Fetal thymus may be the organ for NK cell maturation, but the in vivo evidences are few, Here, by analyzing NK cell receptor, we present that NK cells develop in fetal thymus and fetal liver and that NK cell receptor appears earlier than the expression CD16 or CD56. Moreover, the finding that the repertoire of NK cell receptor is different between fetal thymus and fetal liver lymphocytes suggests that the environmental factors may influence the NK cell receptor repertoire during NK cell maturation.
Killer Cells, Natural* ; Liver* ; Lymphocytes* ; Thymocytes* ; Thymus Gland

A recent study has proposed that dimethyl sulfoxide (DMSO) increases the number of HL-60 cells in high cell density conditions by inhibiting cell density-dependent apoptosis. We observed that dimethylformamide (DMF), a DMSO-related polar compound, also increased the concentration of HL-60 cells. The effective dose range of DMSO and DMF was 0.5-1% and 0.2-0.6% respectively. DMF, like DMSO, inhibited density-dependent apoptosis of HL-60 cells. The flow cytometric PKH26 cell proliferation assay showed DMSO and DMF actively increased cell division. However, the difference in the distribution of cell cycle phase was not noted between the control and the DMSO- or DMF-treated HL-60 cultures. Finally, DMSO and DMF stimulated HL-60 growth even in low density conditions. These results suggest that DMSO and OMF at appropriate concentrations increase the number of HL-60 cells by both apoptosis inhibition and cell division augmentation.
Apoptosis ; Cell Count ; Cell Cycle ; Cell Division ; Cell Proliferation ; Dimethyl Sulfoxide* ; Dimethylformamide* ; HL-60 Cells* ; Humans*

Chronic granulomatous disease (CGD) is one of congenital immunodeficient disease and a rare X-linked or autosomal recessive disease characterized by recurrent life- threatening infections and granuloma formation. We observed clinical features, laboratory findings and genetic subgroups of 33 children who were diagnosed with chronic granulomatous disease in the Department of Pediatrics, Seoul National University Children's Hospital. There were 23 males and 10 females. Activated NBT test of all patients revealed 0% positive cell and mothers of 15 patients had 25%- 75% normal neutrophils in the activated NBT test. According to the result of activated NBT test and family history, the ratio of X-linked and autosomal recessive inheritance was 2:3. There was a significant difference for the age at onset of the first infection in the different genetic subgroups. The X-linked group had the mean onset at 1.98 months of age and autosomal recessive group had a mean onset as late as 3.82 months (p<0.05). The most common type of the first infection was lymphadenopathies (41%) and other infections were skin pustules, fever, perianal abscess, pneumonia and chronic diarrhea. However, the age at diagnosis was not significant in the different genetic subgroups. Lymphadenitis (27%) was the most common infection, and pneumonia, gastrointestinal tract infection, skin infection were also common. The most common infectious agent was Candida sp. (5%) and other microorganisms involved were BCG, coagulase-negative staphylococcus, S. aureus, K/ebsiella pneumoniae, Aspergi/lus sp., and Enterococcus faecium. Chronic condition associated with CGD were hepatomegaly (59%), splenomegaly, and anemia of chronic disease, underweight, and lymphadenopathy. The leukocyte count of patients at diagnosis was within normal limit except in three patients and leukopenia was not observed in any of the patients. The humoral and cellular immunity and complement system were normal, but the level of Ig E in four patients was elevated. Early diagnosis of CGD can be made by suspicion if there are lymphadenitis after BCG vaccination and recurrent pyogenic infections under the first year of age. Though progression in the treatment of CGD, like gene therapy, is concerned, genetic counseling and prenatal diagnosis by carrier detection and molecular genetic analysis is thought to be necessary.
Abscess ; Anemia ; Candida ; Child ; Chronic Disease ; Complement System Proteins ; Diagnosis ; Diarrhea ; Early Diagnosis ; Enterococcus faecium ; Female ; Fever ; Gastrointestinal Tract ; Genetic Counseling ; Genetic Therapy ; Granuloma ; Granulomatous Disease, Chronic* ; Hepatomegaly ; Humans ; Immunity, Cellular ; Korea* ; Leukocyte Count ; Leukopenia ; Lymphadenitis ; Lymphatic Diseases ; Male ; Molecular Biology ; Mothers ; Mycobacterium bovis ; Neutrophils ; Pediatrics ; Pneumonia ; Prenatal Diagnosis ; Seoul ; Skin ; Splenomegaly ; Staphylococcus ; Thinness ; Vaccination ; Wills

D variant of encephalomyocarditis (EMC-D) virus causes diabetes in susceptible mice by direct infection and cytolysis of pancreatic beta cells. cDNA covering the major outer capsid protein (VP1) of EMC-D virus was cloned into Mycobacterium bovis bacillus Calmette-Guerin (BCG). None of the SJL/J male mice, immunized with live recombinant BCG-VP1, became diabetic when challenged with highly diabetogenic EMC-D virus. But the control mice inoculated with normal BCG or rBCG transformed with vector alone developed diabetes in the same challenge. VP1-specific antibodies including neutralizing antibodies were markedly increased as time went on and reached to the maximum titer at week 10 after a single immunization. The plateau of the titer lasted longer than following 4 weeks. Guinea pigs immunized with the live rBCG-VP1 showed strong delayed type hypersensitivity (DTH) to the VP1of EMC-D virus. It means that the live rBCG-VP1 elicit efficient humoral and cell-mediated imrnune responses against EMC-D virus, resulting in prevention of virus-induced diabetes in susceptible mice.
Animals ; Antibodies ; Antibodies, Neutralizing ; Bacillus ; Capsid Proteins ; Clone Cells ; DNA, Complementary ; Guinea Pigs ; Humans ; Hypersensitivity ; Immunization* ; Insulin-Secreting Cells ; Male* ; Mice* ; Mycobacterium bovis*

Presentation of antigen in a suitable form to lymphocytes is prerequsite for the initiation of primary immune response. Dendritic cells (DC) provide an effective pathway for presenting antigens to lymphocytes in situ. The aim of this study was to establish a dendritic cell-line from human tonsils and to investigate the changes in surface phenotype during culture. Immunohistochemical studies using various surface markers indicated that cultured DC were follicular dendritic cells (FDC) from human tonsils. Cultured DC showed typical dendritic morphology at early stage of culture. Their shape changed into fibroblast-like cells over culture time. Surface phenotype study suggested that cultured DC were distinct from human fibroblast. Antigenic pattern of FDC was changed during culture; I-ILA-DR antigens decreased and HJ2 antigens increased with aging of culture. Functional characteristics of human tosillar FDC will be investigated in the future.
Aging ; Dendritic Cells ; Dendritic Cells, Follicular* ; Fibroblasts ; Humans ; Lymphocytes ; Palatine Tonsil ; Phenotype

Present study was aimed to investigate the immunological activities of the 47 kDa protein antigen from Treponema pallidum and conducted on 12 patients with syphilis (early, late, spontaneously healed, congenital and treated patients) followed by therapy. Peripheral blood mononuclear cells (PBMC) were obtained three times from each patient, on admission before the initiation of therapy, 1 and 6 months later. Eleven (96.7%) of the patients prior to therapy, showed depressed lymphoproliferative responses to the 47 kDa antigen (stimulation index <4) by 3H-thymidine incorporation assay. However, these T cell responses were seemed to be transient because most of the patients (63.6%) exhibited significantly higher lymphoproliferation after therapy. Before therapy, PBMC from spontaneously healed syphilis patients resulted in significantly increased gene expression of IFN- and proinflarnmatory cytokines, such as TNFa, IL-1B and IL-6, in response to the 47 kDa. Patients with late latent and late congenital syphilis exhibited lower IFN-r and proinflammatory cytokine mRNA expression than spontaneously healed syphilis group did. After therapy, IFN-r and proinflammatory cytokine mRNA expressions were gradually reduced in these groups. On the other hand, IFN- and proinflammatory cytokine gene expressions were considerably depressed in early syphilis patients, but these patients went on to express prominent IFN-r and proinflamrnatory cytokine mRNA with treatment. These data suggest that the pattern of cellular immune response in response to the 47 kDa antigen may be involved in the evaluation of the clinical course and outcome of syphilis followed by therapy.
Cytokines ; Flow Cytometry ; Follow-Up Studies* ; Gene Expression ; Hand ; Humans ; Immunity, Cellular ; Interleukin-6 ; RNA, Messenger ; Syphilis ; Syphilis, Congenital ; Treponema pallidum* ; Treponema*

LPS and IFN-r induce nitric oxide synthase in macrophages and the resultant NO causes apoptotic cell death in the activated macrophages. NO production and apoptosis were inhibited by N-monomethyl L-arginine (NMMA), a competitive inhibitor of NO synthase. To study the role of BCL-X proteins, RAW 264.7 cells were transfected with the expression vectors with human bcl-Xl or bcl-Xs cDNAs, respectively. Stable transfectants were selected and confirmed by RT-PCR. NO production in response to LPS and IFN-r caused apoptosis in RAW 264.7 cells and vector transfected control cells within 24 hr. Both NO production and apoptosis were inhibited by N(G)-monomethyl L-arginine (NMMA). In contrast, bcl-Xs transfectant appeared slightly susceptible and bcl-X(L)< transfectant appeared slightly resistant, although NO production was similar to control cells. These results suggest that bcl-X proteins play roles in both positive and negative regulation of apoptosis induced by NO.
Apoptosis* ; Arginine ; bcl-X Protein* ; Cell Death ; DNA, Complementary ; Humans ; Macrophages* ; Nitric Oxide ; Nitric Oxide Synthase

Genes of cancer-associated testis antigens (CTAs) are expressed in various cancer tissues. In order to use CTAs as cancer diagnosis marker, we developed molecular method for detection of CTAs transcripts in tissue. In order to know the applicability of DNA of cancer-associated testis antigens (CTAs) on cancer diagnosis, molecular diagnostic methods for detection of gene expression of melanoma antigen gene (MAGE), GAGE, and B melanoma antigen (BAGE) was studied. After comparing DNA sequences of CTAs, S1/AS1 and S2/AS2, GAGE-S/ GAGE-AS, and BAGE-S/BAGE-AS primers were designed for the detection of MAGEs, GAGEs and BAGEs, respectively. The gene expression of CTAs in cancer cell lines, head and neck cancer tissues, ovary cancer tissue, and peritoneal cells of gastric cancer patients were investigated by reverse transcription-polymerase chain reaction (RT-PCR) using these primers. The MAGEs, GAGEs and BAGE genes were expressed in 8/8 (100%), 5/8 (62.5%) and 1/8 (12.5%) of head and neck cancer tissues, respectively. The gene expression of MAGEs were also detected in 8/10 (80%) of ovary cancer tissues and in 9/10 (90%) of peritoneal cells of gastric cancer patients in RT-PCR test using S1/AS1 primers. The results of this study suggest that molecular diagnosis method using CTAs genes, especially RT-PCR using S1/AS1 primer combination, is useful for diagnosis of cancer and it will be used for the prediction of cancer progression or regression and metastasis in future.
Base Sequence ; Cell Line ; Diagnosis* ; DNA ; Gene Expression ; Head and Neck Neoplasms ; Humans ; Melanoma ; Neoplasm Metastasis ; Ovarian Neoplasms ; Pathology, Molecular ; Stomach Neoplasms ; Testis*

Human seminal plasrna (HSP) is mixture of secretion derived from various glands associated with male reproductive tract which comprises approximately 80-90% of the volume of normal ejaculate. The present study was undertaken in an effort to explore the effect of HSP pretreatment on the production of IL-1B, TNF-a and IL-12, in mice, and to investigate if HSP may cause to induce active systemic anaphylaxis (ASA) in mice. In addition, effects of HSP pretreatment on contact hypersensitivity to trinitrochlorobenzene (TNCB), antibody response to polyvinylpyrroridone (PVP), a thymus-independent antigen and on ASA induced by egg albumin (OVA) were also studied in this study. For the experiments of contact hypersensitivity, antibody response and cytokine production, mice were pretreated i.p. daily with 0.3ml of HSP or sterile saline alone (control) for 3 consecutive days before antigen sensitization or lipopolysaccharide injection for the cytokine induction. For the experiments of OVA- induced anaphylaxis, mice were pretreated by a single s.c. injection of HSP 0.3ml per mouse before sensitization. For induction of ASA in mice by HSP, a group of mice were sensitized i.p. 2 consecutive days with 0.3ml of HSP and one day with 0.3 ml of HSP plus 2x10(9) B. pertussis and 1.0 mg of alum (schedule A) or another group of mice were sensitized i.p. with a single i.p. injection of 0.3 ml of HSP with 2x10' B. pertussis and 1.0 mg of alum (schedule B). All sensitized and unsensitized control mice were challenged i.v. with 0.2ml of HSP 14 days after HSP sensitization, and mortality were observed. It was found that HSP pretreatment inhibited the production of IL-lB, TNF-a and IL-12, and also inhibited OVA-induced ASA, contact hypersensitivity to TNCB and anti-PVP antibody production. Interestingly, ASA was induced by HSP irrespective of the applied sensitization schedule. Taken together, this study may provide the direct evidences that HSP may inhibit the production of IL-1B, TNF-a and IL-12 and this may be the first to show the induction of ASA by HSP in mice.
Anaphylaxis* ; Animals ; Antibody Formation ; Appointments and Schedules ; Dermatitis, Contact ; Humans* ; Interleukin-12 ; Male ; Mice* ; Mortality ; Ovum ; Picryl Chloride ; Semen* ; Whooping Cough

Capsaicin, the pungent principle of hot peppers, is a neurotoxin that depletes unmyelinated primary sensory neurons (polymodal nociceptors) of neuropeptides like tachykinins. However, the role of capsaicin-sensitive sensory nerve in the production of cytokines, penicillin V (PEV)-induced active fatal anaphylaxis and other immune responses is not yet fully established. Neonatal mice were pretreated s.c. with a single injection of 10 ug of capsaicin per mouse in volume of 20 ul within 5 days of age. Using 5-8 week old mice pretreated as neonates with capsaicin, the capsaicin- pretreated and vehicle-treated control mice were examined for various parameters of immune responses described above. For the induction of active fatal anaphylaxis with PEV, 8 week old mice pretreated as neonates and age-matched capsaicin- untreated control mice were sensitized i.p. with 500 ug of PEV-ovalbumin conjugate plus 2*10(9) B. pertussis and 1.0 mg alum and challenged i.v. with PEV-bovine serum albumin conjugate 14 days later. It was found that neonatal capsaicin-pretreatment significantly enhanced contact hypersensitivity to TNCB and hemagglutination response to SRBC, but significantly inhibited the proliferation response of rnurine splenocyte to Con A and LPS. Interestingly, neonatal capsaicin pretreatment significantly inhibited the intensity of PEV-induced active fatal anaphylaxis and decreased the mortality due to anaphylactic shock. It also significantly inhibited LPS- induced production of cytokines such as TNF-a, IL-1B, IL-6, IL-10, and IL-12. The capsaicin-pretreatment also resulted in an inhibition of the activation of NF-kB. Taken together, these data showed for the first time that neonatal capsaicin-pretreatment significantly inhibited an antibiotic (PEV)-induced anaphylaxis and production of various cytokines, and suggest that capsaicin-sensitive primary sensory nerve may play an important regulatory role in active fatal anaphylaxis and cytokine production, thus potentially presenting tools for immune intervention. In particular, the data presented also indicated the possibility to selectively down-modulate cytokine production and NF-kB activation may offer a broad application for therapeutic intervention in neuroimmunological diseases and other pathological situations.
Anaphylaxis ; Animals ; Capsaicin* ; Cytokines ; Denervation* ; Dermatitis, Contact ; Hemagglutination ; Humans ; Infant, Newborn ; Interleukin-10 ; Interleukin-12 ; Interleukin-6 ; Mice ; Mortality ; Neuropeptides ; NF-kappa B ; Penicillin V ; Sensory Receptor Cells ; Serum Albumin ; Tachykinins ; Whooping Cough