OBJECTIVETo investigate the value of combined determination of neutrophil CD64 and procalcitonin (PCT) in the early diagnosis of neonatal bacterial infection.

METHODSAccording to discharge diagnosis, 37 neonates with bacterial infection were divided into sepsis (n=15) and ordinary infection (non-sepsis) groups (n=22). Twenty-one neonates without infection who were hospitalized during the same period of time were enrolled as the control group. Venous blood samples were collected immediately after admission. Flow cytometry was used to measure the serum level of neutrophil CD64. Chemiluminescence and immune transmission turbidimetry were used to measure the serum levels of PCT and CRP respectively.

RESULTSThe sepsis group had higher serum levels of neutrophil CD64, PCT, and CRP than the control group (P<0.01), the ordinary infection group had a higher serum level of neutrophil CD64 than the control group (P<0.01), and the sepsis group had higher serum levels of PCT and CRP than the ordinary infection group (P<0.01). The areas under the ROC curve (AUC) of neutrophil CD64, PCT, and CRP in diagnosing bacterial infection were 0.818, 0.818, and 0.704 respectively, and the AUC of combined neutrophil CD64 and PCT was 0.926. A combination of neutrophil CD64 and PCT had a sensitivity of 97.29% and an accuracy of 89.65% in the early diagnosis of neonatal bacterial infection.The sensitivity and accuracy were higher than those of a combination of CRP and neutrophil CD64 or PCT as well as neutrophil CD64, PCT, or CRP alone for the early diagnosis of neonatal bacterial infection.

CONCLUSIONSThe combined determination of neutrophil CD64 and PCT can improve the sensitivity and accuracy in the diagnosis of neonatal bacterial infection, which helps with early identification of bacterial infection.

Bacterial Infections ; blood ; diagnosis ; C-Reactive Protein ; analysis ; Calcitonin ; blood ; Early Diagnosis ; Female ; Humans ; Infant, Newborn ; Male ; Neutrophils ; chemistry ; ROC Curve ; Receptors, IgG ; blood

BACKGROUND: Human neutrophil antigens (HNAs) are involved in autoimmune and alloimmune neutropenia and transfusion-related acute lung injury. The HNA-1 system is important in immunogenetics, and allele frequencies have been described in different populations. This study investigated the frequency of FCGR3B alleles encoding HNA-1a, HNA-1b, and HNA-1c among Thai blood donors and compared these frequencies with those previously reported for other populations. METHODS: Eight hundred DNA samples obtained from unrelated healthy blood donors at the National Blood Centre, Thai Red Cross Society, Bangkok, and the Blood Bank, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand, were included. Samples were simultaneously typed for each FCGR3B allele using an in-house polymerase chain reaction with sequence-specific primer (PCR-SSP) technique. RESULTS: The frequencies of FCGR3B*1, FCGR3B*2, and FCGR3B*3 alleles in central Thai blood donors were 0.548, 0.452, and 0.004, respectively; only FCGR3B*1 and FCGR3B*2 alleles were found in northern Thai blood donors (0.68 and 0.32, respectively). Compared with other Asian populations, central Thais had higher frequencies of the FCGR3B*2 allele (P<0.001), while the frequencies of the FCGR3B*1 and FCGR3B*2 alleles in northern Thais were similar to those previously reported in Taiwanese and Japanese populations. In contrast, the frequencies of the FCGR3B*1 and FCGR3B*2 alleles in the northern Thai population were statistically different from those observed in central Thai, Korean, German, and Turkish populations. CONCLUSIONS: FCGR3B allele frequencies were significantly different between central and northern Thai blood donors. Our in-house PCR-SSP method is a simple, cost-effective, and convenient method for FCGR3B allele detection.
Asian Continental Ancestry Group/*genetics ; *Blood Donors ; DNA/analysis ; DNA Primers/chemistry/metabolism ; GPI-Linked Proteins/genetics ; Gene Frequency ; Genotype ; Humans ; Polymerase Chain Reaction ; Receptors, IgG/*genetics ; Thailand

OBJECTIVETo study the clinical value of the expression of neutrophil surface CD64 in the diagnosis of community acquired pneumonia in children.

METHODSNinety-eight children with community acquired pneumonia were recruited into the study and were classified into three groups according to pathogene: bacterial pneumonia (n=48), viral pneumonia (n=29) and Mycoplasmal pneumonia (n=21). Twenty healthy children were enrolled as controls. The bacterial infection group was subdivided into mild infection (n=36) and severe infection groups (n=12). The levels of peripheral blood neutrophil CD64 were measured using flow cytometry. Dynamic changes of C-reactive protein were also detected for each patient.

RESULTSThe CD64 index and CRP levels in the bacterial pneumonia group were significantly higher than in the other three groups (P<0.05). The CD64 index in the severe bacterial infection group was significantly higher than in the mild group (P<0.05). After antibiotic treatment, expression of CD64 in the severe bacterial infection group decreased significantly (P<0.05). The CD64 index was positively correlated with CRP value (r=0.545, P<0.01). ROC curve analysis showed that the threshold of CD64 and CRP was 2.8 and 8 mg/L respectively. Specificity of CD64 index (90%) was much higher than CRP (74%).

CONCLUSIONSThe determination of peripheral blood neutrophil CD64 contributes to the early diagnosis of pulmonary bacterial infection and the evaluation of anti-infection effect.

C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Community-Acquired Infections ; blood ; diagnosis ; Female ; Humans ; Male ; Neutrophils ; chemistry ; Pneumonia, Bacterial ; blood ; diagnosis ; ROC Curve ; Receptors, IgG ; blood

Establishing Epstein-Barr virus(EBV)-specific cytolytic T lymphocytes(EBV-CTLs) from peripheral blood mononuclear cells(PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma(NPC). In the current study, we performed ex vivo expansion of tumor-infiltrating lymphocytes(TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody(OKT3), recombinant human interleukin(IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+ T cells, a variable percentage of CD3+CD8+ and CD3+CD4+ T cells, and less than 10% of CD3-CD16+ natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.
Biopsy ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; Cells, Cultured ; Herpesvirus 4, Human ; immunology ; Humans ; Immunotherapy, Adoptive ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; pharmacology ; Interleukin-4 ; metabolism ; Lymphocytes, Tumor-Infiltrating ; immunology ; virology ; Monocytes ; pathology ; Muromonab-CD3 ; pharmacology ; Nasopharyngeal Neoplasms ; immunology ; pathology ; therapy ; virology ; Receptors, IgG ; analysis ; T-Lymphocytes, Cytotoxic ; immunology ; virology ; Tumor Necrosis Factor-alpha ; metabolism

Neutrophils play an important role in the human immune system for protection against such microorganisms as a protozoan parasite, Trichomonas vaginalis; however, the precise role of neutrophils in the pathogenesis of trichomoniasis is still unknown. Moreover, it is thought that trichomonal lysates and excretory-secretory products (ESP), as well as live T. vaginalis, could possibly interact with neutrophils in local tissues, including areas of inflammation induced by T. vaginalis in humans. The aim of this study was to investigate the influence of T. vaginalis lysate on the fate of neutrophils. We found that T. vaginalis lysate inhibits apoptosis of human neutrophils as revealed by Giemsa stain. Less altered mitochondrial membrane potential (MMP) and surface CD16 receptor expression also supported the idea that neutrophil apoptosis is delayed after T. vaginalis lysate stimulation. In contrast, ESP stimulated-neutrophils were similar in apoptotic features of untreated neutrophils. Maintained caspase-3 and myeloid cell leukemia-1 (Mcl-1) in neutrophils co-cultured with trichomonad lysate suggest that an intrinsic mitochondrial pathway of apoptosis was involved in T. vaginalis lysate-induced delayed neutrophil apoptosis; this phenomenon may contribute to local inflammation in trichomoniasis.
Animals ; *Apoptosis ; Cells, Cultured ; Female ; Humans ; Membrane Potentials ; Mitochondrial Membranes/physiology ; Neutrophils/chemistry/*immunology ; Receptors, IgG/analysis ; Trichomonas vaginalis/*immunology

OBJECTIVETo explore the change of serious abdominal traumatic patients' cellular immunity and the effect of Astragalus Injection (AI) on it.

METHODSSixty-three serious abdominal traumatic patients were randomly assigned into two groups, the conventional group and the treated group, patients in the conventional group were given conventional treatment, while others in the treated group were given conventional treatment as the basis, with AI 20 ml was added into 250 ml of 5% glucose solution given through intravenous dripping, and then on the first day and 14th day, their T cell activated antigens as well as that of 10 healthy subjects were monitored.

RESULTSOn the first day, in the conventional group and treated group, the levels of CD(3)(+), CD(4)(+), CD(4)(+)/CD(8)(+), CD(16)(+), CD(69)(+) and CD(3)(+)/homologous leucocytic antigen-DR (HLA-DR(+)) were apparently lower than those in the healthy group (P < 0.05), while the CD(8)(+) was significantly higher than that in the healthy group (P < 0.05), and there was no significant difference between the conventional group and the treated group (P > 0.05); on the 14th days, the levels of CD(3)(+), CD(4)(+), CD(4)(+)/CD(8)(+), CD(16)(+), CD(69)(+) and CD(3)(+)/HLA-DR(+) of the treated group got closed to healthy subject value, and got even higher than those of conventional group (P < 0.05); CD(8)(+) got close to that of healthy subjects, while obviously lower than that of conventional group (P < 0.05).

CONCLUSIONAfter serious abdominal trauma, cellular immunity lowered, auxiliary use of AI was beneficial to the restoration of cellular immunity.

Abdominal Injuries ; immunology ; Adjuvants, Immunologic ; therapeutic use ; Adult ; Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; Astragalus membranaceus ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunity, Cellular ; Lectins, C-Type ; Lymphocyte Activation ; Male ; Middle Aged ; Phytotherapy ; Receptors, IgG ; analysis ; T-Lymphocyte Subsets

This study was to explore the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on neutrophil morphology, function and phenotype in patients with acute leukemia undergoing chemotherapy. Neutrophil morphology was observed under microscope with oil immersion; phagocytotic function was examined by measuring the amount of hydrogen peroxide produced by neutrophil; chemotaxis was analyzed by agarose method; oxidative burst was analyzed by flow cytometry using immunofluorescence technique; neutrophil phenotype was analyzed by flow cytometry and immunofluorescence techniques. The results showed that after rhG-CSF administration, the increased "toxic" granulation, vacuoles and Döhle bodies were observed in neutrophils of patients with acute leukemia. Compared with normal control, the functions of phagocytosis, chemotaxis, oxidative burst of neutrophil were impaired after chemotherapy, while these functions were enhanced and returned to normal level or even to be exceeded after administration of rhG-CSF. In patients with acute leukemia the neutrophil presented significantly higher expression of CD64 and CD62L than that in normal control, and a mild increase of CD64 expression and significant increase of CD62L expression were found in patients after rhG-CSF treatment. No modifications of CD16, CD32, CD14 and CD11b expression were detected in these patients before or after G-CSF administration. It is concluded that rhG-CSF administration can modify the morphology, function and phenotype of neutrophils in the patients with acute leukemia undergoing chemotherapy, and these modifications of neutrophil behavior may be supposed to be a reason for the enhancement of organism anti-infection ability.
Acute Disease ; Adult ; Aged ; Chemotaxis ; drug effects ; Female ; Flow Cytometry ; Fluorescent Antibody Technique ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Immunophenotyping ; L-Selectin ; analysis ; Leukemia ; blood ; drug therapy ; pathology ; Male ; Middle Aged ; Neutrophils ; drug effects ; immunology ; pathology ; Phagocytosis ; drug effects ; Receptors, IgG ; analysis ; Recombinant Proteins ; Respiratory Burst ; drug effects

To investigate the changes of donor's peripheral blood immunocytes after mobilization with medium-dose recombinant human granulocyte-colony stimulating factor (rhG-CSF), the amounts of immunocytes in peripheral blood cells and the immunocyte components of donor peripheral blood mononuclear cells (PBMNC) in 12 healthy donors were detected by flow cytometry before and after mobilization with rhG-CSF 10 microg/(kg.day). The results showed that the median amounts of peripheral blood leukocytes before mobilization was 6.25 (4.7-7.8) x 10(9)/L, for lymphocytes it was 2.07 (1.63-3.10) x 10(9)/L, and for monocytes it was 0.163 (0.078-0.414) x 10(9)/L. In the fifth day after mobilization, the median amounts of peripheral blood leukocytes was 37.47 (24-72.57) x 10(9)/L, and for lymphocytes it was 3.22 (1.46-5.31) x 10(9)/L, and for monocytes, it was 1.2 (0.706-3.627) x 10(9)/L. The average amount of leukocytes after mobilization was 6.26 +/- 2.14 multiple of that before mobilization (P < 0.01), and the median amounts of lymphocytes after mobilization was 1.45 +/- 0.76 multiple of that before mobilization (P < 0.05), and the amount of monocytes after mobilization was 7.48 +/- 4.41 multiple of that before mobilization (P < 0.01). The median percentage of CD3(+) T lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization. The ratio of CD4(+)/CD8(+) before mobilization was 1.27 +/- 0.46, while 1.36 +/- 0.51 after mobilization. The median percentage of CD4(+)CD8(+) T lymphocytes was 0.41% [(0.16-1.51)%], and 0.49% [(0.09-2.0)%] after mobilization. The median percentage of CD16(+)CD56(+) NK cells was 13.98% [(4.08-25.08)%] versus 16.65% [(12.06-33.05)%] after mobilization. The median percentage of CD3(+)CD16(+)CD56(+) NK-T cells was 2.75% [(0.37-6.38)%], but 3.13% [(0.46-5.95)%] after mobilization. The median percentage of CD20(+) B cells was 9.28% [(5.97-16.33)%], while 9.94% [(7.36-20.41)%] after mobilization. The median percentage of CD14(+) monocytes was 12.48% [(3.54-19.35)%] versus 29.52% [(16.51-36.76)%] after mobilization. The percentage of CD3(+) T lymphocytes, CD4(+)CD8(+) T lymphocytes, NK cells, NK-T cells and B lymphocytes in PBMNC did not change markedly before and after mobilization with middle-dose rhG-CSF. The ratio of CD4(+)/CD8(+) did not change significantly (P > 0.10). The percentages of CD14(+) monocytes in PBMNC after mobilization increased up to 2.87 +/- 1.51 higher than that before mobilization (P < 0.05). It is concluded that the changes of the CD14(+) monocytes after mobilization with rhG-CSF may be involved in graft rejection and graft versus host disease after allo-PBSCT.
Adolescent ; Adult ; Antigens, CD20 ; analysis ; Blood Donors ; CD3 Complex ; analysis ; CD4-Positive T-Lymphocytes ; cytology ; drug effects ; immunology ; CD56 Antigen ; analysis ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; immunology ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Monocytes ; cytology ; drug effects ; immunology ; Peripheral Blood Stem Cell Transplantation ; Receptors, IgG ; analysis ; Recombinant Proteins

UNLABELLEDThe purpose of this study was to evaluate the effects of cellular immunity activation on P58(+) cells expressing killer cell inhibitory receptor (KIR) and their regulatory function on cellular immunity, and provid theoretical data for preventing graft-vers-host disease (GVHD) in stem cell transplantation therapy. The mononuclear cells from human peripheral blood were incubated with IL-2, Con A and Lipostin (LP) for 72 hours. The KIR expressing cells, P58.1(+) and P58.2(+) cells, were analyzed by flow cytometry. The results showed that the percentages of CD3(+), CD4(+), CD8(+), CD16(+)CD56(+), P58.1(+) and P58.2(+) cells were greatly increased after treated with IL-2, Con A and LP, separately or in combination, and the percentages of above cells in combined treatment groups were higher than those of single stimulated groups, especially the percentage of cells in the IL-2 + LP group was significantly higher than those in IL-2 and LP singly treated groups.

IN CONCLUSIONIL-2, Con A and LP possess the ability to induce the expression of KIR and stimulate proliferation of P58.1(+) and P58.2(+) cells while to activate the celluar immunity response, the expression of P58 gene may be regulated by the activation of cellular immunity.

Adult ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD56 Antigen ; analysis ; CD8 Antigens ; analysis ; Cell Count ; Cell Division ; drug effects ; Concanavalin A ; pharmacology ; Flow Cytometry ; Humans ; Interleukin-2 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Receptors, IgG ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR ; Receptors, KIR2DL3

OBJECTIVETo investigate the relationship between subsets of lymphocytes and between its activated status and the clinical manifestations in patients with PNH, and to unfold immunological mechanism in the pathogenesis of PNH.

METHODSThe peripheral blood mononuclear cells (PBMNC) from 18 PNH patients and 20 controls were separated into two subpopulations using anti-CD(59) monoclonal antibody combined with goat-anti-mouse IgG immunomagnetic beads. CD(3)(+), CD(4)(+) and CD(8)(+) lymphocyte subsets were detected by flow cytometry. In 6 newly diagnosed patients, phenotypes associated with T cell activation such as CD(28)(+)/CD(4)(+) or CD(8)(+) cells, CD(8)(+) CD(38)(+) cells, and HLA-DR(+)/CD(4)(+) or CD(8)(+), and NK (CD(3)(-) CD(16)(+)) cells were detected in the peripheral blood.

RESULTPatients with PNH showed significantly increased CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) ratio as compared with controls (1.22 +/- 0.51 vs 0.86 +/- 0.27, P < 0.05), and the CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) ratio in CD(59)(-) PBMC was higher than that in CD(59)(+) PBMC (2.31 +/- 1.56 vs 0.62 +/- 0.27, P < 0.05). The ratios of CD(4)(+) CD(28)(+)/CD(4)(+) markedly decreased and CD(8)(+)HLA-DR(+)/CD(8)(+) increased.

CONCLUSIONPatients with PNH appear to have abnormalities in their lymphocytes. Increased ratios of CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) and HLA-DR(+) CD(8)(+)/CD(8)(+) lymphocytes as well as declined ratio of CD(4)(+) CD(28)(+)/CD(4)(+) lymphocytes might be involved in the pathogenesis of PNH.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Adult ; Aged ; Antigens, CD ; analysis ; CD28 Antigens ; analysis ; CD4 Antigens ; analysis ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8 Antigens ; analysis ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Female ; Hemoglobinuria, Paroxysmal ; blood ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; Lymphocyte Subsets ; cytology ; immunology ; Male ; Membrane Glycoproteins ; Middle Aged ; Receptors, IgG ; analysis