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H Nariuchi

Publications and source records attributed to H Nariuchi.

At least 127 records · Page 7Linked to original sources

Different effect of LPS-induced macrophage factor on antibody responses to TI-1 and TI-2 antigens.

Effect of macrophage culture fluid (MF) on thymic independent (TI) antibody responses was examined. MF potentiated antibody responses of spleen cells to dinitrophenyl (DNP)-Ficoll and DNP-liposome, TI-2 antigens, but not to trinitrophenyl (TNP)-BA and TNP-LPS, TI-1 antigens. The enhancing effect of MF on the anti-DNP-Ficoll response was dose-dependent. Neither T cells nor macrophages were required for MF to exert the effect, suggesting that MF works on B cells directly. B cells modulated by MF in their antibody responses were indicated to be in mature B-cell subset for the following reasons: (i) the cells were in (CBA/N x BALB/c) F1 female but not in F1 male mice; (ii) the cells bore the receptors for C3 on their surface. MF was indicated to exert the enhancing effect on the antibody response by modulating the proliferation and/or early events in differentiation of B cells and not by promoting antibody secretion. The active component of the MF was indicated to be Interleukin 1.

Animals↗

Responses of B cells with or without C3 receptors to polyclonal B cell activators.

Responses of B cells with or without receptors for C3 (CR) to polyclonal B cell activators (PBA) were studied. Mouse spleen cells were incubated with sheep red blood cells (SRBC) coated with antibody and complement to form rosettes, and they were separated by Ficoll-Hypaque density sedimentation into populations depleted of and enriched with lymphocytes bearing CR (CRL). These 2 populations were cultured with lipopolysaccharide (LPS), purified protein derivative of tuberculin (PPD), or dextran sulfate (DxS) and assayed for anti-TNP PFC. The CRL-depleted population responded well to LPS, poorly to PPD, and it showed practically no response to DxS, whereas the CRL-enriched population seemed to respond poorly to LPS but well to both PPD and DxS. The low responsiveness of the cRL-depleted population to PPD and DxS could not be explained by a shift of time-kinetics, by the dose-response profile of the responding cells, or by the depletion of adherent cells. Suppressor T cells did not take part in the reduced responses, since the treatment of the population with anti-Thy 1.2 plus complement could not restore the responses. These results indicate that B cells with CR [CR(+) B cells] respond well both to PPD and DxS, whereas the cells without CR [CR(-) B cells] respond poorly to PPD and DxS. It was difficult to evaluate the low responsiveness of CR(+) B cells to LPS because of the high background PFC of the cRL-enriched population.

Animals↗

Fractionation of IgG, IgG2a, IgG2b and IgG3 immunoglobulins from mouse serum by protein A-sepharose column chromatography.

Normal mouse serum was fractionated on protein A-Sepharose column by the elution with citrate buffer of different pH. At pH 8.0 IgG of all subclasses were retained on the column and main portion of IgG1, IgG2a, IgG3, and IgG2b were eluted at pH 6.0, 5.0, 4.0 and 3.0, respectively, as assessed by Ouchterlony immunodiffusion. Pure IgG1 and IgG2b were obtained by elution at pH 6.0 and 3.0, respectively, from normal serum. The identity of IgG1 globulin was confirmed by its antibody activity in chromatography fraction of anti-sheep red blood cell antiserum and also by its inability to fix complement. IgG3 globulin was eluted at pH 4.0 and also at pH 5.0. The identity of IgG3 was confirmed by the formation of M-bow-like precipitin line in gamma region of immuno-electrophoresis. The affinity of IgG3 to protein A was reduced by the treatment with 2-mercaptoethanol (2-ME). From 2-ME treated normal serum, therefore, pure IgG2a could be obtained at pH 4.0, although the recovery was low. Otherwise, IgG2a was always eluted with other subclass, IgG1 or IgG3.

Animals↗

Specificity of an anti-murine B cell serum.

An antiserum (ABS) specific for murine B cells was prepared in rabbits by immunization with lymph node cells from nude mice as reported previously (1). To make the antiserum specific for antibody-forming cell precursors (AFCP) extensive absorption was required with mouse serum, erythrocytes and thymus cells. Even after ABS was no longer cytotoxic for thymus cells, further absorption with thymus cells was necessary to make it ineffective for plaque-forming cells (PFC). The correlation between cells bearing C3- or Fc-receptors and the cells bearing antigen(s) to which ABS reacts was studied. In the residual spleen cells treated with ABS and complement, there were some EAg and EAmCm rosette-forming cells. There were also some spleen cells reacting with ABS even after depletion of C3- and/or Fc-receptor-bearing cells. These results show that the antigen(s) to which ABS reacts could be a marker on B cells different from C3- or Fc-receptors. Spleen cells reacting with ABS were compared with cells bearing C3- or Fc-receptors various days after birth. The cells sensitive to ABS increased in spleens earlier after birth than those with C3- or Fc-receptors.

Absorption↗

Antigen specific receptors on murine B cell lineage. 1. On antibody forming cells in various stages of immune response.

In this study it was examined whether or not antigen specific receptors are on direct and indirect plaque-forming cells (PFC). Spleen cells from mice primed both with horse red blood cells (HRBC) and sheep red blood cells (SRBC) were incubated with HRBC to make rosettes and fractionated on Ficoll-Hypaque density gradient. Both direct and indirect HRBC-PFC were depleted from the interface fraction. The depletion was antigen specific and inhibited by the preincubation of spleen cells with antiserum against kappa- or mu-chain for direct PFC and with antiserum against kappa- or gamma 1-chain for indirect PFC. The depletion was not due to the rosette formation mediated by Fc-receptors which might be on antibody forming cells, because PFC were not eliminated by the density gradient sedimentation after incubation of spleen cells with SRBC coated with anti-SRBC IgG antibody in our experimental conditions. Our results show that the antigen specific immunoglobulin receptors are on the cells producing IgG antibody after primary or secondary immunization as well as on the IgM antibody-forming cells even in late stage of immune response.

Animals↗

Antigen specific receptors on murine B cell lineage. 2. Different effect of rosette-forming cell depletion on IgM and IgG antibody forming cell precursors in primary and secondary response.

Spleen cells from unprimed mice or those primed with horse red blood cells (HRBC) were depleted of rosette-forming cells (RFC) with HRBC by the Ficoll-Hypaque density sedimentation, and the cells were examined in the adoptive transfer system whether they could raise IgM or IgG antibody-forming cells (AFC) after an immunization with HRBC. When spleen cells were pooled from unprimed mice, the response to HRBC of those depleted of RFC with HRBC (HRBC-RFC) was decreased to about a half in both IgM and IgG AFC. On the other hand, when spleen cells were from mice primed with HRBC, the response to HRBC of those depleted of HRBC-RFC was decreased dramatically to 1/20 of that of original cells in IgG AFC, but it was decreased to about a half in IgM AFC. In the time course of the response to HRBC of RFC-depleted spleen cells from mice primed with HRBC, an early IgG response was abolished but the late one was as high as that of untreated spleen cells. These results suggest that the depletion of RFC is most effective on the depletion of direct precursors of the secondary IgG AFC.

Animals↗

Mouse spleen cell populations involved in the mitogenic response to protein A.

In this paper, we examined the cellular components involved in the mitogenic response of mouse spleen cells to protein A. The mitogenic response was almost abrogated by the removal of macrophages. The abrogated response could be restored by the addition of peritoneal exudate macrophages from syngeneic mice. The reconstitution experiments were carried out for studying the populations of lymphocytes responding to protein A. In these experiments, spleen cells were passed through a Sephadex G-10 column and then they were separated into nylon-wool-adherent and nonadherent cells. The nylon-wool-nonadherent cells were used as T cells. The nylon-wool-adherent cells were used as B cells after they were treated with anti-BAT and complement. These results indicated that both B and T cells responded to protein A, and T-B cell cooperation could possibly play some role in the response.

Animals↗

Effect of lipopolysaccharide (LPS) on the thymocyte response to PHA: strain difference.

Thymocytes responded well to PHA in terms of [3H]-TdR incorporation if they were precultured in the presence of LPS, whereas fresh thymocytes or thymocytes pre-cultured in the absence of LPS responded poorly to PHA. The PHA response of thymocytes pre-cultured with LPS for several hours was similar to that of fresh spleen cells in the kinetics and dose-response profiles. The effect of LPS was found on thymocytes from BALB/c.Cr, AKR/Jms, and DDD/1 mice, but was not observed on those from C3H/HeJms and C57Bl/6J mice, indicating that there is a strain difference in the PHA responsiveness of LPS-pre-cultured thymocytes. In contrast to thymocytes, the response of spleen cells to PHA was enhanced in both C57Bl/6J and BALB/c.Cr mice by pre-culture with LPS.

Animals↗

Antibody forming cell precursors among glass-adherent peritoneal exudate cells.

Among glass-adherent peritoneal exudate cells (gaPEC), induced by an inoculation of 1% glycogen solution, about 4.5% were classified as antibody-forming cell precursors (AFCP) on the first day of culture by means of anti-mouse B-cell antibody (anti-B-Ab). They proliferated and differentiated into IgG-forming plasma cells when cultured with antigen and thymic RNA in vitro. Pretreatment of PEC with anti-B-Ab and complement suppressed the formation of plasma cells. AFCP had receptors for IgM-antigen complexes and for complement, both of which were independent of Ca++ and MG++ and resistant to treatment by pronase or phospholipase C. Cells bearing detectable receptors for EA (IgM) an EAC diminished by the 6th day when gaPEC were cultured with thymic RNA, but persisted longer in cultures without thymic RNA. The same percentages of cells demonstrated tartrate-resistant acid phosphatase activities but were devoid of esterase. Twenty to thirty percent of anti-B-Ab sensitive cells ingested latex particles. The proliferation kinetics of IgG-forming cells were studied through the 21st day of culture by means of peroxidase-labeled antibody staining methods.

Animals↗

Studies on the adjuvant effect of water-in-oil-in-water (w/o/w) emulsion of sesame oil. 2. Mode of action of the w/o/w emulsion.

The water-in-oil-in-water (w/o/w) emulsion showed potent adjuvant effect on the antibody formation to thymus-dependent antigens, but not to thymus-independent antigens. In hapten-carrier system the priming with carrier in the w/o/w emulsion enhanced more effectively the carrier specific helper function than did the priming with carrier in free solution. The cells responsible for the helper function were radioresistant. In the adoptive cell transfer system, the w/o/w emulsion was shown to enhance helper cell function. It is discussed that our w/o/w emulsion exerts the adjuvant effect by enhancing helper T cell activity.

Adjuvants, Immunologic↗

Studies on the adjuvant effect of water-in-oil-in-water (w/o/w) emulsion of sesame oil. 1. Enhanced and persistent antibody formation by antigen incorporated into the water-in-oil-in-water emulsion.

Water-in-oil-in-water (w/o/w) emulsion developed in our laboratory is as effective as water-in-oil (w/o) emulsion of Freund's incomplete adjuvant (FIA) in the stimulation of antibody formation. The emulsion is prepared by redispersion of water-in-sesame oil emulsion of an antigen solution in phosphate buffered saline with emulsifier, Tween 80. The emulsion can be stored at 4 degrees C for at least 3 months without any evidence of change in the adjuvanticity and in the w/o/w state. Even a single injection of bovine serum albumin (BSA) in the w/o/w emulsion elicited a high antibody response in mice over the period of almost whole lifespan. 10 microgram BSA in w/o/w could stimulate antibody formation up to 2(12) in hemagglutination titer, while the same dose in free solution did not elicit any detectable antibody. The tissue reactions caused by the w/o/w emulsion at the injected site and in the regional lymph nodes were much less prominent than those by FIA.

Adjuvants, Immunologic↗