped patients. All had been on prolonged enteral feeding of a copper-deficient diet and all had abnormalities related to the copper deficiency. Two of them had leukopenia, with or without macrocytic and normochromic anemia. After the oral administration of copper, the clinical and laboratory observations including neutrophil counts and serum copper and ceruloplasmin levels showed a complete recovery. The dietary copper requirement was estimated to be approximately 20 micrograms/kg/day for these patients, based on the correlation between the dietary copper intake and the levels of copper in the serum.

The polyvalent protease inhibitor, trasylol, inhibits DNA synthesis of mouse lymphocytes by an indirect mechanism

Abstract By the use of incorporation of radiolabeled thymidine, uridine, and leucine into mouse lymphocytes, the inhibitory effect of the protease inhibitor, trasylol, on antigenor mitogen-induced lymphocyte triggering was studied in vitro . DNA synthesis, as well as RNA and protein syntheses, were effectively inhibited by 0.3–2.5 × 10 −7 mol of trasylol when responses were induced by homologous antigen, allogeneic cells, phytohemagglutinin, or endotoxic lipopolysaccharide of Escherichia coli . The inhibitory effect of trasylol was reversible. On the contrary, DNA synthesis by nonadherent spleen cells was hardly inhibited by the inhibitor when the cells were stimulated with a relatively large amount of concanavalin A. Antigen-induced DNA synthesis by non-adherent lymph node cells was enhanced by the culture supernatant of macrophages. This helping effect of macrophage supernatant was effectively inhibited either by soluble or insoluble trasylol. These results suggest that the inhibitory action of trasylol on lymphocyte triggering may operate indirectly to interfere with the helping action of macrophages on lymphocytes.

Anti-neutrophil antibodies in patients with nutritional copper deficiency

Ten patients with nutritional copper deficiency were studied in terms of neutrophil counts and anti-neutrophil antibodies (ANA). In four patients with severe or moderate copper deficiency, the production of ANA was positive and two patients with a severe deficiency had neutropenia. After copper supplementation, ANA titres became negative or were reduced in all patients and neutrophil counts reverted to normal in two patients. It thus appears that copper deficiency is linked to the production of ANA, a condition which partly responsible for neutropenia.

ENHANCEMENT OF PHYTOHEMAGGLUTININ-INDUCED DNA SYNTHESIS OF MOUSE THYMOCYTES AND T-LYMPHOCYTES BY A NEUTRAL PROTEASE OF POLYMORPHONUCLEAR LEUKOCYTES

Mouse thymocytes were poorly triggered in vitro with phytohemagglutinin (PHA) to induce DNA synthesis and the response was markedly enhanced by culture supernatant (SUP) of polymorphonuclear leukocytes (PMN). The PMN factor was nondialysable, heat‐labile and precipitated with 65% ammonium sulphate. Its molecular weight was approximately 19,000 on sephadex G‐75. The sephadex fraction had a proteolytic activity on 3H‐acetyl hemoglobin at neutral pH. The protease seemed to be a chymotrypsin‐like enzyme on the basis of inhibition profile using various protease inhibitors. The thymocyte‐helping activity in the protease fraction was absorbed by affinity columns of protease inhibitors. The PMN‐protease also enhanced the DNA synthesis by antigen‐stimulated lymph node cells and by PHA‐stimulated T‐lymphocytes, but not by LPS‐stimulated B‐lymphocytes. Only the lymphocytes already been stimulated with antigen or mitogen received preferentially the helping action by PMN‐protease. The helping activity was effectively absorbed by PHA‐stimulated thymocytes but not by nonstimulated thymocytes. These evidences seems to suggest that acceptor sites for the protease, newly developed on lymphocyte surface after stimulation, may play an important role in the enhanced DNA synthetic response. ACTA PATH. JAP. 27: 857‐868, 1977.

Characterization of antineutrophil antibodies in patients with neutropenia associated with nutritional copper deficiency

Six antineutrophil antibody (ANA)-positive patients with copper deficiency were classified into two groups; those with (group A, n = 3) and those without (group B, n = 3) neutropenia. The percent binding of ANA for normal peripheral neutrophils was similar in both groups (83.5 +/- 7.2 vs. 79.1 +/- 10.5%). The percent binding of sera to cultured promyelocytic leukemia cells (HL-60) was increased in group A (from 3.7 +/- 3.2 to 12.2 +/- 2.3%) but not in group B (from 65.3 +/- 21.7 to 40.7 +/- 6.3%) after stimulation of HL-60 with DMSO. The stimulated HL-60 cells expressed CD 16 and CD 11b antigens. In the presence of monoclonal antibody for CD 16, the titer of ANA was nil in group A and unchanged in group B. Thus, ANA of patients with neutropenia may recognize mainly the CD 16 antigen, the Fc gamma receptor III of neutrophils.

A family of selective immunodeficiency with normal immunoglobulins : possible autosomal dominant inheritance

We report here our findings in two Japanese siblings who experienced recurrent bacterial and viral infections since early infancy. Recent symptoms included diarrhoea, conjunctivitis, rashes, headache, sore throat, joint pain, vomiting and vertigo, all similar to those seen in toxic shock syndrome, except for shock. These symptoms improved following gammaglobulin treatment.Staphylococcus aureus with coagulase type IV was continuously isolated from nasal smears producing toxic shock syndrome toxin-1 (TSST-1). Serum antibodies did not or only poorly responded to TSST-1, diphtheria toxoid, varicella virus and rubella virus, whereas total and subclass levels of serum immunoglobulin and in vitro DNA synthesis of lymphocytes stimulated by TSST-1,Staph. aureus, varicella vaccine and mitogens were normal. In the family, ten other members in three generations (five males: five females) including the mother had similar clinical symptoms. Thus, the disease may be inherited in an autosomal dominant fashion.

Immunologic studies of peripheral blood in a child with hypogammaglobulinemia. Suggested mechanism for the development of malignant B-cell lymphoma

A case of a 7‐year‐old boy with common variable hypogammaglobulinemia who developed B‐cell‐type non‐Hodgkins lymphoma is reported. Immunologic studies of his peripheral blood before the development of lymphoma revealed: (1) although peripheral T‐cell and B‐cell counts were normal, serum IgG and IgA levels were remarkably reduced; (2) DNA synthesis in response to phytohemagglutinin‐P (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM) stimulation were decreased; (3) DNA synthesis in response to EBV was enhanced; (4) in vitro IgG production with the patients peripheral blood lymphocytes was significantly depressed; and (5) helper and suppressor activities of the patients T‐cells did not differ from that of normal controls. Six months after the investigation systemic involvement of malignant lymphoma appeared. The lymphoma was diagnosed as a lymphoblastic diffuse one. Lymph node cell marker analysis revealed that the lymphoblasts had surface μ and k chain, cytoplasmic IgM, and HLA‐DR antigen. Blastogenesis of B‐cell series may be suppressed by a feedback mechanism with the B‐cells and antibody. In the current case, impairment of such a mechanism with defect of immunoglobulin production might finally induce malignant B‐lymphoid proliferation.

Irradiated fetal thymus transplantation in a patient with combined immunodeficiency with predominant T cell defect

A 6 month old boy was diagnosed as a case of combined immunodeficiency (with predominant T cell defect by previous classification). His T cell count was decreased, his B cell count in peripheral blood was increased, his serum IgG level was decreased, his serum IgM level was normal and the thymus was not evident on CT scans and magnetic resonance imaging. Administration of the thymus hormone, thymosin, led to a partial recovery of T cell function without normalization of the T cell count. At age 26 months the patient received an irradiated thymus transplantation from a 16 week old female fetus. After the transplantation, the T cell count (mainly CD4+ cells) increased by 50–70%. A mild graft‐versus‐host reaction (GVHR) occurred and several immunosuppressants were prescribed. Chromosome analysis showed that the T cells have both 46 XY and 46 XX karyotypes while the B cells have the 46 XY karyotype alone. His cellular immunity (skin tests, DNA synthesis, mixed lymphocyte reaction, cytotoxic activity and natural killer cell function) and his serum IgG level remained low. However, being on regular r‐globulin therapy and oral anti‐fungal drugs, he is now living normally with almost no trouble at age 6 years and 3 months. This case showed that irradiated thymus transplantation might be a useful method when an adequate donor for bone marrow transplantation is not available. The unexpected observation that the increased T cells were mainly CD4 may be related to the mild GVHR and the clinical improvement.