• Title/Summary/Keyword: B lymphocytes

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Characterization of B Cells of Lymph Nodes and Peripheral Blood in a Patient with Hyper IgM Syndrome (Hyper IgM Syndrome 환자에서 얻은 림프절 및 말초혈액 B세포의 특성)

  • Kim, Dong Soo;Shin, Kyuong Mi;Yang, Woo Ick;Shin, Jeon-Soo;Song, Chang Hwa;Jo, Eun Kyeong
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.128-136
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    • 2003
  • Purpose : Hyper IgM syndrome(HIGM) is characterized by severe recurrent bacterial infections with decreased serum levels of IgG, IgA, and IgE but elevated IgM levels. Recently, it has been classified into three groups; HIGM1, HIGM2 and a rare form of HIGM. HIGM1 is a X-linked form of HIGM and has now been identified as a T-cell deficiency in which mutations occur in the gene that encodes the CD40 ligand molecule. HIGM2 is an autosomal recessive form of HIGM. Molecular studies have shown that the mutation of HIGM2 is in the gene that encodes activation-induced cytidine deaminase(AID). Recently, another rare form of X-linked HIGM syndrome associated with hypohydrotic ectodermal dysplasia has been identified. We encountered a patient with a varient form of HIGM2. To clarify the cause of this form of HIGM, we evaluated the peripheral B cells of this patient. Methods : The lymphocytes of the patient were prepared from peripheral blood. B cells were immortalized with the infection of EBV. Cell cycle analysis was done with the immortalized B cells of the patient. Peripheral mononuclear cells were stained with monoclonal anti-CD40L antibody. Total RNA was extracted from the peripheral mononuclear cells. After RT-PCR, direct sequencing for CD40L gene and HuAID gene were done. Immunostainings of a lymph node for CD3, CD23, CD40, Fas-L, bcl-2, BAX were done. Results : The peripheral B cells of this patient showed normal expression of CD40L molecule and normal sequencing of CD40L gene, and also normal sequencing of AID gene. Interestingly, the peripheral B cells of this patient showed a decreased population of G2/mitosis phase in cell cycles which recovered to normal with the stimulation of IL-4. Conclusion : We suspect that the cause of increased serum IgM in this patient may be from a decrease of G2/mitosis phase of the peripheral B cells, which may be from the decreased production or secretion of IL-4. Therefore, this may be a new form of HIGM.

Effect of Glutathione S-Transferase Polymorphisms on the Antioxidant System (Glutathione S-Transferase 유전적 다형성이 항산화 체계에 미치는 영향)

  • Jeon, Gyeong-Im;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.708-719
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    • 2007
  • Glutathione S-transferase genotypes GSTT1, GSTM1 and GSTP1 were characterized in 104 healthy male and female subjects and compared with parameters of oxidative stress at the level of DNA and lipids, with antioxidant enzymes, and with plasma antioxidants in smokers and non.smokers. Of the 104 subjects studied, 57.4% were GSTT1 present and 47.6% were GSTM1 present. The GSTP1 polymorphisms a and b were represented as follows: a/a, 75.5%; a/b, 21.6%; b/b type, 2.9%. The GSTT1 null genotype was associated with decreased glutathione in erythrocytes and elevated lymphocytes DNA damage. GST-Px was higher in GSTT1 null compared with GSTT1 present type. The homozygous GSTP1 genotype was not associated with any antioxidant status or DNA damage. The difference in plasma ${\alpha}$-carotene and erythrocytes GSH-Px and GST activities between smokers and non-smokers was detected in the GSTT1 null genotype. Plasma ${\gamma}$-tocopherol and ${\beta}$-carotene decreased significantly in smokers having GSTM1 null genotype. When GSTT1 and GSTM1 were combined, plasma lycopene and erythrocyte GST were reduced in smokers in both null types of these genes. As for GSTP1 genotype, plasma ${\alpha}$-carotene and erythrocytes GSH-Px decreased significantly in smokers with GSTP1 b/b, while erythrocytes GSH-Px activities decreased in smokers with GSTP1 a/b. The different ${\beta}$-carotene level between smokers and non-smokers was seen with both GSTP1 a/a and a/b genotype. It seems that polymorphisms in the phase II metabolizing enzyme glutathione S-transferase may be important determinants of commonly measured biomarkers.

Efficient Transduction with Recombinant Adenovirus in EBV-transformed B Lymphoblastoid Cell Lines

  • Kim, Hye-Jin;Cho, Hyun-Il;Han, Yoon-Hee;Park, Soo-Young;Kim, Dong-Wook;Lee, Dong-Gun;Kim, Jee-Hoon;Shin, Wan-Shik;Paik, Soon-Young;Kim, Chun-Choo;Hong, Young-Seon;Kim, Tai-Gyu
    • BMB Reports
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    • v.37 no.3
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    • pp.376-382
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    • 2004
  • The Epstein-Barr-transformed B lymphoblastoid cell lines, LCL, which express antigens, are potential antigen-presenting cells (APCs) for the induction of cytotoxic T lymphocytes in vitro. However, transfecting LCL with subsequent selection by antibiotics is notoriously difficult because the plating efficiencies of LCL are reported to be 1% or less. Therefore, this study investigated the optimal conditions for increasing the transduction efficiency of a recombinant adenovirus to LCL for use as a source of APCs. The transduction efficiencies were < 13% (SD $\pm$ 2.13) at a multiplicity of infection (MOI) of 100, while it was increased to 28% (SD $\pm$ 9.43) at an MOI of 1000. Moreover, its efficiencies to LCL that expressed the coxsackie adenovirus receptor were increased to 60% (SD $\pm$ 6.35) at an MOI of 1000, and were further increased to 70% (SD $\pm$ 4.56) when combined with the centrifugal method. The cationic liposome or anionic polymer had no effect on the transduction efficiency when compared to that of the centrifugal method. These results may be used as a convenient source of target cells for a CTL assay and/or autologous APCs for the induction of the in vitro CTL responses that are specific to viral and tumor antigens.

Anti-cancer Effects of Cultivated Wild Ginseng Herbal Acupuncture in C57BL/6 Mice Injected with B16/F10 Cells and Reproductive Toxicity by Doxorubicin (B16/F10세포를 이식한 C57BL/6 생쥐에서 산삼약침의 항암효과 및 Doxorubicin에 의한 생식독성 완화효과)

  • Yoon, Hye-Chul;Kim, Ho-Hyun;Kwon, Ki-Rok
    • Journal of Acupuncture Research
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    • v.23 no.1
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    • pp.105-120
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    • 2006
  • To investigate anti-cancer effects of wild ginseng herbal acupuncture and mitigation of anti-cancer drug when taken concurrently, cancer cells from B16/F10 melanoma were injected intraperitoneally in C57BL/6. After inducing cancer, anti-cancer effects and mitigation of reproductive toxicity of Doxorubicin were evaluated. 1. For changes in weight, Doxorubicin treated group showed significant decrease, and administration of wild ginseng herbal acupuncture didn't cause any weight change. 2. Volume of tumor was significantly reduced in Doxorubicin teated group. Wild ginseng herbal acupuncture groups showed slight decrease but insignificant compared to the control group. 3. For hematological evaluation, Doxorubicin only group's reticulocytes were significantly decreased compared to the control group, and Platelet Count was significantly increased. Wild ginseng herbal acupuncture group showed significant increase of Neutrophils and significant decrease of Lymphocytes compared to the control group. 4. For histological evaluation of the tumor, necrosis occurred in a wide range in the Doxorubicin treated group. Wild ginseng herbal acupuncture didn't cause much histological changes. 5. For histological evaluation of the testis, seminiferous tubules of the control group suffered severe damage on epithelial cells. When wild ginseng herbal acupuncture was administered concurrently, damage on the seminiferous tubules was significantly inhibited compared to the Doxorubicin only group. 6. Diameter of seminiferous tubules and spermatogonia count were insignificant between the experiment groups. 7. For BrdU positive reaction of testicle tissue, Doxorubicin only group failed to show any reaction of spermatogonia, but spermatocytes and spermatids showed slight positive reaction. When wild ginseng herbal acupuncture was treated concurrently, much greater positive reaction was made but similar to that of the control and normal groups. 8. For observation of changes in BrdU spermatogonia count of the testicle tissue, Doxorubicin only group didn't show any positive reaction, and relative increase was shown in the group with concurrent administration of wild ginseng herbal acupuncture. 9. For observation of TUNEL positive reaction cells of the testicle tissue, no significant changes were witnessed in all the experiment groups.

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Effect of Jeseupwiryeongtang-gagam on Atopic Dermatitis-like Skin NC/Nga Mouse Induced by Dermatophagoides Farinae Crude Extract (제습위령탕가감이 집 먼지 진드기 추출물로 유도된 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Na, Dong-Gyu;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.22 no.2
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    • pp.81-114
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    • 2008
  • Objectives : The purpose of this study is to investigate the effect of Jeseupwiryeongtang-gagam(JWTG) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of JWTG on AD, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells(PBMCs), splenocytes, draining lymph node(DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results and Conclusions : In vivo, clinical skin severity score were significantly lower in JWTG group than control group. IgE, IL-6, $TNF-{\alpha}$, IgM, IgG2a and IgG2b levels in serum were decreased remarkably in JWTG group than control group and $IFN-{\gamma}$ production, secreted in Th1 cell were increased by JWTG. After experiment ended, we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+$$CD69^+$, $CD4^+$$CD25^+$ and $CD49b^+$) by flow cytometry. It resulted that total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as normal and $CD3^+$$CD69^+$ were decreased significantly compared with control group in isolated DLN and PBMCs from NC/Nga mouse and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by JWTG. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E) and toluidine staining (mast cells marker) and obtained results that JWTG were effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration). Ear thickness was decreased significantly than the control group and the size of inflammatory lymphocytes cells(ILC) and plasma cells(PC) in DLN were also decreased.

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Histopathological Studies of Mice after Administration of Radioactive Iodine($^{131}I$) (방사성동위원소옥소(放射性同位元素沃素)($^{131}I$)가 "마우스"의 주요장기(主要臟器)의 병리조직상(病理組織像)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Ro, Chae-Song;Lee, Mun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.1 no.2
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    • pp.85-97
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    • 1967
  • Histopathological changes of various organs of the mice after intra-peritoneal injections of radioactive iodine ($^{131}I$) were experimentally observed. Sixity healthy female mice, weighing average 25 gm, devided into 6 groups, were used. The various doses of $^{131}I$ were injected intraperitoneally at different intervals. The histopathological changes after these treatments were observed in organs such as thyroids, parathyroids, livers, kidneys and gonads. Following were the results; 1) Thyroid: In the group A given $^{131}I$ with a single dose of $10{\mu}C$ per gm body weight, it was observed that the protoplasms of follicular epithelial cells were destroyed, the nuclei were expanded or dissoluted, showing pyknotic changes of nuclei and vacuolizations of protoplasms. In the group B given $^{131}I$ with a single dose of $5{\mu}C$ per gm body weight, hyperemias, hemorrhages and hyaline degenerations in the whole area were observed. In the group C given $^{131}I$ with 3 doses of $2.5{\mu}C$ per gm body weight every week, the thyroid parenchyms were destroyed and epithelial cells of varing size were observed in the fibrinous tissues. In the group D given $^{131}I$ with 6 doses of $0.5{\mu}C$ per gm body weight every week, some destroyed follicles and new borne follicles were observed. But the histopathological changes resemble the follicles of the normal thyroid gland. In the group E and F given $^{131}I$ with 8 and 10 doses of $0.2{\mu}C\;and\;0.01{\mu}C$ for each group per gm body weight every two days, both pyknotic changes of nuclei and cytoplasmic vacuolizations of the follicular epithelia, hypertrophies of follicles and abnormal irregular follicular structures were observed, and in the group F, lymphocytes appeared around the thyroid glands. 2) Parathyroid: In the group A, hyperemia, proliferations of connective tissues, karyorrhexes and vacuolizations were observed. In other experimental groups, no particular pathological change was observed. 3) Liver: The degnerative changes and acute or chronic inflammatory changes were observed in proportion to the amount of $^{131}I$ injected. Atrophies of the liver cells, dilatations of sinusoids, hyaline degenerations and necrotic pictures were observed. 4) Kidney: In the group A, congestions and infiltrations of mononuclear cells and granulocytes were observed around the cortical arteries, and in the group B, the degenerative changes of cortexes, and, in the group C and D, hydronephrotic changes were observed respectively, and hyaline degenerations were partially observed. 5) Gonad: In the group A, the follicles were degenerated. The ova in the follicles showed irregular figures. The changes in the group B were almost the same as in the group A, but the changes were mild. In the group C, the destructions of whole ova, the hypertrophies of ovarian follicular membranes and pyknotic changes of nuclei were observed. In the group D, the pathological changes were similar to that of group C, but mild in the grade. In the group E, almost none of ovarian follicular fluid was observed, and in the group F, the tissue pictures were almost similar to that of the normal group.

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Reconstructed Adeno-Associated Virus with the Extracellular Domain of Murine PD-1 Induces Antitumor Immunity

  • Elhag, Osama A.O.;Hu, Xiao-Jing;Wen-Ying, Zhang;Li, Xiong;Yuan, Yong-Ze;Deng, Ling-Feng;Liu, De-Li;Liu, Ying-Le;Hui, Geng
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4031-4036
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    • 2012
  • Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

Effect of sodium butyrate on performance, immune status, microarchitecture of small intestinal mucosa and lymphoid organs in broiler chickens

  • Sikandar, Arbab;Zaneb, Hafsa;Younus, Muhammad;Masood, Saima;Aslam, Asim;Khattak, Farina;Ashraf, Saima;Yousaf, Muhammad Shahbaz;Rehman, Habib
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.5
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    • pp.690-699
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    • 2017
  • Objective: This study aimed to examine the effect of sodium butyrate (SB) on growth performance, immune status, organs weights, and microarchitecture of lymphoid organs and small intestine. Methods: A total of 120, 1-d-old broiler chicks were distributed into the following four treatment groups: corn-soy based basal diet (BD) without supplement (control), or the same BD supplemented with 0.1 g/kg zinc bacitracin (ZnB), 0.5 g/kg SB (SB-0.5), or 1.0 g/kg SB (SB-1), respectively. Six birds/group were killed on d-21 and d-35, and samples were collected. Results: Cell-mediated immune response at 48 h post-Phytohemagglutinin-P injection, and antibody titer against Newcastle disease vaccine and sheep red blood cells on d-35 was noted higher (p<0.05) in SB-1 compared to ZnB and control. Lower (p<0.05) feed conversion ratio (FCR) was attained by the supplemented groups. Thymus and spleen weighed more (p<0.05) in SB-1, and bursa registered more (p<0.05) weight in both SB groups compared to control. On d-21, areas of thymus medulla and spleen germinal centers were noted higher (p<0.05) in SB-1 group. The villus height and villus surface area increased (p<0.05) in duodenum and jejunum in both SB groups on d-21, and in SB-1 on d-35, respectively compared to ZnB and control. On d-21, number of goblet cells containing mucins of acidic nature increased (p<0.05) in all the segments of small intestines in SB-1 group compared to control, and on d-35 in ileum compared to other groups. Conclusion: In conclusion, SB improved growth performance and immunity as well as modulated morphology of lymphoid organs and gut mucosa in broiler chickens.

Detection of Canine Lymphoma by the Amplification of Antigen Receptor Gene Rearrangements (재배열 항원 수용체 유전자 증폭을 통한 개 림프종의 진단)

  • Yu, Do-Hyeon;Li, Ying-Hua;Lee, Jong-Hyun;Noh, Dong-Ho;Song, Ru-Hui;Lee, Mi-Jin;Choi, Ul-Soo;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.26 no.5
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    • pp.419-422
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    • 2009
  • We performed the PARR (PCR to detect antigen receptor rearrangements) test on DNA isolated from twelve archival canine cytological slides including nine lymphoma, two reactive lymphocytes and one sample from Ehrlichia canis infected dog. As a result, our PCR control gene, $C{\mu}$, was successfully amplified from all of the DNA samples. Six out of nine lymphoma samples showed a clonal rearrangement of immunoglobulin gene whereas three samples did a clonal rearrangement of T cell receptor gamma ($TCR{\gamma}$) gene. However, we observed no visible or clear bands from PCR conducted using our antigen receptor rearrangement primers on DNA from a reactive lymphoid cell proliferation used as a negative control. False-positive amplification in $TCR{\gamma}$ gene was observed only in one sample from E. canis infection. The use of archival cytological specimens demonstrated in this study offers potential advantages for cost-effective specimen acquisition and efficient high-fidelity DNA analysis.

Anti-inflammatory Effects of Hataedock Extracted from Coptidis Rhizoma and Glycyrrhiza Uralensis on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse (황련-감초 추출물을 이용한 하태독법이 NC/Nga 생쥐에서 유발된 아토피 유사 피부염에 미치는 항염증 효과)

  • Cha, Ho-yeol;Ahn, Sang-hyun;Jeong, A-ram;Cheon, Jin-hong;Park, Sun-young;Choi, Jun-yong;Kim, Ki-bong
    • The Journal of Internal Korean Medicine
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    • v.36 no.4
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    • pp.486-497
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    • 2015
  • Objectives Hataedock is an orally administered herbal extract treatment for newborn babies that dispels toxic heat and meconium gathered by the fetus. The purpose of this study was to evaluate whether Hataedock alleviates inflammatory skin damage in AD (Atopic Dermatitis)-induced NC/Nga mice through regulating and maintaining the skin barrier and anti-inflammation effects.Methods We established an AD model in three-week-old NC/Nga mice through the repeated application of DNFB (dinitrochlorobenzene) on days 28, 35, and 42 after Hataedock treatment was orally administered. We identified changes in the skin barrier and anti-inflammation effects through the histological and immunohistochemical changes of TNF- α, NF-κB p65, iNOS, COX-2, and apoptotic bodies.Results Skin damage and angiogenesis were mitigated in the HT (Hataedock) group. Damage to the intercellular space of the stratum corneum as well as hyperplasia, edema, the infiltration of lymphocytes, and the increase of capillaries decreased in the HT group. Our results suggest that Hataedock treatment significantly down-regulated levels of TNF- α by 38% (p<0.001) and of NF-κB p65 by 70% (p<0.001). But Hataedock up-regulated apoptosis by 183% in dermatitis-induced skin.Conclusions These results suggest that Hataedock alleviates AD through diminishing the various inflammatory cytokines in skin lesions that are involved in the initial steps of AD development. It might have potential applications for the prevention and treatment of atopic dermatitis.