PubMed Health⌕ Search

Biomedical subjects

C Hiramine

Publications and source records attributed to C Hiramine.

At least 37 records · Page 2Linked to original sources
PubMed1991

Proliferation of an athymic mouse-derived T-cell clone on thymic stromal cells with interleukin-2.

An athymic mouse-derived CD4+8+ T-cell clone, N-9F, was established. It expresses both full length gamma and delta T-cell receptor (TcR) mRNA. N-9F clone was not maintained by interleukin-2 (IL-2) alone but required another soluble mediator(s), contained in concanavalin A-stimulated splenocyte culture supernatant, for its proliferation. By culturing N-9F on thymic stromal cells, [3H]thymidine incorporation was retained and expression of IL-2 receptor (IL-2R) was induced. This phenomenon was also observed on thymic stromal cells from H-2 allogeneic mice, but not on other cell types such as splenic adherent cells or fibroblasts. After addition of recombinant IL-2 into the N-9F culture with thymic stromal cells, N-9F showed enhanced IL-2R expression and greatly proliferated. The inability to detect any soluble factors in thymic stromal cell culture supernatant suggests that this interaction is mediated by direct cell contact between T and thymic stromal cells. Because a CD2-negative subclone, N-9.23, also proliferated on thymic stromal cells, there might exist a type of molecule other than CD2/LFA3 or TcR/MHC involved with thymic stroma and T-lymphocyte interaction.

Animals↗
PubMed1990

Differential effects of a single dose of cyclophosphamide on T cell subsets of the thymus and spleen in mice: flow cytofluorometry analysis.

Sequential changes in the distribution of lymphocyte subpopulations of the thymus and spleen in BALB/c mice (male, 8-weeks old) during 2 weeks after a single i.p. injection of cyclophosphamide (CY, 200 mg/kg body weight) were studied mainly through the use of CD4, CD8 and CD3-epsilon markers together with single- or two-color flow cytofluorometry. In the thymus on Day 2 after CY treatment, a marked decrease in the number and proportion of PNAhi, CD3- and CD4+CD8+ double positive (DP) subpopulation was observed in parallel with a marked reduction in the thymus weight, cortical area and total thymocyte number. This phenomenon might be associated with the decrease in the percentage of thymocytes at the S phase of the cell cycle on Day 1 and 2 after CY treatment, owing to the depletion of the rapidly dividing cells in DP subset. There was a significant reduction in the number and proportion in CD4+ single positive(SP) subset on Day 7. The cell number of CD8+SP subset continued to decrease during Day 7 to Day 14. This contrasted with the behaviors of DP, CD4+SP and CD4-CD8- double negative (DN) subsets in which a considerable recovery was attained by Day 14. The spleen from CY-treated mice showed a marked decrease in the DN subset and surface immunoglobulin-positive cells, perhaps B lymphocytes, in both the percentage and the absolute cell number on Days 2 and 7, which paralleled the marked reduction in its weight and total cell number. The absolute cell numbers of CD4+SP and CD8+SP subsets in the spleen were also reduced on Days 2 and 7. The reduction of the CD4+SP/CD8+SP ratio was found in the thymocytes on Days 2 and 7 but not in spleen cells. Our results suggest that the principal target cell population of CY is the PNAhi, CD3- and DP immature cortical thymocytes as well as splenic B cells. The sustained decrease in the number of CD8+SP thymocytes after CY treatment might be in part relevant to the potentiating effect of CY pretreatment on immune responses.

Animals↗
PubMed1990

Establishment of a murine thymic epithelial cell line capable of inducing both thymic nurse cell formation and thymocyte apoptosis.

A thymic epithelial cell (TEC) line (B/c. TEC-L1) was established from a normal thymus of a 4-week BALB/c mouse. The B/c. TEC-L1 had an epithelial morphology showing a contact-inhibited cobblestone-like arrangement with occasional desmosome-like structures at the adjacent cellular membranes. B/c.TEC-L1 cells showed positive staining for desmosomal glycoprotein, cytokeratin, thymosin alpha 1 beta 3, and I-Ad, and MHC class I antigens. The doubling time was 24 hours, and the chromosome number ranged from 52 to 78 with the mode of 70. Coculture of B/c.TEC-L1 cells with syngeneic, peanut agglutinin-agglutinated (PNA+) thymocytes in suspension at 37 degree C was followed by the formation of TEC thymocyte rosettes, after which the reconstitution of thymic nurse cells ensued. At 4 degrees C, PNA+ thymocytes bound to the B/c.TEC-L1 cell but did not form thymic nurse cells. PNA- thymocytes, although to a lesser degree than PNA+ cells, bound to the TECs at 37 degrees C, but at 4 degrees C few cells bound to the TECs. Allogeneic thymocytes also bound to the TECs at 37 degrees C. When the PNA+ thymocytes were cultured on the B/c.TEC-L1 monolayer, the small ones chiefly adhered on the surface of the TECs, while underneath the TECs the relatively large thymocytes (including cells in mitosis) predominated. Although the PNA- thymocytes bound to the surface of the monolayer within a few hours after coculture, by 24 hours nearly all cells disappeared. It is presumed that the thymocytes creeping underneath the B/c.TEC-L1 monolayer and those enveloped within the thymic nurse cell reconstituted in the suspension culture; both may be placed in circumstances analogous to the thymic microenvironment, wherein immature thymocytes appear to contact TECs directly and to be exposed to higher concentrations of thymic hormones and other soluble factors. Additionally, cell death in the PNA+ thymocytes was also observed in the coculture with B/c.TEC-L1 cells. The PNA+ cells revealed the morphological changes termed "apoptosis" characterized by chromatin condensation and nuclear fragmentation.

Animals↗
PubMed1989

Differential effect of cyclosporine in vivo on the distribution of T cell subsets in the thymus, spleen, and lymph nodes.

Cyclosporine treatment of BALB/c mice (at a dose of 20 mg/kg every other day for 3 weeks) caused a remarkable reduction in the PNA-, L3T4+Lyt-2- subset of thymocytes. A significant reduction of the L3T4+Lyt-2-subset was also observed in both the lymph node and spleen cells of CsA-treated mice, though the degree of the reduction was lower than that in thymocytes. Both lymph node and spleen cells from CsA-treated mice showed a significant increase in the percentage of Thy-1.2 negative, L3T4-Lyt-2- cells (perhaps B cells). Thymocytes from CsA-treated mice showed a reduction in the in vitro proliferative responses to Con A and PHA. On the other hand, there was a slight, but not significant, decrease in the responses of lymph node cells to Con A, PHA and LPS. Spleen cells from CsA-treated mice showed a significant reduction in the responses to Con A and PHA, though the degree of the reduction was lower than that of thymocytes. There was a significant decrease in the proliferative response of spleen cells to LPS. These results suggest that CsA affects both thymus and spleen cells in vivo, preferentially impairing the L3T4+Lyt-2- subset (helper T cells or their precursors) within the thymus. The lymph node cells seem to be relatively spared from the in vivo effect of CsA compared with cells in the thymus and spleen.

Animals↗
PubMed1989

Enzyme-linked immunosorbent assay (ELISA) for detecting antisperm antibodies in mice with testicular autoimmunity.

We developed an ELISA for measuring antisperm antibodies in the mouse by using serum samples obtained from mice immunized with murine testicular antigens in complete Freund's adjuvant (CFA) as well as from mice rendered vasectomized. Sperm antigens used were syngeneic epididymal spermatozoa and two types of soluble, murine testicular antigens prepared in our laboratory. This study deals with a) the sequential changes of antisperm antibody levels following immunization; b) determination of immunoglobulin classes of these antibodies; c) a correlation between the absorbance values and the endpoint titers of antisperm antibodies; and d) comparison of endpoint titers of antisperm antibodies detected by ELISA with those by immunoperoxidase staining method in immune and nonimmune sera. It is suggested that serum dilution as high as 1/800 or more is required for detecting antibody titers of immune sera, because nonimmune mouse sera reveal a definite, although low, level of absorbance value at a serum dilution of 1/400 or less.

Animals↗
PubMed1989

The effect of cyclosporine on murine thymic epithelial cells--an immunohistochemical study.

The authors recently demonstrated that cyclosporine (CsA) treatment of mice caused a remarkable reduction of L3T4+Lyt-2- subset in the thymocytes as well as the striking decrease in the size and cellularity of the thymic medulla. To elucidate the underlying mechanism of such thymic alterations, an immunohistochemical study on the thymic epithelial cells was made in mice receiving CsA. In the medulla, a marked reduction of Ia-positive, keratin positive and Ulex europeus agglutinin 1 (UEA 1) binding epithelial cells was observed. CsA also induced a reduction of anti-keratin stainability of cortical epithelial cells and of Ia antigen expression by them. The results of this study indicate that CsA is not only directly impair thymocyte function but also may act on epithelial cells in the thymus. It is suggestive that CsA may affect T cell differentiation and maturation via alteration of Ia-positive thymic epithelial cells.

Animals↗
PubMed1988

Abnormal distribution of T cell subsets in the thymus of cyclosporin A (CsA)-treated mice.

Cyclosporin A (CsA) administration to normal, unprimed BALB/c mice at the dose of 20 mg/kg every other day for 3 weeks markedly reduced the thymus weight as well as the size and cellularity of the thymic medulla with relative preservation of the cortex. Area of the medullary compartment (the ratio of the area of medulla to area of cortex) of the CsA-treated mice decreased to about one half of that of the solvent alone-treated mice. Through the use of monoclonal anti-L3T4 and anti-Lyt-2 antibodies together with two-color flow cytofluorometry, we demonstrated that CsA caused a remarkable reduction of L3T4+ Lyt-2- subset and only a slight increase in L3T4+ Lyt-2+ subset in the thymus. There was no significant difference in the percentage of L3T4- Lyt-2+ subset between the CsA-treated and solvent-treated groups. Additionally, thymocytes from CsA-treated mice showed a marked reduction of in vitro proliferative responses to concanavalin A and phytohemagglutinin. The above mentioned results imply that in normal, unprimed mice CsA may preferentially impair L3T4+ Lyt-2- subset (helper T cells and their precursors) residing in the thymic medulla whereas is much less effective against L3T4+ Lyt-2+ subset present in the cortex and L3T4- Lyt-2+ subset.

Animals↗
PubMed1985

In vivo effects of cyclosporin A: abrogation of the induction of experimental allergic orchitis and sparing of the generation of suppressor cells.

Cyclosporin A (CyA; 10 mg/kg/day) administration for 14 days, starting on the day of immunization with testicular antigen (TA) in complete Freund's adjuvant (CFA), almost completely abrogated the induction of experimental allergic orchitis (EAO), delayed skin reactivities to both TA and purified protein derivative of tuberculin (PPD), and antisperm antibody response in strain 13 guinea pigs when examined 2 weeks after immunization. The unresponsive state induced was of a transitory nature. After cessation of CyA treatment, EAO-eliciting capability and cellular immune responsiveness recovered in relatively short time, while restoration of humoral immune responsiveness was delayed. Transfer of lymph node cells taken from CyA-treated, EAO-suppressed animals at 2 weeks postimmunization into normal syngeneic recipients inhibited the induction of EAO and delayed skin response. The suppressor cell activity appeared to be at least in part antigen-specific. It seems likely, therefore, that the profound suppression of EAO by CyA could be explained as due to the additive effect of the inhibition of helper/inducer T cell generation and the sparing or activation of suppressor cell populations.

Animals↗
PubMed1984

Adoptive transfer of suppression of experimental allergic orchitis with lymphoid cells from antigen-pretreated guinea pigs.

The injection of lymph node cells or spleen cells, obtained from strain 13 guinea pigs rendered unresponsive to experimental allergic orchitis (EAO) by pretreatment with testicular antigen (TA) in incomplete Freund's adjuvant, into normal syngeneic recipients markedly prevented the development of EAO, especially of the interstitial inflammatory cell response, which was expected to occur 2 weeks following orchitogenic challenge with TA in complete Freund's adjuvant (CFA). The suppressive effect of thymus cells from the same donors was much less prominent. The inhibition of EAO by suppressor cells was specific for the relevant antigen TA. In such EAO-suppressed animals delayed skin reaction to TA was suppressed, whereas antisperm antibody formation was not impaired. The active suppressor cells residing in the lymph nodes had characteristics of T lymphocytes, in that they did not adhere to the plastic dish surface and nylon wool and in that they formed rosettes with rabbit erythrocytes. B lymphocytes from the same animals did not have detectable suppressive properties. Lymph node cells from protected donors that had been treated with a single dose of cyclophosphamide (CY) 3 days before cell transfer were unable to transfer unresponsiveness to EAO. The results suggest that the immune prevention against EAO is explainable at least in part by the generation of CY-sensitive suppressor T lymphocytes with the capacity of inhibiting development of effector T cells after antigenic stimulation and that suppressor cells that mediated unresponsiveness to EAO may also regulate the cellular hypersensitivity to TA.

Animals↗
PubMed1983

Antigen-presenting capacity of guinea pig B lymphocytes in T cell proliferative response in vitro.

The capacity of normal (unprimed) B cells and keyhole-limpet-hemocyanin(KLH)-primed B cells to present the antigen KLH to KLH-primed T cells in the guinea pig was examined with in vitro assay of lymphocyte proliferative response. Antigen-pulsed B-lymphocyte population, extensively devoid of macrophages (M phi), induced the proliferative response of KLH-primed T lymphocytes, although less effectively than the peritoneal M phi. The antigen presentation was antigen-specific. There was no substantial difference between the magnitude of T-cell response induced by KLH-primed B-cell population and that stimulated by unprimed, normal B-cell population. The antigen-presenting capacity of the B-cell population was abrogated by pretreatment with anti-guinea pig immunoglobulin (Ig) or anti-I-region-associated (Ia) antigen antiserum and complement, whereas it was not affected with anti-guinea pig M phi antiserum and complement. From studies using strain 2 and strain 13 guinea pigs, histocompatibility requirement between antigen-pulsed B cells and antigen-reactive T cells was suggested: B lymphocytes, as well as M phi, did elicit proliferative response to specific antigen, KLH or purified protein derivative of tuberculin (PPD), in syngeneic, but not in allogeneic, T cells. These results suggest that the Ia-positive, surface-Ig-bearing guinea-pig B lymphocytes - not only KLH-primed B cells but also unprimed B cells - are capable of presenting antigen to primed T cells in a major histocompatibility complex(MHC)-restricted fashion.

Animals↗
PubMed1982

Suppression of experimental allergic orchitis and cellular immune response in the guinea pig by pretreatment with testis antigen in incomplete Freund's adjuvant.

Pretreatment of guinea pigs with homologous testicular antigen (TA) in incomplete Freund's adjuvant (IFA) markedly reduced both the incidence and severity of experimental allergic orchitis (EAO) when animals were subsequently challenged with TA in complete Freund's adjuvant (CFA) and examined at 2 weeks after challenge. The pretreatment rendered lymph node lymphocytes, particularly T cells, specifically unresponsive to the subsequent orchitogenic challenge as judged by antigen-induced in vitro lymphocyte proliferative response and blastogenic factor production, whereas antisperm antibody formation was not affected. The suppression induced was of a transitory nature, lasting 3 weeks after orchitogenic challenge, but between 4 and 5 weeks after challenge the development of EAO and aspermatogenesis became evident in parallel with the restoration of cellular immune responsiveness. Cyclophosphamide treatment 3 days before orchitogenic challenge abolished the preventive effect of pretreatment with TA in IFA. Transfer of serum obtained from pretreated animals failed to prevent actively sensitized recipients from developing EAO, which does not favor a possible occurrence of serum blocking factors in protected animals.

Animals↗
PubMed1982

Blastogenic factor production by guinea pig B lymphocytes stimulated with thymus-dependent antigen.

The purpose of the present paper is to determine whether antigen-stimulated B lymphocytes could produce blastogenic factor (BF) in the system of guinea pigs (strain 2) sensitized with keyhole limpet hemocyanin (KLH) or ovalbumin (OVA), known as thymus-dependent antigen, and, further, whether macrophages (M phi) are required for antigen activation of B cells to produce this lymphokine. Additionally, the effect of BF on normal T and B cells was evaluated. B lymphocytes were purified by double nylon wool adherence, plastic dish adherence. Sephadex G-10 column passage and sedimentation of E rosette forming cells on Ficoll-Isopaque. KLH- or OVA-primed B lymphocytes as well as T lymphocytes could produce BF when stimulated with specific antigen in vitro, and M phi were required for antigen activation of both T and B cells to produce BF. The optimal BF production was observed at a T or B cells: M phi ratio of 1:0.1 (in the system of KLH sensitization) or 1:0.5 (in OVA sensitization). BF production by B cells appeared to be less than that by T cells. BF produced by B cells was able to stimulate both normal T and B cells to proliferate; the proliferative responses of normal B cells to both B-cell-derived BF and T-cell-derived BF were significantly greater than those of normal T cells to these BFs. Readdition of M phi was not required for the T or B cell response to BF.

Animals↗
PubMed1980

Augmentation of guinea pig T lymphocyte proliferative response to antigens in the presence of purified B cells.

The effect of supplemental B lymphocytes on the antigen-induced in vitro proliferative response of highly purified lymph node T cells was studied in two immune systems of complete Freund's adjuvant (CFA) and testicular antigen (TA) sensitized guinea pigs. When the T lymphocytes obtained from animals 2 weeks after CFA sensitization, though unresponsive to purified protein derivate of tuberculin (PPD) by themselves, were cultured together with mitomycin C-treated autologous B (Bm) lymphocytes (1:0.5 ratio), they responded well to PPD. The enhancing effect of Bm lymphocytes was less than that of mitomycin C-treated macrophages (M phi m) from the same animals. Analogous helper effect of Bm lymphocytes on the T cell proliferation was observed also in TA-sensitized animals. Additionally, it is noteworthy that the PPD-induced proliferation of T lymphocytes at 2 weeks after CFA sensitization was absolutely dependent on M phi and B lymphocytes, whereas T lymphocytes at 10--14 weeks exhibited a low but significant response by themselves, although their response to PPD was markedly augmented with supplemental M phi m or Bm lymphocytes.

Animals↗
PubMed1980

Lymphocyte proliferative response in vitro and its cellular dependency in guinea-pigs with experimental allergic orchitis.

The development of cellular hypersensitivity was measured by the proliferative response of lymph node cells in vitro and a skin test to partly purified testicular antigen. There was a slight stimulation of lymph node cells and of the delayed cutaneous response at 1-2 weeks after sensitization with testicular antigen in Freund's complete adjuvant and then, after a decline in response, there was a pronounced peak at 6 weeks after sensitizaton. The early response coincided with the onset and active inflammatory stage of the effect on the testis, while the major peak was consistent with a state of aspermatogenesis. The proliferative response of unseparated lymph node cells could not be attributed to the function of T or B cells acting alone, although at 6 weeks after sensitization B-cell populations did become slightly responsive to antigen. The response of T lymphocytes was enhanced by the presence of B lymphocytes or peritoneal macrophages at 2 and 6 weeks after sensitization, but B cells did not appear to interact with macrophages. We conclude that T cells are essential to the proliferative response of lymph node cells during the induction of experimental allergic orchitis in guinea-pigs.

Animals↗
PubMed1979

Macrophage requirements of PPD-induced blastogenic factor production by T and B lymphocytes in the guinea-pig.

An attempt has been made to characterize the cell type which releases blastogenic factor(s) (BF) on purified protein derivative (PPD) stimulation in complete Freund's adjuvant (CFA)-sensitized guinea-pig system and, further, to examine whether macrophages (Mphi) are required for antigen activation of T or B lymphocytes to produce this lymphokine. Both T and B lymphocyte populations from guinea-pigs 2-3 weeks after CFA sensitization (early-stage T and B cells) failed to produce significant amounts of BF in response to PPD, whereas both the populations from 10 to 14 weeks after sensitization (late-stage T and B cells) acquired the ability to produce BF by themselves. It is noteworthy that readded autologous Mphi not only functioned well to augment BF production by the late-stage T and B cells but also completely made up for the inability of the early-stage T and B cells to produce BF. From these data it is reasonable to conclude that in the guinea-pig sensitized T and B cells can both release BF on exposure to PPD and that Mphi either reconstitute or augment the BF synthetic response of T and B cells.

Animals↗
PubMed1979

Effects of desensitization on different kinds of in vitro PPD-induced response of lymph node lymphocytes from tuberculin-sensitive guinea pigs.

Guinea pigs rendered hypersensitive to tuberculin 2 to 3 weeks (early stage) or 10 to 14 weeks (late stage) after sensitization with complete Freund's adjuvant could be completely desensitized by a single or double injections of a sufficient amount of PPD (purified protein derivative). Lymph node cells from such desensitized animals 48 to 72 hr after the challenge showed a considerable reduction of the ability to produce blastogenic factor and skin reactive factor upon PPD-stimulation, whereas the macrophage migration inhibitory factor activity remained still unaffected. As regards antigen-induced 3H-thymidine incorporation in vitro, lymph node cells from animals desensitized in the late stage after sensitization showed no substantial reduction in the degree of enhanced DNA synthesis upon PPD-stimulation, although the desensitization in the early stage resulted in a significant loss of this activity. These results suggest the possibility that desensitizing challenge does not bring about a uniform and regular effect on every lymphocyte subpopulations relevant to different functions and, in addition, indicate that there are some exceptions to the compartmentalization concept of antigen-reactive lymphocytes.

Animals↗