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Preliminary assessment of correlation between T-lymphocyte responses and control of porcine reproductive and respiratory syndrome virus (PRRSV) in piglets born after in-utero infection of a type 2 PRRSV

  • Cha, Sang-Ho;Bandaranayaka-Mudiyanselage, Carey;Bandaranayaka-Mudiyanselage, Chandima B.;Ajiththos, Dharani;Yoon, Kyoung-Jin;Gibson, Kathleen A.;Yu, Ji-Eun;Cho, In-Soo;Lee, Stephen S.;Chung, Chungwon J.
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.9-16
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    • 2018
  • A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, $CD3^+CD4^+$, $CD3^+CD8^+$, $CD3^+CD4^+CD8^+$ T-lymphocytes, and PRRSV-specific interferon $(IFN)-{\gamma}$ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages ($CD172{\alpha}^+PRRSV-N^+\;PAM$) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of $CD172{\alpha}^+PRRSV-N^+\;PAM$ (p < 0.05) as well as with macroscopic lung lesion (p < 0.01). Plasma and tissue viral loads had significant (p < 0.05) negative correlations with $CD3^+CD4^+CD8^+$ T-lymphocyte percentage in PBMC. Frequencies of $CD3^+CD8^+$ and $CD3^+CD4^+$ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads. $IFN-{\gamma}$-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002). However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

FLOW CYTOMETRIC ANALYSIS OF LYMPHOCYTES IN NORMAL AND INFLAMED PULP (유세포분석기를 이용한 정상치수조직과 염증성 치수조직 내의 임파구 분포에 관한 연구)

  • Kim, Seon-Ah;Bae, Kwang-Shik;Im, Seong-Sam
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.374-387
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    • 1997
  • The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.

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Effect of negative therapy at back meridian points on blood gas components and immune functions in male college students (배부(背部) 경혈(經穴)에 부항요법(附缸療法) 시술(施術)이 남자대학생(男子大學生)의 면역기능(免疫機能)에 미치는 영향(影響))

  • Oh, Jae-Keun;Kim, Sung-Soo
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.75-83
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    • 1999
  • To investigate the effects of negative therapy at back meridian points on blood gas components and immune functions in male college students, this study was conducted on treatment types(abdomen group and back group) at three sampling times (before, post-2 wks and post-4 wks) by using $2{\times}3$ factoral design. Blood gas $components(pH,\;PCO_2,\;PO_2,\;HCO_3^-,\;O_2SAT,\;BE)$, red blood cell, hematocrit, hemoglobin, white blood cell and subsets(neutrophil, basophil, eosinophil. lymphocyte, monocyte), total T cells, helper T cells, suppressor T cells, Th/Ts ratio, total B cells, serum immunoglobulin levels (IgG, IgA, IgM, IgD, IgE), Cytokines(Interlukin$-1{\beta}$, -2, -4, 2 receptor, -6 and ${\gamma}$-interferon), NK cells were measured. Collected with data were analyzed statistically by repealed measured ANOVA. The pattern of change between two groups for hematocrit, hemoglobin, suppressor T cells, interleukin-6, ${\gamma}-interferon$, NK cells at post-2 weeks and BE, lymphocyte, basophil at post-4 weeks was significantly different(p<0.05) And also the pattern of change over time for ${HCO_3}^-$(2 wks vs 4 wks), WBC, neutrophil, lymphocyte(0 wks vs 2 wks and 2 wks vs 4 wks) was significantly different(p<0.05). In summary, these data suggest that negative therapy at back meridian points had an effect on blood gas components and immune functions in male college students because practicing negative therapy at back meridian points was not associated with changes of all blood gas components and immune factors but associated with changes of BE, hematocrit, hemoglobin, WBC. neutrophil, lymphocyte, interleukin-6. ${\gamma}-interferon$, NK cells.

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Inhibitive Effects of Meju Extracts Made with a Single Inoculum of the Fungi Isolated from the Traditional Meju on the Human Leukemia Cell Line (전통 메주에서 분리된 단독균으로 제조한 메주추출물의 혈액암세포에 대한 저해효과)

  • Han, Jung;Kim, Hyun-Jeong;Lee, Sang-Sun;Lee, In-Seon
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.312-317
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    • 1999
  • In order to study the antitumoral effect of meju extracts, which was made with a single inoculum of the microorganism, the cytotoxicity effects on several human leukemia cells such as promyelocytic leukemia cell (HL60), histiocytic lymphoma cell (U937) and acute T-cell leukemia Jurkat cell, and lymphocyte were analyzed by MTT assay. Twenty one microbes, mainly fungal genera, were isolated from Korean traditional mejus of different regions. From those collected isolates, meju was manufactured and extracted with 80% methanol, respectively. Meju methanol extracts exhibited low activites in cytotoxicity tests on HL60 cell, but high antitumoral effects of meju methanol extracts were shown on U937 and Jurkat cells. Meju methanol extracts made with a genera of Mucor, Absidia and Aspergillus showed prominant cytotoxic activities, especially. However all these extracts had no inhibitory effects on the cell growth of lymphocyte under the same conditions.

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Risk Factors of Syme Amputation in Patients with a Diabetic Foot (당뇨발 환자에서 Syme 절단술의 위험 인자)

  • Gwak, Heuichul;Kim, Jeongyo;Kim, Youngjun;Lee, Jeonghan;Lee, Woonseong
    • Journal of Korean Foot and Ankle Society
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    • v.20 no.2
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    • pp.78-83
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    • 2016
  • Purpose: This study examined the factors affecting the treatment of diabetes mellitus foot patients who had undergone a Syme amputation. Materials and Methods: This study included 17 patients diagnosed with a diabetes mellitus foot and who had undergone a Syme amputation from January 2010 to January 2014. Some of the risk factors (age, body mass index [BMI], disease duration, smoking, ankle brachial index [ABI], HbA1c, serum albumin, total lymphocyte, C-reactive protein [CRP], and serum creatine) that affect the successful Syme amputation were analyzed. Results: The healing rate of a Syme amputation was significantly higher when the lymphocyte count was above $1,500mm^3$ (p=0.029). The factors affecting the surgical outcome according to multivariate analysis were HbA1c and the BMI (p=0.014, p=0.013). Regarding reamputation, there was a significant difference with HbA1c, lymphocyte, and BMI (p=0.01, p=0.03, and p=0.01). No significant differences were observed with age, disease duration of diabetes mellitus, smoking, ABI, serum albumin, CRP, and serum creatine. Conclusion: The HbA1c level, BMI and total lymphocyte count are risk factors that must be considered for successful Syme amputation in patients with diabetic foot disease.

Enhancing immune responses to inactivated foot-and-mouth virus vaccine by a polysaccharide adjuvant of aqueous extracts from Artemisia rupestris L.

  • Wang, Danyang;Yang, Yu;Li, Jinyu;Wang, Bin;Zhang, Ailian
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.30.1-30.15
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    • 2021
  • Background: New-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects. Objectives: In this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles. Methods: The mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination. Results: AEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4+ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months. Conclusions: These findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.

Estimation of environmental effects and genetic parameters for somatic cell score, stress and immunological traits in Holstein cattle (젖소에 있어서 원유 중 체세포수, 스트레스 및 면역물질에 대한 환경효과와 유전모수 추정)

  • An, Byeong-Seok;Seo, Guk-Hyeon;Gwon, Eung-Gi
    • Journal of Animal Science and Technology
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    • v.48 no.1
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    • pp.9-14
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    • 2006
  • Milk yield and its quality traits determine the dairy enterprise profitability and sustainability. Milk quality traits including somatic cell counts (SCC) is an upcoming economic challenge for dairy farming community in Korea. This study estimated the effect of parity, stage of lactation (early, mid and late lactation) on SCC, stress (blood cortisol) and immunity (blood IgG, lymphocyte and neutrophil) traits, their heritabilities and genetic correlations between them. SCS and blood neutrophil count were significantly affected by both parity and stage of lactation, however; IgG was affected by only stage of lactation, and blood cortisol and lymphocyte were not affected by both factors. The SCS has shown increasing trend with the parity, however; the difference between first and second parity, second and third parity were not significant. The SCS in early (≤90 days) and late lactation (181≤days) were higher than that of mid lactation (91~180 days). Cortisol concentration in blood was lowest in fourth parity, however; the differences among the first three parties were not significant. The IgG was higher in fourth parity compare with first parity however; all other comparisons were noted non-significant. The IgG concentration was significantly higher in early lactation than those of mid and late lactation. The blood lymphocytes were decreased with increasing parity however the differences beyond second parity were not significant. The neutrophils were increased with the increasing lactation stage however; the difference between early and mid lactation was not significant. Although heritability of SCS was still lower, but it was meaningful value (0.09) and may be considered to improve milk quality. The genetic correlations between SCS and cortisol (-0.96), and lymphocyte (-0.76) were highly negative. Heritability of cortisol was low, however genetic correlations between cortisol and lymphocyte (0.79) was highly positive. IgG with medium heritability was correlated negatively with lymphocyte (-0.88) and neutrophil (-0.98). Lymphocyte was lowly heritable and highly correlated with neutrophil concentration (0.87).This study suggested that cortisol, IgG, lymphocyte and neutrophil being positively genetically correlation with somatic cell score could be used as alternative traits to enhance milk quality in Holstein cattle. Further studies are warranted to estimate genetic relationships between immunological and production traits to increase the genetic merit of Holstein cows for milk yield, to improve animal health and economic viability under intensive management system.

The effects of Galgŭnhaegit'ang and Won's-Galgŭnhaegit'ang for Taeŭmin on the immune response (태음인(太陰人) 갈근해기탕(葛根解肌湯)과 원특상(元特上)의 갈근해기탕(葛根解肌湯)이 면역반응(免疫反應)에 미치는 영향(影響))

  • Park, Seong-Sik;Ko, Byung-Hee;Song, Il-Byung
    • Journal of Sasang Constitutional Medicine
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    • v.9 no.1
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    • pp.315-337
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    • 1997
  • In order to compare the dffects of $Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisusebowon(東醫壽世保元)and Won's(元)-$Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisasansinpyun(東醫四象新編)" on the immune respone, Sprague-Dawley male rats were used and randomly divided into four groups. Normal group was under normal condition, Control group was injected i.v. with 2mg/kg Methotrexate(MTX) on the 9th day and 11th day after sensitization with SRBC on the 5th day, $Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of $Galg{\breve{u}}nhaegit'ang$ and Won's-$Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of Won's-$Galg{\breve{u}}nhaegit'ang$ by oral during eighteen days. In the 9th day and the 11th day after oral feeding with medication, MTX was injected in tail of rats in order to reduce immune function. Leukocyte count, lymphocyte ratio, lymphocyte count of spleen, lymphocyte count of bone marrow, contact hypersensitivity to DNFB, morphologic change of thymus cell, and electropherogram of serum protein were estimated and compared according to the group. The results are as follows : 1. Before and after MTX injection, leukocyte(WBC) count was increased signigicantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 2. Before and after MTX injection, lymphocyte ratio was not significantly different in Won's-$Galg{\breve{u}}nhaegit'ang$ group and in $Galg{\breve{u}}nhaegit'ang$ group compared to control group. 3. The lymphocyte count of spleen was increased significantly in $Galg{\breve{u}}nhaegit'ang$ group compared to control group and Won's-$Galg{\breve{u}}nhaegit'ang$ group. Won's-$Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 4. The lymphocyte count of bone marrow was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group and $Galg{\breve{u}}nhaegit'ang$ group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 5. Contact hypersensitivity was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to other group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control groups. 6. In the morphologic change of thymus cell, control group compared to normal group had a indistinct boundary between cortex and medulla and lymphocyte cell density of thymus was low. $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group had a definite boundary between cortex and medulla and lymphocyte cell density of thymus was high. 7. In the SDS-PAGE electropherogram of serum protein, Won's-$Galg{\breve{u}}nhaegit'ang$ group had a wide band of nearby 25,000 Dalton, and which meant IgG generated more actively. Considering this results, $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group have an effect on the depression of immune function induced by MTX, and especially Won's-$Galg{\breve{u}}nhaegit'ang$ group has an significant effect than $Galg{\breve{u}}nhaegit'ang$ group.

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The Role of Lymphocyte Compartment and Cytokine in Coal Workers Pneumoconiosis (진폐증환자에서의 임파구 분획 및 싸이토카인의 역할)

  • Lee, Jung-Yeon;Yoo, Kwang-Ha;Ahn, Hae-Ryon;Kim, Sung-Ryul;Lee, Hae-Woon;Ahn, Cheol-Min;Kim, Hyung-Joong
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.3
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    • pp.241-250
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    • 2002
  • Background : Coal-worker's pneumoconiosis(CWP) is characterized by a chronic inflammatory lung reaction associated with macrophage accumulation in the alveolar spaces. CWP is usually divided into two stage : simple pneumoconiosis(SP) where there are a limited number of fibrotic lesions remain limited, with radiological opacities smaller than 1cm and progressive massive fibrosis(PMF), which is characterized by the development of a perifocal extensive fibrotic response of the lung and severe alterations in pulmonary function. In this study, the lymphocyte compartment and cytokine were evaluated by measuring the serum levels in the control, SP and PMF groups. Materials and Methods : The coal workers selected for this study were employees(patients?) of the Tae-Baek and Dong-Hae hospital. All were men, 45-76 years old and the mean duration of their exposure to coal dust was 23.2 years in the lymphocyte compartment and 24.3 years in the cytokine checked group. According to X-ray examination results, the patients were classified into either one of the SP, PMF categories. The normal controls examined were 26-70 years old men. The serum cytokine levels were estimated by using an end point enzyme immunoassay technique. Results : T lymphocyte, helper and suppressor T cells were highly related to pneumoconiosis in this study. A statistically significant decrease in the number of suppressor T lymphocytes was observed in the simple pneumoconiosis patients and at the same time, there was an increase in the lymphocyte index. Howevere, there was no statistically difference in the serum cytokines levels among the SP, PMF and control groups. Conclusion : T lymphocyte, helper T, and suppressor T cells may be highly related to the development of CWP compared to the control group particularly in the early stage of pneumoconiosis. The changes observed in the immunological system in patients with pneumoconiosis may lie at the bottom of the pathogenesis of fibrosis. Further study is needed to evaluate the lymphocyte compartment as a marker for pneumoconiosis development in the early stage.

The Relationship between Cell-mediated Immunity and Subtypes of Lymphocyte in BAL Fluid and Peripheral Blood in Patients with Pulmonary Tuberculosis (폐결핵 환자의 말초 혈액 및 기관지 폐포세척액내의 임파구 아형과 세포성 매개면역과의 관계)

  • Uh, Soo-Taek;Cha, Mi-Kyung;Lee, Sang-Moo;Kim, Hyun-Tae;Chung, Yeon-Tae;Woo, Jun-Hee;Kim, Yong-Hun;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.4
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    • pp.334-342
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    • 1992
  • Background: The activated T lymphocyte by inhalaed mycobacterial antigen may evoke cell-mediated immunity in patients with active pulmonary tuberculosis. These activated lymphocyte may influence the response of tuberculin-purified protein derivative (PPD) in skin test. But occasionally, anergy to PPD appear in patients with pulmonary tuberculosis in spite of active stage. Thus we evaluated the effect of change of subtypes of lymphocyte in bronchoalveolar lavage fluid (BAL) and peripheral blood on anergy to PPD in patients with active pulmonary tuberculosis. Method: We performed tuberculin skin test and flow-cytometry analysis of lymphocytes obtained from BAL fluid and peripheral blood in 11 healthy normal volunteers and 20 patients with active pulmonary tuberculosis. Results: 1) The composition of lymphocyte significantly increased in patients with active pulmonary tuberculosis when compared with that in healthy control ($25.2{\pm}4.8$ vs $6.5{\pm}1.3%$, p<0.01), but composition of monocyte significantly decreased ($69.6{\pm}5.7$ vs $89.2{\pm}1.4%$, p<0.05) in analysis of BAL fluid. 2) There were no differences in compositions of cells in BAL fluid between responders and no-responders to PPD. 3) The compositions of CD3 (+), CD4 (+), CD3 (+) IL-2R (+), CD3 (+) HLA-DR (+) significantly increased in BAL fluid when compared with those in peripheral blood in patients with active pulmonary tuberculosis. But the composition of CDS (+), CD4/CDS were not different between BAL fluid and peripheral blood. 4) There were no correlations between response to PPD and compositions of cells and lymphocyte subtypes in BAL fluid and peripheral blood in all patients with tuberculosis, responders, and no-responders, respectively. Conclusion: From these results, we suggest no direct relationship between compositions of inflammatory cells in bronchoalveolar lavage fluid and we could not rule out the possibility of compartmentalization of activated lymphocyte involving in anergy to PPD in skin test in patients with active pulmonary tuberculosis.

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