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I Nakashima

Publications and source records attributed to I Nakashima.

At least 235 records · Page 13Linked to original sources
PubMed1984

Ontogeny of the erythrocyte-dependent IgM antibody responses to cell membrane antigens.

Our previous experiments characterized the T-cell independent type 2 B-cell responses to cell membrane antigens that are controlled by two donor cell types with different antigen-presenting (AP) activities. We here report about the ontogeny of this novel type of responses with special reference to the mutual relation of the development among two AP activities and their acceptor functions. The responses of mice to H-2d antigens on allogeneic cells and hapten (fluorescein isothiocyanate) antigens on syngeneic cells were examined in parallel. The positive AP activity displayed by red blood cells (RBC) for induction of anti-hapten responses was fully developed in the fetus, although H-2d antigens on the RBC for induction of anti-H-2d responses was immature in mice under 7 days old. In contrast, the negative AP activity displayed by spleen cells (B cells) for inhibition of the RBC-dependent anti-hapten and anti-H-2d responses was first developed in mice about 3 weeks old. The B cell functions accepting the positive and negative AP activities were also matured by that time. The possible significance of these findings in the physiology and pathology of the unique responses was discussed.

Age Factors↗
PubMed1984

Paradoxical effects of LPS on the T-cell-independent type 2 anti-H-2 alloantigen antibody responses to allogeneic erythrocytes.

Murine allogeneic red blood cells (RBC) induce primary IgM antibody responses to H-2 alloantigens T-cell-independently (TI). In this study we showed that bacterial lipopolysaccharide (LPS), which should activate B lymphocytes polyclonally, could not trigger an anti-H-2d plaque-forming cell response. We then demonstrated that administration of LPS (on days 0-1) with allogeneic RBC suppressed the response of mice to H-2d, while giving LPS 4-6 days after RBC augmented the response. In contrast, LPS did not enable allogeneic spleen cells to induce an anti-H-2d response. Additional experiments showed that the allogeneic RBC behave as a TI class 2 antigen. It was concluded from these results that allogeneic RBC display a peculiar activity that exclusively triggers a TI type-2 B cell response that cannot be initiated by LPS and is modulated by LPS in an abnormal fashion. The possible significance of this finding in the mechanism of occurrence of natural H-2-specific IgM alloantibodies in aged mice is discussed.

Animals↗
PubMed1984

Adjuvant activity of Klebsiella O3 lipopolysaccharide: comparative study using defined uniform salt forms.

Previously we showed that Klebsiella O3 lipopolysaccharide (KO3 LPS) is much more potent than other kinds of LPS including Escherichia coli O127 LPS (EO127 LPS) in adjuvant activity in augmenting antibody response and delayed-type hypersensitivity to protein antigens and in the ability to enlarge the regional lymph node. Various defined uniform salt forms, the triethylamine, sodium, potassium, ammonium, tris(hydroxymethyl)aminomethane, and calcium salt forms, of KO3 LPS and EO127 LPS were prepared by removing basic materials present in LPS preparations by electrodialysis and neutralizing the electrodialyzed LPS preparations with various kinds of alkali. The triethylamine salt form showed the best solubility and consisted of the smallest granules and, on the other hand, the calcium salt form showed the lowest solubility, compared with the natural form and the other uniform salt forms. Even if the natural forms of KO3 LPS and EO127 LPS were converted to the defined uniform salt forms, adjuvanticity of KO3 LPS and EO127 LPS in augmenting delayed-type hypersensitivity to ovalbumin and the ability to enlarge the regional lymph node did not significantly differ from those of the respective natural forms. From these results it is concluded that the difference in strength of the adjuvanticity between KO3 LPS and EO127 LPS is not due to the difference in their salt forms, solubility or physical state. Moreover, there were no significant differences in lethal toxicity for mice by the intraperitoneal route among the natural form and all the uniform salt forms of KO3 LPS tested.

Adjuvants, Immunologic↗
PubMed1984

Cellular mechanism of primary anti-Thy-1 antibody responses in vitro induced by uniquely immunogenic thymocyte antigens.

Thy-1 antigens are the only cell membrane antigens known to be able to induce primary antibody responses in vitro. We have shown that antigens from the thymocytes of mice and rats were highly immunogenic in cultures of murine spleen cells for the induction of Thy-1.1-specific plaque-forming cell responses, whereas antigens from other tissues, including brains and bone marrow, were poorly immunogenic, if at all. The thymocyte-specific Thy-1 immunogenicity was carried by disrupted cell membranes, and the specific activity for inducing responses was closely linked to Thy-1. We then tried to determine the mechanism of anti-Thy-1 antibody responses in vitro that were induced by the uniquely immunogenic thymocyte antigens. The thymocyte Thy-1 antigens behaved as T cell-independent class 2 (TI-2) antigens: they induced responses in athymic nude mice but not in CBA/N mice with a B cell defect. The apparent TI-2 responses to thymocyte Thy-1 did, however, require Thy-1+ cells in the responder, similar to anti-DNP-Ficoll responses. The full development of the anti-Thy-1 responses required the participation of splenic adherent cells (SAC). Nevertheless, the mechanism of the SAC dependency of anti-Thy-1 responses did not involve antigen presentation to lymphocytes by antigen-pulsed SAC, which contrasted with the finding that the presentation of antigen by live SAC to lymphocytes was indispensable for responses to DNP-Ficoll. The poor Thy-1 responsiveness of SAC-depleted spleen cells was fully restored by the addition of soluble factors (IL 1-like molecules) released from SAC into the culture, which did not replace the SAC-requirement of responses to DNP-Ficoll. It was concluded from these results that Thy-1 or Thy-1-linked structures on thymocyte membranes have an intrinsic activity to directly signal either TI-2 B cells or immature T cells, or both, for activation in the presence of soluble factors released from adherent accessory cells. This conclusion is discussed in relation to a hypothetical view that the thymocyte Thy-1 would physiologically mediate cell-to-cell interactions among special subsets of lymphocytes under thymic influence.

Animals↗
PubMed1983

High-grade tumor-specific immunity induced by L1210 leukemia variants obtained from the culture of L1210 cells fused with Lesch-Nyhan fibroblasts.

A highly immunogenic variant of the murine L1210 leukemia cell (L1210/LN-1) for generation of tumor immunity has been obtained from culture of L1210 cells originally fused with human Lesch-Nyhan fibroblasts. In L1210/LN-1 cells, no human chromosomes were identified and chromosomes M1 and No. 1 carried by the parent L1210 cells were missing. L1210/LN-1 cells displayed an intermediate morphology between L1210 cells and Lesch-Nyhan fibroblasts. Most of the CDF1 mice that were inoculated with less than 2 X 10(6) L1210/LN-1 cells survived over 60 days without evidence of tumor, whereas the original L1210 cells killed all the mice tested in about 2 weeks. When inoculated with more than 5 X 10(6) L1210/LN-1 cells, CDF1 mice developed tumor. The CDF1 mice which rejected 2 X 10(6) L1210/LN-1 cells were protected very effectively against challenge of otherwise highly aggressive 1-5 X 10(5) L1210 leukemia cells; 40 out of 43 primed mice tested survived for 60 days or longer after the tumor challenge. Even the CDF1 mice primed with irradiated or mitomycin-treated L1210/LN-1 cells survived against a challenge of 10(5) L1210 leukemia cells. They were, however, not protected against P388 leukemia or Meth A sarcoma, indicating that the immunity was specific to L1210 leukemia. The immunity induced by L1210/LN-1 cells was transplantable by immune spleen cells into syngeneic recipients. Thus, the L1210/LN-1 cells we obtained seem to be very useful as an immunogen for generation of high-grade tumor-specific immunity against highly malignant L1210 leukemia.

Animals↗
PubMed1983

Studies on the immune response in chickens. IV. Generation of hapten-specific memory and absence of increase in carrier-specific helper memory in antibody response to sheep red blood cell antigen and its hapten-conjugate.

The role of carrier recognition in the hapten-specific antibody response of chickens to hapten (TNP) conjugate of sheep red blood cells (SRBC) was studied. The anti-TNP antibody response to the challenge with either TNP-SRBC or DNP conjugate of Salmonella enteritidis (DNP-Sal) of chickens previously primed with TNP-SRBC was definitely higher than that of unprimed chickens. Injection of TNP-chicken red blood cells, which did not provoke any significant primary anti-TNP response, also induced the memory for secondary anti-TNP response to the challenge with TNP-SRBC or DNP-Sal. The TNP-specific memory generated by stimulation with TNP-SRBC was adoptively transferable by spleen cells into cyclophosphamide (CY)-treated X-irradiated syngeneic recipients. On the other hand, the SRBC-specific memory for augmentation of anti-TNP response to the challenge with TNP-SRBC was never detected in chickens previously primed with either any of a wide range of doses (10(4)-10(8)) of SRBC, solubilized SRBC or SRBC fixed with formaldehyde, nor in CY-treated X-irradiated chickens receiving SRBC-primed spleen cells. The anti-TNP response of chickens to the challenge with TNP-SRBC was rather suppressed by the SRBC preimmunization. The passively administered anti-SRBC chicken serum also suppressed the response to TNP-SRBC. The carrier-specific suppressor activity generated by SRBC stimulation was not adoptively transferable by spleen cells. It is suggested from these results that the mechanism of amplification of the anti-hapten antibody response of chickens to hapten-SRBC conjugate is primarily mediated by the hapten-recognition, and carrier-recognition is more important for suppression rather than for amplification of the anti-hapten response, although the inductive mechanism for anti-hapten response itself should somehow involve the carrier recognition.

Animals↗
PubMed1983

Intact and subcellular form of thymocytes of rats are exceptionally immunogenic in mice for inducing anti-Thy-1 T cell-independent class 2 antibody responses.

Results of the present study show that the primary anti-Thy-1.1 antibody response to rat antigen in Thy-1.2 mice is induced exclusively by thymocyte antigen. Thy-1 antigens of brain and bone marrow, which expressed much Thy-1 antigen, were poorly immunogenic if at all. Brain Thy-1 antigen considerably inhibited the immunogenicity of thymocyte Thy-1. Moreover, we found that subcellular form of thymocytes induce as high antibody responses as intact thymocytes do. The subcellular thymocyte Thy-1 antigen behaved as TI-2 antigen, inducing a good response in athymic nude mice but not in CBA/N mice with a B cell defect. The significance of these findings is discussed in relation to the possible physiological activity of Thy-1 or Thy-1-linked molecules on thymocytes specifically mediating lymphocyte differentiation.

Animals↗
PubMed1983

Control of T-cell-dependent antibody responses of mice to rat antigens by donor cell types.

Administration of rat red blood cells (RBC) into mice induced a rat antigen-specific T-cell-dependent primary antibody response that was detected by a plaque assay using rat RBC as a target. This response was not induced by rat thymocytes or rat spleen cells that should share rat-specific antigens with rat RBC. We then demonstrated that rat spleen cells but not rat thymocytes, which were administered with rat RBC, partially inhibit the action of rat RBC for induction of the anti-rat response. This inhibition required live donor FcR+ cells, and seemed to be specific to rat antigens common to RBC and spleen cells. The findings supported the idea that the control by donor cell types, which was originally shown to work for T-cell-independent antibody response [3,5], should be effective for T-cell-dependent response beyond the species barrier.

Animals↗
PubMed1983

Evidence for the erythrocyte as the principal antigenic cell type that triggers primary IgM antibody responses to H-2D alloantigens.

Antigenic requirements for the induction of T cell-independent primary splenic IgM antibody responses (plaque-forming cell responses) to H-2Dd alloantigens were studied. Results show that some functional activity or structural property of the donor cells is required for immunogenicity, because antigens are not active in subcellular forms. An unexpected finding was that allogeneic red blood cells were exceptionally highly immunogenic, and any lymphoid tissues including purified macrophages and tumor cell lines that were not contaminated with red blood cells were virtually nonimmunogenic. The definite role of red blood cells in donor tissues as immunogens was confirmed by water or ammonium chloride treatment that abolished immunogenicity, as well as by phenotyping of the immunogenic cells with antisera. Thus immunogenic cells were positive for erythrocyte-specific and H-2D antigens and negative for Thy-1, Ig, and NK-1. The possible roles of erythrocytes in induction and regulation of transplantation immunity and in B cell activation in general are discussed.

Animals↗
PubMed1983

The Klebsiella O3 lipopolysaccharide isolated from culture fluid: structure of the polysaccharide moiety.

In a series of our earlier studies, the O3 antigen isolated from culture supernatant of Klebsiella pneumoniae strain Kasuya (O3:K1) (KO3) was shown to exhibit very strong adjuvant activity in mice. KO3 obtained was homogeneous in analyses by either gel filtration or ultracentrifugation. Its molecular weight determined by ultracentrifugal analysis was greater than 2 X 10(6). It contained 37.9% C, 6.20% H, 0.24% N, and less than 0.1% P. KO3 was degraded into the polysaccharide moiety and lipid moiety (about 20%) by hydrolysis with 1% acetic acid at 100 C for 1 hr. The molecular weight of the polysaccharide moiety obtained by the hydrolysis was 16,200 as determined by the Somogyi-Nelson method. Chemical analyses using methylation analysis and Smith degradation as the principal methods indicated that the polysaccharide moiety consisted of a mannan which has a pentasaccharide repeating unit of alpha-mannosyl-1,3-alpha-mannosyl-1,2-alpha-mannosyl-1,2-alpha-mannosyl-1, 2-alpha-mannose joined through alpha-1,3-mannosyl linkages. The number of repetitions was less than 20. The fact that minor components such as 2-keto-3-deoxyoctonate and glucose were detected suggests the presence of a core oligosaccharide, but its precise structure is unknown.

Antigens↗
PubMed1983

Adjuvant activity of Klebsiella O3 lipopolysaccharide: no contribution of proteins to the expression of adjuvant activity.

Previously we found that Klebsiella O3 lipopolysaccharide (KO3 LPS) isolated from culture supernatant of strain Kasuya (O3: K1) or its decapsulated mutant strain LEN-1 (O3: K1-) exhibited very strong adjuvant activity in augmenting antibody response and delayed-type hypersensitivity to protein antigens in mice. The preparation of KO3 LPS after deproteinization by four cycles of treatment with chloroform-butanol (5: 1) usually contained a small percentage of proteins and a definite amount of another antigen which was destroyed by heating at 100 C for 1 hr. This antigen proved to be derived from type 1 fimbriae which are responsible for mannose-sensitive hemagglutination of guinea pig erythrocytes. The preparation of KO3 LPS isolated from culture supernatant of the strains which did not produce type 1 fimbriae exhibited strong adjuvant activity similar to that of the preparation from those which produced them. The preparation of KO3 LPS treated with hot phenol water which is known to remove lipid A-associated proteins exhibited a similar strong adjuvant activity. The preparation of KO3 LPS after extensive deproteinizing, two cycles of pronase treatment followed by ten cycles of treatment with chloroform-butanol, no longer contained detectable amounts of proteins and the fimbrial antigen, but this preparation also exhibited similar strong adjuvant activity. Moreover, there was no difference in strength of the adjuvant activity between the preparation of KO3 LPS isolated from culture supernatant and that isolated by the phenol method from bacterial cells. The present study demonstrates that the strong adjuvant activity of the preparation of KO3 LPS does not depend in any way on proteins contaminating the preparation.

Adjuvants, Immunologic↗
PubMed1983

Virulence for mice of Klebsiella strains belonging to the O1 group: relationship to their capsular (K) types.

Eighty-two Klebsiella O1 strains belonging to various K types were tested for virulence for mice by intraperitoneal inoculation. They comprised 49 strains newly isolated from various clinical specimens, 31 reference strains, and 2 laboratory strains. Of 9 Klebsiella O1:K2 strains, 7 were highly virulent inasmuch as their 50% lethal doses per mouse were less than 10 CFU, whereas the other 2 strains were avirulent even when encapsulated just like the virulent strains as revealed by the quellung test. Klebsiella K2 reference strain B5055 and strain Chedid, which were maintained in vitro for a long time, were highly virulent. Of 8 Klebsiella O1:K1 strains, 2 showed moderate virulence, and the other 6 strains had relatively low or no virulence. We found no definite correlation between the virulence of Klebsiella K1 and K2 strains for mice and the kind of clinical specimens in which they originated. All of the Klebsiella O1 strains tested belonging to K types other than K1 and K2 showed very low or no virulence for mice.

Animals↗
PubMed1983

Experimental autoimmune hepatitis in mice after immunization with syngeneic liver proteins together with the polysaccharide of Klebsiella pneumoniae.

Experimental autoimmune hepatitis could be produced in SMA mice by monthly injections of syngeneic liver homogenate or liver-specific lipoprotein together with the polysaccharide of Klebsiella pneumoniae 03:K1 as a powerful adjuvant. Using a gel-diffusion technique, antibodies reacting with liver-specific lipoprotein and liver-specific membrane lipoprotein were detected in approximately 50% of sera from the immunized mice after an 8-mo period. After a full immunization schedule, 60%-80% of the livers of the sensitized mice developed infiltration of mono-nuclear cells consisting mainly of lymphocytes and plasma cells in portal areas, frequently associated with focal necrosis of hepatocytes. Moderate-to-severe piecemeal necrosis of hepatocytes appeared in 10 of 59 animals. However, a gradual decrease in the morphologic severity was observed 3-6 mo after cessation of injections. The transfer of splenic cells from animals with the damaged liver led to a hepatitis in recipients that was characterized by portal infiltration with mononuclear cells and by necrosis of liver parenchymal cells seen on day 14 after cell transfer. The suppressor cell activity determined by the ability of concanavalin A-activated cells to suppress blast transformation of splenic cells of normal SMA mice was significantly decreased (p less than 0.05) in mice immunized with a mixture of liver-specific lipoprotein and the polysaccaride of Klebsiella pneumoniae compared with mice immunized with the polysaccaride of Klebsiella pneumoniae alone or the polysaccaride of Klebsiella pneumoniae plus kidney lipoprotein.

Animals↗
PubMed1982

Control of primary IgM antibody responses to H-2 alloantigens by antigen-bearing live B lymphocytes.

Alloantigen-bearing (H-2d+) peripheral red blood cells, but not red cell-depleted H-2d+ spleen cells, induce primary IgM anti-H-2d plaque-forming cell responses. In this study it is reported that the primary antibody responses to H-2d+ peripheral red blood cells can be markedly suppressed by a subpopulation of H-2d+ spleen cells when they are injected simultaneously or a few days before injection of red blood cells. This suppression was antigen (H-2d)-specific, did not depend on T cells of either the donor or the recipient, and strictly required live donor cells. An energy-dependent action of the donor cell cortex and some proliferation of donor cells in the recipient seemed to be involved in the mechanism of suppression. The donor-suppressor cell type was largely present in the spleen but not in the bone marrow and thymus, and was present in the spleen of athymic nude mice. The suppressor cells displayed the properties of B lymphocytes: they adhered to the nylon wool but not to glass, were of relatively low density (rho less than 1.09), and were surface Ig+, Ia+, Fc receptor-positive but Thy-1-. H-2d+ suppressor-donor B lymphocytes might directly signal to antigen-specific recipient B cells competing with the signal provided by H-2d+ red blood cells for the B cell activation.

Animals↗
PubMed1982

The donor cell type controls anti-hapten (fluorescein isothiocyanate) primary antibody response to hapten-modified syngeneic cells.

Hapten (fluorescein isothiocyanate, FITC)-sensitized syngeneic red blood cells (FITC-RBC) are exceptionally active for induction of anti-hapten primary antibody response, and FITC-modified syngeneic spleen cells depleted of RBC (FITC-SSC) are not immunogenic [4]. The present study has demonstrated that FITC-SSC injected simultaneously with FITC-RBC inhibit partially the anti-FITC response to the latter. Either the immunogenicity of FITC-RBC or the response-inhibiting activity of FITC-SSC was increased as the concentration of hapten-sensitizing cells was raised from 0.005 mg/ml to 2 mg/ml. The inhibition of anti-FITC response by FITC-SSC strictly required live donor cells, but was not dependent on T-cell activity of either the donor or recipient. Neither FITC-thymocytes nor the FITC-T-cell-rich fraction of SSC showed a definite activity for inhibition, whereas the FITC-B-cell-rich fraction of SSC acted very effectively. These results suggest that the primary anti-hapten antibody response to hapten-modified syngeneic cells is primarily controlled by antigen-bearing live donor cells of different cell types.

Animals↗