Effect of neonatal thymectomy on murine small intestinal intraepithelial lymphocytes expressing T cell receptor alpha beta and "clonally forbidden V beta s".

Neonatal thymectomy NTX performed on Day 3 of C3H mice causes over a 50% reduction in small intestinal intraepithelial lymphocytes (IEL) expressing the alpha beta T cell receptor (TCR) when analyzed at 10 weeks of age. Furthermore, this reduction is most notable in alpha beta TCR IEL expressing the CD8 alpha homodimer and no Thy 1 marker. This is in direct contradiction to the present theory that alpha beta TCR IEL expressing these markers are thymic independent.

Characterization of a Rana catesbeiana lectin-resistant mutant of leukemia P388 cells.

Sialic acid-binding lectin (SBL-C) from Rana catesbeiana eggs inhibits the growth of tumor cells such as P388 and L1210 leukemia cells (K. Nitta et al., Cancer Res.

Autoreactive and heat shock protein 60-recognizing CD4+ T-cells show antitumor activity against syngeneic fibrosarcoma.

A CD4+ heat shock protein (hsp) 60-recognizing autoreactive T-cell line (BASL1) and clone (BASL1.1) were examined for their antitumor activity against major histocompatibility complex class II- syngeneic Meth A fibrosarcoma (Meth A), which was immunofluorescently stained with monoclonal antibody specific for hsp 60. In in vitro proliferative assay, BASL1.

Protective effect of ren-shen-yang-rong-tang (Ninjin-youei-to) in mice with drug-induced leukopenia against Pseudomonas aeruginosa infection.

When 200 mg/kg of cyclophosphamide (CY) or 5-fluorouracil (5-FU) were given subcutaneously to mice, severe reduction of leukocyte numbers in the peripheral blood was observed on day 4 and from day 4 to day 8 after the treatment, respectively. Daily administration of ren-shen-yang-rong-tang (Japanese name: Ninjin-youei-to, NYT), (1 g/kg/day) and recombinant human granulocyte-colony stimulating factor (rhuG-CSF, 2 micrograms/mouse/day) either from day 0 to day 4 after treatment with CY or from day 0 to day 8 after treatment with 5-FU accelerated the recovery of peripheral leukocytes. Administration of NYT and rhuG-CSF inhibited decreases in the number of colony forming units in the spleen (CFU-S) in drug-treated mice.

A new subpopulation of intestinal intraepithelial lymphocytes expressing high level of T cell receptor gamma delta.

The murine intestinal epithelium contains T cell receptor (TcR) gamma delta-bearing T cells in high frequency. In the present report, we showed that TcR gamma delta-bearing intestinal intraepithelial lymphocytes (IEL) from C3H/He (H-2k) mice can be divided into two subpopulations based on TcR expression level; a subpopulation with a remarkably high level of TcR expression and a subpopulation with a moderate level of TcR expression, designated as TcR gamma delta hi IEL and TcR gamma delta mod IEL, respectively. In flow cytometric analysis, the TcR gamma delta hi IEL expressed a high level of TcR (mean fluorescence channel 518) when compared with the TcR level of TcR gamma delta+ T cells in other lymphoid organs (mean fluorescence channel 88).

Effect of a traditional Chinese medicine, bu-zhong-yi-qi-tang (Japanese name: Hochu-ekki-to) on the protection against Listeria monocytogenes infection in mice.

Effects of Bu-Zhong-Yi-Qi-Tang (Japanese name: Hochu-ekki-to) on the resistance against Listeria monocytogenes were observed in ICR mice orally administered this medicine daily for 10 days. Survival rates were increased by the pretreatment in mice inoculated i.v.

Age-associated increase in number of CD4+CD8+ intestinal intraepithelial lymphocytes in rats.

A significant number of CD4+CD8+ T cells were detected in intestinal intraepithelial lymphocytes (IEL) of various strains of rats including Wistar, WKA, BN, LEW and F344. The site of the CD4+CD8+ population in IEL increased with age in all strains we examined. Most IEL bearing CD8 expressed no CD5 antigen in young rats, while all CD4+CD8+ IEL and some of CD8+ IEL in aged rats were of CD5+CD45RB- phenotype.

The stage of negative selection in tolerance induction in neonatal mice.

In flow microfluorometry analysis of thymus and lymph node cells of C57BL/6(I-E-,Mlsb) mice rendered neonatally tolerant to (C57BL/6 x AKR/J)F1 (I-E+,Mlsb/a) lymphoid cells, both CD4+ and CD8+ cells showed a striking reduction in the number of V beta 6+ cells capable of recognizing Mlsa in the context of major histocompatibility complex (MHC) class II molecules, indicating that clonal deletion of V beta 6+ cells by Mlsa antigen occurs just at a stage of immature V beta 6lowCD4+CD8+ thymocytes. On the other hand, the number of V beta 11+ cells capable of recognizing I-E was markedly reduced in CD4+ cells, but CD8+ cells showed only partial (20%) reduction of such a population. The clonal deletion of V beta 11+ cells by I-E may begin at the transitional stage from V beta 11lowCD4+CD8+ to V beta 11highCD4+CD8- single-position cells, and V beta 11lowCD4+CD8+ cells differentiating to V beta 11highCD4-CD8+ cells seem to be resistant to clonal deletion.

Failure of T cell receptor V beta negative selection in murine intestinal intra-epithelial lymphocytes.

The intestinal intra-epithelial lymphocytes (IEL) are divided into several subsets on the basis of expression of T cell receptor (TCR) alpha beta and gamma delta, intensity of Thy-1 expression and expression of Lyt-3 chain. To investigate the differentiation pathway of the IEL, we examined the repertoire of V beta segments of T cells in the IEL in BALB/c (H-2d, MIs-1b2a) or AKR/J (H-2k, MIs-1a2b) mice. Among freshly isolated IEL, an appreciable number of T cells bearing V beta 3 or V beta 11, which recognize MIs-2a- or MHC IE-encoded molecules respectively, were detected in BALB/c mice.

Self-reactive T cells are activated by the 65-kDa mycobacterial heat-shock protein in neonatally thymectomized mice.

To elucidate the mechanism of autoimmune disease in neonatally thymectomized (NTX) mice, we have investigated the responsiveness of the self-reactive T cells which have not undergone clonal deletion in such animals. Consistent with a recent report (Yuuki et al., Eur.

Effect of methyl esterification of isoglutamine residue in 6-O-acylated muramyl dipeptides on expression of immunopharmacological activities.

The effect of methyl esterification of the isoglutamine residue in 6-O-acylated muramyl dipeptide (MDP) on some biological activities was investigated. Methyl esterification influenced more or less the expression of all activities tested. The adjuvant and colony stimulating factor (CSF)-inducing activities of 6-O-acylated MDP analogs carrying a 3-hexadecanoyloxytetradecanoyl [C14-O-(C16)] group were stronger than those of the corresponding analogs carrying a 2-tetradecylhexadecanoyl (B30) group.

Structural and biological comparison of lipopolysaccharides (LPS) from Bordetella species.

Lipopolysaccharides (LPS) isolated from Bordetella pertussis (Bp), B. parapertussis (Bpp), and B. bronchiseptica (Bbs) were analysed for their chemical composition, molecular heterogeneity, and immunological and biological properties.

Clonal anergy in self-reactive alpha/beta T cells is abrogated by heat-shock protein-reactive gamma/delta T cells in aged athymic nude mice.

Although T cells proliferate and differentiate primarily in the thymus, athymic nude mice contain an appreciable level of T cell receptor alpha/beta and gamma/delta T cells, suggesting the existence of the extrathymic pathway in the development of both T cells. Recent studies with nude mice indicate that clonal deletion of self-reactive T cells does not occur extrathymically. In the present study, we have investigated the responsiveness of self-reactive T cells differentiating along an extrathymic pathway in aged BALB/c (H-2d, Mls-1b2a, I-E+, 7-8 month old) nude mice.

Biological properties of lipopolysaccharides from Bordetella species.

Biological activities of lipopolysaccharides (LPS) extracted from Bordetella pertussis, B. parapertussis and B. bronchiseptica were compared with those of Escherichia coli LPS.

Stimulation of all T cells bearing V beta 1, V beta 3, V beta 11 and V beta 12 by staphylococcal enterotoxin A.

To determine the molecular mechanisms of T cell stimulation by staphylococcal enterotoxin A (SEA), we examined the expression of T cell receptor (TcR) V beta on the T cells from four strains of mice stimulated in vitro with SEA, using flow cytometric analysis for the number of T cells bearing V beta 3, V beta 6, V beta 8, V beta 11 and RNA blotting analysis for the amount of transcripts of V beta 1, V beta 5 and V beta 12. The number of T cell blasts bearing V beta 1, V beta 3, V beta 1 or V beta 12 were increased in the T cell blasts proliferating in vitro in response to SEA in C57BL/6 mice. In AKR/J mice, which contain few V beta 11- or V beta 12-bearing T cells due to a tolerance to the self-MHC class II IE-antigens, T cells bearing V beta 1 or V beta 3 responded to SEA.

Biological properties of lipopolysaccharides isolated from Bordetella.

Biological activity of lipopolysaccharides (LPSs) separated from Bordetella, i.e., B.

Protective effect of saikosaponin A, saikosaponin D and saikogenin D against Pseudomonas aeruginosa infection in mice.

Effects of saikosaponins and their genins on nonspecific resistance against Pseudomonas aeruginosa and Listeria monocytogenes infections were investigated. When mice were administered intraperitoneally (i.p.

[Adoptive immunotherapy in patients with medulloblastoma by LAK cells].

Medulloblastoma is one of the most popular malignant brain tumors in children. It accounts for about 15% of all pediatric brain tumors. Radiochemotherapy has prolonged the 5-year survival rate up to 60-85% for patients with medulloblastoma.

Immunopharmacological activities of 2-keto-3-deoxyoctonic acid-(alpha 2----6)-linked 4-O-phosphono-D-glucosamine derivatives carrying N- and 3-O-acyl substituents.

The immunopharmacological activities of 2-keto-3-deoxyoctonic acid (KDO)-(alpha 2----6)-linked lipid A-subunit analogs, 4-O-phosphono-D-glucosamine derivatives carrying N- and 3-O-acyl substituents, were compared with those of the corresponding analogs without KDO, GLA-27, GLA-47, and GLA-60. Among the analogs tested, GLA-60, a 4-O-phosphono-D-glucosamine carrying N-3-hydroxytetradecanoyl and 3-O-3-tetradecanoyloxytetradecanoyl groups, exhibited the most intensive activities in terms of mitogenicity, adjuvanticity, and mediator (tumor necrosis factor and colony-stimulating factor) induction. Binding (alpha 2----6) of KDO to GLA-60 failed to enhance the activities.

Enhancement of immunoglobulin G responses in mice against hepatitis B virus surface antigen, influenza virus hemagglutinin vaccine, and tetanus toxoid by 6-O-acylated muramyl dipeptides.

The adjuvant activity of chemically synthesized 6-O-acylated muramyl dipeptides (MDP) was tested in aqueous form. The activity was assessed by determining immunoglobulin G (IgG) titers in sera of mice immunized with hepatitis B virus surface antigen, influenza virus hemagglutinin (HA) vaccine, or tetanus toxoid with an enzyme-linked immunosorbent assay. Administration of 6-O-acyl-MDP analogs with antigens induced marked enhancement of primary and secondary IgG antibody responses and maintained high antibody levels for at least 7 weeks.

Activation of murine peritoneal macrophages by saikosaponin a, saikosaponin d and saikogenin d.

Macrophage activation by saikosaponins and saikogenins was investigated and compared with that by other saponins and macrophage stimulants. Saikosaponins a and d induced a marked cell accumulation in the peritoneal cavity when administered intraperitoneally. Among saikosaponins and saikogenins tested, saikosaponin d significantly activated peritoneal macrophages in terms of enhancement of phagocytic activity, increased level of cellular lysosomal enzyme (acid phosphatase), induction of cytostatic activity and expression of Ia antigen on the cell surface.

Immunostimulatory activity of 1-O-acylated muramyl dipeptides, with or without a 6-O-phosphoryl group, in aqueous form.

Immunostimulatory effects of 1-O-acylated derivatives of N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP) methyl ester, with or without the 6-O-phosphoryl group, on augmentation of IgG antibody response against influenza hemagglutinin (HA) vaccine, in vivo macrophage activation and enhancement of non-specific host resistance against Pseudomonas aeruginosa infection were investigated. The activities were tested intraperitoneally (i.p.

Effect of chemical modification at C1 of the glucosamine backbone of lipid A-subunit analog GLA-27 on manifestation of immunopharmacological activity.

The effect of chemical modification at the C1 position of GLA-27, 4-O-phosphono-D-glucosamine carrying N-3-tetradecanoyloxytetradecanoyl [C14-O-(C14)] and 3-O-tetradecanoyl (C14) groups, was investigated. Replacement by SH or S-acetyl groups of the OH group resulted in the enhancement of mitogenicity but gave rise to a reduction, in macrophage-stimulating ability such as induction of tumor necrosis factor and enhancement of phagocytic and cellular acid phosphatase activities. Bisphosphorylation at C1 and C4 resulted in a slight decrease in mitogenicity or almost complete loss of the macrophage-stimulating ability.

Importance of fatty acid substituents of chemically synthesized lipid A-subunit analogs in the expression of immunopharmacological activity.

The immunopharmacological activities of chemically synthesized lipid A-subunit analogs, 4-O-phosphono-D-glucosamine derivatives carrying different N- and 3-O-linked acyl groups, were investigated. None of the synthetic compounds tested exhibited any detectable pyrogenicity at a dose of 10 micrograms/kg. Weaker lethal toxicity in galactosamine-sensitized mice was detected at 1 microgram per mouse for all the synthetic compounds except GLA-58.

An adoptive immunotherapy of patients with medulloblastoma by lymphokine-activated killer cells (LAK).

An adoptive immunotherapy of 6 patients with medulloblastoma by lymphokine-activated killer (LAK) cells is described. They were from 2 to 9 years in age and had cerebrospinal fluid (CSF) dissemination of the tumours. All patients underwent the whole-neuraxis irradiation and chemotherapy.